National Academies Press: OpenBook

Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1 (1982)

Chapter: Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.

« Previous: Appendix H: Evaluation of Data from Short-Term Testing of Anticholinergic Chemicals-- by Virginia C. Dunkel, Ph.D.
Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Suggested Citation:"Appendix I: Digest Report--Anticholinergic Chemicals-- by Henry Wills, Ph.D.." National Research Council. 1982. Possible Long-Term Health Effects of Short-Term Exposure to Chemical Agents: Volume 1. Washington, DC: The National Academies Press. doi: 10.17226/740.
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Below is the uncorrected machine-read text of this chapter, intended to provide our own search engines and external engines with highly rich, chapter-representative searchable text of each book. Because it is UNCORRECTED material, please consider the following text as a useful but insufficient proxy for the authoritative book pages.

APPENDIX I 1)IGEST REPO RT--ANTICHOLIN.ERGI C CHEMICALS Introduc tion Es ters of Tropic Ac id Studies in Animals Human Data from General Li terature Human Oata f rom Edgewood Arsenal Summary Esters of Benzilic Acid Introduc tion Data from Experiments with Animals 3-Quinuclidinly Benzilate (BZ or EA 2277) Diethylaminoethy 1 Benzilate Effect s on Han Human Data f rom Edgewood Arsenal S',nmary Esters of Phenylcyclopentylglycolic Acid tJ-He thyl-4-piperidinyl-( pheny~cyclopentyl)-glycolate (EA 3443) Di tran (CS 4297) 3-Qu inuc lidinyl-( pheny~cyc lopentyI) -glycolate ~ EA 3167 L -2-a lpha-Tropinyl-L-( pheny~cyclopenty] )-glycolate c i s-2-He thy 1-3-quinuc lidinyl-( pheny~cyclopentyI)-glycolate (301, 060) Esters of Phenylisopropylglycolic Acid N-bie thyl-4-piperidinyl-(phenyIlsopropyl~-glycolate (EA 3834 4-~1-Me thy I-1-l, 2, 3, 6-t e trahy dro pyri dinyl ) -phenyli sopro pyl- glycolate ~ 302, 668 ~ Esters of 1-Propyny~cyclopenty~glycolic Acld N-~* thy1-4-piperidyl-(phenylcyclobutyl)-glycolate (EA 3580) - Hi sc ellaneous Es ters 302, 282 302, 537 WIN 229 9 Nonester Anticholinergic Compounds ~ Benzetimide (CAS 14051-33-3) blepiperphenidol

"TAB' Mixture Di scussion No te: The chemical nomenc lature may dif fer f rom that given in the master file. The EA or other number serves for t~entification of the compound in the master file. e

APPENDIX I DIGEST REPORT ANTICHOLINERGI C CHEMICALS by J. lIeary Wills, Ph.~. INTRODUCTION The Board on Toxicology and Environmental Health Hazards of the National Research Counct1 provided 58 reports that are related to the administration of anticholinergic compounds to human volunteers by personnel ant contractors of the Chemical Corps Medical Laboratories at Army Chemical Center, Md., and its successor organizations at Aberdeen Proving Ground, Edgewood, Maryland . Those report s recorded studies of the responses of humans, mostly ~en, to 19 individual substances and to a mixture- of atropine, benactyzine, and the oxime, trimedoxime (THB-4~. Two of the 19, benzetimide (Dioxotrine) and mepiperphenidol (Darstine), are N-all~ylated derivatives of piperidine, but the other 17 are esters. Anticholinergic substances not mentioned in these reports, but said to have been administered also to volunteers, are N-methy ltropiny~pheny~cyelopenty~glycolate, 3-quinuclidinyI-~-hydroxypheny~cyclopentylacetate, and N-die thy laminoe thy lphenylcyclopenty~carboxylate . The ~ 7 esters represented in the 58 reports mentioned above belong to seven series of compounds: esters of tropic acid, of benzylic acid, of phenylisopropy~glycolic arid, of phenylcyclopenty~glycolic acid, of 3-quinuc lidinol, of ethyl-4-piperi:dinol, and of N-diethylaminoethanol. Esters of tropic acid occur in nature and have a comparatively long history of use as drugs. Atropine ~ I'L-hyoacyamine) and its optical isomers, the D- and L-hyoscyamines, anti scopolamine (L-hyoacine) and its optical isomer, D-hyoscine, have been obtained from such planes as deadly nightshade (Atropa belladonna) ~ jimsonweed (Datura stramonium), henbane (Ryoacyamus niger), horsenettle ~ Solanum , carol~inen~e), and various species of Scopolia. The L isomers of both esters are more potent than the D isomers. Both esters have peripheral and central anticholinergic activities; i.e., they interfere with the actions of acety~choline at peripheral ne~roeffector junctions on smooth and cardiac muscles and on cells of glands that produce external secretions and at synapses between some neural elements. Scopolamine is especially potent in the lest regard. whereas atropine and L-hyoacyamine are especially effective in blocking the action of acety~choline at peripheral neuroeffector junctions. Even at neuromuscular June tions on skeletal muscles, which atropine usually is considered not to affect significantly, a sufficiently high concentration (10~4 M) of this drug in vitro caused a shortening of the duration of end-plate currents ~ I) . I l

Quaternization of these alk~Ioide removes much of their ability to penetrate into the central nervous system and to affect its functions. However, the same variation in chemical structure increases the weak ability of these materials to interfere with the action of acety~choline at the motor end plates of skeletal muscles. Toxic psychoses and delirium from ingestion of atropine and acopola~ne have been Anon for many centuries; descriptions of the effects of these drugs antedate by long times the recognition of their mechanism of action. For example, henhane was reco~e-nded in the Ebere papyrus of about 1550 B.C. for the relief of abdominal distress. After -the first isolation of atropine from plant material in IB32, the actions of this group of alkaloids were identlfiet rapidly. By 1875, Ringer (2) vea able to give the following description (excerpted) of the actions of atropine belladonna): - Be lladonna employed either internally or externally checks or even suppresses the secreelon of the glance. This at least is true of the marry, sudoriparous and salivary glands, and possibly of other glands. les influence on the secretion of the submaxillary glance has been fully worked out. This gland reeei~res branches from the chords ~cympani nerve which is endowed with two sets of fibres, one acting immediately on the cells, the other causing the blood-vessels to dilate, being vaso-inhibitory. Belladonna acts through the nerves distributed to the cells, for after the injection of atropia, if the chords timpani nerve is irritated, the Vessels of the submaxillary gland become distended as usual, but the gland does cot secrete. The paralyzing effect of atropia is antedated by physostigma, for after the injection of physostigma, irritation of the chorda tympany causes the gland to secrete. Dropped into the eye, applied to the skin in its neighborhood, or taken by the stomach, preparations of belladonna very speedily produce extreme dilatation of the pupil. This is one of the most characteristic effects of belladonna. ~ full dose of belladonna produces great dryness of the tongue snd roof of the mouth, extending down the pharynx and larynx, inducing consequently cone difficulty in swallow) - , with hoarsenese, and even d" cough; and large dose will sometimes induce dryness of the Schneidarian membrane, and dryness of the conjunctive, with much injection. Belladonna often relieves colic of the intestines; and is especially serviceable in the colic of children. After a considerable dose of belladonna, the face becomes much flushed, the eye bright, d", and injected, the pupil dilated, the sight dim and hazy, while the power of accommodation in the eye for I 2

distance in lost. The mind and senses are peculiarly affected. The ideas, at first rapid and connected, become incoherent and extravagant; there is often decided delirium, with pleasing illusions. Sometimes the patient is possessed with constant restlessness, keeps continually moving, and cannot be quieted. A kind of somnambulism is occasionally observed; thus cases are recorded where, under the inf luence of belladonna ~ the patient for a long time performs the movements customary to his occupation; thus, it is narrated of a tailor that he sat f or hours moving his hands and ares as if sewing and his lips as if talking, but without uttering a word. The delirium may be furious ant dangerous, requiring the patient to be restrained; nay, it is recorded of one poisoned by this drug that so violent did he become that he was ordered to be confined in a mat-house . The f irst effect of belladonna on the pulse ts to increase its quickness, fullness, and force to the extent even of 50 Lo 60 beats in the minute.... Meuriot is of opinion that belladonna paralyses the peripheral branches of the vagal nerve, and by this means accelerates the heart 's action. Ringer had this to say about the ef facts of hyoscyamus ~ scopolamine): Thus it produces dryness of the mouth and throat, dilatation of the pupil, presbyopia, lightness and swimming in the head, delirium and hallucinations, a drunken gait, and often a strong desire to fight. Sometimes there is aphonia, and often sleepiness, with oppressive disagreeable dreams. ~ red rash has been observed after large doses. The pulse at first is much lessened in frequency but soon recovers itself, sometimes becoming even quicker than before the medicine was taken . Hyoseyamus is generally used to produce sleep when opium disagrees. It has also been employed in neuralgia. It was only af ter the role of acetyJ.choline in the functioning of the autonomic nervous system, in neuroeffector transmission to the various entities innervated by that system and to skeletal muscles (3-~), and in synaptic ~cransmission within some areas of the central and peripheral nervous systems (12-3~7) had been demonstrated that the proximate mechanism of action of atroplne, scopolamine, and other compounds of similar activity could be understood. These compounds seem to attach to the same receptors to which acety~choline usually attaches itself and thus to interfere with the ability of acety~choline to produce changes in the properties of cellular membranes. The actions of atropine and Scopolamine to block synaptic transmission within some areas of the central nervous sytem and Lo induce c oma were applied to the treatment of I 3

mental disease by Forrer ( 18) and Goldner ( l9) . In 1950, Forrer published an account of a study of 16 schizophrenic patients treated intramuscularly with large doses of atropine sulfate on 3 days of each week for a long period. The doses of atropine sulfate were increased gradually during a course of treatment from 20~32 mg to 212 ma, to maintain the achievement of unconeciousness, with hyperreflexta and development of the Babinaki sign, despite a gradually developing tolerance to the Snug. In 1957, Hiller et al. (20) reviewed the cases of 206 mental patients treated in this general way, includlag 148 who had been ill for 2 ye or more before treatment. At the time of that report, the technique was to inject atropine sulfate at 3:00 a.m., so that the patient would recover sufficiently from the effect of the drug to be able to eat lunch and take part in organized activities in the hospital during the afternoon and evening of the same day. Four treatments were given each week, instead of the three that had been given initially. The desired state of coma was produced ult}`in 30~60 mic of intramuscu3.ar injection, and spontaneous recovery, with complete clearing of the sensorlum. required 6-9 h at ter the in Section. - Of 206 patients treated, Il5 (55.~) were considered to have improved moderately or markedly, 60 (29.1Z) to be unimproved, and 31 (15.01) to have imp roves only slightly. Six months after completion of a course of injections, 115 patients (55.8Z) were atoll improved moderately or markedly. In a series of about 500 patients treated with large doses of atropine, with about 10,000 individual inductions of coma, there was one death (21~. This fatality was attributed to the replacement, without notification of the physician, of the specially trained nursing and technical personnel on the treatment ward with persons who were Scat thoroughly familiar with the procedures for caring for comatose patients, and particularly the procedures for controlling febrile reac tions . The patient died of uncontrolled hyperthermia. Goldner ~19) reported that he had used tntraeuscular toses of 5-50 mg of scopolamine in treating beneficially an unspecified number of mental patients, but then had switched to atropine because of lts greater a~railabllity. He reported on a group of 20 patients treated with atropine-toxicity therapy, 13 (65%) of whom were considered to have benefited moderately or markedly. Goldner also gave a limited comparison of the results of electrocon~rulai~re therapy (ECT) and of a ~ropine-toxicity therapy, stating that several patients who had not benefited from ECT improved on substitution of atropice-induced coma. Dada et al. (22) reported on 51 psychoneurotic and psychotic patients treated with intramuscular inJectione of 10-220 mg of atropine sulfate every other day during a period of 3-7 ok. The average dose was 30-50 tog. The longeat period of observation mentioned in the paper was 6 mot Of the 51 patients, 28 (S4.9%) seers considered to have improved detectably. The proportion that improved among the psychotic patients (53.3Z) was smaller than that among psychopathic and I 6

psychoneurotic ones (66. 7Z). The greatest alterations in symptoms and signs of the patients' diseases were in psychomotor excitement, hallucination, euphoria, anxiety, compulsion, and delusion. dada _ al. stated that trepro~rement, when it occurred, began with the third to the seventh injection and increased with later injections. If no improvement occurred by the tenth injection, it was not likely t O appear. The authors concluded that atropine-toxicity therapy i s a relatively sat e type of shock therapy, but stated that it was successful in only about 53.3% of their psychotic patients and that its efficacy probably had been overestimated by Schwarz (23~. Schwarz had concluded that 25: of 155 cases of psychoneurosis and psychosis had improved markedly with atropine-toxicity therapy and that another 32% had improved moderately . Hiller et al. (20) described the phenomena experienced by patients given large doses of atropine to induce a toxic coma: The various stages are passed through quietly and comf ortably . Neurologically , there are in order: progressive muscular incoordination, decreased pain sensitivity, and hyperreflexia with the development of the Babinski sign. On the psychological side, we observed clouding of the sensorium, disorientation, loss of time-space relationship, distortion of perception by illusions and hallucinations, confusion, and coma which proceeds to but does not go further than the early fourth stage in one and one-half hours af ter the drug is admini stered . Although the sensorium is markedly clouded, there is no cessation of psychological phenomena . Patient s undergoing treatment indicate by their actions, and occasionally by word, that psychic processes continue even during the coma. Illusions seem to be experienced, inasmuch as patients attempt to pick up the bedclothing with their f ingers and later report that they saw, and attempted to pick up, flowers, bugs, snakes, et cetera. Conversations may be held with absent persons--indicative of an hallucinatory state. With both atropine-induced i llusions and hallucinat ions, highly signif icant past events in the patient's life seem to provide the mayor psychic stimuli. Behavior is correlated with psychological phenomena. Circumoral movements, including sucking and illusory "eating" and 'smoking," are commonly observed. When a sufficient degree of motor coordination i s re tained to accomplish the necessary movement, a progressive, coordinated, and purposeful series of events may of ten be observed. For example, a pantomime of striking a match, lighting a cigarette, and subsequently of smoking that cigarette is not infrequently observed. Affective lability and corresponding rapid alternation of facial expressions suggestive of pain, sadness, querulousness, and euphoria are commonly observed . This clinical experience indicating that even large and temporarily incapacitating hoses of atropine and scopolamine had no persistent deleterious effects on the health of human I 5

beings made it seem that the administration of other' possibly more incapacitating, anticholinergice to huff volunteers was not unethical or immoral if two conditions had been fulfilled. These were that a compound to be administered to volunteers had been tested adequately in experimental animals to determine that the probability of its having persistent ill effects on health was small and that the volunteers had been informed as fully as possible of the possible effects of the c ompound . In the cases of compounds that hat been administered to man previously, the effects could be described quite accurately; if the only previous recipients had been experimental animals, the description of possible effects had to be somewhat hypothetical but usually could be re ~sonably accurat e . Before a discussion of the specific groups of anticholinergic compounds, it may be worth while to have a general view of the procedure of the experiments carried out wit}` human volunteers at Edgewood Arsenal and followed also by contractors. The following description is a composite from two sources: a report from Edgewood Arsenal (24) and a report by a contractor (25~. Test subJec ts were selected in two stages. Groups of possible volunteers were acquainted with the general nature of the experimental program through a briefing, during which they could ask questions about the general conditions and procedures of the studies. Those who volunteered to take part in the experimental program after this general briefing were given thorough medical examinations, including psychologic or psychiatric assessment. The volunteers considered acceptable for a particular experiment were then given a specific briefing and an opportunity to volunteer to take part in that study. The studies were carried out in air-conditioned areas from which hazards of accidental injury hat been eliminated as thoroughly as possible (e.g., '~lass panels in doors were replaced with wood, electric outlets and cables shielded, ant sharp corners padded) . Beds and toilet, messing, and recreational facilities were provided within the research areas. The research was conducted under sur~reillance of qualified physicians, who administered all in Jections or oral doses of the test substances. Men volunteers were exposed to inhalation of vapors or aerosols, the concentrations of these substances in the air Lo be breathed by the sub jects were established by appropriate personnel. Breathing of the contaminated air by the ~ro1unteers was observed and supervised by physicians. After an exposure to an agent, the volunteers were observed continuously f or 8-24 h, and then intermittently for up to 14 d. In a few cases, followup studies were made 6 ma or more af ter the exposure. I 6

ESTERS OF TROPIC ACID This group contains f ive of the substances said to have been administered to volunteers, but only three of these are mentioned in the 58 reports related to studies of the effects of anticholinergic compounds on human beings: aeropine, scopolamine, and methylscopolammonium. The ether two substances in this group that were administered Ho volunteers are D-hyoscyamine and methylatropinium. Atropine and scopolamine appear, respectively, in 18 and 11 reports, whereas methylscopolammonluss salts appear in only three. Atropine and scopolamine differ only in the basic moiety esterified with tropic acid. Scopolamine and the hyoscines have an oxygen bridge inserted between the two carbon atoms in the dihydropy~role ring of tropine that are not adjacent to the nitrogen atom, forming scopine, whereas atropine and the hyoscyamines have no such bridge. The facts that atropine is a racemic mixture of D- and L-hyoscyamines, that scopolamine is L-byoscine, and that the L f ores of both hyoscyamine and hyoscine have much greater biologic effectiveness than the D forms mean that atropine sulfate, with only 50~- of its active isomer, has only about 60Z as much active material per unit of weight as scopolamine hydrobromide, despite the addition of an oxygen atom in the bridge in the latter molecule. STUDIES IN ANIMALS Several relatively recent reviews of the actions and potencies of the solanaceous alkaloids are available (26-32~. Generally, they give a great deal of information about the medically useful or undesirable actions of the compounds without dealing with their toxicity in any quantitative manner. Table I-1 contains a sugary of information on the LDso doses for experimental animals of the f ive compounds in the group of esters of tropic acid, collected from a variety of published sources (33-41) and from internal reports of Bristol Laboratories, Lederle Laboratories, Maltby Laboratories, the Squibb Institute for Medical Research, Strasenburgh Laboratories, the Up John Company, and Sterling-Winthrop Research Institute . Albanus (42) studied the ef fects of subcutaneous in deco ions of 13 anticholinergic compounds on central and pert pheral cholinergic activities in the tog, recording such factors as the appearance of ataxia and of what he called "obstinate progression' (i.e., the failure of the animal to pull hack af ter encountering an obstacle in its path), the light reflex of the iris, salivation, ant heart rate. The compounds administered to dogs included a eropine, scopolamine, and a methylatropinium salt. A dose of atropine at 0.1 mg/kg produced no significant alterations in the dogs, but a dose of 0.2 mg/kg produced ataxia in three of four dogs and increased their heart rate by 76: ~ A dose of 0.3 mg/Icg mate five of five dogs ataxic ant causes three of five to engage in obstinate I 7

progression. ~ dose of 0.5 mg/kg rendered seven of seven dogs both at.axic and out of contact with the environment,, as indicated by obstinate progression. This dose also abolished the light reflex of the iris, blocked salivation between 45 and 180 sin after the dose, and increased heart rate by 891. Scopolamine was considerably more active than a tropine in this series of experiments, a dose of 0.03 mg/kg producing ataxia in six of six dogs and increasing heart rate by 24%. A dose of scopolamine at 0.05 mg/kg rendered four of four dogs ataxic and caused them to engage in obstinate progression. This dose also resulted in abolition of the light reflex of the iris, paralysis of salivation between 45 and 135 min after the dose, and a 5 9X increase in heart rate. The methylatropinium salt was much less effective than either of the onetime alkaloids. A dose of 2.5 mg/kg produced ataxia in none of four dogs. A dose of 10 ~g/kg induced ataxia in f ive of eight dogs and obstinate progression in three of eight. It abolished both the light reflex of the iris and salivation between 45 min af ter the dose ant the end of the period of observation, at 315 win after the dose. Doubling that dose induced both ataxia and obstinate progression in four of four dogs. The only study of the subchronic toxicity of members of the group of esters of tropic acid f ound is one by Boyd and Boyd (43) with atropine. Groups of rabbits were given daily intramuscular injections of atropine sulfate at 44-118 mg/kg for 100 d. The daily dose required to cause death within 100 d was 78 + 5 mg/kg, whereas that required to cause death within 10 d was 127 + 9 mg/kg. The Boyts obtained a relationship between the number (y) of dally injections to produce death and the size of the daily dose (x): log y ~ 2. 35 - 0.343 log x. The principal sublethal ef f ec ~ ~ not iced i n the rabbi t ~ were ~ in the approximate order of their appearance): increased intraco10nic temperature, decreased drinking, decreased production of uri ne, and decreased growth. Three other papers were related to a special type of subehronic toxicity induced by atropine: alteration of cells, embryos, and fetuses exposed to the chemical. Ishidate ee al. (44) exposed cultures of fibrob1asts from Chinese hamsters Lo atropine sulfate at 2SO ug/ml in saline for 48 h. At the end of the period of incubation, I: of the cells were polyplold. Of cells examined for chromo~omal aberrations after incubation with atropine for 24 h, FEZ had gaps and breaks in their chromosomes. These percentages of polyplol~y and of chromosomal aberration were only slightly greater than those in cultures to which plain saline had been added. Examination of atropine sulfate for carcinogenicity in animals by the method specifies in Survey of Compounds Which Have Been Tested f or Care inogenic Ac tinily (U. S . Public Health Service Pu blication 149 ) and f or mutagenicity in bacteria yielded no evidence that atropine had either activity. I 8

Butros (45) explanted 21- to 22-h chick blastoderms onto agar-containi~ substrates with atropine at 40~100 g/~1 for 24 h. Blastodems exposed at 40-8O,ug/~1 had alight but dose-related injury to the developing neural tube. Those exposed at 90 ug/mI hat arrested development with pyccosis and slight cytolysts of the cells of the brain wall and with somites that were smaller than noneal and that hat slightly opaque cells. Blastoderms exposed at 100 ~g/ml had their growth arrested and their brain walls wrinkled and folded, the cells evincing early cytolysis. In some of these blastoderms, the neural tube was completely cytolyzed; in others, eve neural tube was open at both its ends. Some 85% of the embryos had normal beating heart tubes; some heart primordia were incomplete and had lateral gaps. Grunf eld and Balass ~ 46) reported that pregnant rats given daily subcutaneous injections of atropine sulfate at 0.1 or 0.5 mg/kg-d from day Il to day IS of pregnancy had low heart rated, hypothermia, and greater than usual slowing of the heart by a dose of pilocarpine for about a week after the end of administration of atropine. The progeny of these rats had tachycardia for 2 wk after birth and mydriasis for a week after their eyes opened. No long-lasting effects on either the tams or the pups were repo reed . Scopolamine, added to the culture medium in a Petri dish at 1 ~g/plate after incubation with microsomes prepared from livers of rats, produced no change in the cultural characteristics of Salmonella typhimurtum strains TA 98 and TA 100 (47~. In agreement with this finding of non~utagenicity of scopolamine for bacteria, a study by Vrba (48) with HeLa cells, human leukocytes, and monkey renal cells revealed that scopolamine hydrobromide at a f ina1 concentration of I% in contact with these cells induced aberrations of the chromatics in the HeLa cells, but not in the monkey renal cells or the human leukocyte s. Vrba concluded that change ~ in chromatics in HeLa cells do not prove that a chemical is mutagenic in man. Campbell and Ramirez (49) gave three pregnant rats daily intraperitoneal injections of scopola~ ne at I. 2 mgJkg. Three other groups of two pregnant rats each were given nothing other than the usual food and water or daily intraperitoneal injections of physiologic saline or scopolamine at 2.4 mg/kg. All experimental procedures began on the tenth day of pregnancy ant were continued UDti1 parturition. Some of the of f spring of these dams, when 120 d old, were deprived of water for 18 h and then placed in a conflict situation in which they were exposed to continuous electric shock of variable voltage whenever they tried to drink. The voltage required to inhibit drinking was recorded. Offspring of the three dams that had received daily doses of scopolamine at I.2 mg/Icg were I 9

said to have tolerated elec tric shock better than those of dams under either of the other regimens. No data were presented . Lavallee (50) collected screening data on scopolamine wi th dogs and monkeys for comparison with the data from experiments with human volunteers performed at or for Edgewood Arsenal. In dogs, the incapacitating dose of scopolamine had been estimated to be less than 100 mg/kg. In monkeys, with only two animals, only a single dose of scopolamine had been used. The incapacitating dose was clearly above that dose, which had been 32 ~g/kg. In lean, the incapacita~cing dose in 50: of the sub Sects was said to be about lO,ug/kg. The agreement between the values for man and for the experimental animals was not good. Indeed, when a series of seven compounds was examined for comparative effectiveness in man and in togs and monkeys, the comparative grading in man did not agree with those in the other animals. - In addition to the more or less standard actions of anticholinergic compounds due to their ability to interfere with the actions of acety~choline on muscarinic receptors (resulting in acceleration of the heart; relaxation of smooth muscles in bronchi and bronchioles, intestinal tract ~ urinary trace, iris and cil.tary body of the eye, bile ducts and gall bladder, and some cutaneous blood vessels; and decrease or abolition of secretion by glands of the gastrointestinal tract and skin), these substances may have pronounced effect!. on the central nervous system, as has been mentioned earlier. A. large amount of research wi th experimental animals has been directed toward elucidation of the basic mechanisms involved in these actions. Much of the research has been oriented around one of the basic tenet s of pharmacology and toxicology: that a chemical entity affects particular structures in the body because they contain particular aggregates and arrangements of molecules and atoms that establish a special affinity between the structures and the chemical (51) . As a corollary, the particular aggregate and arrangement of atoms in a molecule of a chemical entity may condition its affinity for a given site, or receptor. Thus, isomerlc fovea of a molecule may have differen~c affinities for a given receptor, as has been found to be true for the ~ and L isomers of l~yoscyamine and hyoscine (38~. Also, fairly small differences between two molecules may make extensive differences in their abilities to react with particular sites in the body. For example, Graham and Lazarus (34) found that methy latropinium nitrate had approximately the same activity as atropine sulfate in preventing cholinergic s simulation of isolated rabbit intestine, but that it was some 3 times as lethal to mice after intraperitoneal injection. They also found that methylatropinium nitrate had a more pronounced inhibitory effect on the rabbit cardiovascular system than atropine sulfate; this I 10

difference between the two compounds certainly contributed t o, and may explain, the greater lethality of methylatropinium nitrate. There are indications that quaternization of atropine tid not alter extensively its affinity for the receptor on intestinal smooth muscle, bloc did increase the affinity for receptors on components of the cardiovascular system and perhaps other structures. Schallek and Smith (52) reported that intravenous injection of atropine at 0.01 mg~kg into togs anestbetlced with thiopental had no detectable effect on their EEGs or their cardiovascular systems. A dose of 0.! mg/kg increased the predominant frequency in the EEGe of three of f ive dogs and induced tachycardia in two of five. A dose of 1.0 mg/kg decreased the predominant frequency of the EEGs in four of five dogs and caused hypotension in one of five. Still higher doses increased these actions ant also induced bradycartia. Similar effects by atropine and scopolamine in curari zed cats ant monkeys had been reported previously (53, 54), and stimulation followed by depre ssion by atropine and scopolamine of maze-runnin8 by rats had been seen by Macht (55~. In 1963, Zvirblis and Kondri tzer (56) f ound that mitochondria isolated from 0.2 g of brain of young adult white rats adsorbed between B.4 ant 12.6: of the [14C]atropine in 4 m1 of atropine solutions with concentrations of 0. 063-0. 500 ,ug/~1. When the mitochondria isolated from 0.4 g of brain were immersed in the same concentrations of atropine, the adsorption was between ~ ~ O and 3~3. 53. If the lower of the last two value ~ i s erroneous ~ i t was obtained with the second lowest concentration of atropine) and the correct value should be about 13.3: between the values for the lowest and the next higher concentrations, the range of adsorption by the larger amount of ~tochondria would be 11.4-13.53. ' Even with substitution of the hypothetical value in the second series, increasing by 100: the mass of brain substance represented by the isolated mitochondria seems to have increased the adsorption of atropine by no more than a mean of about 131. Aggregation of ~itochondria in the higher concentration of these organe3~les may have reduced the effective surface for adsorption of atropine. In these experiments, Zvirblis and Kondritzer found also that 3-quinuclidinyl benzilate was adsorbed by mitochondria 2. 6-3.3 times as avidly as was atropine. In 1973, Far row and O'Brien (57) performed a study somewhat similar to that of Zvirblis and Kondritzer, using a tropine and muscarone as the adsorbates and various fractions isolated from a homogenate of rat brain as the adsorbents. They found that both adsorbates--but not decamethonium, dimethyltubocurarine, or nicotine--were bound reversibly on micochondria in a crude fraction. Muscarone was bound to the extreme of only about 0.37 time the binding of atropine. Atropine bound especially to fractions that consisted of presynaptic ant postsynaptic I 11

membranes, membranes of uncertain origin, ~icrosomes, and synaptosomes. It bound also to homogenates of liver and to ~itochondria prepared from liver, but not to homogenates of lung or kidney. Muscarone hat a similar pattern of binding to various fractions, except that in did not bind to mitochondria from liver. Hiley and Burgen (58) used [3H] propy~benzilyI- choline mustard as a ~ode! muscarinic agonist to examine the concentrations of muscarinic receptor in various regions of dog brain. Synaptoso~es and membrane. prepared from the head of the caudate nucleus, the proreal gyrus, the uncinate gyrus, the suprasyIvian gyrus, the lingual and suprasplenial gyri, and the anterior sylvian gyrus had the largest uptakes of the model agonise, whereas those prepared from the dorsal and ventral halves of the lumbar spinal cord, the subcortical white matter, the pes pedunculi, the corpus callosum, and the cerebellar cortex had the smallest uptakes. Hiley and Burgen (S8) measured also the choline acetylase and acety~cholines~erase activities of selected areas of the brain, in an attempt to determine whether the concentration of muscarinic receptors in a location was related to either of these enzyme activities. The amount of the labeled propylbenzily~choline mustard bound was no well correlated with either the choline acetylase or the acetylcholines terase ac tivity; there was a modes t correlation between binding of the model agonist by synaptosomes and membranes prepared f rom nine areas of brain and the choline acetylase activities of the same areas, but it was far from precise. Burgen_ al. (59) determined that byoscine was 10 times as active as atropine in vitro in reducing the uptake of [3~] propylbenzilylcholine mustard by synaptosomes from rat cerebral cortex. A meehylatropinium salt was only slightly less active than atropine, but had a very flat dose-action curve: an increase in its concentration by a factor of 10 increased the percent change in uptake of the propylbenzily~choline mustard only from 64Z to 671. These investigators also report et that the muscarinic receptor in cerebral cortices of the mouse, the guinea pig, the dog, the pig, and the macaque behaved very similarly t o the ~ in the rat . In 1974, the finding of Zvirblis ant Kontritzer (56) of a particularly large affinity of the muscarinic receptors in brain for 3-quinuclidinyl benzilate was applied to the assay of the quantity of muscarinic receptor in a sample of tissue (60, 61~. Intramuscular injection of atropine 30 mitt before intravenous injection of f 3H] 3-quinuclidiny! benzilate decreased the binding of the quinuclidinyl benzilate in various regions of the brain. Although the corpus striatum had the highest uptake of the labeled substance, the cerebral cortex was not f ar behind . The perc ent decreases by a tropine in the uptake of the ligand were approximately equal for the corpus striatum and the cerebral cortex. The hippocampus, I 12

which came nest in magnitude of bensllate binding, was also next in magnitude of the change in binding of the benzilate caused by atroplce. Indeed, in the six~embered series of corpus striatum, cerebral cortex, hippocampus, bypo thalamus, pons~edulia oblongata, and cerebellum, there was complete agreement in rank order for benzilate bluding and reduction by atropine of benzilate binding. Admintstration tO rats of mecamylamine, phenobarbital, or L~d~hydroxyphenylalanine before latra~renous injection of [38] 3-qulnuclldicyl benzilate had no effect on benzilate binding in the six areas of the brain. In these experiments, the highest concentration of labeled benzilate bount to protein was in the microso~l fraction of a homogenate of whole brain, and the next highest was in the mitochondrial fraction, the protein of the microsomal fraction binding nearly 2.5 times as such of the label as that of the ~ltochondrial fraction. The protein of tbe nuclear fraction bound only about one-twelfth as much of the labeled benzilate as that of the microsomal fraction and about one-fifth as much as that of the mitochondrial fraction. The muscarinic receptor substances) in the rat brain appears to be fairly generally distributed in membranes of subcellular components and to be localized especially in the corpus striaeum, the cerebral cortex, ant the hippocampus, the last of these three binding nearly 3 dices as high a concentration of labeled benzilate as the next most active structure in such binding, the hypothalamus. Because both muscarinic anticho3.inergle agonistic drugs (seopolamlne, isopropamide, and atropine) and muscarinic egotistic drugs (oxotremorine, acety~choline, methacholine), but not nicotinic anticholinergic drugs (mecamylamine, bexamethoniu=, and D-tubocurarioe) or nico~inic agonistic drugs (~imethy~pheny~piperazinium and nicotine), strongly inhibited the binding of labeled benzilate by homogenates of rat brain, binding of this compound was proposed as an indicator of the presence of muscarinic receptors in particuJ-ar brain structures. In the nine~embered series of corpus striatum, cerebral cortex, hippocampus, superior-inferior colllcull, pons~idbrain, thalamus, hypothalamus, cer~rlcal con-medulla oblongata, and cerebellar cortex, Yamamura and Sayder (61) found, as had Hiley and Bergen (58) with binding of labeled propylbenzily~choline mustard, that binding of labeled benzilate was not well correlated Smith either the choline acetylase or the cholinesterase activity of tI2e particular region' but was slightly better correlated with choline acetylase activity than with that of cholinesterase. Farrow and O'Brien (57) had concluded that binding of labeled atropine to various subcellular components of the rat brain could not be accounted for by binding to acety~cholinesterase. Farrow and O'Brien (57) had found also that scopolamine was a much more potent inhibitor of the binding of labeled atropine to synaptic membranes than a I 13

variety of cicotinic anticholinergic and ago~atic drug e; the next most active inhibitor used by these investigatore had been carbamy~choline chloride, found by Yaaamura and Snyder (56) to be less effective than acety~choline and me thachollne in reducing binding of labeled 3-quinuclidicyl benzilate. In later work. Sayder's group (62,63) used autoradiography to determine more precisely the locations of binding of [3ll]3-quicaclidiny! benzilate within the various regions studied earlier. In the cerebral cortex, the most ac tive areas of benzilate binding were the occipital cortex and the cortex of the cingulate gyrus. The cortex in the pirlforoa area was less active in this regard, and that of the frontal pole still lese so. In the hippocampus, three strata of the archipallium were fount to have approximately equal abilit:tes to bind 3-quinucl141ny} benzilate, but the white matter of the alveus had much lese activity of this sort. The striate body also had high biding activity for this I{gand, which was particularly striking in the autoratiographs because of the comparatively low binding activity of the adjoining globus pallidus. Phallic and hypothalamic nuclei had 1~w binding activi ties f or 30qulnuclidiny! benzilate similar to those of the inferior and superior colliculi. Still lower in activity in binding this ligand were cerebellar cortex, medulla oblongata-pons, and the cerebral peduncles. Nerve tracts-~e.g., the optic chiasma, the cervical spinal cord, and the corona radiate -had the lowest binding activities evaluated by Snyder et al. On the basis of the research outlined in the preceding paragraph, one would expect anticholinergic compounds to produce changes in affect and motor and autonomic regulatory activities. Krieger (64) has found that a 408e of atropine thee was sufficient to block the circadian increase in the plasma concentration of 17-hydroxycorticosteroids during the night did Doe prevent stimulation of secretion of 17-hydroxycorticosteroids by the adrenal cortex after administration of ACTS or of stressore, such as insulin and piromen. Thus, the effect of atropine seems to have been exerted not on the adrenal cortex, but rather on the pituitary, the controller of activity by-the adrenal cortex through modifiable release of ACTS. Eaton ant Rang (65), building on indications in previous papers (66~69) that atropine antagonized the action of acety~choline in tissues by combining reversibly with specific receptors, found evidence of the existence, in the longitudinal muscle of the small intestine of the guinea pig, of two binding sites for atroptne with different capacities for it. The equilibrium constant for binding atropine amounted to slightly more than S.Ol x 10~7 M, or about 1, 180 pmol/g of muscle. For methylatropinium 8alt8, binding seemed to be to one site with reasonably def ini te binding propert ies and to an ill~definet series of other sites with much higher 1 14

equilibrium constants. Methylatropinium was found to be about 2.34 times as active as atropine in blocking the receptors in this smooth muscle from access by acety~choline. Yamamura and Sayder (70) found that these muscarinic receptors resembled those in brain in having an affinity for [3H]quinuclidinyl benzilate that was greater than that for atropine, was the same as that for scopolamine, and was slightly less than that for methylatropinium salts. Scopolamine was about 3 times as active as atropine and about 1.7 times as acquire as a methylatropinium salt in preventing contraction of the longitudinal muscle of the ile~. The activity of 3~quinuclidinyl benzilate in this regard was the same as that of the methylatropinium salt. Sayers ant BUrki (71) determined the activities of a group of 13 psychoactive compounds, includlug atropine, in four. different test systems, among which was assay of interference with binding of labeled 3-quicuclidiny! benzilate to a homogenate of rat brain. The agreement between interf erence with binding and interf erence with stimulation by acety~choline of isolated guinea pig ileum was good . The agreements between i-~terf erence with benzilate binding and interference with production of tremor in mice by intraperitoneal in Section of a standard dose of oxotremorine or dilatation of the pupil of the eye of the mouse were less good. Witter et al. (72) compared the distribution in the rat brain of tritium-labeled atropine sulfate ant methylatropinium nitrate inJec ted intraperi toneally at 10 mg/kg. The rats were killed 30 gin after the injections. Livers, kidneys, and brains were collected. Neither a tropine nor the qua~ernized alkaloid was bound to a very great extent in basal ganglia, but both were bound rather extensively in pons-pyramidal tract, preoptic area, and cere bellum. The principal disagreements between the localizations of the two compounds were in the septum, whe re a tropine was bound relatively poorly and the quaternized alkaloid was bound comparatively well (atropine was still bound at nearly twice the concentration at which the methylatropinium radical w88 bounty, and in the hypothalamus and the cortex. In the last two areas, the alkaloid was bound comparatively extensively, whereas the atropinium radical was bound comparatively poorly. In all regions of the brain, atropine was bound to an extent that was 1.9-3.5 times that to which the atropinium compound was bunk. In both the liver and the kidney, as well as in plasma' there was a higher concentration of the atropicium radical than of atropine itself. There seems to be no general decrease in the ability of the atropinlum ton to move in the body, therefore, but rather a relatively poor ability of that ion to pene bate to or into ache cells of the central nervous system. This difference between the alkaloid and its quaternized form may be related simply to the lipoid-rich nature of the central nervous system. I 15

In 1952, Mlchaella et al. (73) reported that atroplne given before or after DFP d ~ not affect acetylchollne concentration in the cortex of rabbit brain ant that DFP increased that concentration by more than 1001. Deaplte this negative report on the Influence of atropine on acetylcholine concentration In the brain, Berry and Storz (74) stated in 1956 thee intraperltoneal iaJecelon of atropine at 6 ~g/kg decreased the mean concentration of acetylcholine in the ret brain from 365 ug/g to 204 ug/g at 30 min after injection. The depreseant effect of atropice on acety~choline concentration in the brain was confirmed by Giarsan and Pepeu in two papers (75, 76). In the first, they reported that intraperltoneal injection of equal doses (50 mg/kg) of atropine and scopolamine into rats produced the same mean percent decreases in acety~choline concentration tn the brain within 10 min after injection. A dose of atropine (400 ~g/kg) that produced alternation of central excitation ant tepreasion, "remora, and convulaiona within 10 win produced a slightly smaller mean depreasion of acetylchollne concentration than the smaller dose. The difference was not significant, however. In the second paper (76), the inveatigators compared the effects on brain acetylcholice concentration In the rat caused by a series of toses of atropine sulfate with those causes by two doses of hyoacice hydrobromide ant one dose of meehylatropiniu~ nitrate at 30 win after in~raperitoneal injections of the drugs. The three lowest doses of atropine surface (1, 5, and 25 eg/kg) produced dose-related mean decreases in acety~choline concentration, but the highest dose (50 ~g/kg) produced a smaller mean decrease than the two next lower doses. ~ dose of methylatroplciue nitrate equlmolar with a dose of atropine sulfate of 2.5 mg/kg resulted in a slightly smaller decrease in the mean acetylcholine concentration than the atropine dose of I.0 mg/\g. A dose of h~oscine hydrobromide equtmolar with atropine sulfate at 2.5 mg/kg vaulted in a lowering of the mean acetylcholine concentration nearly midway between those resulting from doses of 5 and IS ~/kg of atropine sulfate. In a ~ tudy of the time course of the decrease in the acety~cholice concentration in the brain in the rat, Giar~an and Pepeu (76) found that the maximal change occurred at about 60 gin after tatraperitoneal injection of hyoacine hydrobro~de and that in Sections of this compound at ~ mg/Icg three times per day for 6 ~ resulted in progresst~e decreases in acety~choline concentration during the first 3 d. Later, the concentration increased slightly, so that at 145 h after the fires injection of hyoacine hydrobrosIide (1 h after the nineteenth injection) the Dean value vea 60X greater than the lowest one, observed at 73 h following the first Inflection (1 h after the tenth injection). The greatest change in acety~choline concentration was in the cerebrum (a decrease of 351), accompanying changes in the rostral ~ 16

mesencephalon and in the caudal mesencephalon, with the metencephalon being below the leered of significance. Aqullonlus en al . ~ 77) studied the ef feces of atropine and of methylatropinium nitrate on the release of acety~choline by the cortex of the ret brain (by vividiffusion) and on motor activity (Jiggle cage). Atropine sulfate and methylatropiMue nitrate at 0.25-10 o~g/kg were injected into one itlgular vetn. Although doses of ~ethylatropinium nt era te of 0.25~0.75 mg/leg produced sli ghtly greater release of acety~choline into the ~ c ollecti~ cups than the same doses of atropine sulfate, the former drug in dose-e up to 10 mg/kg produced a maximal increase in motor activity of 2.33 times basal activity, whereas atropine sulfate produced an increase in motor activity of 6.75 dimes basal activity. By using atropine and ~ethylatropinium labeled with trirlum, Aquilonius et al. (77) found that injection of equimolar doses of the two compounds into a jugular vein resulted after 30~60 min in nearly equal concentrations of the labels from the two compound in cot only the brain and cerebrospinal fluid but also the plasma. Despite the fairly marked differences between the fractions of the injected toses of the two compounds tha~c entered the cerebrospinal fluid and brain, the fractions of the doses that entered the fluid in the collecting cups were not very different after 30 mini after 60 min. they were somewhat more different, but perhaps not significantly so. To explain the appearance of the methylatropinium compound in the vi~ridiffualon cup and in the cerebrospiDa fluid, the investigators adopted the suggestion of Szerb ~ 78) that. such charged compounds as ache qus~cer~zed alkaloid may pass f rom the blood into the subarachnoid fluid and both seep into a vividiffusion cup and cause superficial layers of cortical cells to release acetylcholine. Following the f indite that septal lesions result in increased concentrations of acety~choline in the hi ppocampus, Rommelepacher and Kuhar ~ 7 9) ~ tudied the ef feet ~ of muscarinic cholinergic agonists arid antagonists on the acety~chollne concentration in the rat hippocampus after lesions of the septum had been produced electrolytically. The J=gs were injected intraperitoneally just before administration of ether for creation of the repeal lesions. The rats were killed 20 min after the lesions had been made. Doses of 5 8~14 i0 mg/Icg of atropine ant scopolamine resulted in decreases (atropine) and increases (acopola~ine) in the mean concentration of acety~chol~ine in the hippocampus. Smaller doses of atropine (0.l and I.0 mg~kg) may have induced increases in the acety~choline concentration; only two rat ~ received each of these doses . Berger et al . ( 80) induced hippocampal after-discharges by electric stimulation of the fornix and f ound that doses of atropine at I-2 mg/kg, pre sumably injected intravenously, doubled the duration of the I 17

af~er-discharges induced by a standard stimulus. This result may have been due to increased release of acetylcholine. Frances and Jacob (~] ~ found that eserine and arecoline, on the one hand, ant atropine am scopolamine, on the other, had oppositely directed effects on the acety~choline concentration in the mouse brain. Beerine and arecoline induced lacreased concentrations, whereas atropine and acopolamine induced decreases. These data indicate that a suboutaneousI, injected dose of scopolamine about one-f if teenth that of atropine administered similarI,r produced the same decrease in acety~choline concentration in the brain as the larger dose of atropine. In the case of scopolamine, the percent decrease in acety~choline concentration was a linear function of the log dose of the alkaloid in the range of do-see used--0.3-3 ag/kg. Atropine at ~ to about 8 agJl~g yielded a linear relationship between percent decrease in acety~choline concentration and log dose. At 10-30 Aging, there was no further effect on acety~choline concentration by atropine, although the maxioul change induced by this compound was only about two-thirds that induced by the largest dose of scopolamine used. There was a Itneas relationship between the percent decrease in acety~choline concentration and activity ( jiggle cage) in rats given scopolamine at 3 mg/kg or atropine at 10-30 ~/kg. Ilerink et al. (82) used septal lesions to modify the motor activi ty of rats, as had Ro~elepacher and Kuhar (79~. An intraperitonesI injection of atropine at 1 eg/\cg in normal rats increased aggressiveness within 5 din, whereas in rats with septal lesions it decreased aggre asiveness. Atropine decreased defecation, an index of emotion. in both types of rate. tJsui and Twahara (83) used freely moving male bars with recording electrodes in the dorsal hippoc~pus and in contact with the aura over the frontal and occipital cortices and found that atropine at 5-30 mg~kg in jected i ntraperi toneally delayed the f ire t appearanc e and shortened the f irst episode of paratoxic sleep. Doses greater than 10 mg/kg usually prevented paradoxic sleep entirely. The regularieg of the theta rhythm f roe the hippocampus was markedly disturbed in paradoxic sleep without rapid eye movements; when paradoxic sleep was accompanied by rapid eye movements, theta activity was increased in both frequency and amplitude. Theta activity associated with body movement was not altered by atropine. The hippocampus seems to have two systems for generating theta activity, one that contains ~uscarinlc cholinergic receptors and is susceptible to modification by atropine, and one that is not susceptible to alteration by atropine. Takeyasu en al. (84) gave male rats two intraperitoneal injections of atropine at 3 mg/kg each day for 4 wk. after which the rats were not given atropine for 5-8 d. At the end of the withdrawal period, the effect of I lS

subcutaneous atropine (1-20 eg/kg) on motor activity was evaluated. Previously unatropinized rats were subJec ted to the same doses of atropine while their activity was monitored. Both the naive and the atropine-adapted rats yielded dose-related curves of increased activity as the dose of atropine was increased; the curve f or naive rats was considerably higher than that for the atroplne-adapted ones. Takeyasu et al. found evidence that the masses of muscarinic cholinergic receptors in the cerebral cortex, the hippocampus, and the corpus striatum were all increased by the repeated daily 608e~ of atroptne. They suggested chase the smal3.er response to atropine in the atropine-adapted rats than in the naive ones might be due to this factor. (Both curves in Figure ~ of the paper by Takeyasu et al. tend to become parallel to the dosage axis at different levels of activity, rather than converging as required by their hypothesis. ~ In dogs, intravenous in Sections of atropine at 0.5-~.5 mg/kg produced a syndrome of decreased alertness and responsiveness to commands, stumbling and slipping, swaying, incoorti&ation and ataxia, splaying of tile hind legs, blundering into obstacles and attempting to progress forward despite the presence of an obstacle, wilting or barking ~ agitated pawing, apparent pursuit of unreal moving objects, collapse, and sleep from which the dogs c ould be aroused temporarily by handling or noise ~ 85) . Recovery from a I~g/kg dose of atropine required about 5 h. Intravenous injections of tetratydro-5-aminoacr:~ine [THAI at ~ mg/kg and yohimbine at O.S mg/kg were found to produce some recovery from the effects of atropine, TEA being more effective than yohimbine [86~. These authors did not find that small intravenous doses of physostigeine OF neostigmine (0.02 mg/kg) were significantly -antagonistic to the central effects of anticholinergic drugs. Because the usual dosage of atropine is small, tts metabolism could not be studied effectively until fairly recently, when methods for labeling the drug with - radioactive isotopes and for measuring the concentrations of the isotopes in biologic samples tee came s~railable. Everesbusch and Gelling (87) seem to have been the first investigators to use i4C-labeled atropine. Their atroplne was prepared bar extraction from Atropa beliaton grown is an atmosphere of Deco:, so that the position of the label was unknown. The most sensitive method for determining atropine had been the mouse eye assay, ~eh which Pulewka (~) and Tonnesen (89 ) were able to determine that only a fraction of a done of atropine administered to man (by mouth or by subcutaneous ~ ejection) appeared unchanged in the urine. Evertabusch and Geiling (90) found that a portion of the label from randomly labeled [14C3 atropine appeared in the expired air as i4CO2. In 1953, Gosselin et al. began an intensive study of the metabolic fate of atropine made by esterifying I 19

tropinol with tropic acid that contained an atom of i4C in the position between the benzene ring and the carboxyl group. ltice were the first animals studied (over 100 were used), followed by rats, a few cats, and finally eight human beings. The original group of in~restlgators separated into three parts bef ore the entire series of studies was completed, ant one of those parts studied the fates of two differently labeled atropinea in man. The first part of the research to be reported was the study of the metabolic fates of labeled tropic acid and atropine in mice ant rats (9l,92~. Hlce and rats given intraperi toneal injections of alpha-~14C] tropic acid at 1 mg/kg excreted in their urine only a single labeled compound that had essentially the same chromatographic characteristics as authentic tropic acid. The excretion of i4C administered in this fore was complete within about 2.5 h in the mice and within slightly more than 3 h i n the rats . When the labeled atropine was administered to both mice and rats by both intravenous and intraperitoceal injections, the cumulative excretion of i4C in the urine by the mouse was always considerably greater than thee by the rat. The label from alpha-~14C3atropine injected intravenously into mice appeared in their urine slightly more promptly and to a somewhat greater extent than that f rom labeled atropine thee had been injected i ntraperi toneally or su~ utaneously or that had been administered by garage. The curve for the clearance of 4C from the body of the mouse with time elapsed af ter su bcutaneous ir~Jec tion of the labeled alkaloid required three simultaneous exponential equations to represent the observations after a period of latency of about 45 min. during which excretion of i4C followed none of the three exponential relationshipa. In addition tO atropine itself, at least three other substances containing i4C appeared in the urine of the mouse. At 30 On af ter intravenous injection of labeled atropine into elce, the structures found to have the highest concentrations of 14C were the gall blaster ant the bile, the urinary bladder, and the duodenum and jedunum. By 4 h after injection, the structures with the highest concentrations of 14C were the gall bladder and the bile, the urinary bladder, and the ileum. At 24 h at ter injection, the structures with the highest concentrations of i4C were the colon and cecum, the gall bladder and the bile, and the stomach. (In all cases, the hollow Discus and its confects are designated by the name of the discus alone. ~ The comparatively high concentration of the label in the stomach 24 h af ter injection may indicate that the mice had ingested some of their feces, the contents of the colon and cecum having high concentrations of i4C at that time. Lo i4co: was found in the expired air. It is clear that the principal routes of removal from the body of the atropine label used by Gosselin et I 20

al. (9l, 92) were the urine and the feces, the ictesti"l route of excretion being comparatlvel~r ~nor. Whatever the mode of administration of the labeled atropine to mice, 80~9OZ of the 14C was excreted in the urine. The consistent presence of the gall bladder am its contents among, the structures with the highest concentrations of the label indicates that the label enters the gastrointestinal tract af ter secre tion by the liver into the bile, which there flows into the small intestine through rhe bile duct and the colon duct. The progressive shif t of the segment of the tntestical tract with the highest concentration of t4C from the duodenum and jeJunum to the ileum and then to the large bowel reflects the movement of a bolus of art especially high concentration of the isotope don the tract during a period of about 24 h. The failure to f ind \4co: in the expired air of the mice indicates that the tropic acid moiety in the at.ropine molecule is Doe metabolized--in accord with the f inding that labeled tropic acid itself was excreted without loss in the urine. The finding of atropine but no tropic acid in the urine of the mice indicates fit hydrolysis of the ester did not occur rapidly. The latter finding was somewhat unexpected because an esterase capable of hydrolyzing atropine is present in the livers of many vertebrate species, although it or a similar enzyme appears in the sera of only a few species ~ 93-95) . his enzyme is also able to hydrolyze L-hyoacyamine, tropinyl benzoate ~ and caramiphen ~ 95) . The rat was found ~ 96) to excrete in its urine substances contalain'8 i4C derived from the labeled atropine with approxima~cely the same paper chromatographic re tartation factors (Rfs) as those found previously in the urine of the mouse. The rat differed from the mouse in exere tiny about 40X of the label in it8 feces, compared with about 10: for the mouse. The guinea pig, unIthe the other mammals studied, exere ted in its urine a large part of the 14C in a form that was probably tropic acid or some similar molecule (Rf slightly below that for tropic acid added to urine from the guinea pig). The majority of the 14C in the urine of the cat was excre ted in the form of atropine. only two other small peaks of radioactivity being evident on paper chro~tograme. The rat and the cat, unlike the mouse arid the guinea pig, excreted almost as much of the label in feces as in urine. In the rat and the cat, the livers were core active both in initial uptake and in maintaining a high uptake until 4 h after intravenous injection of labeled atroplne than those of the mouse and the guinea pig. Increased transf er of the isotope f rom the blood into the bile by the liver in the f order two species probably explains its greater fecal excretion by these animale than by the other two species. Despite the quantitative differences in excretion of labeled atropine by different routes in the mouse and the rat, Gabourel and Gosselin ~ 97) proposed that atropine ~ 21

follows the same metabolic pathways in the rat as in the mouse. They studied mice given intravenous injections of alpha-~14C]atropine and found that approximately 25: of the dose was excreted in the urine as atropine, more than 50Z as glucuronides, and the remainder as intermediate oxidation products and, probably, tropic acid esters with altered tropyl mol.etles. Although they were unable to prove that the aromatic ring of the tropic acid moiety was hydroxyla~ed, they proposed that bydroxylation of this rising, initially at the 4-position and then at the 3-position, resulted eventually in the production of monoglueurocides and diglucuronides. However, Philli peon et al. (98) found that Incubation of aeropine or hyoscine with a preparation of ~crosomes from guinea pig liver led first to the dealkylation of the nitrogen atom of the tropy] or the scopyl moiety and sequentially to production of the N-oxides and N-hydro~cynoratropine or ~ hydroxynorhyoacine. It seems possible that conjugation with glucuronic acid from uridine-5- dlphospho~alpha-I)-glucuronic arid under the catalytic i nf luence of urldine diphosphate glucuronyl transf erase occurred in the course of the oxidative series of reactions mentioned earlier and resulted 1c the formation of an N-glucuronide of noratropine. Such a reaction is known to occur with Secondary aromatic atones, such as Dora ~ropine. Werner and Schmidt ~ 99 ~ studied the types of chemical reactions undergone by [~4C]atropine introduced into mice, rata, guinea pigs, rabbits, cats, and monkeys. Their general conclusion was ~hat, although various species differed quantitatively in the relative amounts of part icular end products of atropine metabolism, the chemical changes undergone by the molecule in the various species were the same: hydrolysis of the ester linkage, hydroxylation of the benzene nucleus, glucuronidation, and. Oxidation to cog. Kalser et al. (100) found that bile of rats into which fi4C]atropine had been injected intravenously might contain up to five peaks of radioactivity of comparatively low Rf: 0.02, 0.08-0.09, 0.16, 0.22~0.23, and 0.68. The subetances represented by Me of 0.08-0.09 and 0.16 were present in the bile in the highest concentrations. No tropic acid or unaltered atropine was f ound in bile. ~tabolites of atropine appeared in the bile Ethic 10 min after intravenous injection of that alkaloid . The plasma of these rats c obtained only unchanged atropine in concentrations great enough to be detectable. Isolated, perfused livers of rats exposed to [~4C]atropine injected into the perfusion fluid excreted about 60Z of the i4C into the bile during 4 h (lOl). All the meeabolites were more polar than a~cropine itself, but they were not identif ted . Winbladh (102) presented data on the tistri button of atropine labeled randomly with 3ll in newborn pups, I 22

pups at 3 ant 6 wk of age. pups at 3 mo of age, ant adult dogs (10 to old or older) . Af ter subcutaneous injection of labeled atropine at 0.5 agJl~g, the isotope W8S removed from the blood plasma much more slowly in the newborn pups than in the adult Impale. The pups of other ages had intermediate half-~io~es for removal from the plasm, the greatest step of difference befog between newborn pups ant those 3 wk old. The half-~mes for removal of the label from the plasma after intravenous injection of labeled atropir~e were not strik~rig,iy different from those measured af ter subcutaneous injections. In newborn pups, the organs that initially took up the highest concentratione of the label were the kidneys and the liver, the kidneys hang a higher concentration. The liver maintained a higher proportion of its early concentration of 3H until ~ h after the lnJec~ion than the kidneys, however. Pups 6 wk old behaves qualitatively like the newborn ones, but in the adult dogs the initial uptake of the label by the liver was conelderably greater ~ by up to aoz' than that by the kidneys. Of the 3H in the urine, 50-9OZ~was in the for of unchanged atropine. In brain, Winbladh (102) found a progressively decreasing concentration of atropine (the only form in which the label was found in brain) as serial samples were taken f rom the cortex to the vicinity of the lateral ventricles. Harri son et al. ~103) used atropine labeled with 3H in the pare po~t~n of the benzene ring in the tropic acid moiety of atropine to follow the disposition of that alkaloid in the rat. They found that the mesa half-eimes for removal of the label from various tissues after lneraperitoneal injection were as follows: plasma, 44 mini heart and kidney, 54 mini liver, 74 mini brain, 76 mini and fat, 101 min. The half-eimes were determined f rom the slopes of the decay curves. Two fairly recent papers on the metabolic fate of atropine in man have been published (104,105~; an excellent scary of the work was presented by Kalser ( 106~ . The two subjects in the first of these papers excreted within 24 h BS: of an intramuscularly injected dose of 2 mg of alpha-~4C]aeropine. A urine sample collected from one of the subjects between l.5 and 4 h after injection contained subetances thee produced four small, interconnected pealce of radioactivity on a paper chromatogram that were apparently removed by incubation of the urine with bacterial beta-glucueonidase. Only three clear peaks of radioactivity appeared on the paper chromatograms of this urine after it had been exposed to glucuronitase. Two of these had Rfs that agreed with those for atropine (the mayor pealc) and for tropic acid (a small peak). The identity of the third peak, with an Rf below that for atropine, was not determined. After alkaline hydrolysis of this sample of urine, the only radioactive substance detected tn the chromatogram hat an Rf similar to that of tropic acid; thus, the tropic acid I 23

portion of the ester seeme not to have been altered metabolically to any important extent. The second paper (105) used Hero samples of [~4C]atropine; one was labeled on the methyl group attached to the nitrogen atom of the tropine motet~r, and the ocher was labeled at the two carbon atoms in the ring structure of the tropine cotety adjacent to the carbon atom involved ic the ester Itr~ge. Two Bale and two female subjects received intramuscular inJectione of 2 mg of one of these labeled atropines. One subject received ladi~ridual injections of each labeled atropine on two different days. When the meth~rI-labele'd atropine was administered, i4CO2 appeared in the expired air. Administration of the other labeled atropine did cot result in the excretion of i4Co:, nor had this substance been found in the air expired bar the only one of the subjects in the fires of these papers (1043 whose expired air was exa=ned. One concludes, therefore, fit the only part of the a~cropine molecule that is subject to complete oxidation is the methyl group on the nitrogen atom of the tropine moiety. As for exeretlon of the label in the urine, the two samples of atropine used by Kaiser arid ~lcLain yielded similar results. There was rapid excretion of i4C during the fires 4 h after injection that was nearly linear with time, but by 16 h after injection the rate of exere tion of 14C in the urine had decreased to a much lower rate that was also Dearly linear With time. The low rate persisted f or up tO 48 h af ter injection in the one subject who received each of the labeled atropines on separate days. Kalser and McLain (105) found that glucuronidation was an important metabolic pathway in the early excre tion of a tropine f rom the body, but that this type of reaction seemed not to be part icularly import ant af ter 4 h f ram the time of injection of atroptile. Referring to the work of Phillipson et al. (98), one may suppose that glucuronidation occurs when noratropine i8 produced by oxidative dealkylation of the nitrogen atom of the tropine moiety of atropine. The paper by Kalser and McLain indicated that excretion of ]4CO2 in the expired air reached a peak about 75 In after the intramuscular injection of atropine and suggested that by 4 h at ter such a dose it Way have been fairly low agate. Possibly, therefore, glucuronidation and oxidative deallglation proceed contemporaneously and in a coupled fashion. The two labeled atropines used by Kalser and McLain (105) yielded the same chromatographically distin8utshable i4C-containing substances in the urine excreted by their subjects. There were four principal peaks of radioactivity, all of which appeared to consist, at least in part, of glucuron1des at the early times of collection of urine. Peaks of radioactivity with Rfe similar to those of labeled atropine and tropine were the most prominent ones in the paper chro~atograme of the ~ 24

urine of these subjects. In samples of urine collected during the first 6 h after 8 dose of labeled atropine, a component of Rf 0.70 seemed to be converted to one of Rf 0.55 by incubation with beta-glucurocidase from beef liver. In summarizing this work on the metabolism of atropine by man, Falser (106) used 'ro~esen's data (89) on the urinary excretion of atropir~e by five subjects to show that there is a f airly broad range of rates of excre tion of atropine; during the 12 h after ingestion of atropine, these f ive subjects exere ted 15. 5-42Z of the ingested dose in their urine. It is not surpristog, therefore, that one subject given labeled atropine intramuscularly exere ted only 35.3: of the injected dose Turing the fires ~ h, whereas the five experiments reported by Kalser and McLain (105) yielded cumulative excretion of injected atropine in urine Turing the same period between about 63X and Il2Z of the inJec ted amount. After omission of the highest value, which came from urine samples that the investigators thought ~gt~t have been handled improperly, there is still a variation between 63: and 833. (Three experiments with three cliff scent sub jects yielded closely similar excretion rates, with a mean of about 80% of the inJec ted dose. A fourth experiment with one of the same subjects yielded the low value of 633.) When the data an 14C still in the body at various times after intramuscular injection of tropine-labeled a tropines were plot ted and analyzed to yield two linear regressions in time, the derived half-ti~es ranged between 1.3 and 2.1 h for the early parts of the curves and between 12.5 and 38.0 h for the later parts. When the data f or the subJec t studied most extensively by Gosselin et al. (104) after intramuscular injection of tropine-labeled atropine were treated similarly, the half-~imes were I.8 and 16.5 h. Apparently, the two types of labeling led to very si~lar estimates of the rate of removal of a tropine f tom the body . Two other studies provide somewhat similar findings. In one (107), coma-producing doses of atropine sulfate were injected intravenoualy, [3~] atropine being mixed with unlabeled atropine to permit a dose of 100 uCi of 3}I to be included in the total dose of atropine sulfate. One psychopath, one epileptic, and 13 schizophrenics were studied. Graph e of data from three patients were presented in the paper. One can received a total dose of 70 fig of atropine sulfate (~.06 mg~kg), another man received 150 fig (2.S mg/kg), and the third patient, a woman, received 270 fig (4.5 mg/kg). All injections were said to have been intravenous, but the curve f or the concentration of ache isotope in the blood of the woman ts like that from a subcutaneous tnJeetion. During the 24 h after Injection, the recoveries of the label in the urine, from the lowest dose to the highest, were 27.1:, 48.0Z, ant 81.~. The concentrations of the isotope in the blood of the two-men decreased rapidly during the first 15 min and adore slowly thereafter; in the ~ 25

woman, the concentration increased to a maximum at about 15 min after the injection and then decreased moderately slowly. In all three patiento' the concentration of the ~ sotope in the blood became zero by or soon af ter 24 h at t er injection. Hayden et al. (108) used a radioi~munoassay method to measure the removal of atropine fro. human plasoaa in four subjects given intravenous injections of 0.32 mg of stroplne. They found a very rapid decrease in the plasma concentration during the first 5 win after injection, followed by ~ slower decrease. By 4 h after injection, the atropine concentration tc the plasma was only about 2.25: of that present 5 min after injection. The initial period of rapidly changing concentration of atropine in the plasma way be a period of mixing in the plasma and equilibration between the plasma and various tissues. A paper by itirtanen et al. (109) presented graphs of concentration of atropine in the serum of two women given 1.3 mg of atropine intravenously. There was an initial very rapid decrease in concentration,, estimated by a radioimmunoassay me tool, followed by a slower decrease . The half-times f or removal of atropine from the blood in these two women were 2.09 and 1.86 h. These times are 8 imilar to those calculated by Kalser and lIcLain (-105~ for the early parts of their curves. The half-times in the paper by Gauzier et al. (107) correspond wore nearly with these calculated by Kalser ant McLain for the later parts of their curves. Because tropic acid was the only substance found after alkaline hydrolysis of urine from subjects given tropate-labeled atropine and a substance that was either tropine or a close relative was the only one f ound in large quantities af ter alkaline hydrolysis of urine from subjects given tropine-labelet atropine, it is apparent that atropine did not undergo drastic metabolic modification in human subjects. Oxidative deall~ylation, with excretion of i4CO2 in the expired air, probably led to the temporary formation of noratropine. This compound may then have been glucuronitated directly or been oxidized further to the hydroxylamine form and then glucuronidated . The f inding that glucuro~dat: ion became a relatively unimportant reaction within about 4-6 h after the injection of atropine into muscle suggests that there is some sort of coupling between oxidative dealkylation of the tropine moiety of atropine and glucuronidation; in other words, glucuronidation may depend on the presence of some particular concentration of substrate, whether that substrate be coratropine or hydroxynoratropine. HUMAN DATA FROM TtIE GENERAL LITERATIJKE Gordon and Frye (27) collected from the literature re port ~ on I] deaths of humans considers d to be due to atropine. Five of these tnvo1,red application of solutions of atropine to the eyes-~one person was thought to have received 1.6 me of.atropine ant another 18.1 ma. I 26

Four deaths were related to ingestion of atropine. One person had taken 198 mg of atropine with 100 me of morphine. ~ 3.5-yr-old child had taken 540 fig of atroplste ant 1,350 mg of phenobarbital. Another person was reported to have taken a daily dose of tincture of belladonna equal to I.2-2.5 mg of atropine for 6 yr before death (probably the highest dose for most of the time); there was nothing in the report to indicate clearly that death was attributable to long-Berm ingestion of atropine (110~. Iwo deaths were connected with Injection of atropine, in one case of 32 ma. On the other side of the picture, Alexander et al. (~) reported that an adult had survived after ingesting 1,000 fig of atropine; Mackenzie and Piggott (112) told of three children weighing 14-24 kg who livet after taking 600 mg of atropice. Report s of cases of serf ous, nonf atal intoxication by tropic acid esters indicate that recovery f ram such episodes is reasonably rapid. For example, Sims (113) reported the case of a 67-yr-old woman who had applied belladonna plasters to her back for a month to relieve backache. ])uring this time, the skin to which the plasters had been applied became excoriated. On the day of admission to a hospital, the patient had disturbed her husband in the early morning by wild and incoherent mutterings. On admission, she had two plasters on her back and was delirious ant flushed. Her pupils were dilated equally and unreactive to light. tier arms were held in flexion and the legs in extension. After removal of the plasters from her backs she regained consciousness withi n about 12 h. Her pupils remained dilated f or more than 36 h, however. She was released from the hospital as f ully recovered, including healing of her skin, on the tenth day after admission. Intoxication by scopolamine with similar signs was reported by Beach et al. (~14~. A 35-,rr-old woman had taken 12 Sominex tablets (equivalent to 3 me of scopolamine). She was in stupor with occasional clonic convulsions. Her pupils were dilated, her mouth was dry, her skin was hot to touch, her pulse was rapid, and she reacted vigorously to minor stimuli. When she was put to bed, her hands and fingers made repetitive plucking mo~remento at the bedsheet. Apparent visual hallucinations, evidenced by avoidance movements, accompanied by screaming and severe disori entat-ion began to abate after 12 h. The patient was discharged on the fourth day after admission to a hospital. Stol1 (26) compared the actions of atropine and methylatropinium nitrate on several functions of the human body. Hethylatropinlum nitrate at 10 ~g/kg produced mean increases (four subjects) in pulse rate twice those produced by the same dose of atropine. In a series of experiments in which subjects were given identical doses (20 ~g/kg) of the two drugs, they were found to have almost identical effects on pulse rate. Hethylatropinium ni bate induces somewhat greater increases in the ~ean- ~ 27 To

systolic and mean diastolic blood pressures and a cocst~erably greater i~cecase in the "cute Produce of the heart than atropice. Atroplne tocr~sed the consumption of oxygen, whereas methyIstropinium nitrate decreased it. HethylatropiMu. nitrate was about t^ce as effective as atropine in increasing skin temperature. Cullumbine et al. (~15) studied the effects of atropine sulfate on health, men. Twenty men each received 1 mg of atropine sulfate by subcutaneous ant intraeuccular infection and 0.5 mg by intravenous injection. Forty men received double doses by the suse three routes of admini stration. The f test sign of action by atropine in many subjects was a slight, temporal decrease in heart rate, followed by a gradual increase. Intravenous injection produced effects more rapidly than the other routes of administration. The Increase in heart rate induced by ~ mg of atroptne sulfate injected intravecousI, Cat stmi tar to, but of shorter duration thud, that t"uced by subcutaneous or intr~uccular injection of 2 - . Subcutaneous injection of atroplne sulfate induced acceleration of the pulse more rapidly, but lese mutiny, than the Awe dose injected intramuscularly. All of 44 men given 5 ~ of atropine sulfate complained of dryness of the mouth and throat, of dizziness, giddiness, or light-headednese, and of difficulty in reading. Other complainto. in order of diminishing frequency, were of difficult urination; of tiredness, lassitude, or sleepinese; of headache or eyeache; and of nausea. After 3 fig of this drug, the only complaint voiced by 12 men so treated was of dryness of the mouth and throat; after 2 ~8 of atropine sulfate, only 37 of 45 subjects complained of dryness of the couth and throat. The most definite objective effect of stropine was an increase tn heart rate. Systolic blood pressure tended to decrease slightly and diastolic pressure to increase. There were also slight, but insignificant, changes in the concentrations of hemoglobin, leukocytes, and blood glucose. Some sub jects had changes in their electrocardiograme that indicated sinus arrhythmia, Peale eniarge~ent, premature a trial contractions, and flattened, coved, or inverted 2 waves. Daily loJectione of 2 fig of atropine repeated for 5 t decreased either the severity of or the subjective response to the changes in secretion by the glands of the mouth and in accommodation of the eyes to light and to nea~cese. Wyant and Dobkin (~16) compared atropine, L-hyoacyamine, and scopolamine for ability to stop salivation induced by intravenous injection of a mixture of carbacho] and epinephrine. Scopolamine was slightly more active in this regard than L-hyoacyamine, and both were considerably more active than atropine. Bachrach ~ Ill ~ surveyed the literature related deco a group of anticholinergic drugs, including atropine and methyIscopola~ontum bromide, and report ed the results of studies of small groups of patients. Atropine was about 4 ~ 28

times as active a stalolytic drug as methyIscopola~onium bromide, but produced more side effects. In a group of Il subjects glares 0.6 eg of atropice orally, nine complained of side effects; blurred vision was the most common of chese; one subject complained of dysuria, and another of nervoueness, dizziness, a" anorexia. 0~, one of eight patients given about 2.4 ag of ~ethyIscopola~ontum bromide complained of blurred vision, and one complained of dy sphagia. Be~xheimer (~) compared a small group of ant! cholinergic truge--including atropine sulfate, scopolamine hydrobro~de, a" methyIscopolammonium bromite--for ability to affect several functional systems of man after subcutaneous injection. Scopolamine hydrobromide (0.2-0.8 ma) was strikingly more active than either methylecopola~onium bromide (0.125~.0 ma) or atropine (0.5-2.0 ma) In inducing dilatation of the pupil and paralysis of accommodation. MethyIscopol~aamon'um bromide was more potent than scopolamine h~rdrobro~ide, which was more potent than atropine, in inhibiting secretion of saliva. The two salts of- scopolamine were equally potent, and more so than atropine, in alowi~ urination. HethyIscopola~ocium bromide was strikingly more potent than atropine sulfate in increasing heart rate. The incidence of drowsiness was 100% with sc opolamine hydra bromide, about 25: wi th methylacopolammonium bromide, and lowest Smith atropine. The effects of methylecopola~ontum bromide and atropine sulfate on heart rate became evident (~-19 ~n) and reached their greatest intensittes (37-40 min) at about the same times, despite the fact that the dose of me thy Isc opolammonium bromide was only 22. 5: thee of atropine sulfate. When equally-effective doses of the c o~pounds were administered, methyl~copolammonium bromide was slower than scopolamine hydrobromide and atropine sulfate in exerting its maximal effect on salivation and on the iri s. The effects of scopolamine bydrobromide on the iris and on accommodation for near vision lasted considerably lounger than those of equal doses of me thy Iscopola~oni~ brogue and atroplne sulfate. Using salivary secretion as the Indicator, tfiratchur ( 118 ~ f ound that atropine and scopolamine were about 2 and 4-5 times as active, respectively, after intramuscular injection as after ingestion. When heart rate was used as the indicator, both alkaloids were about twice as active after intramuscular injection as after ingestion. When the secretion of sweat was used as the indicator, however, ingested acopolamine was nearly as active as that in jected intramuscularly, whereas atropine was about twice as active when ingested as when injected. In a combination of literature review and reports of personal experiences with patients, Schweitzer and Mark (120) summarized the effects of atropine on the intracardiac conduction system and the myocardium. The chronotropic effects of atropine are the best-kno~ ones I 29

on the heart, constating of an early and brief (a few minutes) sio.wing of the heart I, most apparent with small doses or with doses adel~letered by routes likely to yield slow entrance of the substage into the body, and a later and predominant increase in heart rate. The predominant action is due largely to blockade of vagal impulses to the heart, but is contributed to by imp roved sinoatrial conduc tion, increased frequency of discharges by the a~rioventricular node, and factIttation of conduction through that note. Atropine may affect the refractory period of atrial muscle and conduction by the His-Tawara bundle, but these actions are uncertain. Atropine may induce aerial fibrillation, atrial ventricular dissociation, ventricular tachycardia, and even ventricular fibrillation. The last result is particularly likely (122,123) when the work of the heart is increased suddenly under conditions of hypoxemia, as when atroplne is administered intravenously to an apneic person who has Bust been overcome by exposure to an inhibitor of cholinesterases and whose heart ~ti}i Moo Despite the reputation of scopolamine for causing amnesia, Hardy and Wakely (124) found that only 13 of 100 patients given subcutaneous injecelo~as of h~occice at about 6.3 ug/l~g and morphine at about 157.2 ug/Icg hat any amnesia of being show a card Just before induction of anesthesia. Only secret of these patients hat complete amnesia of preoperative events. When atropine at about 9.4 ,ug~kg was substituted for acopolamice in the preanesthetic Fixture, only one of 100 patients had partial amnesia of belong show a card before induction of anes thesis. Der~rent and Karacan (125) examined the effects of intramuscular injections of atropine sulfate (15-60 ug/kg), acopolamine hydra bromide (~.5-6 ~g/kg), and methyIscopola~ocium bromide (~.5O6,ug/kg) on rapid-eye~ovement (REM) sleep and its neurogenic correlate, noc turns] pe"le tumescence (NPT) . The drugs were injected 2 h after onset of sleep. Fifteen healthy male volunteers between the ages of 21 and 35 were used. Scopolamine at 6 ~g/l~g and atropine at 60 ~ag/kg suppressed both REM sleep and HPT. Quaternized scopolamine had neither of these effects. Ostfeld et al. (126) gave oral doses of 10 mg of atropine sulfate tn 100 ~l of water to 10 subjects and recorded the rate of salivation, the grades in the seven moods of the Clyde wood Scale, the elec troencephalogram, and BEG arousal induced by single and repetitive flashes of light before and after drug administration. Atropine i greased heart rate by 60: and tecre aged the amount of saliva produced during ~ min by 6BX. Five subjects complained of mild difficulty in urinating. These effects were ~ximal.at 2 h after atropine was ingested, as were those on BEG arousal by f lashes of light . The last effects consisted of decreasing the duration of the arousal induced by single flashes ~ by 60. 6Z) and repeated I 30

flashes (by 57.~). There was a 81ig~t tendency for arousal by repeated flashes of light to be affected before that by at ngle flashes. In the spontaneous BEG recordings, waves of lower amplitude and longer duration (5-8 per second) replaced the noneal waves at 8-12 per second, beginning at about 30~60 min after atropine ingestion. Obser~ratlon of spontaneous behavior and evaluation of mental status a" mood revealed that atropine shifted behavior in the direction of reduced energy, decreased voluntary speech and motion, reduced attention span, impaired memory of contemporaneous events, and drowsinese. The changes in the spontaneous BEG and 1c behavior appeared at about the same time. Ostfeld ant Aruguete (127) performed a similar study in which 54 volunteers were subjected to subcutaneous injections of 150~800 us of scopolamine hydrobromide. The lowest dose induced moderate brad~cardia, but decreased sali~ratlon. The - highest dose completely blocked the secretion of saliva and seemed to induce sleep, hallucination, and mental disorientation more frequently than the dose of 10 eg of atropine sulfate used by Ostfeld et al. (126~. The larger J08e8 were associated with decreased ability to pert one taske requiring close attention. Comrein et al. (128) report et that intravenous injection of physostigaine sulfate at 40 ~g~kg into six patients in coma as a result of administration of atropine was capable of decreasing the depth of the coma, but that the duration and extent of the chase were variable. The treatment could be repeated, however. In a later paper, the same group of investigators (129) report et that treatment with physostigmine in six cases of attempted suicide with anticholinergic preparations could be monitored with BEG recordings, using BEG alerting as a signal of effect. When the preparations used included overdoses of drugs other than atropine or other anticholinergic subota~es, physostig~ne had no effect. They pointed out that therapy for anticholinergic coma with phy~ostigeine was 3-imite.1 by the shore period of lto activity and by its secondary effects, including premature ventricular contractions and convulsant acti~ri By. Hollender _ al. (130) reported on a woman who complained of auditory and visual hallucinations after taking two Somicex tablets. It developed that she had also taken ch10rpromazine, which has tisti~t anticholinergic activity, and a~triptyline. which has the greatest anticholinergic activi ty of any of the colon tricyclic antidepressants. The woman' ~ pupils were markedly dilated and f iced and did not constrict when drops of ~ FEZ solution of pi10carpine were applied to the eyes. Eventually, she vas found to have been using eyedrops of tropicamide, a potent but nonpersistent mydriatic. Shortly after admission to the hospital, fiche woman had a seizure. This was thought to have been of psychogenic origin; there were no recurrences. With no I 31

drug treatment and prevention of any access to drugs by the patient, her disorientation cleared rapidly. (She claimed, however, that hallucinations persisted during her stay in the hospital of 2 wk. but the investigators considered that she was probably attempting to prolong her hospitalization. ~ Aucamp and Meyer (131) reported that well-controlled epileptics may experience sudden exacerbation of their seizure s if they begin taking drugs with anticholinergic activity (a. in some antacid preparations) or antihistaminic activity (as in preparations for amelioration of colds and influenza). Moskovitz et al. (132) analyzed the hallucinations experienced by a group of 88 patients, 32-84 yr old, wi th Idiopathic parkinsonism for an least 6 mot The therapeutic regimens when these patients began to hallucinate had included anticholinergic drug e with or hi shout amantadine ~ 13 . 9% ), alone ~ 23 . 3: ), and wi th amantadine or an~cichol~nergic drugs or both ~ 62. S: ~ . The hallucinations had been visual for 87.IZ of the patients, auditory f or 35 . 5X, and tactile for 4 0 B:; 25. B: had both visual and auditory hallucinationsO The hallucinations were nonthreat.ening for 72e 72 of the patients and threateni.ng for 27. 3% ~ The investigators believed tthat anticholinergic drugs probably were the principal factors in giving, rise to hallucination. Holland et al. (133,134) gave men intramuscular injections of sterile water, 2 me of atropine sulfate with 500 mg of N-me thy 1-2-hydroxyiminome thy lpyri dinium methyl sulfonate (P2S), 750 mg of P2S, 750 fig of P2S with 2 fig of atropine sulfate, or 2 me of atropine sulfate. The initial rate of absorption was Judged by the time after injection required for attainment of the maximal reduction in heart rate. Five volunteers were used to examine the effects of water, 750 fig of P2S, 750 fig of P2S with atropine sulfate, and atropine sulfate alone; seven were used to determine the effects of 750 me of P2S with atropine sulfate and atropine sulfate alone; 10 were used to compare the effects of SOO fig of P2S with atropine sulfate and atropine sulfate alone. EEGs were recorded af ter the subJec to had reclined quietly on beds for an hour. The injection was then give" -into the upper outer aspect of the thigh, and EGG recording continued for about 2 hr. The air temperature was 21-24°C. All 22 volunteers had control ECGs recorded on day I. After 2 or 3 &, volunteers ~ to 12 were given 2 fig of atropine sulfate; 5 ~ later, they received a mixture of 2 mg of atropine sulfate and 750 mg of P2S. Five of these volunteers were given sterile water 2 t af ~ er the injec tion of the Mixture of a tropine and oxide; a day later, these fire recel~red 750 fig of P2S. The 10 remaining volunteers received injections of 2 me of atropine sulfate and 5 ~ later of 2 mg of atropine sulfate and 500 fig of P2S. I 3;2

The heart rates of the volunteers given no injection, injections of sterile water, or injections of 750 ag of P2S had closely similar variations with time. The graphs of heart rate after injections of 2 mg of atropine sulfate, of 2 mg of atropine sulfate and 500 mg of P2S, and of 2 mg of atropine sulfate and 750 ~8 Of P2S followed closely similar courses with time after injection. These were distinctly different from the first set of lines. One can conclude that 750 mg of P2S alone had no significant effect on heart rate and that addition of either 500 or 750 mg of P2S to 2 fig of atropine sulfate had no striking effect on absorption of atropine from a site of intramuscular injection. The initial rate of absorption of atropine seeme to have been increased slightly by both doses of P2S (mean time to Maxims] bradycardia was decreased from 18.7 min to 12.9 min by 500 mg of P2S and from 15.3 ain to 12.2 -~n by 750 fig of P2S). Martln (135) followed up this work by comparl~g the effects of intramuscular injections of 2 fig of atropiac sulfate alone and mixed filth 750 fig of P2S on heart rate and sweating in nine exercising men. These volunteers worked at 25°C and a relative humidity of 65-751 until their heart rates had risen to \~0-~30 beat/ and then got injections of atropine alone or of atropine and P2S. The exercise was continued for an additions! period. The dose of atropine increased the mean heart rate at the end of the period of exercise by 34.4: and decreased the mean rate of sweat production by 36.[X. Addition of P2S to atropine had no important effect on these changes. The General Practitioner Research Group (136) surveyed the effects of drugs in women in the United Kingdom during the first trimester of pregnancy. Of 661 p ~ gnant women who took drugs of some sort Turing the first trimester, 57 (~.61) had either elacarriages, stillbirths, neonatal deaths, or abnormal children. Antihistamines were involved in 38 of these, barbiturates - in 10, and female sex hormones tn 10. Antthistamines had been used by 76.9: of the pregnant women, barbiturates by 9.2:, ant female sex hormones by 9.~. Thus, wouen who used barbiturates or female sex hormones Turing the fires trimester of pregnancy were the most likely to hare one of the f our types of reproductive accidents considered. ~ pregnancy that respited in a blind infant had involved the use of scopolamine during the eighth week and of promethazine during the ninth week. Scopolamine was the only tropic acid ester used by ~ pregnant woman who delivered an abnormal child irt this survey. Hellman et al. (137) studied the reliability of the response of the fetal heart to administration of atropine to a pregnant woman, the fetal heartbeat being recorded and analyzed by a computer as an indicator of transplacental transfer. Intravenous injection of atropine sulfate at 22 ug/kg durlDg a 2-min period to 28 normal pregnant women resulted first ir1 slowing of fetal I 33

heart rate and then tachyeardia in 26 cases. In two cases, there was no apparent transf er of atropine to the fetus, although both women delivered apparently normal infants. Mellin (138) studied the consequences of the use of drugs during the first trimester of pregnancy by 3,200 pregnant women seen at the Col~bia-Presbrterian Medical Center in New York during 1953-1957. Of these 3,200 pregnancies, 266 were recorded as resulting in malformed infants. Two control groups were established by selecting 266 record ~ immediately preceding and 266 immediately after each record relating to a Informed infant. Drugs were used duri ng the f irat trimester by 53Z of the women who delivered malformed babies, whereas 50.9: + I.9Z of the women in the control groups had used drugs during the first trimester. The difference is not significant. However, when fetal and neonatal deaths among the offspring of women who used drugs during the first trimester and of those who did not were seated separately, only 6.9: of the of fspring of the latter group died in hero or soon after birth, whereas 9.5X of the offspring of the drug°users died before or soon after birth. Atropine was found to have been taken by one woman whose infant died soon after birth among 13 offspring of women who had used this drug during the cri tical period of embryogenesis; three of the other 12 babies of these women were malformed. Of three women who had taken scopolamine during the f irat trimester j two delivered apparently normal children. There is no way of determining from the paper what other drugs the women who used these two anticholinerg to substances may beve taken, except that one of seven babies with both congenital heart disease and another unspecified malformation was born to a woman who had used chlorpromazine, di~enhydrinate, meperidine, morphine, and secobarbital sodium in addition to scopolamine . Hellman and Fillisti (139) extended the study by Bellman et al. (137) of placental transmission of atropice in norma~en by studying 24 pre-eciamptic women, three with ecl~paia, I] with chronic hypertension, and 16 with diabetes. Sixteen of the 24 pre~ecia~ptic patients, tHO of the three ecl~amptic patients, and four of the 24 records on the 3~6 diabetic women gave no evidence of transf er of a tropine to the fetus. Iwo of the negative records on diabetic mothers can be paired with one each from the same woman that provided evidence of transplacental transf er of atropine. Another of the negative recorto pertained to a worn who had yielded three records indicatlug placental transmission of a tropine to the fetus. The general conclusion from botb studies (137,139) was that the atropine Heat of placental function is not sufficiently precise to indicate whether placental function is normal in any given case. Quilligan (140) pointed out that comparison of blood from the umbilical veins and arteries of placentas I 34

of six women who received I.0~.5 ~ of atropine and of 13 women who received no atropine revealed that the Biblical blood from the atropine-treated women had lower POPS than that f rom the control patients, al though the acid-base relationships of the umbilical blood from the two groups were not particularly different. Kivalo and Saarikoski (141) studied tra~placental transmission of atropine by injecting [38jetropine intravenously (0.5,ug/kg) into 2S women undergoing delivery by Cesarean section. Samples of blood were tarn from the an~ecubital vein of the mother and fro. the umbilical vein and artery. The tnvestigatore found that the concentration of the label in blood from the umbilics1 art ery was about 50X that in blood from the umbilical vein. At ~ and 5 min after injection, the concentrations of 3H in the blood from the umbilical vein were 12X and 93:, re spective-ly, of those in the maternal vecous blood at the same times. Atropine seems to cross the placenta rapidly . Onnen et al. (142) estimated the concentrations of atropine in blood samples by bioassay- on isolated ileum from guinea pigs stimulated by acety~cholice. They gave rapid intravenous in Sections of atroplne sulfate to 28 pregnant women at 12.5 ~g/kg and found -that tachycardis in the fetus started 5-10 gin after Injection, at a time ~n the initial rapid decrease in the atropine concentration in the mother's blood had been replaced by a comparatively low rate of decrease. In another experiment, they gave 45 women in labor atropine injections and took samples of maternal and of mixed cord blood at birth. They found that 5-15 min after injection the ratio of the mean concentration of atropine in cord blood to that in maternal blood was I.2 ~ 0.5. This paper also indicates that atropine is transferred across the placenta within matter of a few minutes. In addition to the effects on the reproductive functions of the female, there has been some study of the effects of tropic acid esters and related compounds on sexual function in the male. One paper on this topic (125) has been mentioned above. Horowitz and Gobs (143) have reviewed the literature on drug-induced sexual dysfunction in the male anti have coariuded that say drug with atropis~e-like effects may interfere with penile erection. (They pointed out that impotence induced by antimuscarinic drugs may occur without reduction of libido and may thus be particularly frustrating to the ~ale. ) X~untila et al. (144) reported finding an unusually high incidence of chromosomal aberration ~ broken chromatics, acentric fragmenes, Bicentric chromosomes, etc.) in lymphocytes from a woman who had been vaccinated against rubella and 3 ~ later had received a variety of drugs, including atropine, in connection with an operation on varicose veins; after the operation, she had been given indomethacin for 10 ~ and furose=de for 12 d. The your woman was considered to be in good health. Lymphocytes ~ 35

cultured from the blood of four ocher women who had been vaccinated "gaicat rubella also contained unusually la On e numbers of broken ehromatida, but did not have the other chromosomal abnormalities. The percentages of aetaphase cells with broken chromosomea from these four women were less (by 3-11%) than that to the blood from the first patient (15X). The inveatigaeora Suggested that the mayor aberratioca in the first patient 'a chromosomea may have resulted from a combined effect of the vaccine and one or more of the 10 drugs that she took during and after her operation. They recommended, there fore, that prescribing drugs Should be avoided, if possible, near the time of vaccination for rubella. (In view of the fact that the inveatigaeors had no knowledge of the state of the patient'a chromosomes before her vaccination and operation, their Suggestion and recommendation aces to be on rather Shaky ground.) Hinich et al. (145) attempted to detect mutagenic products in the urine of patients who were taking a large variety of drugs, including atropine, as a part of their therapeutic regimens in the hospital. The urine was tested with and without the addition of a preparation of microsomea from the livers of rata that had been given a 1X solution of phenobarbital iD place of drinking water for 6 t before removal of the livers. Minich et al. fount no evidence of mutagens in the urine of the patients given atropine. HUMAN DATA FROM EDGEWOOD ARSENAL Grob ee al. (147) compared the effects of atropine administered intravenously ant intramuacula~-ly. Four Subjects were used. Two received injectiona of 2 mg of atropine sulfate by the intramnacular route only; one of these aubJecta received only one injection, and the other, two. A third subject received l me of atropine aulfate intravenously on one occasion, 1 me of atropine aulfate intramoacularly on another occasion, and 2 mg of atropine Sulfate intramnacularly on four other occasions. The fourth Subject received 1 me of atropine aulfate intravenoualy on two occasions and 2 me of atropine aulfate intramuacularly on three occasions. Atropine aulfate by either route of adeiniatration induced an increase in heart rate, an increase in akin resiatance, an increase in pupil size, dryness of the skin, and an increased sensation of dryness of the mouth. In general, the effects appeared earlier after intravenous than after intra~uacular injection. Prior administration of sufficient TEPP to increase Sweating ant salivation and to induce anorexia and mild nausea slightly delayed the onset of the effects of atropine and slightly reduced the extent of those effects. Marsulli and Cope (148) gave 40 men single injections of 1 fig of atropine intramuscularly, 28 men two doses of 1 me of atropine 90 win apart, and 20 men single doses of 3 me of atropine. The last group of Subjects contained men who hat received the two toses of 1 mg of atropine at least 2 wk previously and the alogle dose of 1 me still earlier. The inJectiona were into a deltoid muscle. Testing of the subjects began 30 mitt after each dose of 1 me of atropine and 60 win after the dose of 3 ma. A battery of 10 tests, including nine ~ 36

for grading visual acuity, stereopals, and other properties of the eye and one for grading cognition, was uses. Four of 25 subjects to had received intramuscular injections of saline reported silght dryness of the mouth, two complained of headache, and one who had co~plalned of headache also mentioned a slight nambaese of the Ilpe. The 40 open given 1ntrasauscular injection of ~ ag of atropine all reported driers of the mouth; I] complained of dizziness, 13 of feeling tired and sleepy, 10 of dryness of the nose, and eight of having difficulty in reading. The 28 men who received 2 mg of atropine all complained of dryness of the mouth; I? complained of feeling tired and sleepy; 12 reported either that they had slept during the day or had gone to bed earlier than usual; nine complained of f eeling incoord1nated. The 20 subjects given 3 mg of atropine all reported dryness of the mouth; I~ had felt tired or sleepy; 14 recalled either that they had slept during the day or had retired early in the evening; seven reported tat their sleep during the night after the injection had been broken by trivia and disturbing dreams. Three men in the latter group reposed seeing objects in unusual colors; a fourth had a visual sure. No true hallucinations were described. The only significant objective charges seen in this study were a decrease in the ability of the eye to accommodate for near vision and tachyeardia. The mean magnitude of accommodation by the eyes of the 20 Den before they were exposed Lo atropine we. 10.3 diopters (148~. After ~ single dose of 1 ma, -the mean magnitude of accommodation was S.5 diopters. When the same subjects had been given two doses of ~ ma, the mean amount of dioptric change was 7.5. After a single tose of 3 ma, the eyes of the same 20 subjects had ~ mean magnitude of accommodation of 4.8 dloptere. This report provided no data to enable the reader to Sludge whe ther the tachyeardia that followed the various doses of a~ropine was dose-related; data on heart resee are prodded odd after the single dose of 3 ag. The paean heart rate in the 20 men before exposure to atroplne was 75.8 beat/. At IS "n after injection, the mean was Il6.8 beate/min. After another 15 min. the mean was Il7.8 beats/ in. Thereafter, the heart rate decreased gradually, the mean at 7-8 h after the injection being 66.1 beats/. The mean was below that during the control period from the third hour to the eighth hour after injection. Norwich (149) used four subjects to study the effects of intramuscular atropine at 0.07~0.12 agog on the "G. personality structure, heart rate, breathing rate, and pupil diameter. line pupil~diaueter increased by a mean of ll4.3I, the heart rare by a ocean of SI.O:, and the breathing rate ~q a mean (for only three subjects) of 48.5%. There was an incresee in the voltage of the predominant frequency in the EEG, impairment of the alerting reaction on opening the eyes, and increased proportions of slow waves and spindles in the EEG. even though the subjects were awake and responsive to spoken co~ande. Ant four subjects found concentration on a single topic difficult, both attention and logical progression of _ ^_

thought being attenuated. Judgment became poor, sensitivity to the environment was' lessened, the grasp of reality was weakened, and droweiness affected all four subjects. Three of the subjects became irritable, although the good of the fourth was predominantly euphoric. The three irritable subjects had disturbed sleep ant fantastic dreams during the night, whereas the fourth slept well. 41} four subjects had come degree of ataxia; two complained of urgency of urination and congestion of the conjunctivae. The most persistent effect of atropine noticed in this study was paralysis of acc~otation, Inch disappeared gradually during 3-4 ~ after injection. Wechsler and Koskoff (150) used n] ne surgical patients to study the effects of doses of atropine between I.3 and 5.85 mg--administered by intravenous, intramuscular, or intracisternal inJection--on cerebral blood flow and metabolism and on neural and cardiac indexes of normal function. One patient received atropine latracisternally on two occasions, I.8 mg the fires time and 4.2 mg the second time. Three patients were given intravenous doses of t.3~.6 ~ of atropine, and five were given intramuscular doses of 2. 0~5 .85 me . The ineracist ernal injections induced no charges in heart rate or blood pressure within 30 min after injection. The intravenous in Sections produced tachyeardia in the three patients so treated and small increases tn mean arterial blood pressure in two of them, whereas the third had a small decrease in blood pressure within 3 min after tnJection. All five patients given intramuscular atropine developed tachycardia, and four of them (tile only ones whose blood pressures were recorded) developed small increases in mean ar~cerial blood pre ssure . Atropine by any route of administration in these studies induced no consistent changes in cerebral blood flow, cere brat vascular resistance, cere bral consumption of oxygen, cerebral arterio~renous oxygen difference, cerebral respiratory quotient, or pH of blood from the Jugular ~rein. The only significant neurologic change after admintatration of atropine was dilatation of the pupil. Only one patient given atropine yielded any indication of an effect on the BEG: a mistral depression of the alpha component. Ilo morbidity was attributable to the atropine. The authors concluded that imidity in the use of atroplne is not warranted . And:rews et al. (151) used nine sub jeces ta a preliminary comparison of the effects of atropine sulfate injected intramuscularly and subcutaneously in wam (35°C, BOO re let ire humidity) a" coo} ( 16°C, ambient relative hu - dity) environments and 24 subjects in a preliaina~ comparison of the effects of inhalation of 2 fig of atropine sulfate as a aunt and as an aerosolized solution in water. Finally, tO subjects were used to repeat the compari son of the ef feces of atropine sulfate administered by the two methods of injection in ware and coo] environments ant to compare the affects of 2 and 5 fig of atropine sulfate inhaled as a dust and an aerosolized aqueous solution in the warm envlro~ent. The report tid not state whether the same subjects were used in more than one part of this study. The results of the study indicated that 2 fig of atropine sulfate had practically identical effects on heart rate and sweating, whether injected intramuscularly or subcutaneously.

The mean lacrease in heart rate after injection of 2 ag of atropine sulfate to the wane environment was 42.7:, whereas that to the cool environment was 31.3%. The mean increase in heart rate after inhalation of 2 eg of atropice sulfate as a dust or an aerosolized solution wee 23.71; the mean increase after lshalation Of 5 ~8 of the drug wee 6~8.11. The increeaea with the two types of diaperalon of atropine sulfate were essentially identical for the two doses used. It is apparent that inhalation of atropine sulfate is not as effective a method of delivery, with heart rate as the indicator, as intramuscular or subcutaneous tn jection. Although tnhalations of 5 me of atropine sulfate latticed an increase in heart rate coo~parable with that induced by the injection of 2 ma, all subjects reported headaches, glddinese, tlretneas, lethargy, ant muscular wea~esa or incoortination after inhaling the 5- mg doses. it took 24 h for all the subjects to feel able to return to their usual acti~ritiea after inhaling the larger dose of atroplne sulfate, ant several reposed visual hallucinations ~ euphoria, and tysuria. The authors concluded that the use of inhalators for self-admi~atration of atropine sulfate would ~ impractical and that intramnacular injection would be the easicat method to teach to nomedical persons. White et al. (152) gave 12 subjects intramuscular doses of atropine sulfate at 22.6-134.0 ug/Icg (D'ean, 50.8 ug/lcg). The mean blood pressure before adminiatration of atropine sulfate was 123/79; after administration, it was 129/91. The mean rate of the radial pulse wee 69 beet s/min before admintatratlon of atropine; within 30-60 ~ after injection, it was 61.5 beats/mini it increased later, reaching a aeon maximum of 100.5 beatsJ~n. The Dean breathing rate before atropiDe wee administered was 15.1Imin; after atropine had been admlaistered, it rose to 17.1/~. The mean diameter of the pupil was estimated to be 4.8 ~ before administration of atroplne; under the influence of that drug, it 1DC~ea8ed to 6.5 mm. Bight of the 12 subjects had extensions of the near point under the influence of atropine, the subjects glveD the two largest doses having the greatest increases. All 12 subjects complained of dryness of the mouth; fits became re stlese; two complained of sensatlo" of heat on their alcins, although they were in an air-cooditioDed room; 10 were obeyed to become drowsy; 10 found that waling ~ straight line was difficult; and five had malaise, headache, vertigo, and slight nausea. These complaints had disappeared by the following morning. Other effects of atropine sulfate seen in the 12 subjects who took part in this experiment, in order of decreasing incidence, were aleepinese, shortened attention span and blurred vision, dreaming, eye irritation, lack of desire for mental work, and url nary urgency. The usual charges in the BEG pattern after ad3niniatration of atropine were a flattering of the record, a decrease in frequency, and intermittent blocking of the alpha rhythm. There was a decrease in alerting on opening the eyes. Five subjects had notable improvement in photic driving after administration of atropine, and fire others had alight improvement. Only one subject had a decrease in re sponse to photic stimulation.

In attempting to deprive a battery of tests useful tn assessing the effects of chemicals on h' - ~n capabilittes, Elkln et a1. (153) gave Il subjects intramuscular tnJections of scopolamine at 12 /kg after the sub.5ec~ce had spent a day gaining familiarity With the test battery. Four subjects received placebo inJectione. me only pare of the test battery that may have been affected by the placebo was ~ nual~exterit~r test in which the subject moved as many blocks as possible in 30 ~ from one hole to an aJ]olr~iDg hole. The number of blocks aoved 0.75 and 2.25 h after the placebo was about 8.3: greater than Ache umber moored before administration of the placebo and 4.25 h after the placebo. Scopolamine decreased both near am far visual acuities' the effect on near visual acuity being much the greaecr. In the manual~exterity test, scopolamine decreased by 38.6: the number of blocks moored, this decrease perelating for 2025 h after the injection of scopolamine and then dIo~nishing gradually. By 8.75 h after the injection, the Humor of blocks moved was only sIlghtly below the baseline measurement. ltean grip strength was lowered from about 56 Ice to about 48 kg by the dose of scopolamine. Scopolamine lowered the mean duration of balancing on a beam on one foot from about 17 to about 5 s. The ocean nuaber of additions performed within 3 min decreased from about 42 to about four. The mean number of digit o that could be recalled after auditory presentation decreased from about 7.l to about 5.~. The mean ability to estimate 10 was altered only sIlghtly by scopolamine in the direction of underestimation. However, the variance of this estimation was increased by about 128Z by scopolamine. The mean reaction time (the subject pressed a switch as soon as possible after receiving a visual cue) increased from 0.22 to 0. 38. The authors concluded that the OSt dramatic chase ~s in near visual acuity. Grip strength, simple reaction tissue, and estimation of elapsed time were affected ODly slightly by acopola~ine. Kitzes et al. (154) analyzed the records of lS subjects who had been given intramuccular injections of atrop1ne at 32-125 ug/kg a" who weighed 57.2-90.0 kg. The results on the Thurber Facility Test (completion of as ~~y additions ss possible during 3 min) were used to grade each subject's response to the drug. No correlation between body size and decrease tn the Number Facility Test was found. Cummings and Craig (IS5) exposed IO men to temperatures up to about 52°C to induce ~ range of rates of sweating and then subjected them to a varies, of doses of atropine sulfate by intravenous injection of various priming doses followed by infusions of various co~centratione of the drug for 13-53 min. The total dose of atropine received by ~ subject varied between zero and I.6 ag; the report was unclear on whether this value is for atropine alkaloid or atroplne sulfate. Higher rates of sweat)", induced by exposure to high environmental temperatures, were found to require larger doses of atropine to inhibit sweating. Sweating recovered rapidly (6-74 ~n) after the end of infusion of atropine sulfate, particularly at the higher teepera~cures.

Kligman and Copelan (156) reported that 13 subjects had been exposed deco atropine. No details or results of the study were given. Crowell and Ketchum (157) used 33 Boreal male volunteers to assess the ability of phy-sostig~ine to antagonize the deliriant activity of scopolamine. The volunteers received intramuscular injections of acopolamine hydrobromide at 24 g/kg; subgroups of the Intoxicated men received physostig~ine salicylate intramuscularly at 50 ug/kg 15, 30, or 90 min after injection of scopolamine. Whose treated 15 min after scopolamine did not have any Immediate reveres] of feelings of fatigue and drowsiness and gradually developed a typical delirium, but at a lower rate than those who had not been given physostig~ine. Phy~ostigmine lowered the heart rate in these men and maintained it on a plateau below thee in the untreated group Until the heart rate in the untreated group decreased to equal that in the treated group 3-4 h after the scopolamine had been administered. Physostig~ine had only a minor and brief effect on the diame~cer of the pupil, but decreased significantly the decrement in the Number Facility Test due to the dose of scopolamine during the 2 h after its admluistration. The subjects treated with physostig~ine 30 min after the injection of scopolamine improved drastically and rapidly in their pert ordnance of the Number Facility Test . These men relapsed into delirium about 3 h after administration of physostig~ine and then recovered at about the same rate as the untreated men. The effects of physostigmine on heart rate and pupil diameter were similar to those in the group treated earlier in the intoxication. The men treated 90 min after injection of scopolamine, af ter development of a full-blown intoxication, improved dramatically and rapidly, the change being noticeable within 10 min of injection of physostig~ine and becoming maximal within 30 min. At that time, the subjects were alert, well-oriented, and logical. Their pert ordnance in the Number Facility Tes~c decreased slightly about I.5 h after the dose of physostig=ne, but did not fall to near the level of performance of the untreated group at that time. The pert ordnances in the Number Facility Test of the untreated group and the group treated with physontigo~ine became essentially identical at about 6 ~ after the dose of scopolamine. The heart rate decreased within 30 min after the dose of physostigeine, but this drug had little effect on pupil size. Baker et al. (158) summarized their studies with 82 volunteers given scopolamine. No details of the experiments or of their results were given, but the individual subjects were said to have reacted quite differently to a given dose of scopolamine. After extensive analysis of the data, the authors concluded that research was needed to identify the factors that cause "subject by treatment" interactions ant that means for controlling for them were necessary for obtaining consistent effects by a given chemical. Krieger (64) examined the effect of indec ted atropine on the circadian ~rariatlon in the concentration of ~ I,1

17-hydroxysteroids in human plasma, using two healthy male subjects. One subject was given a subcutaneous injection of 3.0 mg of atropine, the ocher a similar injection of 3.5 ma. Three other subjects took oral doses of 2.5 mg of atropine. None of these subjects gave any evidence of an effect on the usual pattern of variation in plasma 17-hydroxysteroide. Me author pointed out that the dose of atropine used in these human studies was only about one-tenth that used in experiments with cats in which atropine had been found to block the nocturnal rise in plasma 17-hydroxysteroid concentration, when administered before the time at which the circadian rise could be expected to occur. Safer (1~59) investigated the possible interplay of sleep deprivation and scopolamine intoxication in producing deterioration of hen abilities. Men were deprived of sleep for one or two nights and were then given scopolamine hydrobromide intravenously at 5 or 10 ~g/kg. Behavior was rated with a checklist. The performances of the subjects in the Number Facility Tests a manual-texterity testy and a vigi lance test were evalua~ced. Eleven subjects were given scopolamine at 10 ug/kg9 10 were deprived of sleep for a night and given intravenous saline, six were deprived of sleep for a night and were given scopolamine aft 10 ug/kg, two were given acopolamine at 5 ~g~k~, four were given scopolamine at ~ ug/kg after being deprived of sleep for a night, and four were given scopolamine at 5 ,ug/kg after being deprived of sleep for two night ~ . The combination of lose of sleep f or one night and Scopolamine at lO,ug/kg had a more than additive effect on performance in the Number Facility Test and considerably more effect on manual dexterity than the scopolamine alone. The combination also produced hallucinations in about 2.7 times as many subjects as the same dose of scopolamine alone. The results with the lower dose of scopolamine were similar Lo those reported above, but less strikingO The men deprived of sleep for two nights became so somnolent after the dose of scopolamine that the tests could not be run. The effects of large lotra~uscular doses of at.ropine on the electrocardiogram were examined by Hayes et al. (160, 161), with six healthy male volunteers. The volunteers were given atropine sulfate at 175 ~g/kg. By 2 win after injection, there was ~ lowing of the heart wi shout any ~ trt king change in the EGG record. At 5 In after the dose, the Pwave was engulfed by the QRS complex, and an A-V nodal rhythm was instituted. This A-V dissociation was temporary, a sinus tachycardia becoming apparent at 6-7 min after the injection and persisting for about 3 h. Four volunteers who had received scopolamine, three who had been given atropine, and nine who had received no chemicals were interviewed and examined at some unknown time af ter their service as experimental subjects (162). None of the subjects f elt that he had suf fared physical or psychic ink ury as a result of serving as a subject. Blood counts for all the former subjects were within the normal range. Three of the former control subjects ant one former subject who had been I 42

given both scopolamine and a Salt of LSD hat up to 10 white cells per high-power field in their urine; that suggested low-grade urinary tract infection. One foneer subject who had been given a~cropine had a mildly positive test for urinary glucose (there are so see ny possible causes of such a situation that no precise meaning can be attached to it). Me of the format control subjects had an increased activity of GOT in his serum, and another had a slightly increased concentration of BUN. No abnormalities of serum cheotistry were recorded for the former subjects who had been given atropine or scopolamine. This study revealed, therefore, no definite evidence of long-term physical or psychic injury by esters of tropic acid. Klapper et al. (163) examined the records of eight subjects who had been given intramuscular injections of atroplne at 62,ug/kg, six who had been given atropine at 104 ug/kg, and six who had been given Scopolamine at Il.8 ~g/kg. The performance of each subject on the Ilumber Facility Tese after injection was compared with his scores on the three Fragility scales and 10 standard scales of the Itianecota Multlphasic Personality Index (+PI). The bent correlations between performance in the Number Facility Test and scores on scales in the HI for the men given atropine at 62,ug/kg were with those in the hypochondriasis and mania scales. The best correlations for the men given atropine at 104,ug/kg were with Scores in the lie and mania scales. For the men given scopolamine at 11.8,ug/kg, the best correlations were with scores in the hypochondriasis and lie scales. If atropine and scopolamine are taken as representative of the group of tropic acid esters, the best indexes of performance in the Number Facility Teat under the lnf luence of such esters are the scores in the lie, hypochondriasis, and mania scales of the ~Pl. After a similar analysis for fire antlcholinergic compound, three other substances with atropine and scopolamine, Clapper en a,. (162) decided that scores in the positive tes~c-tal~ng attitude, hypochondriasis, and mania scales of the MMPI were poolti~rely related deco performance in the Number Facility Test. Sidell et al. (164) reported that intravenous injection of the potent cholinesterase inhibitor, S-( diisopropylueinoethyI)~ethylmethy~phosphonothioate, at T.5-1.7 ,ug/kg into men 1.5 h after intramuscular injection of acopola~ne hydrobromide at 24,ug/kg was effective in reversing the perf ormance and cognitive decrements produced by the scopolamine. If the anticholinesterase was administered 30 min after scopolamine, the initial therapeutic benefit was less than when it had been given at 90 min. This anticholinesterase had a more durable effect than physostigmine, its antagonistic activity per8iStitig as long as the agonistic activity of scopolamine, whereas that of physostigmine had been found by others to last for only 2. 5-3 h. Ketchup et al. (24,165,166) reported the results of intramuscular injection of atropine sulfate at 32-175 ~g/kg, of intravenous injection of scopolamine hydrobromide at 5-24 ug/kg, of intramuscular injection of Scopolamine hydrobromide at 5-24 ~g/kg, of intramuscular injection of Scopolamine methy~bromide at 5-30 ug/kg, and of attempts to find ~ 43

aneagonisto of intoxication with these chemicals. The research involved 127 individual experlmente. The EDsos of aeropine sulfate to initiate the following effects to recognizable extents were: poorly coordinated, 89 ~g/kg; attention span shortened, 95 ~g/kg; stumbling gait, 106 ~g/kg; ~e8~1eS8De888 126 ug/kg; temporal confusions 130 ~gtkg; inability to obey apple orders. 13S ~g/kg; and haliuci~ation9 169 ~g/kg. Although the ratios between the EDsos of atropine and of the other anticholinergic compounds for the production of the different effects varied somewhat, intravenously injected scopolamine hydrobromide was about 8.7 times as active as atropine sulfate and, when injected intramuscularly, about 7.5 times as effective as atropine sulfate. Scopolamine methy~bromide was about 4 times as active as atropine sulfate, except that no dose of the foneer substance produced as much as a 90: decrease in performance in the Number Facility Teat, so comparative potencies had to be assigned on the basis of less than- Biaxial actions by atropine and scopolamine. Therefore, scopolamine methy~bromide was only about 53X as potent as the hydrobro~aide. Intramuscular in jections of scopolamine hydrobromide at 24 ,ugJkg and of atropine sulfate at 175 ug/kg induced approximately equal decrements in performance in the Number Facility Test, except that the maximal effect after scopolamine was attained in about half the time required to reach that - after atropine. Intravenously injected Scopolamine hydrobromite produced its 3`axio~a1 effect even more rapidly than that injected intramuscularly. Scopolamine methy~bromlde was slower in exerting its maximal tachyeardial effect than Scopolamine hydrobromide, but no information about the rapidity of its action on performance in the Number Facility Test was provided in any of the three reports (24,165,-166~. The increase in heart rate induced by scopolamine methy~bromide was greater than those induced by the same amounts of Scopolamine hydrobromide or atropine sulfate. The decreases in performance in the Number Facility Test induced by atropine or scopolamine were antagonized rapidly by intramuscular injections of physostigmine at 15-60 ug/kg, but the antagonistic effect wore off after 3-4 h. Neostigmine had no immediate effect on performance in the Number Facility Test, but may have Screamed slightly the rate of spontaneous recovery 5-3~0 h after the intramuscular dose of atropine sulfate. With respect to ache tachyeartia induced by atropine, neostigoIine may have exerted a prompter and more potent bradyeardiac action than physostig~ine. Physosti~mine also antagonized the decrement in performance on the Number Facility Te St i educed by scopolamine . Me organophosphorus anticholInesterase compounds, OFP and Sarin, had some antagonistic effect against the decrement in performance in the Number Facility Test induced by scopolami ne, but these effects were less striking, although more persistent, than those of physostign~ne. Tetrahydroaminoacridine, also an inhibitor of cholinesterases, had some antagonistic activi ty against the decrease in performance in the her Facility Teat due to scopolamine. Intramuscular injection of chlorpromazine after a ~ 44

8~1 608e of scopolamine hydrobromite (8 ug/kg) increased the decrement in performance in the Number Facility lest due to the scopolamine. When 50 mg of chlorpromazine was injected intramuscularly after intramuscular scopolamine hydrobromide at 8,ug/kg, the resulting decrease in the score in the Amber Facility Test approached that induced by scopolamine l~ydrobromide at 24 Agog alone, although chlorpromazine bar itself induced only a alight decrease in performance. Perphenazine, another derivative of phenothiazine, had effects similar to those of chlorpromazine. Stroll (167) reported that absorption of atropi-ne from an intramuscular site of injection in man was prolonged when it was mixed with a 30: solution, but not when mixed with an 18: solution, of pralidoxlme chloride. Because absorption of atropine was also delayed when it was mixed with an 8.5: solution of sodium chloride, Sidell suggested that the delayed absorption with 30Z pralido~cime chloride may have been due to osmotic effects,-rather than to any specific effect of the oxime. However, as pointed out -by Sidell, although absorption of atropine from the mixture with 8.5: sodium chloride was delayed, eventually this mixture caused the same mean increase in heart rate in the six volunteer subjects as atropine alone at the aame time after injection. In contrast, neither mixture of atropine with pralidoxime chloride accomplished this; the mixture of atropine with the I8X solution came closer than the mixture with the 30X solution to achieving the same increase in mean heart rate as atropine alone. Therefore, pral1doxime may have had a dose-related effect on the total absorption of atropine. Whether this is due to a simple osmotic effect on the absorption of atropine, with some of the alkaloid held at the site of injection by unabsorbed pralidoxime chloride in solution, or to come more specific effect of the oxime is Dot clear. Sidell ( 168) studied the effectiveness of intravenous administration of physostig~ine as an antagonist of the central ~ Number Facility Test) and peripheral (heart rate) actions of atropine and acopola~ ne . The report simply reiterated the effectiveness of physostigmine as an antagonist of the central alkalolde and demonstrated that repetition of ad-~nlatration of physostigeine after disappearance of the effect of a first dose renewed the antagonism of persistent central actions of the alkaloids, but, like the first dose of physostig~lne, had only a minor effect on the tachyeardia due ~ o the alkaloids. actions of these two SUMMARY ~- This review of the general literature portrays the tropic acid esters with which this report is concerned as compounds that are not particularly lethal in single or repeated doses. The members of the group that are tertiary amines may affect psychic activity in comparatively small doses, but there is fairly rapid adaptation to repeated doses. Removal of the compounds from the body, so far as it has been studied, seems to be comparatively rapid and to occur by a combination of urinary and fecal excretion and metabolic transformation. The ~ As

transport of Intact atropine through the tubular epithelium of the chicken is by the cation transport mechanism and i. reduced by drugs and ehemicale that inhibit that transport process: quitllne, choline, cocaine, and cyantne dyes (146~. Symptoms and signs of the action of the tropic acid esters on h''man subjects include complaints of mouth dryness and decreased secretion of saliva; increase in heart rate, possibly preceded by a short period of decrease In heart rate; dryness and flushlog of ache skin' particularly in the "blush area"; difficult urination; ataxia; disorientation; hallucination; delirium; dilated pupils, possibly nonreactive to light; prostration; and coma. The greatest hazard to life arises from suppression of the ability to secrete sweat, which can give rise to fatal hyperthermia if body temperature is not controlled artificially during hot weather or strenuous activity. Another source of hazard to life arises from the effect of the compounds other than scopolamine and its qua ternary form in accelerating heart rate and factlitating Intramural conduction and transmission of impulses. These actions may result in serious arrhythmias up to and including ventricular fibrillation. The quaternary amine forge of the tropic--acid esters in which we are interested are more active in some respects than the tertiary amides, as far as peripheral actions are concerned. This is especially true with respect to actions on nicotlutc effecters in ganglia and striated muscles . The quaternary amine s penetrate into the central nervous system poorly, but, once there, affect muscarinic effecters ta the same way as the tertiary amines. The anticholinergic compounds that are esters of tropic acid are not known to have any important mutagenic, teraeogenic, or carcinogenic actions. There has been little study of long-tem toxicity of these compounds, however, so one cannot say categorically that they have no carcinogenic activity. The rapidity with which atropine is removed from the body, although it cannot be taken as proof of nos~carcinogenicity, does Agitate against a high probability of such an action by this compound. ESTERS OF BENZILIC Ale The master file of the Board on Toxicology ant Environmental Health Hazards lists four compounds that are esters of benzilic acid. One of these compounds seems to be an isomer of another~-both are benzilic acid esters of- tropine. Of the 12 reports that concern administration of compounds of this group to volunteers, either at Edgewoot Arsenal or at contractors' facilities, Il are related to the results of studies ulth quinuctidiny! benzilate and three with tropinyl bencliate (these three reports are concerned exclusively with the ~ isomer of the cater) . Accordlugly, the literature review in this section centers on these two compounds insofar as possible, and especially on quinuclitinyl benzilate. The first compound made in this group of esters of benzilic acid seems to have been N - iethylami-noethy] benzilate (benaceyzine), whose synthesis was reported in 1938 by ~ 46

Horenseein and Pahlicke (169~. The a~rnthests of tropiny! benzilate was described in 1952 by ~omatka et al. (1703. In 1952, Sternbach and Kaiser (171) reported methods for producing several bicycllc alcohols, including 3-quinuclidinol. These alcohols were then used to produce esters with organic acids (~72), including benzilic acid. The esters were ex~ined for their ability to induce relaxation of isolated rabbit intestine that was made to contract by acety~choline; the esters of 3~quinuclidino] were found to have greater spasmolytic activity than the corresponding esters of diethyla~inoethanol and other bicyclic alcohols. The 3~quinuclidinyl ester of dipheny} acetic acid was prepared in its ~ and ~ foma. Me levorotatory ester was found to have most of the activity, the dextrorotatory ester having only very slight activity. The toxicities of the isomers were stated to have been identical, but no data on toxicity were reported. A paper by Randall et al. (173) gave the intravenous LD,os for the mouse of the L isomer as 29.5 mg/kg and of the I) isomer as 27 mg/kg. The synthesis of ester. of 2-tropanol was descri bed by Archer and Bell (174) in 1961. In 1971, Atkinson and Mc~tchie (175) reported methods for synthesizing all four of the isomers of 2-tropanol. The chemical literature seems not to reveal when the production of L-2-tropinyl benzilate actually occurred, but it was probably in 1971-1972. Randall et al. (173) assesses the potencies of 3~quinuclidinyl benzilate hydrochloride ant N-methyl-3-quinuclidinium benzilate bromide in several different activities. 'rhe LDsos of 3-quinuclitiny! benzilate for the mouse were about 1.46 and 2.04 times those of the quaternized compound of ter intravenous ant intraperi toneal injection, respectively. In producing relaxation of isolated intestinal Smooth muscle stimulated by acetylcholine, 3~quinuclidinyl benzilate was somewhat more effective than a tropine. me 3~quinuclidinyl benzilate had Rome activity in relaxing spasm of intestinal smooth muscle induced by histamine and was quite effective in antagonlsin8 that induced by barks. It was also effective in preventing contraction by bronchial smooth muscle on exposure to aerosolized methacholine and in overcoml~g the bradyeardia and hypotension induced bar injection of acety~choline or by electric stimulation of the vagal nerves. A dose of 2 mg/kg prevented the response of the cat nictitating membrane to electric stimulation of the cervical sympathetic nerve. The chemist who made 3-quinuclidiny] bencliate took about 0.5 mg of the compound and experienced not only dryness of the mouth and mydriasis, but also a sensation of weakness in the knee s. De spite having recel~red several subcutaneous doses of neostigmine, he Slept f itfully that night and spoke incoherently during waking periods. On the next morning, he was still unsteady on his feet, and his pupils were dilated. By the second morning after he tad taken the compound, all symptoms except those due to persistent mydriasis had disappeared. He revealed that he had felt confused arid that t ime had seemed to pass very slowly. Two other employees involved in work with 3~quinuclidinyl benzilate had similar llinesees, with apparent hallucination in one patient (176~.

In addition to those three occupational exposurea, two of 8~ p8tieDt8 given 0~1 mg of 3-quinuclidinyl benzilate three times a day as a treatment for parkinsonism hat nocturnal confusion (176~. STUDIES IN ANIMALS In an attempt to explain these confusional states9 Schallel: and Smith (177) studied the effects of fire spasmolytic substances, including 3~qutnuclidinyl benzilate and its quaternized font, on the electroencephalogram, the electrocardiogram, and the blood pressure of the dog. Intravenous injection of 3~quinuclidiny! benzilate at 0.01 mg/kg decreased the dominant frequency of the BEG of two of fire dogs; a dose of 0.} mg/kg enlarged to three the number of dogs in which the dominant frequency of ache EEG was decreased. Larger doses, up to 10 mg/kg, did not further enlarge the proportion of dogs with a decreased dominant frequency. The dose of 0.l mg/kg induce-d bradyeardia in one of five dogs. A dose of 10 mg/kg induced bradycardia In all five dogs ant resulted in death due to cardiac arrest in No of five. The only dose of quaternleed 3-quinuclidinyl benzllate that induced any alteration in the BEG was 10 mg~kg, which resulted in a decrease in the dominant frequency in the EE(; of one o f f ive dogs . Thi ~ dose also induced bradyea rdia in three of five anlmala snd hypotension in all five. No toga given thi ~ dose of Note thyI-3~quinuclidinyl benzilate bromide died . Intravenous doses of 0.l and I.0 mg/kg caused tachycardia in one of five and two of five dogs, respectively. Before proceeding with a discussion of the effects of 3~quinuclidinyl benzilate, it may be worth while to look an the available information on the lethality of single doses of the benzIlic acid eaters with which we are concerned. Table I-2 is based on information supplied by Boffman-LaRoche, Inc., the Sterling~inthrop Research institute, and a Ember of investigatore ~36,173,176,178 IB9). The "r~zilic acid esters of both diethyla~noethano] (179, I8O,1SS) and tropine (182,183) were found to be capable of overcoming spasms of "ooth muscle induced by acety~choline and of inducing relaxation of smooth muscle, such as the circular muscle of the iris. Both compounds had some local anesthetic activity, both produced changes in the EEGe of experimental animate similar to those induced by atropine, and both had slight antthist~ninic activity. I.isunicin (~87) reported that tropiny} benzilate increased the effectiveness of other antispasmodic drugs, such as tropiny! diphenylacetate (tropactne), is antagonizing the tremor induced by subcutaneous nicotine. Larsen (181) fed rats diets containing either 0.01Z or 0.05: of the benzilic acid ester of ttethylaminoethano} from 28 t of age until spontaneous teeth. Another group of rats was fed the basal diet. The rats fed benzilic acid ester gained weight slightly faster than those fed the normal ties, but tied younger than the controls (means of 24 ant 26 mot compared with one of over 29.5 mo). No striking pathemas in the animals due to the benzilic acid ester were reported. I 48

3-QUINUCLIDINYI. BE=ILASE ( BE, EA 2277) McNamara (184) reported that rats placed into at3~0apheres contalai~g 3-quinuclitinyl benzilate tispereet as a tust, as a thermally generated smoke, or as an aerosol of a 1: solution of the benzilic acid ester in methyleae ttehlorite for concentration-time (Ct) products up to 29,508 mg.min/~3 experienced no permanent effects from the exposures, except occasional teethe randomly distributed with respect to intensity of exposure. One of four guinea pig a exposed to a Ct product of 10~123 m8.min/~3 and two of four guinea pigs exposed to a Ct product of l4~748 mg~min1~3 tied. Mice exposed to an aerosolized I: solution of the ber~zilic acid ester in methylene dichloride at Ct products up to 14,137 mg.minIm3 did not experience any clearly dose-related mortality, but 69 of 70 mice exposes to thermally generated awoke of 3~quinuclidinyl benzilate at Ct products of 8, 213-9, 331 mg.min/~3 ties. McNamara (184) reported that tally subcutaneous doses of 3~qulnuclidinyl benzilate at up to 150 uglkg on 5 days of each week for 3 wk induced no evident Bights of toxicity in mice and guinea pigs. Dogs given daily intravenous doses of lOO,ug/l~g for 14 consecutive days had the same LDso as togs that hat not been pretreated, but the time between injection of the daily dose and the appearance of ataxia increased from 4 min to 14 win during the period of pretreatment. In dogs trained in a conditioned-escape routine and then given grated intravenous doses of 3-quinuclidinyl benzilate at up to 12~5 ~g/kg, there was no effect on performance. A dose of 25,ug/kg resulted in failure to escape by four of four dogs. The same report (~84) a~arized data on- the effect of intraperitoneally injected 3~quinuclidinyl bencllate on spontaneous motor activity of the rat. When the data were plotted and the curve extrapolated to zero change in spontaneous activity, the maximal dose that could be given without altering spontaneous activity seemed to be about 0.12 mg~kg. No rata were glares tally injections of saline, however, so a decision on what portion of the observed changes was ateri butable to the compound, rather than to the procedure, is not possible. Studies of the absorption of 3-quinucIldinyl benzilate from solutions applied in a =xture of alcohol and cresol to the clipped ·kln of five species of aware, summarized in the came report (~64), found that doses of up to 500,ug/kg produced acacia in the cat after about 30 min and in the dog after about 55 min. No ataxia was produced tn the goat or the monkey (probably rhesus) by the highest dose; the highest dose applied JO the skin of the rabbit was lOO,ug/Icg. Doses of 50.ug/kg applied to the skin of the rabbit, of 100 ug/kg applied to the skin of the cat, of 250.ug/kg applies to the skin of the mookey, and of 500,ug/kg applied to the skin of the dog and the goat were the lowest 608e8 found to have effects on the eyes. In all species except the monkey, recovery from the ocular effects was stated to occur overnight. In the monkey, the ocular effects apparently persisted for 2 d or more.

In the tog, an intravenous dose of lO.ug/kg produced a mean increase in four dogs of about 163: in heart rate, which decreased slowly Turing art obeer~ration period of up to 6 h, but was stitI evident at the end of that period. By 24 h after the dose, the heart rate was back to normal. A dose of 12.5,ug/kg, but not one of half that amount, decreased by 20X the mean time Turing which dogs would run on a treads. None of the experiments s''~marized in this re pore (184) seeme to have involved any persistent effect of exposure to 3-quinucIttiny! benzilate in surviving antmale. Neither tit the report of Wal (186) mention any conalstent toxic effects of this compound. Ketchum et al. (~89) reported that rats and dogs with whole-boty exposures and rabbits and monkeys with head-only exposures to clouds of 3-quinuclidinyl benzilate dispersed in the field at wind speeds of 8-ll ~ph developed such effects as ataxia, dyspnea, mydriasis, c~rcloplegia, tachycardia9 sedation, hyperactivity, and nasal stuffinese. The most persistent of these effects was cycloplegia, which persisted in three of five monkeys for more than ~ do ^st other effects disappeared within 48 h after exposures. No study of long-tem administration of 3-quinuclidinyl benzilate to experimental animals has been found. The closest approaches to such a study are I-yr studies of the results of feeding the methyl bromide of 3-quinuclidinyl benzilate to rats in the diet (190) and of savaging dogs 5 d/wk with the same quaterna~y amine sale of 3-quinuclidiny} benzilate (191~. RBLS were fed diets containing the quaternary amine salt at concentrations calculated to yield daily doses of zero, 5, 25, and 50 mg/kg. Dogs were garaged with the quaternary salt at zero, I, 5, and 25 mg/kg. In both experiments, no evidence of dose-related toxicity was seen. The livers of the male dogs that had received tally doses of 5 and 25 mg/kg seemed to be larger than those of the controls (3.0X and 3.2: of body weight, respectively, verius 2.7: for the controls), but were normal in structure on necropay and in function (glutamic-pyru~ric transaminsee and alkaline phosphatase activities in serum and Bromsuiphaleln retention within normal lignite). Although the compound used in these two studies was sot 3~quinuclidiny! benzilate and probably was absorbed from the intestinal trace lese rapidity and less completely than than copout, it contained the same combinations and arras~gemento of atoms. Therefore, the negative findings with respect to organ and tissue damage in the two studies may indicate that Pillar experiments with 3~qulnuclidinyl bensilate would result is no chemically induced pathemas. me great avidity of aitochondria from rat brain for 3-quinuclidiny} benzilate has been mentioned previously (56~. Lareson et al. (192) recorded the infrared spectra of seven esters of benzilic acid, of eight esters of 3-quinuclidinol, and of atropine and scopolamine and attempted to correlate the relative a~crengtho of the intramolecular hydrogen bond with the threshold doses of the various compounds in producing paychotomimeelc effects in toga. The data of Albanus (42) on the psychotomimetic activities of anticholinergic compounds were used for the comparison. Of the esters of benzilic acid, I 50

the moat active wee 3-quinuclidinyl benzilate. According to Albanua's data, it wee 50 times as active as tropinyl benzilate. Four other eaters of 3-quinuclidinol were as active as 3-quinuclldlnyl benzilate, but none wee more active. Larsaon et al. were not able to detect any correlation between the strength of the hydrogen bond, as derived from their spectra, and the biologic activities of the compounds estimated by Mbanua. Doga given subcutaneous doses of tropinyl benzilate at 0.50 mg/]cg became ataxlc after a mean elapsed time of 16 min and obstinately progressive after a mean of 19 mini ataxia disappeared after a mean elapsed time greater than 283 ein, wheresa obatluate progresaion lasted only a mean of 244 min. Doga given close a of 3-quinuclldinlrl benzilate at 50 ,ug/kg became staxic after a mean of 36 min and became obstlastely progressive after a mean of 42 min. These two effects disappeared at mean times of greater than 313 and greater than 305 min. respectively. Tropinyl benzilate in a dose 10 times that of 3-quinuclidinyl benzilate produced toxic effects somewhat more rapidly than the latter, but these effects were leas durable than those of the smaller dose of 3-quinuclidinyl benzilate. Lavallee (50) reported that doses of 3-quinuclitlnyl benzilate at 10 and 18 ug/kg had no progressive effects on performance of a mul~ciple-stimulus conditioned-avoidance response by dogs, but doses of 24 and 32 ug~kg decreased the number of correct responaea progresaively. Monkeys given doses of 32 and 56 ug/kg may have made a few more errors in a vioual-discrimination avoidance task than monkeys given no compound, but the difference wee not statistically significant in the absence of control data. Cats given intravenously injections of 3-qu$nuclidinyl benzilate labeled with 3H in the 3-position of quinuclidinol were used to determine the distribution of the label in the brain and brain stem (193). Taking 304 counts/min.mg in the nervous tissue as the halfway point between the highest ant the lowest concentrations measured, the areas of the brain that retained the greatest concentrations of the label were motor cortex, sensory cortex, caudate nucleus, lateral gesticulate, and medial gesticulate (167, 140, 121, 107, and 101 counta/min.mg, respectively). The arena that retained smaller concentrations, In order of decreasing concentration of the label, were thalamus, hippocaolpua, hypothalamus, medulla oblo~ata, colliculi, cerebeliar cortex, pyramids of the medulla, cerebral white matter, and cerebellar white matter. Both cholicomio~etic and anticholinergic compounds were found (193) to reduce the retention of the label in some arena of the brain, the most strongly affected areas varying with the compound. The cholinomimetic compounds used were tetrahydroamlsoacridine (THA) ant S-diethylaminoethyldiethylphosphorothioate (TEIRAM)' both inhibitors of chollneaterases. The anticholinergic compounds used were N~ethyl-4-piperidinyl- (phenylcyclopentyl)-glycolat. (EA 3443) and 302,028 (not identified except as an analogue of 3-quinuclidinyl benzilate). The last compound had a much Lore ~ C1

general action than any of the oehere used, dloplacl~g the label of 3~quinuclidiny! benzilate from mose of the sampled areas of brain and brain stem. In the cases of both EA 3443 and 302,028, dloplacement of the label was related probably to the subetitutlon of one intoxicant by another, so that neither of these two anticholinergic subeeances would be practical antagonisto of intoxication induced by 3~qulnuclidinyl benzilate. A~tagontem eight reasonably be expected from THA, which partly displaced the label from motor cortex, lateral gesticulate, medulla, and cerebellar cortex; antidotal activity by TETRA+~hich displaced the label from lateral geniculate, thalamus, and hippocampus--~s less likely. No attempts to assess antidotal effectiveness were made in this study. Z~irblis and Ko~ritzer (56) found that a mitochondrial fraction prepared from rat brain adsorbed 3-4 times as much 3-quinuclidiny] benzilate as atropine in the physiologic range of pa. Me optimal pa for adsorption of 3~quinuclidinyl benzilate was about 8.0, whereas that for atropine was about 9.7; at these pils, the adsorption of the two anticholinergic compounds was about the same. TEA decreased the adsorption of the benzilate by mitochondria9 but a ratio of 100 ~ (T~: 3-quinucItdi"l benzilate) was required to reduce the adsorption by 501. Both Ca2+ and po 2- decreased the oaltochondrial adsorption of 3~quinuctidiny} bencllate, the effect of Ca2+ not being particularly large and requiring an estimated ratio of 30,000:1 (Ca2~:3-quinuclidiny! benzilate) to decrease adsorption of the benzilate by 50%. With phosphate a ratio of about 21.5 :] (pO42-: 3-quinuclidiny! benzilate3 would be expected to decrease adsorption of the benzilate by 503. Yamamura et al. (194) studied the adsorption _ _ of Il on particles in homogenates of various regions of monkey brain after incubation with [-3~13-quinucildiny! benzilate in solution tn phosphate buffer at a pil of 7.4. The regions that were the most acrid adsorbers of the benzilate, in order of decreaslag uptake, were putamen, caudate nucleus, occipital cortex, cingulate gyrus, postcen~cral gyrus, hippoca~pus, amygdala, precentral gurus, pyriform cortex, frontal cortex, superior colliculi, thal~mus, and inferior colliculi. This distribution of adsorptive sites among various parts of monkey brain differs from that for uptake of 3H from intravenously injected [3~13~quinuclidinyl benzila~ce by cat brain (193~. The two studies are in agreement that the caudate nucleus ts one of the ~O8t important areas of localization of the label in the fom of [3813 - uinuclidinyl benzilate. Aura et al. found that uptake of the label by various areas of brain had a tendency to be correlated with choline uptake 8~4 with choline scetylase activity. Yamamura and Snyder (195) found that interruption of the septal hippocampal tract, and elimination thereby of cholinergic afferents to the hippocampus, reduced by about 70X the activity of choline acetyIase in homogenates of the hippocampus, but did not alter the binding of [3H] 3~quinuclidinyl benzilate by the particles of such homogenates. They proposed, as a possible explanation, that I 52

presynaptic muscarluic sites that are innervated by the septal hippocampal nerve fibers do not adsorb the labeled bencllate9 but that postaynaptic muccarinic sites Just across the synaptic gap fro. the presynaptic ones do adsorb it. Another possibility mentioned He that there may be no presynaptic muscari~c sites in the hippocampus. The ATPase in microsomes prepared from whole rat brain, from rat cerebral cortex, and from cerebral cortex of rats that had received 3~quinuclidinyl benzilate intraperitoneally at mg/kg an hour before they were killed was found (196) to be enzymatically competent in all cases and not to be inhibited significantly by the addition of 3-quinuclitiny} bensllate in vitro. Furtheneore, slices of guinea pig brain cortex incubated in vitro with 10~4 M 3~quinuclidiny} benzilate had the same patterns of lose of I+ a" uptake of Nan as slices incubated without the benzilate. Inasmuch as ASPase is related intimately to the active transport of the monovalent cation in nervous tissue, the experiment with the brain slices gave an additional indication that AIPase activity is not altered by 3~quinuclidinyl benzilate. Jovic and Zupanc (197) measured the effects of 3~quinucItdiny! benzilate on oxygen consumption by rats and oxygen uptake by slices of rat cerebral cortex and medulla oblon,gata. Subcutaneous injections of the benzilate at 4-15 mg/Icg decreased oxygen consumption by a mean of 12.~; there was no clear dose~effect relationship. Rats whose oxygen consumption had been increased by a mean of 33.5: by a small dose of soman (25 ug injected subcutaneously) had the increase limited to about 2X by a subcutaneous dose of 3-quinuclidinyl benzil~ate at 15 mg/kg. Slices of cerebral cortex and medulla oblongata stimulated by KC] at a final concentration of 100 mM increased their oxygen uptake by respective means of 59.2X and 41.7X (197~; inclusion of 5 x 10~4 M 3-quinuctidlnyl benzilate tn ache medium bathing the slices reduced-the stimulated oxygen uptake to 16.3Z for cerebral cortex ami to 3.6% for medulla oblongata. When rats were tiered 3-quinuclidinyl benzilate subcutaneously at 5-20 mg/kg a" were killed ~ h later for pre - ration of slices of cortex and medulla oblongata, there were dose-related decreases tn oxygen uptake by both tissues stimulated by inclusion of KC} in the fluid surrounding the slices. Frances and Jacob (~] ~ reported that intraperi toneal injections of 3~quinuclidinyl benzilate into mice at 0.3-3.0 mg/kg decreased the concentration of acety~choline that could be measured in the brain. There was a linear relation between the decrease in acety~choline concentration and the logarithm of the dose of benzilate. The line for this relationship with 3-quinuclidiny! benzilate was close to and nearly parallel with that with scopolamine. Aqullonius et al. (77) found that intravenous 3-quinuclid1nyl benzilate at So ug/k8 produced about the came iOBS of acety~choline from the rat cerebral cortex as atropine at 750 ug/kg. Intravenous 3-quinuclidinyl benzilate at 0.15, 0.2, and 0.5 mg/kg produced greater initial increases in rat ~ 53

locomotor activity than atropine at 1.3, 2, ant 10 mg/kg. The immediate responses to the benzilate were not so well esintalned as those to atropine. ~ the benzilate dose was increased from 0.l Lo 0.2 ~/kg, locomotor activity increased rapidly to an arose maximal value that was about 9.3 times the control Prague. The concentration of acetylcholine in rat hippocampal elesue was tacreased slightly (10.33) by intraperitoneal 3-quinuclidinyl benzilate at 0.l mg/kg (79~; after a dose of 5 mg/kg, the acety~choline concentration was reduced by 44.5X, compared with the mean concentration found in control rats. When septal lesions were created, the mean concentration of acetylcholine in the hippocampus increased by about 47.4%. As in intact rats, administration of 3-quinuclidinyl benzilate at 0.1 mg/kg increased the mean concentration of acetylcholine in the hippocao~pua, in this case by 5.3%. Unlike the result in intact rats, administration of 5 mg/kg resulted in a further increase in the mean concentration of acetylcholice in the hippocampus, to 16.6% above that in control rats. Shis last result was the converse of that found with atropine, but was similar to that found with scopolamine. HeyerhUffer ~198) fourid that L-3-quinuclidinyl benzilate was about 20 times as active as the Disomer in dogs in producing staxia, tach~cardia, and obstinate progression ant in reducing salivation. Babes et al. (199) reported that rats given doses of 0.8, 2.0, and 5.0 mg/kg of 3~quinuclidinyl ben21late (by an unidentified route) had dose-related increased in spontaneous motor activity. Physostigelne (0.2 mg/kg), amobarbital (20 mg/kgy, and chlorpromazine (2 mg/kgy, all given by unidentified routes before the benzila~ce, held the spontaneous motor activity even below the control values. Kabes (200) examined the effects of 3-quinuclidiny! benzilate on the abilities of rats to learn a condltlonet~escape teak and, once trained, on performance of the task. Doses of Oe5. 1, and 2 mg/kg of the benzllate hastened acquisition of the conditioned response in a dose-related manner' but a dose of ~ mg/kg had different effects on performance of the task in relation to the proficiency of the animal in its performance before the benzilate was administered. Rats that were poor performers (more than 2= errors in control sessions) and some that were good performers (iese than 20: errors in COIltrOi sessions) Id their pert omance improved by the benzi late, whereas o eher good performers had their performances worsened. Babes (201) examined the effects of 3-quinuclidinyl benzilate on beagles. There were dose-related responses to doses of 10 and 50 ~g/kg in autonomic functions (heart rate, pupil diameter, flow of saliva, etc.), motor functions (motor coordination, tremor, ataxla, etc.), and central nervous system functions (alertness, orientation, reaction to external stimuli, etc.~. Physostig~ine (l mg/kg) aborted all these types of action. Routes of administration were not stated; physostig~ine was said to have been injected, but the site was not stated. ~ 54

Fused et al. (202) used isolated rat je ju~ and atria to examine the effects of 3-quicuctidinyl benzilate on tissue response to chollnerglc agonists. The benzilate alone had no effect on isolated de junum, but, when applied before a stimulant concentration of furtrethoM~ (10-7-10-4 by, it shifted the dose-response curve for furtrethonium to the right, tee., a higher concentration of furtrethociue was required tO produce a given response in the presence of 3~qulnuclidIny} benzilate than in its abeeme. The application of 3~quinuclidinyl benzilate at a range of concentrations (3-30 x 10~9 M) to isolated atria had no effect on the contractione of either spontaneously beating or electrically driven atria (202~. hen the atria were subjected to the influences of both furtrethonium and 3-quirtuc3.idiny] benzilate, the dose-effect curve for the former was shifted to the right. This was similar to the result with isolated Jejune, the molar ratio of furtrethontum to 3-quinuclidinyl benzilate being about 3.3 x lob :~. Herink et al. (82) injected 3-quinuclidinyl benzilate at ~ mg/kg into the peritoneal cavities of normal rats and rats with electrolytic septal lesions. In coneal rats, there was no significant effect on aggressiveness, but defecation was decreased. In the rat. with septal lesions, both aggressiveness and defecation were reduced by 3~quinuclidiny! benzilate (the latter heir affected to the greater extent). Poe] (203) trained female rats to run in a circular runway for a reward of a drop of chocolate paste. After comple~eion of training, the rats were given intraperitoneal injections of 3~quinuclid1nyl benzilate at 0.l, 0.3, I, 3, or lO mg/kg and were placed in the runway 20 min later. The benzilate induced a marked increase in the time required for ache rat to leave one reward area and begin to seek a second reward (latency); the increase in latency was dose-related. There was a dose-related increase tn the time required for the rat to traverse the circular runway (rusming time), but the slope of the regression line for the relationship of running time to the dose of 3~quinuctidiny! benzilate was much smaller than that for the relationship of latency to dose. These results were interpreted as tndicating that the benzilate intenelffed a conflict between a desire to remain in the ~rici"ty of a prior reward and a desire to obtain ~ later reused, coupled with some interference with motor function similar to that found by yerhUffer (198) and Kabes et al. (199,200~. Bell and Gershon (86) found that intravenous injection of 3~quinuclidiny! benzilate at 50 ug/tcg into dog e induced intoxication that persisted for more than 5 h. Intravenous in Section of I, 2, 3, 4-tetrahydro-9~P'n4 coactldine (Tactile or THA) at ~ mg/kg within 30~50 min after administration of the benzilate induced recoverer in only two of nine dogs, but similar treatment 2-3 h after the benzilate had been injected was successfully antidotal in seven of seared dogs. Yohimbine, which has cholinomimetic and sympatholytic actions (204), injected intravenously at 0.5 mg/kg within 30-50 gin after the benzilate had antagonistic activity in four of nine dogs. When yohimbine was not administered until 2-3 h after the benzilate, I 55

. it antagonized the intoxication in only two of seven dogs. Piperoxan, also an adrenergic blocking agent, was mentioned as being capable of antagonizing intoxication with 3-quinuclidinyl benzilate, but no details of stupor of its action were given. Fusek et al. (205) reported that Tacrine t~ibited both acety~cb~olinesterase and butyry~chollnesterase in dog lunge ant heart and slightly inhibited acety~cholineste-rese in the brain, especially in basal ganglia. Most of the change in the acety~cholinesterace activity of the brain was found to be in the ~icroso~l frac~cton. Butyry~cholinesterese in the bodies of ganglion cells was said to be completely t~hibited in all regions of the brain. These authors found that an 1atramuccular dose of tacrine at 10 ag/k~ completely abolished within 2 h the signs of intoxication induced by an tneramuccular injection of 3 - uinucl1diny] benzilate at 50 Aging. Sram ee al. (206) se pore ed the results of cytogenetic analyses of spermatozoa from mice that had been given intraperitoneal 3-quinuclidiny! bencllate in single {rt~ections at 40 ag/kg or 15 dally doses at 10 mg/kg, of bone Barrow cells from mice that had received 15 daily doses at 2 or lO mg/kg. and of head I - phocytes that had been exposed in vitro at 10-4-lo-3 it. may looked also for reverse mutations in yeast cells exposed in vitro at 5 x 10~4 M and 5 x lo-2 It, for host~ediated transformations in yeast cells injected into the peritoneal casualties of mice that were then given 50 or lOO o~g/kg subcutaneously, and for dominant-lethal effects in mice after single intraperitoneal injections at lO or 40 mg~kg or repeated tn jectione--~S at 2 mg/kg, 15 at 10 mg/kg, or 5 at 20 mg/kg. The yeast cells exposed to 3~quinuclidiny} benzilate in vitro gave evidence of weak mutagenic activity. The only other positive results were in bone marrow cells of mice given doses of 10 mg/kg or more. Sram et al. (206) concluded that, because the frequency of chromosome] breaks was not dose-related above the threshold dose for the occurrence of breaks, the breaks were probably due to systemic toxic activity by tise benzilate, rather than to any specific mutagenic effect, and that the co~pou" has no serious genetic risk f or ear. In another paper, Srae (207) examined the production of chromosocal abnormalities in bone marrow from mele Chinese hunters given single intraperitoneal injections of 3-quinuclidiny} benz-ilate at 0.l, I, 10, 20, 40, or 60 mg/kg, dissolved in dieet~l sulfoxide. Control hasIstere received dimethy! sulfoxide alo - . Doses of 1 mg/kg or more induced core chr~oso~l gaps and breaks per cell than the solvent. Doses of I-40 mg/kg yielded apparently dose-related increases in the incidence of chro~os~al abnonealities: 0.012 per cell with ~ Aging, 0.064 per cell with 10 mg/kg, 0.224 per cell with 20 mg/l~g, and 0.292 per cell filth 40 mg/kg. The highest dose, 60 mg/lcg, produced only 0.076 per cell. Sram considered, on the basis of the count Irish the highest dose, thee the tacidence of chromoso~l gaps and breath was Doe dose-related, but that is questionable for the doses between ~ and 40 eg/kg. Ilot~ever, as Sram pointed out, chromosomal gaps and breaks seem to be evidence of toxic damage to the individual, rather than of heritable change, so they probably do noe indicate a serious genetic risk for man. ~ 56

Hughes et al. (208) gave 10 beagles intravenous 3-quinuclidinyl benzilate at 3 mg/kg twice a week for 3 wk. Flve control dogs were given 0.1 N EC1, corresponding in Solve and pa to the solution of bencllate at 0.1 m1/kg. Some anleals from each group were killed 72 h after Ache last dose of benzilate, and the others were killed 60 d after the last dose. No lealons attributable to bencllate were found in the testes of the dogs that had been given that compound. DIE~NOET~L MNZI~TE The only other member of the group of esters of benzilic acid on whose effects there is a considerable body of information is diethylamis~oethy! benzilate. One review of its pharmacology has been mentioned previously I; it conceded the general pharmacology of the drug as a peripherally effective muscarinic blocking agent. In addition to having cholinergic spasmolytic activity, diethylaminoethg.l benzilate also has mydriatic effecti~rences, is able to antagonize ba~um-induced contraction of intestinal smooth muscle, has local anesthetic properties, and has an ar~ttarrhy~heic action on the heart similar to that of quinidine. In 1955, a series of papers established that diet~lamir~oethy! benzilate affects the functioning of the nervous system in the cat (209), the rat (210), and man (211~. Ber8er et al. (212) stunted the toxicity of slide doses of diet~laminoethyl benzilate in mice and monkeys and its general actions on several functional sys-teme. In the monkey, an intravenous dose of ~ mg/kg resulted in mydriasis and some decrease in voluntary movement. ~ dose of 2 mg/kg induced ataxia with occasional convuist~re Jerks. A 608e of 6 ~/kg induced intermittent clonic convulsions in one monkey; the convulsive state lasted for about 10 min and was followed by a depressed state lasting for over an hour. Papillary dilatation, the effect of longest duration, sometimes pereleted for as long as 20 h. Dtethylaminoethyl benzilate injected intraperitoneally into mice with hexolbarbital at 100 mg/kg (212) prolonged anesthesia due to the hexolbarbital: benzilate at 10 ag/kg tacreased the duration of anesthesia by 35.9 min (1243) and at 2 0 mg/kg increased 1 t by 68. 3 man ( 2 361 ~ . When diet~la~noethy! benzilate was administered to mice by mouth at 24 mg/kg and the mice were then subjected to what was usually a nonlethal con~Igenic electric shock. 50: of the mice died. The only antIo~uscarinic effect detectable in these mice was mydriasis. This dose of diethylam4noe~ r! benzilate Aid Dot Prolog the late=y to onset of confusion after delivery of the electric shock; a larger dose, 37 ag/kg, did prolong the latency significantly. Where difference agonists (acetylcholine, 5-hydroxy~rypeamine, and histamine) were used (212) to stimulate the smooth muscle in isolated rat colon, diethylaminoethyl bencllate had some ability to antagonize the stimulant action of all three agonists. Its ability to block the action of acety~choline was about 20 and 100 times tts activity against the stimulant actions of 5-hydroxytryptamine I 57

ant histamine, respectively. Diethyla~inoethyl benzilate did not increase the urinary excretion by the rat of 5-hydroxyindoleacetic acid, but did ~agr~fy the pressor action of epicephrine and reduced ies Logo. The compound appears, therefore, rtot to affect the metabolism of serotoMn, but to alter in some way the relation between epinephrlne and its receptors. Diethylao~inoethyl benzilate had no significant effect on the knee-Jerk or flexor reflex in the cat (212~; when injected into curarized cats, it altered the predominant frequency of the BEG from 40 per second to 8-15 per second and induced enlarged discharges. The changes in the potentials recorded from the brain were seen in records from both cortical and aubcortical reglona. Electric arousal consequent to peripheral or thala~c stimulation of cerebral afferent fibers - s blocked by diethylaminoethy} benzilate, but recruitment of cortical neurons by stimulation of the thaws was not affected. Giarman and Pepeu (75) reported in 1962 that diethyla-'noethyl benzilate differed from atropine in that a dose equimolar with atropine at 50 mg/t~g (which reduced the concentration of acety~choline in rat brain by 33%) caused no significant change in the brain's concentration of acety~choline. Both atropine and diethylaminoethyl benzilate were said to induce mild excitation, however. In contradiction of this report, Frances and Jacob (~) published a graph indicating that intraperitoneal atropine at ~ to 7 or 8 mg/kg had a greater effect in lowering the brain aceeg~Icholine concentration in the mouse than doses of diethylaminoethy] benzilate in the same range, but that the benzilate at 25-30 mg/kg induced a greater lowering of the acety~choline concentration than atroplne. McColl ant Rice (213) found that intraperitoneal injection of mice with diethylaminoethyl benzilate at 10 mg/kg 15 In before similar administration of tremorine increased the ED50 of tremorlne from 5.8 to 38.0 mg/k8. Such an effect can be explained as being due to depletion by the benzilate of the store of acety~cholice in the brain cells. In 1964, Jacobsen published a review of the literature through 1962 on the actions of dic~hylaminoethy! benzilate (214~. It reported that oral doses of this benzilate of up to 80 mg had been tolerated by man, but that dally doses of about 90 mg had given rice to mild arrhythmias and cyanoale. The compound had been found to enter the rat brain tc unaltered form. When 14C-labeled diethylaD~noethyl benzilate had been administered to rats, SS: of the label had been recovered in the urine within 24 h. Nothing was know at that time about the nature of the metabolizes. The compound had been found to inhibit oxt~ative ph-osphorylatio" in brain cells in vitro, and, at high concentrations (10~3 M), to inhibit monoamine oxidase. In man, doses of 40-200 mg of this benzilate had been found to reduce excretion of 5-hydroxyiMoleacetic acid to zero. Thus, the compound interferes in some way with the utilization or metabolism of serotonin, and presumably with the function of serotonergic synapses. In addition to having peripheral antimuscarinic activity, graded by various 1 58

investigators as between 0.01 and 0.33 that of atropine, diethylaminoethy! benzilate had been found to be particuiar3~y potent in blocking tranemission through the ascending reticular system; it had also been found to affect the descending reticular system, but not to be useful in treating parkinsonism, because of its other central effects. Diet~riaminoethyl benzilate Inhibits the metabolism of barbiturates and potentiates their central depressant activity. In addition to inducing hyperactivity in mice and rats and reducing the reactions to stress in rats and cats, diethylaminoethyl benzilate had been found not to affect sham rage, to inhibit f fighting between rats exposed to electric shocks, and to decrease the rate of learning of complicated task (tasks requiring dlecrimination between Stimuli and the making of different responses to different stimuli) in mice, rats, and cats. Berger et al. (80) reported that diethylaminoethy} benzilate was particularity effective in increasing the duration of after-~chargea by the cat hippocampus after a weak, brief electric shock, being 40~100 times as active as atropine in this regard. As to its antispasmodic action in vitro and Its ability to block salivation induced by intracranial injection of 80 us of acety~choline chloride into a mouse, dieth~riaminoethy! benzilate was 4X and 77X, respectively, as potent as atropine. Shit ov (215) fount that benacryzine and two other anticholinergic compounds increased the action of magnesium sulfate on the BEG of the rabbit in proportion to the activity of the compounds in activating cholinesterase. Banshchikov and Stoliaro~r (216) reviewed a group of substances that had been found to be capable of evoking mental disturbances. All the substances have marked anticholinergic act ivity, and diethylaminoethyl benzilate was among them. The others are 8iel's compounds, JB-31B, JB-329 (Dieran), and JB-336. The authors (216) stated that usual medicinal doses of diethylaminoethyl benzilate not infrequently produce mental changes--retardation or stoppage of the current of thought, a feeling of emptiness, forgetfulness, shortened attention span, sensations of heaviness and altered shape of the Itebe, apathy, sluggishoese, impression of isolation from the en~lronment, hostility ~ apprehenston, horror, amiety, am l~ypochondriasis. A relatively brief period of these effects ~ghe be followed by a period of euphoria if more than 5 me of the benzilate had been taken. The fires report of a psychotomimetic episode due to diethylaminoethyl benzilate in a huff being was credited to VoJeechovsky (217) by Banshchikov and Stoliarov (216~. This investigator hs~ published in 1958 a case report on a female physician who had accidentally taken about I.4 g of the bencllate. The woman experienced visual hallucinations and brief delirium. Intentional ad=~stratione of diethylaminoethyl benzilate to human subjects had shown (218) that the psychotomimetic state lasted for 4-12 h and was accompanied by a decrease in urinary excretion of 5-hydroxyi~doleacetic acid. These authors stated that I 59

diethylaminoethyl benzilate had been oxidase only slightly lese strongly than paychotomlmetic state waned, the urinary excretion of 5-hydro~qindoleacetic acid increased to such an extent thee the 24-h excretion of this degradation product of aerotonin was within coral Iteits. It had been found ta the saee experiment that the urinary excretion of 17-ketoateroita wee markedly reduced during the period of psychosis and that the activity rebounded at about the same time as the urinary excretion of 5-hydrox~indoleacetic acid. In the case of the 17-ketosterolds, however, the rebound increased the total daily urinary excretion of 17~tosteroide by nearly 501. Etelson et al. (219) used t381diethyl~inoethyl benzilate to study the absorption, distribution, and metabolic fate of this benzilate in the rae. me substance used was randomly la beled . Af t e r I ntraperi t oneal i nJe c t ion of benz flat e at about 82.5 o~gtkg, 38 was detectable in the blood at a fairly high concentration about 12 .1n after injections the blood concentration was highest about 36 win after injection. At 5 h after injection, the 3E co~en:ra~cion was about 2SZ of its peak. The authors estimated the half~eime of the label in the blood to be about 80 min. If one calculates (frock data in the paper) the time required for removal of half the peak 3 concentraelon from the blood, that time is found to be about Ill min after the injection. The authors' value of 80 min for the half-time would require the concentration of labe! in the blood 5 ~ after injection to be about 6.7X of the peak concentration, whereas it was actually about 28: of the peak. The estimate of ll4 min is bearer to be a beeter value for the half-eime. Within 24 h after injection, 44.8Z of the label was recovered in the urine and 41.62 in the feces and the gsatrointestinal tract ant its contents. The carcass contained 14.2Z of the label, of which the greatest amount was in the liver. The expires air contained about 0.3% of the 3H, in the form of water. Me urine contained the unchanged compound, e thy laminoethyl benzilate, ant benzi kc acid. Apparently, considerable portion of the diet~riaminoethy1 benzilate had been hydrolyzed to release beDzilic acid, which accounted for 54.4: of the label in the urine. ~ smaller portion of the original compound hat been monodealkylated to eehyl~tnoethyl benzilate, which accounted for 13~82 of the label in the urine. This desll~ylation may have contributed the small amount of the label fount tn the water of the expired air. The fate of the tiethyl~aminoethanol that bust have been released by hydrolysis of the original compound remains uncool. ~ study of the hydrolysis in vitro of the original benzilate at a pE of 7.4 during incubation at 37°C revealed that approximately 50: of the compound unte~en~c spontaneous hydrolysis within 5 h. Gram (207) gave Chinese hamatere diethylam'noethyl benzilate tntraperitoneally at 0.l, I, ant 10 mg/kg. The animals were killed 24 h later, ant the cells of their bone marrow were examined for chroo~oso~l aboor~slities. The lowest dose of the benzilate did not give rise to any detectable chromosomal abcomalities; ~ mg/kg resulted in a 50: increase ~ 60 fou" to inhibit monoamine i oroniazid . As the

in the occurrent e of chromosomal gaps and breaks in the bone marrow cells. The highest dose (10 mg/kg) induced a 325X increase in the incidence of gaps and breaks in chromosomes of bone marrow cells. The same dose of 3-quinuc3-idinyl benzilate had resulted in only a 107: increase in the incidence of chromosomal gaps and breaks; thus, diethylam~noethy] benzilate, despite its lower anticholinergic activity, was more toxic cytogenetically than 3-quinuclidiny} benzilate. EFFECTS ON MAN Davies ( 2 20) summary zed the effects of tiethylaminoethy] benzilate at various doses on human beings. Moderate doses--1-4 mg-~characteristically produced a sense of divorce between outer reality and emotional reactions. Larger doses-~4-8 mg--produced a more distinct detachment: muscles were felt tO be-relaxed and limbs were sensed as awkward, enlarged appendages with blunted sensations. The ability to concentrate was felt to be diminished, and thought processes were slowed. Mental vacuity was common, and reaction to external stimuli was slow. Continuity in thinking was Jolt, 80 that people who had taken doses In this range frequently satd something like: "I have forgotten what t wanted to say." Complaints of mild palpitation and of dryness of the mouth might be made. The cost common complaint among Il0 patients treated with die~hylaminoethy! benzilate was of a loss of ability to concentrate and a feeling of detachment from the people and objects in the immediate en~rlrooment. All doses were given by mouth, the highest dose administered to this group of people being 16 mg/t for several months. No delayed effects or effects that persisted after benzilate administration was stopped were reported. Raymond and Lucas (221) studied the effects of diethylaminoethyl benzilate on a group of 43 outpatients and 10 normal subjects. Patients given daily oral doses of the drug of ~ mg three times a day (t.i.~.) complained of no side effects. About half the patients who took 2 me t.~.d. experienced side effect., particularly a sensation of limb heaviness. Most patients given. 3 fig t.i.d. complained of aide effects, including a feeling of heaviness or rubberiness of the lege. ataxia and clumsiness, difficulty in readlag small print, poor ability to maintain continuity of thought or attention, giddiness, diarrhea, anxiety ~ and drowsiness. The 10 normal subjects were given single subcutaneous injections of 5 mg and then had their EEGe recorded. Five had markedly reduced alpha rat - S in their EEGs. Complaints mate by these subjects after they lad received the drug, in order of decreasing frequency, were weakness; headache, lightheadedness, and a f eeling of heaviness of the limbs; drowainess; and increased complexity and coloration of photic pattern and facial numbness. Complaints expressed only by some of the 10 subjects were dry mouth, tif f icu3~ry in expre Being thought a, f ear approaching panic, sensation that the floor was distant or seemed to slope' huskiness of voice, heaviness of the shoes, alight nausea, and sweat) ng .

Kinrose-Wright and Hoyer (222) reported the results of giving dleehylaminocchyl benzilate to 42 patlenta. They agreed with others that tally close a of 1 mg producet no effects that could be distinguished from those of a placebo. Side effects were infrequent with tally tosea of lesa than 4 me. With daily doses of 4-8 ma, there were complainta of dryness of the mouth, papillary dilatation, alight palpitations, and difficulty in concentration. Daily doses of more than ~ mg resulted in such complaints by half the patlenta, with the adtitlon in some of nausea, anorexia, and constipation. Patients given tally doses of 12 mg or more were affected also by feelings of heaviness of the limbs and of mental confusion, emotional lability and feelings Of unreality, drowelnese, dizziness, and stasis. Me patient developed a ~aculopapular reah. Vo~techovsky et al. (223) summarized their trials of administering doses of up to 75 mg of diethyla~inoethyl benzilate to 17 healthy volunteers. The state of toxic psychosis--resembling a comblcation of psychoses due to atropine intoxication, to chronic alcoholic damage of the brain stem (Koreakov'a ayntrome)$ and to chronic alcoholic deliriu~-lasted for 4-6 he They found no 8O.atiC complications after doses of up to 75 ma. These investigatora Suggested that the combination of anticholinergic activity ant ability to inhibit monoamine oxidase (resulting in the demonstrated laterference with the metabolism of serotonin and possibly of catecholamlne transmitters as well) explains the phenomena of intoxication by this ben211ate. Rickels ee al. (224) reported the findings in an experiment in which 52 patients were given tablets containing fig of diethylaminoethyl benzilate and were instructed to take Five tablets a day, but were allowed to decrease that to three tablets a day if it seemed desirable and, for abort times, to two a day. The group of patients studied conslated of approximately equal numbers of psychiatric patients (mildly to moderately depressed neurotic outpatients) and patients seen in general medical practice. A placebo group (45 people) of similar composition received tablets of identical appearanc e and with the same inetructions. The side effects experienced by the subjects were recorded 2 ant 6. wk after begiQ"Dg ingestion of the tablets. The ale subjects who hat taken a tailor average of two or fewer tablets for a hawk period were excluded from the final compilations of data. Hean intakes of the two types of tablets were not stated, but would be expected to be about 4.5 tablets a day on the basis of reported intakes of similar tablets containing other bugs involved tn the same experiment. Of the 45 persons who took the place ho f or 2 wk. 23 (5L:)complatnet of site effects; 15 (33Z) complained of sedation. Of the 52 persons who took the "nzilate for 2 wk. 26 (50%) complained of site effects; 15 (29%) complained! of sedation. Of the 30 persons who took the placebo for 4 wk. 17 (S 7X) reported site effects; 11 (37Z) complained of sedation. Of the 26 persons who took the benzilate for 4 wk. 10 (3BX) mentioned side effect e; 4 (153) complatnet of sedation. It ts apparent that the persons who ingested tiethylaminoethy} benzila~ ~ had no greater incidence of side ef feet ~ than those who ingested the placebo.

Sidell (225) surveyed studies of the effects of 3-quinuclidinyl benzilate on human subjects carried out by Edgewood Arsenal or its contractore between 1960 and 1969. The initial studies were by and on four members of the research staff, who took the compound to become familiar with its effects, so that the' could describe to potential volunteers the experiences that they might have as experimental subjects and could devise appropriate means for measuring functional impairment and safeguarding the subjects from physical hare. Sitell was able to identify 314 other subjects who had been given 3~quinuclidinyl" benzilate at the Biomedical Laboratory at Edgewood Arsenal or at other sites. In addition, he reported that there had been 24 cases of accidental exposure to 3-quinuclidinyl benzilate among employees of Edgewood Arsenal. Of the 26, 14 had no effects other than ~ydriasis; four others complained of dry mouth, fatigue or sleepiness, photophobia, and inability to accommodate for near Vi8~0~; 8iX had difficulty in concentrating on a topic, slowed thought processes, and ~~d confusion. No long-ter~ effects were re port ed . In 1964 and 1969, field tests had been conducted with 3~quinuclidinyl benzilate. As a part of each test, eight volunteers were exposed to mo~tored aerosolized benzilate and were then assigned mode] military tasks to perform. ~ contractor had performed a study of the effect of exposure to thi s benzilate on the ability of trained pilots to fly airplanes, using the Link flight simulator and other appropriate measures of performance. The procedures used in 23 studies with 3-quinuclidinyl benzilate were outlined bar Sidell (225), who also summarized the effects of the compound on man and stated that tt is about 20 and 3 times as potent as atropine and scopolamine, respectively, in altering functions of the central nervous system. When equipoten~c doses were used, the effects of 3~quinucI1dinyl benzilate lasted about 6 and 9 times as long as those of atropine and scopolamine, re spectively . Sldell's description of the effects induced by 3~quinuclidinyl benzi'ate is as follows: At low doses, the effects include a dry mouth, decreased gastric mobility, inhibition of sweating, an increase in heart rate, papillary dilatation and lose of ac~o~odation, mild sedation and mental slowing. At high doses these effects are intensified. There are marked disturbances of function at all le~rele of the central nervous system: motor coordination, attentiveness and con~erol of thought and lear"ng processes all decline. Confusion, restlesenese, impalement in perception, interpretation, ant memory span, poor judgment and deficient insight are all features of this syndrome. True hallucinations are present. If the dose is quite high the subject may become stuporous or even comatose for several hours. I 63

Freedman (226) reported the fintinge of a research project designed to determine whether small amounts of 3-qutnuclldinyl benzilate modify undesirably the ability of experienced pilots to perform the tasks involved tn flying an airplane. The IS pilots chosen to take part in the study, from a panel of 61 potential subjects, were 2S~45 yr old (mean, 38 ye) and had flying times of 200~5,000 h (2eean, t9800 h). A Link flight simulator and 14 tasks, including the Number Facility Test, were used to assess the competence of the subjects before and at laterals after intramuscular injection of 3 - uinuctidicyl benzilate at I-4 ~g/l~g. These taclts yielded a total of 3S items that were graded in cycles during each test period. Practically no effects followed doses of 0.5 and I.0 ug/kg. After doses of 2,ug/lt,g, there were slight decrements tn performance in some of the tasks, a deflate increase tn heart rate, a decrease in systolic blood pressure, and a slight increase in diastolic blood pressure. The subjects given 4 ug/kg all made lower scores on the performance tests and exhibited definite functional changes, including increased heart rate, dizziness, drowsiness, mydrtasis, and difficulty in accommodating the eyes for near vision. Most of the subjects had increased diastolic blood- pressure and complained of dryness of the mouth. The tests associated with the I.ink flight simulator were the first to demonstrate effects of the 4-.ug/l~g dose; especially ~triRlng changes occurred in vigilance and in judgments of altitude, headinge, air speed, and glide paths. These decrements appeared between ~ and I.5 h after injection. By 3 h, half the subjects court not have completed ~ mission involving simply taking off and landing an airplane. None could have completed a moderately involved mission. The abilities of the pilots to perform the standard tasks presented to them decreased further up to the end of the obeer~stion period, 6 h 40 min after the benzilate had been administered. In July 1962, it was eked (227) that exposure of a human population to a Ct product of 3-quinucItdinyl benzilate of Il8 mg.min/oa3 would result tn incapacitation of 30Z of the population, a Ct product of 170 mg.mln/m3 50: of the population, and a Ct product Of 347 mg.~3 84Z of the population. Ketches (228) satirized the experiments with 3~quinuclidinyl benzilate performed between August 1960 and July 1963 with human volunteers. The report included an appendix, prepared by Claude McClure, Jr., on the cheeisery and biochemistry of 3~quinuclidiny} benzilate. The outatanding i~fomation derived from this appendix ts that the brain was the only organ of mice found to contain 3H 48 h after intraperitoneal injection of [3813~quinucl-idinyl. benzilate. Perfusion of this benzilatc through the livers of rats had lubricated that the liver destroyed the compound quickly, 90: of the compound having disappeared from the perfusate within 30 min and less than 0.2% of the compound being found unaltered in the bile. The products were not arrows. Several hypotheses of biochemical mechanisms of action of 3-quinuclidiny} benzilate were presented, but no fis" support for any of them was available. I 64

The main report concerned 290 exposures of 215 subjects other than staff members to 3-quinuclidinyl benzi3.ate by intravenous injection (1~.31), latramuccular injection-~39.31), ingestion (12.41), inhalation (21.7X), and application to the skin ~14 . 8JI ) . Intravenous and intramuscular indectione were considered to be equal in effectiveness, although changes occurred more rapidly after intravenous than after intramuscular injection. By ingestion, 3~qutnuclidiny! benzilate was about 0.9 times as effective as it was by injection. Inhaled 3~quinuclidinyl benzilate was abut 0.6 times as effective as it was by injection. Application to the skin in solution in benzy! alcohol resulted eventually Rafter 24-36 h) in a group of effects representative of the responses to an injection of about one-se~renth the amount. Estimates of the doses required to be inhaled (expressed as Ct. mg.min/m3) to produce various effects were as follows: mild incapacitation (moderate dilatation of puplle and slight blurring of vision, minimal incoordiDation, some slowing of thought) ~ 66-124; moderate incapacitation ~ hallucination, confusion, unorganized hyperactive ty ~ incoherent speech, shortening of memory and attention span), 102-152; and severe incapacitation (stupor or coma, possibly preceded by a period of agitation, followed after 10~1~5 h by prolonged hallucination and inappropriate behavior), Il0-165. The LD50 for a man was estimated to be 5.7-6.7,ug~kg. Repeated tally exposures of volunteers to intramuscularly injected 3-quinuclidinyl benzilate at 1 )ug/kg yielded co evidence of cumulative effects, but 2 ,ug fig induced clear signs of cumulation. One of 12 volunteers glares daily doses of I-2 ug/kg appeared to develop tolerance. Repetition of a second administration of 3-quinuclidiny} benzilate 2-3 wk after an initial dose induced a more rapid and more pronounced development of intoxication than ache f fret dose; that suggested a BeDSitiZation Deck in-the response o the second dose. Tetrahydroaminoacridine (TEA) had been found to decrease heart rate when tachycardia was present, to increase secretion of saliva, to increase sweating and lower core temperature If le had become high, to reduce Secular rigidity, and to improve, at least temporarily, the mental status of an intoxicated subject. THA seemed not to antagonize sleepiness and might have promoted it. Physostigo~lne, given orally at 2-6 mg/h, caused nearly total remission of toxic effects within about ~ h after administration of the benzilatc. Physostigorine seemed to be a more complete antagonist of the effects of 3-quinuclidiny! benzilate than was THA. In 1965, the Directorate of Hedical Research of the Chemical Research and Development Laboratories, Edgewood Arsenal, issued a booklet (229) on the management of casualetes from 3-quinuclidinyl benzilate. The booklet gave the following sequence of effects to be expected is persons who had received large doses: in l-4 h, tachycardia, dizziness, ataxia, vomiting, dry mouth, blurred vlaion, confusion, and sedation progressing to stupor; in 4-12 h, inability to respond I 65

effectively to the environment or to moire about; and in 12-96 h, increasing activity, rando's unpredictable behavior' and gradual return to normal 48-96 h after exposure. The booklet recommended treatment of casualties wl th intramuscular injection of 3 mg of physostigmine salicylate, followed by another such dose 40 min later, if necessary. Ilalatenance by oral doses of 2-5 fig of pl~ysostig.~ne salicylate every I-2 h was suggested. With improvement in mental status, the frequency and alze of the maintenance doses can be reduced gradually. Heart rate was recommended as an indicator of the adequacy of therapy. me pulse should be between 70 and 80 beats/min when good control has been achieved. If the heart rate falls below TO beate/min, the frequency or magnitude, or both, of the maintenance doses of physostigeine sallc~rIste shoult be reduced, but the patient should be observed carefully for possible reversion to toxic delirium. Peripheral effects of physostig~ine overdoses (profuse sweating, chary skin, abdominal cramps, vomiting, muscular twitching, tremor, and weakness) may be antagonized by sm811 doses of ~eth~riatropinium nitrate or, if that is not available, of atroptne itself. Fizzes and Vancil (230) reported the rcoulto of a study to determine the intramuscular dose of 3~quinuclidiny} bensilate that constitutes the minimal effective dose (~) for ~ mea, here defined an the smallest-dose that produces a decrease of at least 25X in performance in the Number Faellity Test. The effects on heart rate were also recorded. Thirteen male volunteers were given intramuscular injections of 3-quinuclidinyl benzilate hydrochloride at 203 or 2.7 ugfkg. One man given 2.7.ug/kg became confused and developed halluclaations. The most common complaints of the other volunteers were of dry mouth, blurred ~risios~, and trowainese. Several developed restlessness, an inability to sleep despite drowsiness, and anorexia. Of six men given 203,ug/kg, one had a decrease in performance in the Number Facility Test of more than 25:, and three had dilated pupils. Lois dose Induced no tachyeardia or hyperther~ia. Of seven men given 2.7 ug/kg, five had decreased performance in the Number Facilleg. Test of more than 25:, three had dilated pupils, two had heart rates greater than 100 beata/min, and one had a blood pressure above 140/90. There were no cases of hyperthermia (the report did not state the en~rtro~ental conditions of the study). The mean heart rate at ear the larger dose was about 92 beate/mln, whereas that after the smaller dose was about 79 beate/min. Kitzes am Vancil estimated that the MED related to performance in the N - ber Facility Test was 2.54 ug~kg, with 9St confidence limits of 2.31 and 2.80 ug~kg, and that the HED for inducing changes in somatic functions (~1on, heart rate, and blood pressure) was about 2.7 ug~kg. Crowell (231) described experiments in which 27 male volunteers were given intramuscular l.-2-alpha-tropiny] benzilate at 1.9-8.6 ~g/kg after a precious unpublished estimate that the intravenous HED was about 3.8 ~g/kg. This benzilate was found to have a quicker onset of effects after intramuscular injection and a briefer duration of action than 3~quinuclidinyl benzilate. Every subject who received a dose I 66

of 2.8 )lg/kg or more experienced dryness of the mouth, blurred vision, and dizziness. Nearly all the subjects given 4.1 ~g/kg or more complained of anorexia, nausea, weakr~ese, cohesion, hallucinations, and ataxia. Host expressed the illusion that their hands and feet were as red as blood. About one-third suffered headaches, and a few vomited. The tntramuscular EI)5o for inductr~g a decrement in performance in the Number Facility Test of at least 25% was about 3.l ug/kg, that for increasing heart rate to at least 100 beato/min was about 5.6 ug/kg, and that for reducing performance in the Number Facility Test by at least 90: was about 5.9.ug/kg. For men given L-2-alpha-tropinyl benzilate at a mean dose of 6.l,ug/kg, the mean duration of severe effects of the compound was 2.5-3.l }I, and the mean. time of onset for reduction of performance in the Number Facility Test by at least 90: was about ~ h. At 10 ~ after intramuscular injection of L-2-alpha-tropl~yl benzilate, no evidence of- any organic damage attributable to the compound could be found. - Crowell's estimate of the Incapacitating dose by intramuscular injection was 6.0 ~ 0.75 ug/kg. Kitzes e t al. ( 154) reported the results of 40 experiments in which volunteers with body weights between 59.1 and 96.3 kg were given 3~quinuclidinyl benzilate at 2.0~.0,ug/kg. These experiments revealed no clear relation- between body weight or ben211~te dose and performance in the Number Facility nest. part of the same study, 10 experiments were performed with volunteers with body weights between 65.4 and 87.7 kg who were given L-2-alpha-tropinyl benzilate at 1. 9-~. 6 ,ug/kg. With ebis be no clear relation between dose and have compound, there seemed to pert ormance in the Number Facility Test, but there may have been a positive relationship between body weight and performance. Men the range of weights of these subjects was divided in half, the upper half of the range lacluded four experiments with men given a mean dose of 5.b. ,ug/kg and having a mean body weight of 85.3 kg, and the lower half included six experiments with men given a mean dose of 4.3 ~g~kg and having a mean body weight of 69.7 leg. Despite the smaller mean dose of the benzilate received by the men in the lower half of the weight range, that half irlcluded all the men rated as being the most severely affected in performance in the Number Facility Test . Wi ah 3~quinucI161ny} benzilate, the ewo halves of the weight range contained close to the same proportions of severely affected men: 45.4: in the upper half and 48.3X in the lower half. Here, also, the mean doses for the two halves of the weight range were similar: 4.3 ug/kg for the upper half and 4. 7 ,ug/kg for the lower half . Ketchum en al. (~89) reported the results of field tests in which two groups of four men each were exposes on different days to clouds of an aerosolized solution of 3~quinuclidinyl benzilate in chloroform. After exposure to the cloud, the subjects were rated on their performances in a variety of tests of physiologic and psychologic properties. Baseline performances in these tests had been recorded after 12-20 practice runs before exposure; the five best scores in the practice runs were averaged as the baseline scores. A try run before the actual experiment incorporated all features of the experiment except the exposure; the scores on the test battery I 67

during the dry run did not deviate significantly from the basellce scores. The conclusions from the study were that the EDso for incapaciteelon by 3-quinuclidiny! benzilate under field conditions is about 60 mg.min/m3 for a asc with a body weight of 75 kg and a minute volume of respiration of 15 To. Intramuscular injections of 2 or 3 mg of physostig~ine salicylate were found to reverse the toxic delirium rapidly; repeated injections every 2 or 3 h or oral doses about one-ehird larger than the intramuscular doses, on the sane schedule, restored and maintained acceptable performance in the test battery and in carrylug out simulated guard duty of a post perimeter or almulated sentry duty in a foxhole under attack by an aggressor. Craig et al. (232) reported the results of exposing 24 volunteers to high environ~enta1 temperatures at various ti~e8 after they had been given intramuscular 3-quinuclidlcy} benzilate at 3, 4' 5, or 6 ug~kg by intramuscular injection. Two series of exposure to heat were performed. In the first, four subjects were exposed to 41°C for 2 h on each of two days and six subjects were exposed to the same temperature during two 2-h periods on one day; on the third day after the first day of exposure to heat, all 10 subjects were given intramuscular injections of the benzilate during the fires of t ~ 2-h exposures to 41°C, separated by 3-h, followed by two similar exposures on the same schedule on the following day and by a single 2-h exposure of six men on the morning of the sixth day of the experiment (four of these men had another 2-h exposure to heat on the afternoon of the same say). In the second series, 14 men were exposed to 41°C for ~ h on day I, to 52°C for ~ h on day 3, and to 41°C for ~ h on day 7, beginning 4 h after an injection of 3-quinuclidiny! benzilate. The effects of the benzilate reached their maximum about h af ter inJecelon. At that time, the heart rate, skis temperature, rectal temperature, and inhibition of sweat secretion were locreased in relation to the dose of bensllate administered. The highest rectal temperature was 380 3°C, an increase of lese than I°C above the mean normal temperature. The disturbance of regulation of body temperature by 3-qulnuclidicy! benzilate under the condition of recumbent rest used tn these experiments should not be hazardous to a health, person. Physical activity during the exposure to heat might. alter this general picture, although a previous study had found that the combination of 3~quinuclidiny! benzilate at 4 ug/kg, exercise, and an en~rtronmental temperature of 46°C produced only a slights' greater increase in rectal temperature than the exercise and environmental temperature without the benzilate. The inhibition of sweating by this benzilate regressed at about half the rate at which that induced by atropine regressed. Klapper et al. (162) reported the results of followup studies on two men who had been given 3~quinuclidinyl benzilate and one who had been given I`-2-alpha-tropinylbenzllate earlier. The first two had histories of intermittent hematuria before their service as volunteer sub Sects; at the times of the followup examinations, they had fewer than 10 red blood cells per high-power field in their urine. One of them reported that he had had a flashback of his experiences under the influence I 68

of the benzilate when drinking alcoholic beverages about 3 wk after exposure. There were no remarks about the man who had received the other benzilate. Clapper et al. (163) examined the records of 24 volunteer subjects who had taken the MALI test and then had been given intramuscular injections of 3-quinuclidiny] benzliate at 2.] (six men), 2.4 (~] men), or 4.5 (seven men) ~g/kg in an attempt to determine the factors of the HOPI that are best correlated wi th the re sponse to the benzilate represented by performance in the Number Facility Test. The dose of 2.] ug/l~g produced a mean decrease in performance in the Number Facility Test of 27%, 2.4 ,ug/Icg produced a mean decrease of 26X, and 4.5,ug/kg produced a mean decrease of 74%. The scales of the ~P! that were best correlated with the individual scores in the Number Facility Test were that for positive test-taking attitude for 2.4 ug~kg and that for hysteria for 4.5,l1g/kg. There were no significant positive correlations for Ache lowest dose. There were significant negative correlations with the schizophrenia and the hypachondriasis scales after that dose, but these negative correlations were not reported for either of the la rger dose s . Sitell et al. (164) found that S-( 2 - iisopropyiminoe thyl ~ me thy~methy~phosphonothioate in jected intravenously at 1. 5-~. 7 ug/Icg af ter intramuscular in jec tion of 3-quinuclidinyl benzilate at 6 ug/kg was effective in inducing rapid improvement in the scores of three volunteers in the Number Facility Test. This finding was similar to that reported earlier in subjects exposed to scopolamine. Sidell (168) reported that four volunteer subjects given 1 Intramuscular in jectlons of 3-quinuclidiny] benzilate at 7 ug/kg had their scores in the Number Facility Test improved dramatically by intramuscular injec~clon of 7 or ll me of physostigmine salicylate followed by oral admintatrations of 24 or 12 ma, respectively, of the same drug during total periods of treatment of 43 and 42 h, respectively. Two other subjects given the same done of the bensllate were treated entirely with oral administration of physostigeine in total doses of 80 and 211 me during periods of treatment of 37 and 71 h, re spectively. The scores in the Ilumber Facillty Test of the last two subjects were better malotained than those of the f irst two. SUM.~Y ~- 3-Quinuclidinyl benzilate, the benzilic acid ester that has been studied the most by Edgewood Arsenal, has effects similar in general to those of the tropic acid esters, but more prolonged. Two other benzilic acid esters have similar actions. In part, the particularly long duration of the central action of 3-quinuclidiny] benzilate may be related to its greater affinity for nervous rl.~ue, and especially for mitochondria within neuronal cells. In agreement with ins especially strong adsorption on mitochondrla' the subcellular organelles concerned principally with the supply of energy to the cell, this benzilate has been found to reduce the oxygen consumption by nerve cells stimulated in various ways. This I 69

ant the other two-benzilates on which some information was found have some activity as local anesthetic agents and as antihistaminic compounds, in addition to their activity as peripherally and centrally effective antagonists of cholinergic agonlats. The only long-ter~ toxicity study of a benzilic acid ester that was found was of the ester with diethyla~inoethanol; no significant panthers were found in rats during thee lifetime feeding study, but the lifetimes of the rats fed the —nzilate may have been shortened somewhat. Feeding of the ester With quaternized 3~quinuclidinol to rats for a year and garaging dogs with this compound 5 d/wk for a year produced no significant pathemas. Both cholinergic and antichollnesterase compounds antagonized the toxic actions of the benzilates. M though 3-quinuclidiny} benzilate has been found Lo have weak mutagenic activity for yeast cells in culture and to produce gaps and breaks in chromosomes of bone marrow cells, it has not been proved to produce heritable changes in mammals. Diethylaminoethy} benzilate was more toxic cytogenetically than 3~quinuclidinyl benzilate, despite the lower anticholinergic activi ty of the former compound. ESTERS OF PHENYLCYCLOPENTYLGLYCOLIC ACID This group of anticholinergic esters contains five compounds, the subject acid being esterified with the following a lchohols: tropine, 3~quinuclidinoJ~, Worthy I-3-quicuclidinol, N~ethyI-4-piperidinol, and a mixture of N~ethyl-3-piperidinol and N-e thy I-3-pyrrolidyImethanol. Of 20 reports on these compounds by personnel of Edgewood Arsenal and its contractors, 10 are on the ester with N~ethyI-4-piperidinol, four on the ester with the mixture of N-ethyl-3-piperidinol and It-ethyI-3-pyrrolidy~methanol, and two each on the other three esters. The literature on the ester with N-methyI-4-piperidinol is entirely from B4gewood Arsenal, whereas more has been written about the ester with N-ethyl-3-piperitinol and N-ethyl-3-pyrrolitylo~ethanol (Ditran) by investigators other than those at Edgewood Arsenal than ~ the ones at that site. Most of the information about the other esters in this group originated at Edgewood Arsenal. The following discussion concentrates on the two ~08t popular esters in this group. Tile information on the lethal activities of the members of this group of esters ts sparse. Even when reports dealing with lethality according to their titles were furnished by the Board on Toxicology and En~riro~ental Health Hazards, the texts provided were brief and uninformative. An example ts a report (233) entitled A Review of the Pharmacology and Toxicology of CAR 301, 060. This compound is the ester of pheny~cyclopenty~glycolic acid and 2~e thyl-3-quinuclidino] . The entire text reads: CAR 301,060, a ~Z-like glycolate, is, on the basis of extensive pharmacological evaluation, a potent anticholinergic compound thee displays marked behavioral ~ 70

and CNS effects. The compound, in Boat. instances, is equal to or more potent than BE, ant has similar temporal parameters. It is exceptionally unique in regard to profound potency in the ACRBL-HAZLETON visual discrimination test. Extensive acute ant subacute toxicological evaluation failed to reveal any extremely profound toxic or pathological effect in any ma for organ system at the doses tested. CAR 301, 060 has a very subatantia~ safety margin in all species tested. Such pap is a waste of paper ant time. Other reports of that ilk (234-236) give ~ little useful inf ormation. The first of these (234) stated that repeated daily latravenous doses of the pheny~cyclopenty~glycolic acid ester of N-methyI-4-piperidino] produced mydrlaels in monkeys, that doses of 0.! mg/kg or more resulted in above-nomal ratios of adrenal:body and testes:body weights, and that the highest dose administered, I.0 mg/kg, resulted in unusually small livers. None of the organs mentioned was considered to have been harmed histologically. The second report (235) concerned the effects of the same compound in the dog. The lowest daily intravenous dose, 0.01 mg/kg, induced mydrias1~; the two higher toses, 0.1 and 1.0 mg/kg, resulted in-ataxia, decreased activity, and dryness of the mouth, in additon to mydriasis. They also induced significant effects on the performance of the dogs in a multiple-stimulus conditioned-avoidance test. The highest dose induced a statistically significant decrease in body weight. At necropsy, no significant effects that could be a ttri bused ~ o the agent were f ound . The third report in thi group (236) described the effects of the phenylcyclopenty~glycolic acid e otter of 3-quinuclidino] on dogs. Repeated daily intravenous doses of 0.l mg/kg produced maximal midriasis -that persisted throughout the study, decreased activity and ataxia, decreased concentration of the urine, and significant changes in the weights of the gonads and kidneys. The lower dose, 0.01 mg/l~g, was not reported Lo have produced any e-ffecto other than mydriasis and decreased activity. The dogs that had been trained in the multiple-stimulus conditioned-a~roidance test displayed a rapid development of tolerance to a daily tose of 0.l mg/kg. N - HE THYL~4 - PI PERID ~ NYL- ~ P HENYLCYCLOPENTYL ~ - GLYCOLATE ~ EA 3 443 ~ N-He thyl-4-piperidinyl-( phenylcyclopentyl)-glycolate, also designated EA 3443, was found (193) to be effective both prophylactically and therapeutically in reducing the adsorption of 3-quinuclidinyi benzilate by the cat's motor cortex, caudate nucleus, and sensory cortex after intravenous injections of the benzilate and the glycolate. When given after the benzilate, EA 3443 reduces the retention of that compound by the medial and lateral geniculates. When it was injected first., EA 3443 reduced the adsorption of 3 - uinuclidinyl benzilate by the hypothalamus and thalamus, in addition to the three parts of the brain first mentioned. Prophylactically administered EA 3443 decreased the adsorption of the benzilate by the I 71

hypophysia, livers lungs, skeletal muscles, spleen, and sciatic nerve. When it wee administered to eats after the benzilate, it reduced retention of the benzilate by skeletal oracles, lunge, sciatic nerve, pancreas, liver, spleen, and heart. When added to the fluids aurrountin~ a segment of isolated guinea pig ileum, EA 3443 reduced afloat to zero the response to acetylcholine. This glycolate hat about the aaoIe activity in this regard as 3-quinuclitinyl benzilate, but the anticholinerglc effect of the glycolate Irma the one more readily reversed by teerabytroaminoacritine. Kitzes and Ketchum (237) reported giving 39 volunteers intramuscular EA 3443 at 1.0-2.7 yg/kg. These tosea induced no hallucinations or abnormal necrologic algna. Dry mouth, blurred vision, and drowainese were the most colon coo~plainta. Mydrisaia occurred to a gild degree. There vea no consistent dose-reaponse relationship for the heart rate, but that rate may have been decreased, rather than increased. me minimal effective dose for reduclag performance in the Number Facility Teat of half the subjects by at least 25X (MEDso) is this study was 1.21 ~g/kg, with 95Z confidence limits of 1.00 and 1.48 ug/kg. The time for onset of these o~luimal effects wee about 8 h, ant the duration of effects was about 4 h. Kitzea et al. (154) found that 80X of a group with a mean body weight of 87.2 kg who had been given EA 3443 at a mean dose of 2.2 ~g/kg suffered degradation of their performancea in the Number Facility Teat. Only 46.7% of a group with a mean body weight of 71.6 kg who had received a mean dose of 2.4 yg/kg suffered degradation of their perfor~ancea in the same teat. In the case of this compound, therefore, body sloe may have conditioned the reaponae. Hart and Balter (238) exposed an unstated number of volunteers to EA 3443 by application to an unidentified area of the akin at 60~120 ug/kg (mean, 68.5 ug/kg). The volunteers were rested with a modification of the Army General Classification Teat before and at 7 wk after EA 3443 exposure. During the exposure, the Nimber Facility Teat was used to determine whether the aubJecta experienced any degradation of their mental abilities. m. criterion for an NED (two of five consecutive scores below 75X of the baseline score) vas met by 71: of the exposed volunteers. A control group of 15 volunteers were tested and reseated 20 d later without any known intervening exposure to a psycho toxic aubetance. The glean score of the control group at the second testing vea 3.2: below that at the first testing. The second testing of the exposed group yielded a mean score for that group that vea 2.4: above that at the first testing. Neitherof these tlfferences is significant. Lavallee (50) reported that the mini~sal intravenous dose of EA 3443 that decreeasd performance by at least 25: in the Number Facility Teat by half the members of a group of volunteers vea 1.2 ug/kg. The correaponting incapacitating dose, the IDso, vas 3.1 ~lg/kg. That for 3-qulnuclidinyl benzilate was 6.6 ug/kg. When EA 3443 was tnJec ted intravenously into dogs trained in a multiple-stimulus conditioned-avoidance teat, a dose of lOO ug/kg decreased the I 72

number of correct responses to visual stimuli by about 28.2X. This decrement had been surpassed by that induced by 3~quinuclidinyl benzilate at 32 ~g/kg. In monkeys derailed in a visual-discrimination avoidance task, intravenous EA 3443 at up to 56 ~g/kg had no significant effects on performance of the talc. The same doses of 3~quinuclidinyl benzilate also had no significant effects on the responses of monkeys in this test. Lavallee concluded that the screening tests with dogs and monkeys were not satisfactory for predicting the relative potencies in man of the anticholinergic compounds tested. K3apper et al. (163) found that the scale of the MNPI that was the best correlated with the degradation of performance of volunteers in the Ember Facility Test after intramuscular doses of EA 3443 (~ix volunteers, I.S,ug/kg; five volunteers, 2.4 ~ug~kg) was that for paranoia. The lowest negative correlation was that between the social introveralon scale and the degradation of performance in the Number Facility Test. These statements were true for both doses of EA 3443; that ts different from the situations with some of the ocher compounds studied in this way. With both atropine and 3~quinuc3.idinyl benzilate, the larger tose produced a degree of degradation of performance in the Number Facility Test that was best correlated with the score in a different scale of the AMPS from the one that yielded the best correlation with the lower dose of the same agent. Armstrong et al . ~ 239) used a best that required water~deprived rats to press four levers in a preset sequence to obtain water. After administration of EA 3443 by an unstated route, the EDso for a minimal mydriatic effect was 37 ~g/kg; that for reduction of the number of drops of water obtained within a given period was 53.2,ug/kg. DI'rRAN (CS 4297) In 195B, Abood et al. (240) reported initial studies of a group of esters of alkylated derivatives of piperidine, i nclutin~g a material that was thought t o be N-ethyl- 3-piperidyI-(pheny~cyclopentyI)~glycolate. This substance was given the trivial name Ditran; it iS know now to be a mixture of two compounds: the pheny~cyclopentyl glycolates of N-ethyI-3-piperid1no] and N-ethyI-3-pyrrolidy~methano] In the proportion 3:7. This substance was not as centrally active (producing hyperactivity in rats) as the methyl hoeIologue, but was less lethal and more potent as a peripheral an~cicholinergic. Ditran was administered to more than 40 human volunteers by Ostfeld et al. (241) and was found to be actively hallucinogenic. Esters of 3-piperidyt" alcohols were found to be much more hallucinogenic than corresponding esters of 4-p iperi dy} alcohols . Abood et al. (242) compared 14 related esters of substituted piperidyl alcohols. They found that the phenylcyclopentyl and pheny~cyclohexy} glycolates were more actively hallucinogenic in human subjects than dipheny1, pheny~prc~py1, or phony} 2-thienyl glycolates. Esters of quaternized piperidinium alcohols were not hallucinogenic. I 73

hydroxyl group on the carbon atom alpha to the carboxyl group of the acid was found to be essential for hallucinogenic activity. Diagram was found no have in intravenous LDso for the rat of 19 mg/kg and one for the souse of 44 mg/kg. The latter value may have been erroneous, inasmuch as Usdin and Efr-on (263) later stated that the intravenous Logo for the mouse was 10 mg/kg. However, Uadin ant Efron may have confused the dose given to human sub Jecta by Aboot et al., which eras 10 mg/man, with the figure for the I.Dso for the mouse after intravenous lnJection. me value reported by Abood et al. by this route accords better with the Logo after intraperitoneal injection in the compilation of Usdin am Efron than that stated by the latter authors. Usdin and Efron gave also intravenous LD,os for the guinea pig and the rabbit of 45 ant i2 ~g/kg, respectively. Intraperitoneal injections of ])ieran into nice and rata yielded LDso eati~atea of 60 and 25 mg/kg, respectively. L;ittle correlation between peripheral anticholinergic activity and paychotogenic potency wee foul by Abood et al. They reported that intraperitoneal injection of physoatigo~ine at 0.5 o~g/kg into rata 5 min before injection by the aame route of Ditra3 at 25 olg/kg prevented mydrisaia and lessened tachyeardia and hypermedia of the tail and Elba, but had no visible effect on hyperactivity or on weakness ant poor coordination of muscles. Gershon and Olariu (244) assessed the ability of Tacrine to antagonize the actions of Dltran on human subjects and compared the effects of Dltran with those of mescaline, Sernyl, and LSD. The moat common complaints of subjects given dosea of Ditran by intramuscular injection or by mouth, or by both routes ~ were of hallucination, confusion, and impaired contact. Gershon and Olarlu found that Tacrine was able to antagonize both the peripheral and CNS effects of Ditran tn man, but was incapable of preventing those due to SernyI. Succinate, which effectively reverses both the peripheral and central effects of Sernyl, had no actlvi ty against latoxlcat' on filth Ultras. Lee authors gave the following typical sequence of signs and sys~ptoma after latraoIuscular lnjectlon of 10-15 fig of Dltran: In 20~30 min. autonomic reactions (dryness of the mouth, slight tachycardia, flushes face, and muscular relaxation); in 45~60 ~ a, central effects (some confusion, speech difficulty, decreased ability to concentrate, slight disorientation, occasional vertigo, and hallucinations); in 2-6 h, maximal central effects (hyperreflexla and ataxia); ant In 5-8 h, abatement of 8y~ptom8 and 8ign8e Blel et al. (245) reviewed the effects produced by a large group of antlchollnerglc compounds in an attempt to develop guidelines for the development of new psychotropic drugs and reached the following general concIusions: only the piperidyl and pyrrolltyl glycolatea that were potent antlcholinerglc compounds exerted profound effect a on ENS functions. However, peripheral antlcholinergic potency did not endow a compound automatically with brain activity. Over 95: of a dose of Ditran is excreted in the urine within 2 h after its adminiatratlon. The largest brain concentrations of several of these subetances were in the hypothalamus and exudate nucleus, I 74

areas related intimately with emotion and mood. Psychotropic anticholinergic compounds had little effect on several. enzyme systems. Ditran is a clinically effective antidepressant agent . An inhibitor of cholinesterases, te trahydroa~noacridine, is capable of ameliorating the psychotic episode induced by anticholinergic paychotogene. Piperidine, trifluoperazine, and several other tranqutItzere were effective in overcoming hyperactivity induced by an ti chol i ne rg i c c ompound ~ ~ ~ ra t ~ . Bell en al. (85) used dogs to study the psychotropic activity of Ditran ant it8 antagonism. Meer developing a rating scale including six indexes of autonomically evoked activity, six of motor activity, and eight of central newous system activity, they were able to present a graph demonstrating that the time courses of each of the ma jor types of effect induced by intravenous Ditran at 0.5 mg/kg had a somewha t ind ividual time course . Mydriasis, tachycardia, and decreased secretion of saliva were apparent within 3 min after injection. Between 3 and 6 min after injection, the pupil response to light became sluggish, and alertness and respos:sivenese to commands began to decrease. Within 3~2-15 a1n after injections incoordination and ataxia had reached a plateau, and the pupi ~ re spouse to light usually had disappeared. Obstinate progression and disturbed behavior (whining or barking, pawing at the floor, and apparently chasing imaginary objecta3 became evident at 30~40 min after injection. By an hour after the injection, most of the dogs had collapsed. Prostration, from which the togs could be roused temporarily by noise or handling, lasted for 15-120 min. The integrated total effect (obtained by action of scores on the various Arts of the rating acale) reached an approximate plateau at about 0.5 h after injection ant rematnet there till about 2.5 h after injection. Approximately complete recovery from the effects of this tose on dogs did not occur until about 5 h after injection. Mydriasis was the last sign of effect by Ditran tO disappear, often befog evident for up to h after injection. Bell e ~ al . ~ 85) f ound that doses of physostlg~ine or neostigmine injected intravenously at 20 ug/kg 30 In after Ditran antagonized the effects on autonomic functions, but did not alter the central actions. Arecoline (150 Aging) restored salivation and heart rate to normal, but had co other detectable antagonistic actions when injected intravenously 30 min after Ditran. Tacrine at l mg/kg, injected intravenously 30 min after Ditran, abolished all effects of Ditran except mydriasis and decreased secretion of saliva. About 40Z of the dogs so treated had relapses about 10 ~n after the dose of Tacrine; repetition of the dose of Tacrine induced complete a" permanent recovery, except for persistent mydriasis. Some derivatives of glycolic acid (phenAnthry~glycolic acids), rphenyImacdelic acid, and benzilic acid injected 30 or 60 min before Ditran shortened by up to 55X the duration of the intoxication due to Ditran, but did not alter the initial severity of its effects. These organic acids exerted less antagonism to intoxication with Ditran when they were I 7S

administered therapeutically than when they were used prophyl.ac tically. Bell and Gershon (86) reported that yohi~bine antagonlsed the ir`toxicatir~g actions of Ditran, but differed from Tacrine in this sort of activity, in that its antagonistic activity was less time~epe~ent than that of Tacrine. The latter had only slight antagonistic activity early in the intoxication and had greater activity when it was administered later after the Ditran, whereas yohimbice had more nearly equal antagonletic activities at various times after the dose of Ditran. Brown et al. (246) gave 48 dogs intravenous Mitral at 0.5 mg/kg and 30 oiin later injections of saline, chlorpromazine, imipramine, or two unidentified substances, IN 1060 and W2045. IN 1060 is Mown also as cyprolidol-, an antidepressant, but an equivalent for W2045 could not be found. Chlorpromazine and i~pramine deepened the intoxication induced by Ditran, whereas IN 1060 snd especially W204S lightened the intoxication. When chlorpromazine and the two other compounds were fed to doge daily for a month before the 8Di~8 were given Ditran, chlorpromazine accentuated the paychotogenic action of Dltran, whereas IN 1060 and W2045 both attenuated the pe~rchotogenic action of DitranO After the montholong dosing with the Iast two antagonists, IN 1060 had a much greater effect on the central actions of Ditran than W2045, whereas the reverse had been true after single therapeutic administrations of the two drugs. For both compounds with troth schedules of administration, the effects on the central actions of Ditran were greater than chose on the motor and autonomic actions. Banshchikov and Stoliarov (216) compared the effects of three of Bie ~ ' ~ co2epounde--JB-318, JB-329 (Ditran), and JB-336~-amon8 themselves and with those of benacryzine and atropine. They also discussed the use of these compounds in the practice of medicine. Ditran had a much less prolonged action than ~-318, which was described as causing loquacity and hyperactivity with an elevated mood that persisted for days, weeks, or even months. In the few people who hat been persuaded to take repeat doses of this compound, resistance to the hallucinogenic action developed, but the mydriatic effect peraleted. The effects of all three of Biel's compounds were qualitatively at - lar and resembled those induced by atropine and benaceyzine. Daily toses of Dit ran were stated so lead to development of resistance to its actions: after 3 wk of daily intake, a dose of 20 mg/kg no longer resulted in mental disturbance. The posthallucinatory euphorlant action of Ditran has been applied to the treatment of patients in depressed states of mind, apparently with some success. Ditran has also been used in treating patients with neuroses and psychoses. Mbanus (42) reported that subcutaneous injection of Ditran at 0.5 mg/kg into ll dogs resulted in the development of ataxia after a mean of 22 In, of obstinate progression after a mean of 31 min. and of complete loss of contact with the environment after a mean of '5 min. Obstinate progression disappeared about 230 min after the injection of Ditran, and ataxia af t er about 280 min. ~ 76

Gershon and Angrist (247) summarized the research that had been done with Ditran and other anticholinergic paychotomieetic compounds and with possible antagonists of their paychotomimeetc activities. Nothing particularity new was reported in this paper. Tacrine is regarded as the base overall antagonist. Sayers and Burki (71) compared the effeceivenesses of 13 psychoactive compounds, including Dieran, in four assays of anticholinergic potency: displacement of 3-qutcuclidiny} benzilate from Me particles in a homogenate of rat brain, antagonism of stimulation of isolated guinea pig ileum by acety~choline, antagonism of induction of tremor by oxytremorine in mice, and production of pupil dilatation in mice. Ditran was graded first in the first two testes third in the oxytremorine test, and second in the mydrissis test. For comparison, atropine was as good as Ditran in the second test, first in the last two tests, and second in the first test. Overall, therefore, Ditran was Judged to be slightly less anticholinergic than atropine. The general conclusion was that none of these tests is seeisfac tory for predicting is Vito effects on brain functions. Kligman and Copelan (156) reported that 16 volunteers had been used for studies with Ditran, but gave no infor~&elon a bout the studies. Ketchum ee al. (24,165,166) gave 22 male volunteers intramuscular Ditran at 50-170 ug/kg and concluded that 150 ug/Icg produced a syndrome of per! pheral parasympatholytic effects, disturbances of basic neuroregulatory functions, al disruption of }sigher integrative functions thee culminated in delirium characterized by restlessness, confused speech, hallucinations, and moment-to-moment variability. Twelve of these volunteers were treated with physosti8~ne and three with Tacri ne after development of toxic psychosis. Both inhibitors of cholinesterases were found to be effective in overcoming changes in the Ilumber Facility Test Induced by Dieran. 3-2UINUCL IDINYL-(PHENYLCYCLOPENTYL )-GLYCOLATE (EA 3167 ) The phenylcyclopentylglycolic aclt ester of 3~quinuclidinol was included in the group of glycolic eaters studied by Lareson et al. (192~. They recorded its threshold dose for producing a paychoto=metic effect in the dog as 50 ug/kg. Five subetances with threshold doses of 10 ug/kg had hydrogen bond strengths (expressed as I°B K, where K is the constant for the equilibrium between free and bonded CO groups in infrared spectra of the glycolates) of 0.~-~.2 (mean, I.01~. Two substances with threshold doses of 50 ~g/kg, one being Dieran, had log R values of 0.80 and O.93; two other substances with threshold doses of 0.5 mg/kg, one being tropinyl benzilate, had log R values of 0.92 and I.2. It is evident that there ts no correlation between the strength of the intramolecular hydrogen bond and the threshold dose for psychotomimetic of feet . Ketchum et al. (248) gave six volunteers tntremuscular 3 - ulnuclidinyi-( pheny~cyclopenty3. )-glycolate by latramuscular injection at 2.4 ug/kg, seven at 2.9 ,ug/kg, and six at 3.4 I 77

ug/kg (the hydrochloride sale was used). Heasurements and observations of the subjects were made hourly for 6 h, bihourly for another 6 h, and every 4 h thereafter until recovery was Judged to be virtually complete. The ME:USo was found to be 2.5 ~ug/kg, and the ID50 was estimated to be 4.1 ~ug~kg. With the latter dose, mild effects were to be expected after 2 h and severe ones after 5 h. Severe effects would be expected to last for about 96 h, and eild ones for about 240 ho Eighteen of the subjects were examined for evidence of organic changes, and 13 were given psychologic evaluations about 6 mo after they had received the compound. Most subjects displayed some charge from baseline performance and behavior when examined 2 wit after injection of the glycolate; these changes usually disappeared by the fourth week after the injection. The effects detected in these earI:r f ollowup examinations included increased irritability; mild impairment of memory, Judgment, or abstraction; mental sluggishness with occasional confusion; ner~rousnese; and tenseness. No positive evidence of organic change was found at 6 mo after the doses; there may have been a s~l increase in ~Q at that time. The profiles in the ~! of seven subjects who were tested 6 mo after exposure, as well as before exposure and a month after exposure, were close to the base3-ice profiles, although a month after exposure they had contained significant Increases in the scores or the hypochondriasis, depre ssion, hysteria, psychasthenia, schi zophrenia, and mama scales. Intoxication with the glycolate was found to be treatable with physostig=ne or Tacrine; because of the I°~8 duration of the effects in some subjects, doses of antagonists may have to be repeated for a rather long time. L -2-ALPHA-~OPINYI.-L-( PHENYLCYCLOPENUL)~GLYCOt^'rE ~ 226, 086 . The ac t10~8 of L-2-alpha-tropinyl- L-(phenylcyclopentyl)-glycolate on experimental animals have been described (188~. This was found to be the most active and the safest (largest ratio of LD,O to MED,o) of the six possible isomers of tropinyl-( phenylcyclopentyl~glycolate by intravenous injection into mice. LDsos by intravenous tnJectione into male mice and rats were 61 + 3 and 65 + 3 mg/kg, respectively; the I.D,os by gavage in the came two species were 98 + 21 and 170 + 53 mg/k~. Depression of general motor activity and depression of respiratory activity were the principal effects observed in the two test species during the estimations of LDsos. A range-fir~din8 test with intravenous injection of the glycolate into monkeys yielded an Ills probably between 10 and 20 mg/kg. In the cat, the LD,o probably is greater than 20 mg~kg. L-2-alpha-TropinyI-L-( pheny~cyclopentyI)-glycolate elicited mydriasts in the mouse after intravenous doses of 10 ,ug~kg or more. One estimate of the MEDso was 5.6 ~ I.5 mg/kg. Msrdriasis became maximal after intravenous~doses of about 0.3 m~g/kg. Other effects that were evident after doses of 10-30 ug/kg were typeracti~lt~r, hyperreactivity to touch, and motor deficits. Respiratory depression became evident I 78

after intravenous doses of 0.3-3.0 mg/kg. General activity became lessened and ptosis of eyelids appeared after doses of about 30-50 ag/kg. Mydriesis was the most persistent effect, which eight last for 3~4 ~ after the highest doses mentioned above. With ogrdriasis as the indicator, this glycolate was about one-third as active when applied to the skin of the mouse as when injected subcuteneousI, (benzy] alcohol was a suitable solvent for application of the glycolate to the skin). Intravenous doses of about 50~100 ug~k8 depressed a poly~ynaptic reflex (ipailateral flexor reflex) in the anesthetized cat, but tit not affect the pateliar reflex. This glycola.te was effective in antagonizing tremor in mice (induced by subcutaneous injection of tremorine at 10 ag/kg), in subcutaneous doses of 10-30 ~g/kg whereas, by the same route of adminiatration, doses of 12-16 ug/kg were effective in inducing ydriasia. The glycolate had a dose-related negative isotropic effect on the isolated, perfused rabbit heart, with only a slight negative chronotropic effect. In the intact anesthetized dog, it hat dose-related negative effects on both heart and mean blood pressure. L-2-alpha- Tropin~ -I.-( phenylcyclopeney1 )-glycolate was act ive prophylactically agatnet 2 Ll)sos of TEE P (2.3 mg/kg ineraperitoneally) in mice pretreated by intraperltoneal injection of benzoquino~um at 0.3 mg/kg (to prevent neuromuscular paralysis by TEPP), its intraperitoneal EDso being 40 ug/kg. Rats given five daily tntravenous injections of L-2-alpha-tropinyI-1~-(pheny~cyclopentyl)-glycolate at 50 mg/kg during ~ d arid rats given intravenous injections 5 d/wk for mo at 10 otg/kg had no changes observable at necropsy, except decreases in the sizes of the spleen and the thymus in the animals given 50 mg/kg. Histologically, both groups of rato had increased numbers of granules in the Paneth cells in the crypts of Lleberk~n in the small intestine. Two of five rats given the largest dose were thought to have alight depletion of thymocytes. Monkeys given dail-'r tntrevenous Injections of L-2-alpha-tropinyI-I.-(phenylcyclopentyl)-glycolate at 0. 4, 2. 0, or 10 mg/kg 5 d/wk for a month had mydriasis with all doses. Tremor a occurred in the animals given the two larger tones, and convulaione in those given the largest. Various extents of involution of the thymes and of increased granule formation in the Paneth cells were seen in animals given each of the doses. Patty changes were seen in the livers of come of the monkeys given the two larger doses. One of five monkeys given 0~4 mg/kg died on the twelfth day, ant one of fire given 10 mg/kg died on the twentieth day. No specific cause of death was identified in either instance, but both angle bud lost more weight than the means of the living members of their groups. The actions of this glycolate in unacesthetized beagles were studied (249~. Groups of four dogs were given daily intravenous injections at 0, 0.8, 6.0, and 20.0 mg~kg 5 d/wic for a month, receiving 21 doses during 29 d. Each group contained two Bales and two feeler. Tree of the dogs given the highest dose were found dead on the fourteenth, I 79

seventeenth, and twer`~-sixth days; the fourth dog in that group was moribund and was killed on the twentieth day. No ani~le from the other groups died during this study. All dogs that received the gI'colate had eidriasis a" reduced activity, the ~ydri-asts persisting eared over weekender In the high-lose group, panting, peralatent anorexia, tremor, ataxia. prostration' fits of running move~nte of the legs (possibly accompanied by barking) in prostrate aureole. and convulsions were seen commonly. These dogs lost an average of at lesat 23.5: of their body weight before death. The togs of the groups given the low and medium doses of the glycolate loce 4.4: and lO.8X, respectively, of thelr original body weight during the administration of the glycolate. The heart rates of the dogs given the glycolate all increased during the first 15 min after administration of the first doses, whereas those of the control animals, given intravenous inJectiona of saline, decreased slightly on the average. After the first weekend, after three injections during the previous week and before injection of another dose of the beRzi]8t6, the heart rates of the anima~a given the two higher doses were still above their baseline valuesO Baseline heart rates were not available for the dogs given the lowest dose, but the heart rates of two of the four dogs in this group on day 6 were above the highest baseline rate recorded for the other 12 dogs in the experiment. OF the twenty-ninth day of the experiment, at 2 h after the last doses of saline or glycolate, the mean heart rates, in comparison with those measured at the same time after the first doses, for the three groups of dogs then alive were as follows: control, 9 beats/min below that on the fires day; low tose, 29 beats/mln below that on the first day; and medley dose, 43 beats/min below that on the first day. The composition of the blood (blood chemistry and ~ hematology) changed significantly only in the group of dogs given the largest dose of the glycolate. Flaal samples of blood were obtained on only two of the four dog e in this group; in both, the prothrombin time was prolonged, the total serum cholesterol concentration was very low, ant the alkaline phosphateae activity in the serum was very high (70 and 198-K-A units/100 m1 vet 4-12 tn baseline measuremente). Both d°8e had other evidence of interference with liver function: increases in the activity of glutamic-oxaloacetic tranea~insse and in the concentration of serum bilirubin in one, and decreases tn the concentrations of albumin and total protein and in the ratio of albumin to globulin in the serum of the other. The fire t of these two animals had a concentration of serum urea nitrogen that was above the noneal ranges so there seemed to be some intctterence with renal function also. The most striking change tn the hematologic picture in these two dogs was the complete eradication of eosinophilic polymorphonuclesr cells from the blood. The total leukocyte counts and heeatocrits were decreased in both dogs; one also had a moderate decrease in e~ythrocyte count. Terminal samples of urine were obtained from the bladders of all four of the dogs in the group given the highest dose of the glycolate. Two were bloody, and three contained bilirubin, I 80

bile salts, and urobilin. M1 four samples contained albumin. The urine of the dog mentioned above as having increased serum urea nitrogen: contained hyaline casts. Elec trocardiographic recordings from the dogs given the medium dose of I.-2-alpha-tropinyI-L-(pheny~cyclopentyI)- glycolate contained suggestione of deepening of the S wave and of elevation-of the T wave. Because baseline recording of the EGG had not been made for these animals, the significance of the apparent changes cannot be assessed. The only significant pathemas found in the dog e at necropsy at the end of the experiment were in those given the highest doses of the glycolate. The livers of these dogs were pale yellow to orange and were coarsely lobular or swollen. Three of these dogs had bile staining of the sclerae and aort as . Two of the dogs had. tarry material in their gastrointestinal tracts, reddish urine in their urinary bladders, and fatty bone marrow. Hemorrhages into the real cortices were found in these two dogs. Microscopic examination of organs from these dogs revealed decreases in the size and number of the islets of La-~~gerhans in one dog from each of the low~tose and high-dose groups and in two dogs from the medium~ose group. In the high-dose group, irritative and degenerative or necrotic changes were also found in the livers, kidneys, bone Marrow, adrenals, testes, epididymides, and gastrointestinal tracts. The livers of all the dogs in this group were markedly fatty, the hepatocytes containing numerous small cytoplasmic vacuoles. Widespread vacuolation of the tubular epithel1~m in the cortical region of the kidney was seen in three dogs; epithelial crescents were seen in Bo~an's spaces in the kidneys of two of these dogs. The bone marrow of all four dogs had shifted tO an indenture type, with increased n''mbere of blast forms of the granulocytic series. 2-~:THYL-3-QUINUCLIDINYL-( PHENYLCYCLOPENTYL) -GLYCOLATE ( 3 01, 060) . Hayes (250) reported the results of a study in which 29 men were given single intravenous doses of CAR 301,060, also calle d ci s -hue thyI-3~quinucl idinyi-( pheny~cyclopentyl ) - glycolate, at 0.5-5.3 ug/kg . They were graded in performance in the Number Facility Test. Two subjects given 5.3 ug/kg became incapacitated (two consecutive scores in the Number Facility Test below 10: of haseline); the mean duration of significant effect was 73 h. These two subjects were disoriented, confused, restless ~ and less coordinated than noneal, spoke disjointedly ant in a slurred manner, had Ispoes of memory, and experienced visual hallucinations. One was episodically paranoid and mildly hostile to the medical personnel in attendance. 80th were drowsy and lethargic. Their heart rates were increased markedly for more than 24 h after injection, but the peak of the tachycardia (+85X) occurred about 12 h after injection. This dose was the only one that resulted in pupil dilatation, by about 2 am. Doses of 2.0 and 2.7 ug/kg produced only alight and irregular increases in heart rate. The heart rate was increased in a dose-related manner when doses of 3.2 ug/kg and I B]

more were admintetered. The lowest dose that produced a significant decrease in the performance of one of four subjects tn the Number Facility Test was 2.7 ~g/kg; 302 ug~kg produced significant decrements in the performances of five of seven men, and 3.S Ug/kg caused significant decrements in the perfoneances of six of eight sub-]ec-~. "yes concluded that the intravenous Also of this glycolate for can Is about 3.0 ug/kg and that, after a dose of 3.2.ug/kg, a significect decrease in cognitive ability occurs about 5 h after injection and last s f or about 7 h. ESTERS OF PIlENYLISOPROPYLGl.YCOLIC ACED Two esters of phenylisopropylglycolic acid are Song 26 antichollnergic subetances studied by Edgewoot Aree~l: Name thyl-4-piperidinyl-( phenylisopropyl )-glycolate (" 3834) and 4-(l~ethyl-1, 2, 3, 6-tetrahydropyridinsrl)-phenyl- isopropylglycolate ( 302, 668) . Now THYL~4- PI PERTI) ~ NYL-( PHENYLIS OPROPYL ) - GLY-COLATE Copelan (251) gave 16 volunteers EA 3834 at 1.0-2~7 ~g/kg intravenoualy and administered the Number Facility Test to them at intervals thereafter. Two groups of two men given I.0 or I.4 ug/kg had their performances in the Number Facility Test unchanged. Some members of each of two groups of four Den given I.6 or 2.0 ug/kg had their performances reduced to below 75% of their baseline performances. Three of four Den given 2.7,ug/kg had their performances reduced Lo below 75: of the baseline. The estimate of the HE:Dso was 1.96 ug/kg, with 95^ confidence limits of 1.45 and 2.66.ug/kg. Significant change in performance in the Number Facility Test usually began at about I.5 h at the highest dose administered (2.7 ug/l~g), the mean time of onset of significant effect on perfomame was I.9 h, and the duration of significant effect was 2.8 he Copelan described the effects of this agent as follows. At 3-15 mitt after injection, the subjects reported that they f elt "high, ~ "light-headed, ~ or "dazed . ~ At about this time, they were unsteady in walking and particularly in turning about . Talking among the subjects became quieter, ant their speech was slightly muffled, but without significant ·lurri~g. A feeling of headless of the eyelids and a tendency to tiplopia became evident at 10-15 min after injection. Sedation, with troweinese and dozing, were noticeable at 15-30 ala. Oce subject (tose group not states) reported an illusion of undulating movement in the glass of a window. Two subjects (dose groups no~c identified) reported hallucinatione of crawling ants ami of mo~rlog strings or hairs on the floor, respectively. The are two subjects recognized that they were hallucinating . There was no conf usion, significant disorientation, or delirium among the subjects in this stuty. dryness of the south' blurred vision, tachyeartis, ant mild reddening of the condunstivae were noted occasionally and might appear within 15-30 min after injection. No abnormal changes in blood and urine were detected 24 ant 48 h ant 8 ant 28 after injection. I 82

Averill et al. (252) summarized studies of the toxicity of EA 3834 in mice, rats, rabblta, cats, togs, and monkeys. Table 3 gives the available aiD8le-dose LDSos for both EA 3834 and 302,668. It t s apparent that the latter is less lethal than the former ln the species for which values on both compounds are available. The values for 302,668 come from Sim (253~. In addition to the I.D,o values stated above, Averill et al. a~arlset the results of studies in which grates doses of EA 3834 hat beer atminiatered to various animals by a variety of routes. In office, intravenous inJecelona at less than lO ug/kg induced mydriaels and those of 32-lOO ug/kg induced unusual sensitivity to being touched, restlessness ~ ant ataxia. Intramuscular doses of lOO mg/kg in tI'e rat indu^.ed paralysis of the leg into which the compound was injected within 5 sin after injection ant Jerking movements of the head, hyperpnea, ant exophthalmos within about 15 "n after injection. Intravenous doses of 0.7 ug/kg induced mydriasis in rabbits, and tachycardia followed doses of lO-lOO ~g/lcg. Ataxia in this species appeared after intravenous doses of 3 mg/kg, ant prostration ant con~rulaion occurred after doses of Il-16 mg/kg. In the cat, mytriasis, decreased activity, increased heart rate, ant ataxia all occurred after intravenous doses of 10~100 ug/kg. ,. The heart rate of the tog was increased and the performance of a conditionet-avoidan~e task was degraded by lntravenoua toses of 10-28 ,ug/kg. Doses of 12-37 )lgJkg reduced physical activity, and those of 25-30.ugthg induced ataxia. Prostration and convulsions occurred after doses of 10-20 mg/kg. Inhalation exposure of dogs to a Ct product of ~ 3834 of 36 mg.min/D~3 resulted In incressea in heart rate. Physical activity was reduces by exposure to Ct products of 40~50 mg.min/m3. 2lydriasts ant ataxia followed exposure to a Ct product of 90 o~g.min/m3. A Ct product of 102 mg.mln/m3, with 95X confidence limits of 82 ant 126 mg.~/~3, causes 50% of the exposed dogs to fail in performance of the conditioned-avoidance task. In the monkey (species not stated), intravenous d08e~ of 6.ug/kg Induced ydriasis. Visual dlscrimlaation was reduced by doses of 7-31 ~ug/lcg. Tachycardia and decreased activity followed doses of about 130 .ug/kg. Prostration and convulsions were produced by dosea of 14-16 ~g/kg. Sitell and Braun (2S4) discussed the results of an experiment tn which two men were given EA 3834 by mouth at 7 yg/kg, four were given 10 ug/kg, three were given 14 yg/kg, and two were given 20.ug~kg. One man given 7 ug/kg and two given 10 pa/kg suffered severe effects (reduction of score In the Number Facility Test to below 25% of the baseline score). M1 the men given 14 ant 20 ug/kg experienced severe effects. Of chose who experienced severe effects, the three with the lower range of doses had a mean dose of 9`ug/]c~g and experienced severe effects at about 2.3 h after drinking a solution of the agent in water. The five Len with the higher range of doses had a mean dose of 16.4 ~gikg and experienced severe effects at Smut 1.4 h after ingestion. The two Den given 7 ug/l~g had a mean score of 47: of their baseline ~ ~3

scores. The four men given 10 ug/kg had a mean score of 40.4%. The five men given a mean tose of 16.4 ug/kg had a mean score of 2.6X. The eight severely affected den had recovery times of 12-16 h (mean, 12.5 h). The two men given 10 aging who did not suffer severe effects had recovery times of 6 and 10 h. The severe effects Iseted 0.5-9.0 h; the mean duration of severe effects for the three den who took a mean dose of 9 g/kg we. 303 h, whereas that of the five men who took a mean dose of 16.4 ~g/kg was 6.4 h. Mild effects (reduction of score in the Number Facility Tese by at least 25: of the baseline acore) for all the severely affected subjects lasted 10-14.5 h (mean' Il.2 h), whereas those in the two men who took 10 Agog without developing severe effects Iseted for 3 and 7.5 h. Mild effects in the two men given 10 ~g/kg who never developed severe effects began 2.5 and 3 h after ingestion. In the eight men who developed severe effects, the mean onset to e for mild effects was I.3 h (95: confidence Ilmits, I.0 and 2.0 h). McCarroll et al. (255) described a study with EA 3834 in which eight enlisted volunteere from the U.S. Arty were divided into two four-an teats, with a ninth servl~g as the leader of both groups. The ninth man had been given intramuccular scopolamine at 23.5 ~g/kg several weeks earlier to familiarize him with the effects that the others would experience later, but he was not given an antichollnergic substance during the actual study. The members of the two te~s were glares directions for perfoneing a simulated =litery ~~ioc in the field and then practiced the Bloc for 3 d. On the fourth day, the tens went through the entire -lesion under the observation of two judges, who graded their performances on a number of items incorporated into the exercise. The exercise included capture a ~ interrogation by a simulated aggressor. On the day of the test, intramuscular injections were given at 9 am., and the exercise was started immediately. It ended at 4 p.m. A battery of sects (heart rate, blood pressure, rectal temperature, Number Facility Test, and behavioral checklist) was administered to each volunteer at 9:30, 10, and Il a.~., noon, and ~ and 4 pa.. The performance of volunteers on this day was graded by the sure Judges who had been present on the control day. One four-man team received EA 3834 at 2 ~g/kg (one man), 4 ug~kg (two men), and ~ ug/kg (one man), for a mean dose of 4.5 g/kg. The second four-a team received 4 ug/kg (one ean), g~kg (two aen), and 12 Gig (one man), for a Bean dose of g/kg. Both teems were considered to be composed of capable, effective soldiers on the control day. On the test day, the team with the lower mean dose was Judged to be otarglaally effective. The man given ~ ug/kg had to be removed from the exercise about 2 h after indectlon. The other members of that team stumbled and fell over - nor impediments in the terrain, lost some ~ their gear. made serious errors in calculating an azimuth with compasses, were unable to remember simple directions or to repeat instructions given just "fore question)", and were unable to operate a field telephone. The man given ~ ,ug/kg cooperated filth the ~ ~4

"enemy" interrogator and tried to follow his instn~ctione without ic~cellectual or physical resistance. The team that received the larger mesa dose became totally ineffective within about an hour. Me man given 12,ug/kg became tccapacitated within about 30 ate and was removed from the exercise. One of the men given 8,ug/kg became incapacitated about 75 min after injections and the other became incapacitated about 141 min after injection. The man in this group given 4 ug/kg was not incapacitated, but the team leader had to prod trio' constantly to get him to accomplish the simplest tasks. For both groups, the correlation between performance in the Number Facility Tese and in the various military' tasks tacluded in the scenario for the field exercise was ~good. The volunteers evacuated f rom the f ield exercise were flushed and had hot, dry skins, parched lips and tongues, Ciliated pupils, tachyeardia, and exaggerated deep tendon reflexes. After treatment with physostigmine salicylate (55 ,ug/kg intramuscularly), these volunteers were restored to lucidity within 20 min. After institution of maintenance doses of physostigmine salicylate by mouth, ache subjects were able to return to the field exercise and to perform their duties capably. When the maintenance doses of physostigo~ne salicylate to the volunteer who had been given EA 3834 at 12 ,ug/kg were dlecontinued after four such 408e8, he became delirious again ~rlehin about 3 h and remained in that state for 5 or 6 h. No abnormalities of renal and hepatic functions were detectable in any subject 24 h and 7 ~ after the injections of EA 3834. Cucinell et al. (256) applied EA 3834 in a solution tn ethanol to the sklne of 11 volunteers on single occasions, of three volunteers on two occasions, anal of two volunteers on three occasions. In all instances, the area of akin to which the solution was applied was about 9 cm2 of the neck at the angle of the Jaw. When the solvent had evaporated, the area of application was covered with a plastic cup or with Teflon taped to the skin. The contaminated skin was washed with alcohol 4 h after the application. Some of the volunteers were placed in a hot room at 40.5°C and 24: relative humidity with a wind speed of 0.5 m/e (slightly more than 1 mph); others remained under unspecified room conditioner Three of the volunteers who received EA 3834 core Can once were exposed to it both tn the hot room and in the regear laboratory rooms; the other two in this category were exposed ossify in the hot room. The amounts of the agent applied to the skin ranged from 0.l to 5.0 mg in the hot room and from 2.0 to 4.0 mg in the regular laboratory rooms. The hydrochiori-de of EA 3634 had no measurable effect on rate of sweating or performance in the Number Facility Test. When EA 3834 base was applied to the skin, the time to onset of inhibition of sweating (38~170 min) in the hot room was related inversely to the amount of the agent placed on the skin within the range 0.~-3.0 ma. The rate of change of the rate of sweat secretion once lshibition of that rate had begun was somewhat less variable (13-42 mgIm2.min2) within the same range of amounts of the agent, but was randomly distributed throughout ~ RR

the ravage. The two extreme values of the rate of change of the rate of sweating came from experiments in which sequential amounts of the agent near the middle of the rate were applied to skin, the larger of the two a~oun~ce producing the "aller rate of charge. Because the earliest time after application of EA 3834 to the stun at which the Number Facility Test was administered to the volunteere tram 4 h, there is no factual basis for ache assumption by Cucinell et al. that there is ~ constant relation-ship between the time of onset of inhibition of sweating a" the extent of decrease tn performance Use the fiber Facility Test. Their proposed expression for estimation of the systemic effect of contamination of s~n - th this agent depends or this spurious relationchip and cannot be considered re liable. 4- ( l ME THYL-1, 2 , 3, 6-TETRAHYDROPYRID INYL) -PHENYL IS OPROPYLGLYCOLA ( 302 ~ 668) Copelan (257) summarized experiments tn which 20, volunteers were glares 302~668 intravenously at loO-406 ,~g/kg and were tested at intervals for ability to perform tn the Number Facillty Test. Hen given doses of t.O-, l.4, and 2~0 ,ug/kg lost lese than 25: of their baseline abilities to perfone the additione. Of four men given 2.7 .ug/kg, two louse at least 25: of their ablileles. Of four men given 3.2 ug~kg, one lost 29% and another logic 241e Four men given 3.8 ~g/kg and two given 4.6 aging had mean decremen~ce of 41: and 50Z, respectively, and all lost at least 25: of their abilities. Karger (258) est1£eated the ID50 of 302~6613 by giving 14 volunteers intravenous doses of 5~0-13~0 ug/kg and administering the Number Facility Test at intervals thereafter. Incapacttatlon was defined as the making of two consecutive scores below 10: of the baseline score. ~ clinical evaluation of incapacitation based on obeervation of the subjects was also made. Me scores on the Number Facility Test yielded an ID,o of 10~l ug/kg, with 95: confidence limits of 9.2 and J~.l ug~kg. The clinical e~raluatlons yielded an estimated ID,o of 9.5 ~g/kg, wish 95X confidence limits of 8. 5 and 3~0. 6 ug /1`g . As a second part of the she study, Merger gave six volunteers 302,668 intravenously at 10 ug/kg and adhered physostig~ine at various times. ~ intramuscular dose of 3 ag of p~costig~ine injected 10 ~n before atoIinistration of 302,668 attenuated the decrement in performance in the Number Facility lest for about 4.5 h after injection. At thee time the performance of two unprotected subjects and of the two prophylac tlcally protected subject s became eseentially identical. When a dose of 3 fig of physostigs~ne was inJec ted in~eramuscularI' 10 gin after intravenous injection of 302, 66S, there was rapid recovery from about 35% of baseline performance to about 921, but perfoneame in the ~ber Facility Test was don to about 20: of baseline by 2 h after the injection of 302,668. When the phycostig~ine was not injected until 45 ~n after the agent, performance in the Number Facility Test rose from zero to about 85: within about 45 min and to a peak of I 86

abou~c 93% within 75 Id. Performance in the test oscillated between 93X and 70: of baseline until about 3.75 h after injection of the agent and then drifted don to about 40: at h. Thereafter, it improved gradually without further administration of physostig=ne; thus, performance in the test had improved to about 60Z of the baseline value by 10 h after initiation of intoxication. Karger concluded that ache best results in maximally incapacitated persons would be obtained by giving repeated intramuscular doses of 2 or 3 ~ of physostig~ne at inter~rale of about 2 h. S1dell ee al. (259) estimated the incapacitating dose of 302,668 inhaled as an aerosol. Performance in the Number Facility Test was the criterion of incapacitation. Nineteen volunteers inhaled aerosols of the agent while seated and wearing an oronasal mask, so that the expired air could be collected and analyzed for unretained agent. Four ranges of retained dose of the agent were recognized: 1.99-7.60 ug/kg, five volunteers; B.3-~.54 ug/kg. four volunteers; 12.36-12.95 ug/kg, six volunteers; and 13.05-~.77 ug/kg, four volunteers. The mean performances in the Number Facility Test of all groups fell below the level for minimal effect (ec ore less than 75X of baseline score), but that for the least group was the only one that fell below the level for incapacitation (ecore Jesse than lOX of baseline score). The mean performance of ache last group was below the criterion for incapacitation from about 0.25 h to about 5.5 h after exposure. That group also underwent marked decreases in its scores on an Orthorator for near and far Derision, in its ability to move pegs in a pegboard, and in its pert omance of a bend-~wlet-touch task, whereas the other three groups hat comparatively minor changes in these four measurement ~ . The initial signs and symptoms induced in these volunteers consisted of tryDe88 of the mouth, drowsiness, and ataxia; the He were f ollowed by hallucinations i market confusion, poor coordination, res£1eseness, and prolonged impairment of acco~oda~elon for both near and far vision with the larger retained mounts. Retention of large amounts (IS pa/kg) sometimes resulted in fine fasciculations of muscles. One subject in each of the two highest ranges of retstned agent vomited, and eight others, tocluding subjects from all ranges of retained agent, complained of nausea. The heart rate was not altered significantly among the volunteers in the three groups with the lowest ranges of retention, but three of four sub Jects in the other group had heart rates above 100 beats/. No consistent effects on blood pressure were observed. Pupil dilatation occurred tn all groups; in some subjects among those with the higher retentions, mydriasis persisted f or 72 h. From the results of these exposures, Sid-ell et al. (259) calculated that the ICtso for a man breathing 10 L/min would be I81 m'.min/m3, with 9S: confidence limits of 159 and 206 mg.min/~. For a man exercising moderately and breathing 15 Livid, the ICtso would be 121 mg.min/~3, with 95: confidence limits of 106 and 137 mg.min/m3. The incapacitating dose of retained agent for 50: of exposed Hen, I 87

the Intro, was calculated to be 13.1 ug/ltg ~ with 95X confidence liD'its of 11.1 and 15.3 ug/kg. With this reesinet dose, the time to onset and the duration of mild effects were expected to be 25 min and 12 h, respectively; the times for severe effects were predicted to be 30 min and 5 h, re spectively . Lavallee (50) collected and graphed data from hear experimente that indicated that 302,668 at cooperatively low 608e8 (below4ug/kg) approached 3-quinuclldinyl Ventilate in activity in decreasing perfoneance in the Number Facility Test, but at higher doses (above 9 Jug/kg) wee only s tightly more active than scopolamine. Extrapolation of Lavallee's graphs suggests that the doses of 302,66B and of scopolamine that would be able to lower performance in the Number Facility Test to zero would be identical. Sim (253) summarized the body of research with 302,668, printed or unpublished, performed tn 1964-1971. The first one-third of the report discussed the chemical and physical properties of 302,668. the last two-thlets summarized the studies performed to determine the actions of 302~668 and several other anticholinerglc subetances tn aniesI and horn subJectsO As shown in Table I'd, 302,668 was cost lethal to the rabbit and least to the monkey after intravenous injection. It was most mydriatic in the rat and least in the monkey. The species with the safest relation between the lethal dose and the mydrlatlc dose vas the rat, followed by the dog. Species with much smaller values of the ratio of LDso to EDso were the mouser the rabbit, the cat, ant the monkey. Although 302,668 had approximately the same actions as other anticholinergic compounds, the only sort of activity at which it excelled was the inception of increased reotor activity in the mouse. The ICtso in humans had" been. found to be 121 mg.min/~3, which was larger than those for 3-qutnuclitinyl benzilate, EA 3580, ant EA 3834. "iton, tiethyI-S- (diethyla~no)-ethy~phosphorothiolate, was mentioned se having been fount to be a sore potent act longer-lasting antagonist of the autonomic effects of 302,668 tn cats than physostigo~ine. ESTERS OF 1-PROPYNYLCYCLOPENTYLGLYCOLIC ACID This group of gircolates includes two compounte: the esters of the subject acid with 3-qulnuclidino] (302,212) ant with ~ methy1-4-piperidinol (302,196~. Copelan (260) gave 25 volunteers 302,212 intravenously at I.0~4.6 ug/Icg. Me initial effect experienced by the subjects was a feeling of--light~eatednese. or of milt -dizzinese. tbat starlet 15-30 min after injection. M though this symptom was reported by the subjects to be eapecially noticeable when chef were starting, it was not associated with orthostatic hypotension. Between 30 ant 60 Fin after injection, the subjects became drowsy. They often fell asleep; whenwallting, they staggered miltly ant often brushes against corridor wallet Speech became muffiet and slow, but was not slurred. Slight impaineent of logical thought ant of performance in the Number Facility Test became evident about an hour after I 88

injection. The doses used in this study did not produce incapacitation. Periphers] effects were limited usually to dryness of the mouth and, after the higher doses, ~1~d blurring of near vision. The compooltions of blood am urine and the removal of Bromsuiphalein from the blood were writhes normal limits at 24 and 48 h, ~ 4, and 4 wk. The HEDso for inducing at least a 25: reduction in the score in the Number Facility Te st was estimated to be 3.3 ug/kg. The latency of the effect with this dose was expected to be about 4 h, and the duration of the effect was estimated to be about 4 h. The latency of the effect chased little, if at all, as the dose was increased, but the duration of the effect increased irregularly, but rather definitely, after the larger doses. KIlgman and Copelan (156) stated that 25 subjects had been exposed to 302,212, but gave no t~or~ation about the experiments. The 25 subjects probably were those used by Copelan ~260). Lavallee (50) collected data from studies of the effects of 302,\96 on humps subjects that enabled him to draw a line relating the effects on the score in the Number Facility Teat to the logarithm of the dose. This ester was lese effective than scopolamine in decresaln8 the ability to perform this test. Sidell et al. (261) injected 302,196 into 10 volunteers intramuscula ~ y at 14-54 ug/kg, bracketing the previously determined intravenous ID50 of 22.5 ug/kg. The ID,o after intramuscular injection (reductional score in the Number Facility Test to nor more than lot of the baseline score) was calculated to be 28.9 ug/kg, with 95: confidence limtto of 12.0 and 6S.5 Agog. Therefore, this ester was nearly 80: as active by intramuscular. as by intravenous in Section. At the IDso, the duration of severe effects was expected to be about 101 ~n and the total duration of effects to be about 263 min. The latency of onset of severe effects was expected to be only about 12 min after intramuscular injection. The EI)so of 302,196 to increase the heart rate by 30 beats/Bin was calculated to be 23.5 ug/kg. The maximal increase occurred at about 15—n, and the heart rate was close to the baseline value by 4-6 h after Intramuscular injection. The mean blood pressure was tacreased slightly, with a time course similar to that of heart rate. Pupil dilatation was slower both in de~relopis~g and in disappearing than other changes that followed intramuscular injection. No abnormalittes of blood and urine were found at 24 h and 7 d after injection. N-HETHYL-4-PIPERIDYI.-< PIlE:NYLCYCLOBUm) -Gt.YCOLATE ~ EA 3SSO) _ Hongrel dogs unselected for sex were given 20 daily intravenous injectione of the hydrochloride of Name ~ hy1 -4~piperidyl-( phenglcyclobuty 1 ) -glycolate ~ EA 3580 0.01, 0.1, or 1.0 mg/kg during a 28-d period (262~. No detectable effects were observed with the lowest dose. Mydriasis, peosis, dry mouth, decreased activity, ataxia, and weakness of limbs were observed after the two larger doses. Tolerance to the compound developed during the study. ~ ~9

A similar e3cperl~ent vea perfoneet t263) with male rats. Hydriasis occurred at the low Lose (0~01 agIlcg~d). Tremors occurred at the two higher doses. The rats given the tHO higher toses hat their ratios of atrena1 weight to body trelght Increased algalficantly. No quantitative information h a been mate available. ~ al~ailar experiment wee perfoneed with the free base in place of the hydrochiorlte (264). The ·~e doses used in the two preceding experlmenta were ad~niatered dally, 5 d/vk, for about a month. M1 doses induced o~ydriasis. Hne body tremor e also occurred. These two obcervaUe effects were the only ones mentioned. Heir appearance was rapid after lnJectlon, their durations were "relatively short,- ant to} were re.'rotucet each day. The groups glares the two higher doses had liver weights and ratios of liver weight to body weight that were significantly larger than those of the control rate. These three estimations of subacute toxicity indicated that a modest number of repeated doses of ~ 3580 in the fore of the free base or the hydrochloride and in amounts of cot more than 10 yg/kB should be safe, although possibly disturbing. Typical antlcholinergic effects would be expected. AghaJanlan et al. (265) tote of giving four volunteers EA 3580 lntramuacularly at 1 ug/kg, seven at t.4 yg~kg, four at I.7 ~g/kg, and five at 2.0 ug/kg. With the production of at least a 25X decrement in performance tn the Number Facility Test as the criterion of a significant effect, the MED'o wee calculated to be 1.47 ug/~g. After a dose of 2 ug/kg, alg~lflcant decrement of performance extended from about 3 to about 12 h after injection; the maximal decrement after this tose (mean of four cases) was to about 60: of baseline performance. All aubJecta experienced drowaineca beginning about 1-2 h after injection and lasting for at lesat 6 h. Some of the subjects had mildly lapalred Intellectual functions, but no delirium or halluclcatlona. None of the subjects had tachycartla, but two subjects developed distinct bradycardia after unstated doses. There was no complete blockage of aweatlag, although sweating wee reduced. Decreeact ability to accommodate for near vision was obacrvet In come subjects, and four subjects (with ucatated doses) hat spears of accommodation. ~ report (266) that preasnta the reaulta of atminiaterin8 two unstated doses of EA 3580 by an unstated route to groups of four men has been mate available. The aubJecta were familiarized with the problem-solvi~g task, which involved deciding whether an array of seven letter and number symbols wee the acme as an array seen previously and then precaing one of two buttons to Indicate whether the two arrays were the same or were different. Sly days after a "seline aeselon following the familiarization aeasion, the subjects severe given ~ 3580 and were tested for ability to perform the task with the duration of presentation that had been found to yield a stable accuracy of 65-75: in the fas~lliarizatlon aeselon and that bed been used in the baseline session. About all that can be derived from: the data presented is that Ed 3580 produced a more marked decrease in the rate of responding than in the accuracy 1 90

of the responses and that the effect was more marked under conditions of extinction (no reward for a correct response) than under those of reinforcement. One group of subjects - e affected by EA 3580 more than the other, but the 408e8 are unknown. Crowell (267) presented an estimate of the incapacitated dose of EA 3580, in the fone of the hydrochloride, for can, based on the results of intramuscular lnJectio~ into 22 volunteers at 2.0-5.3 ugJkg. The criterion of incapacitation was reduction of the score in the Number Facility Test to not more than lob of the baseline score for three consecutive scores. Doses of 2.0 and 2.7 ug/k~g incapacitated none of nine subjects. One of five subjects given 3.8 ug/kg was incapacitated. All eight subjects given 4.5 and 5.3.ug/t`8 were incapacitated. The IDso was calculated to be 3.9,ug/l~g, with 95X confidence limits of 3.6 and 4.3,ug/l~g. The duration of incapacitation after the II),o was about 90 Ad; the duration of severe effects af ter that dose was estimated to be about 5 h. The time to onset of severe effects was about 2.5 h. No adverse effects of single doses of EA 3580 of the magnitude used in this study on the liver, kidneys, or blood-foming organs were detected. me effects on the volunteers were ~ imilar to those of other anticholinerg ic subetances. By 24 h after administration of the largest dose (5.3 ug/kg), the subjects were able to care for themselves. Complete recovery did not occur until 28-52 h after injection, without apparent relation to the dose. Kitzen _ al. (154) found that two groups of men with average body weights of 83.} and 66.7 kg that had received similar doses of EA 3580 (3.l and 3.2 ug/kg, respectively) suff ered significant decrements in their scores in the Number Facility Test in 45.5: and 53.~: of the cases, respectively. It seems probable that body weight played no important role in conditioning the response. This puts EA 3S8O in a class with 3~quinuclidinyl benzilate and distinguishes it from EA 3443 and tropinyl benzilate. EA 3443 seemed to have a positive correlation between body weight and response, whereas tropiny} benzila~e seemed to have a negative correlation between body wet ghc and re sponge . lIart and salter (2 38) f ound that volunteers exposed by t ~ respiratory route to EA 3560 at 4.9-~.6.ug~kg and tested before and at intervals up to 5 wk after the exposure with a modified A='r General CIassifica~cton Test gave no evidence of persistent effects from the exposures. Baker (268) compared the responses of ~rolunteere to EA 3580 adminletered as a dose per man or as a dose per unit of body weight. Four indicators of effect were used: accommodation for near vision, are-hand steadiness, dynamic flexibility, and manual dexterity. The general conclusion was that the use of the dose per unit of body weight may increase variance, rather than control for extraneous sources of variations when the purpose of a study is to establish the effects of a substance itself. The use of multlple-regreselon equations was suggested as an approach to the establishment of definitive luformation on effects of chemicals. ~ 91 the

Klutzes et al. (269) exposed 16 volunteers to an aerosolized 2% solution in acetone of EA 3580 in the fone of the free base and subjected them to a baste" of tests before and after exposure. The men were exposes to three ranges of concentration of the ester: seven were exposed to concentrations corresponding to doses of 4.9-5.9 ug/k8 (mean, 5.4 ug/kg), 8iX to 408eB of 6.4-13.4 ug/kg (meat 9.l ug/kg), and three to doses of 17.0-18.6 ug/kg (Bean, 17.9 yg/kg). The lowest range of doses produced no hallucinatione or hostile or irritable behavior. me middle range of doses produced hallucinations in four of the six subjects and m11d hostile behavior in one. The highest range of doses produced hallucinations and hostile or irritable behavior in all three subjects. In one of these subjects, the degree of hostility induced was considered severe, whereas it was "~d in the other two. US the subjects in the last group were incapacitated within 30 min after exposure. M] became delirious, a~ one semicomatose. All the subjects in this group and two of those exposed to the middle range of doses were thought by phyelciana who were not privy to the details of the experiment to require treatment by infection an. oral a4~i"stration of physostigmine. The toxic delirium disappeared within about 15 min after the beginning of therapy. The [D50 of aerosolized EA 3580 was calculated to be 8.4 ug/kg, with 95% confidence limits of 5.8 and 12.2 ug/kg. The time to onset of incapacitation was about 2 h, and spontaneous recovery from severe effects required about 9 h from their inception. The effects induced by inhalation of aerosolized EA 3580 were the aame as those induced by latramuscular lnJectlon of this agent. To judge by comparison of the retained ID50 for the aerosolized material and the ~50 estimated bar Crowell (267) for the intramuscularly injected compound, the inhaled compound seems to have only 46.4X of the activity of the inJec ted compound . Baker (270) added to the previously mentioned volunteers given fixed doses of EA 3580 tn the fore of its hydrochiorIde ( 2 68) 1 6 ~rolunteere given intramuscular irt~ectio" of the estimated IDso for a subject of average weight; 16 other volunteers were control subjects. Nine measures of perforate. were used. Each volunteer was tested five times Turing a 25-h period after injection of agent or placebo. The indicatore of effect of EA 3580, in order of decreasing sensitivity, were manual dexterity, accommodation for near vision, accommodation for distant vision, am-ha~ steadinese, estimation of elapsed time, short-term memory, Uranic flexibility, a" static strength. Baker et al. (158), tn a sugary report, stated that performance in a series of tests had been shown to be correlated with gracious personal traits and that changes in these performances caused by EA 3580 were clearly differentially related to the particular subject being tested. If the personal traits of a subject were talcen into account, a significant portion of the subject-related variance could be compeneated. The opinion was expressed that in this way generalizations about effects of substances that would be I 92

independent of the special group of sub Ject ~ used in the research could be generated. Allen and Safer (271) compared the temporal characteristics of intoxication induced by intramuscular injections of 0.25 mg of EA 3580 in ache bydrochiorlde form is men resting in the laboratory and in men considerably more active in connection with field testing. me subjects in both aituations developed their initial deficits in performance in the ember Facility Test at the same rate, but the subjects in the f ield test experienced a somewhat aver deficit and returned toward comal more rapidly. The duration of marked mental incapacitation among the subjects in the field test was about six-teethe that among those tn the laboratory. Lavallee (50) used log lose-effect graphs to demonstrate that EA 3580 was nearly as active as EA 3443 and considerably more active than 3-quinuclidiny} benzllate and f our other anticholinergic subetances in bringing about decrements in performance in the Number Facility Test. In contrast with the sequence of decreasing potency in hen subjects of EA 3443, EA 3580, ant 3-quinuclidiny] benzilate, the sequence in dogs performing a multiple-stimulus conditioned-avoidance task ~s 3-quinuclidinyl benzilate, EA 3443, and EA 3580. In monkeys performing a ~risual~iscrimination and avoidance task, the sequence was EA 3S8O, EA 3443, and 3-quinuclidinyl benziiste. In a similar task involving visual discrimination and escape by monkeys, the sequence of effectiveness of the two numbered compounds was EA 3580 and EA 3443. It He clear that none of the screening tests with experimental animals had reproduced the sequence of potencies found in the hen test. The author of the report suggested that groups of animals larger than the two of each species required for the screening tests might increase the reliability of the results obtained with laboratory animals~ Klapper et al. (163) fount that, for six men given a mean EA 3SB0 dose of ,.8,ug/kg by intramuscular injection, the scale of the PI yielding the greatest correlation with their performances in the Ember Facility Test was that for hysteria. For seven men glares a mean dose of 3.5 ug/kg, the scale with the best correlation was that for mania. For the lower dose, the most negative correlation was with the paranoia scale; for the larger dose, the gore negative correlation was with the depression scale. Sidel1 et al. (164) found that intravenous injection of S-(dlisopropylaminoe thyme thy~methylphosphonothioate Into men intoxicated with sufficient EA 3580 to decrease performance In the Ember Facility Test to l5% or less of their baseline values removed the decreases promptly and nearly completely. Oral administration of the same phosphonothioate was ineffective. The doses of the theraupetic compound were cot stated; oral admiMatration of a different dose of the phosphonothioate might be effective. Arm~tro~ et al. (239) stated tat the ED,o for ~ 3580 in decreasing the number of rewards obtained by rats trained in a sequential-response task was 37.1 ~g/kg. The ED50 for producing the least detectable mydriasis in rats was 16-30 g/kg . I 93

MISCELLANEOUS ESTERS 302,282 Kligman and Copelan (156) reported that the 1-eethyl-4-piperldyl ester of phenyl-(3-.e thylbut-1-yn-3-enyl)- glycolic acid (302,282) had been administered to 20 Den. The results were reported by Copelan (272), who gave 24 volunteers intravenous doses of 1~0~605,ug/kgo Doses of 3.8 ~gJkg and less of this agent had only alight effects on performance in the Number Facility Test by the 12 men given these doses. ~ dose of 4.3 ~g/kg decreased the test scores to below 75: of the baseline scores for three of four men. A similar result followed doses of 5.4 ~g/kg in four men. Doses of 6.5 ug/kg reduced ehe scores of all four men given them to below 75X of the baseline scores ant reduced the score of one man to 93. The mean score for this group was 30.5X of the baseline scores. The time to onset of a minimal effect after the HED50 of 4.4 ug/kg was estimated to be about 30 men, and the duration of the minimal effect was estimated to be about 30 h. The peak effect usually occurred about 2 h after injection. The initial symptom was elld -dizziness.- Heaviness of the eyelids was a part of the initial experience in many subjects. Mild unsteadiness on standing and walking began between 5 and IS min after injection. At the same time, speech became muffled and slowed. Sedation, with drowsiness and dozing, started about 15-30 min after injection. About 30 Din after injection, degradation of performance in the Number Facility Test began. A few subjects hat visual ha1J.ucinations, and one had auditory hallucinations also. Dryness of the mouth or throat was reported by some subjects given the lowest toses and by all subjects given doses of 3.8,ug/kg or more. Hild blurring of near vialon and occasional urinary frequency began after doses in the upper part of the MEDso range, but never affected all men tn a dosage group. Mydriasis was not apparent. Nausea was rarer Con June tiV8} reddest frequently "came evident within the f irst hour after injection. No significant alteratione is blood and urine were found at 24 and 48 h' ~ d, and 4 wk after injection. Saf er (2 73) gave eight volunteers intravenous doses of 302 ~ 282 at 7. 5-13.5 ,ug/kg . Three of the sub Sects became anxious after injection and two of them became agitated, restless, and passively resistant to experimental procedures. One subject vomited and later developed mild myoclonus, hyperactive reflexes, and indications of effects on the pyramidal tracts (positive Chaddock's reflex and possibly positive Babinski's sign). One subject had a Factually detected arrhythmia, which was not detectable in a later recording of the ECG. All the subjects had increased alkaline phosphatase tn their serum 24 h at ter injection; one subject whose baseline activity of alkaline phosphatase (12 g-A unite) was at the upper limit of noneal had that acti~ri ty increase to IS.2 K-A units at 24 h after injection of 13.5.ug/kg. On the seventh day af ter in Section, the active ty of allcallne ~ 94

phosphatase in his serum was 11.5 K-A uilit8. "~line phosphatase activity in one other subject rose from 7.9 to 12.4 K-A units at 24 h after injection and was 8.4 K-A units on the seventh day after injection. ~~s subject also received a dose of 13.5 ug/kg. Two of the subjects had 5-10 white blood cells per high-power field on the seventh day after injection. No other abnormalities tn blood and urine were reported. Safer (273) concluded that the ineraver~ous 11350 of 302,282 is 12.5,ug/kg, but that the compound abould Doe be administered to human subjects at doses greater than Il.5 ~g/kg because of the untoward changes mentioned a - ve. 302, 537 Brown (274) stated that the ester of 2-propeny~cyclopenty~glycolic acid with 3-quinuclidinol (302,537) is a potent antimuscarinic compound with profound effects on the central nervous system and with a potency comparable with that of BA 3580 and somewhat greater than that of 3~quinuclidis~y! benzi late . The times to onect and extinction of effects were unusually short with this agent. It was Judged to have no marked toxic or- damage-producing effects in any organism or organ system studied. lye doses and organ systems used were not stated. The organisms studied were the rat, the dog, and ache monkey. The blood-forming organs, liver, and kidneys were said to have been normal after administration of the substance. Pre sumably, other organs also were examined for pathemas. Leib (275) Stated that he gave 302, 537 intravenously at l.0~7.5,ug/Icg to I~ volunteered However, Figure ~ of 08 report contains 21 points at doses of 1.0~5.6 ug/Icg, no explanation for this discrepancy was glares. Bight men given doses of 2.0 ug/kg or less had scores in the Number Facility Test greater than 75X of their baseline scores. One of three men given 2.8 ug/kg had his score in that test decreased to slightly below 75X of his baseline score. me 10 men given 4.0~5.6 ~g/kg had their scores in the Number Facility Test reduced to below 75X of their baseline scores, but the lowest score represented tn Figure ~ of the paper was 25:, for a subject given 4.8 ,ug/leg. The intravenous M!Dso Of 302,537 was estimated to be 3 . 3 ,ug/kg, with time to onset of minimal effects of 3 h. After a dose of 4.0 ~g/kg, the time to onset of minis - ! effects was 2.6 h; but after a dose of 7.S,ug/kg, the time to onset of Visional effects was only 12 ~ n. The duration of minimal effects after the largest dose was 43 h. 008eS of 5.6 ~g~kg and more were reported to cause hallucination and delirium accompanied by restlessnese and anxiety. In the one graph for a man given a dose of 7.S,ug/kg, incapacitation, indicated by reduction of score in the Number Facility Test to below 10% of his baseline score, occurred about 90 man after the injection. Intramuscular injection of 3 ma of physostigo~ne about 7 h after the injection of 302, 537 induced a marked, but temporary, improvement in the subJect's performance in the Number Facility Test. Tmprovement lasted for about 2 h. Repetition of this dose hours later, after some ~ 95

spontaneous improvement hat occurred, had much the same sort of effect as the firat dose, but may have tocreased the rate of spontaneous recovery. t~lN2299 Fine _ al. (276) found that the ester of cyclopentyI-2-thieny~glycolic acid and die thylaai noe thanol (WIN 22993 antagonlced the muscarlaic (lowering of blood pressure of the dog by intravenous injection of acety~choline at 5-10 g/kg), the nicoti~c (increase in blood pressure by intravenous injection of acety~choline at 300~400 ug/kg into an atropinized dog), and the central nervous system (production of large-voltage cortical discharges in unanesthetized curarized cats by Sarin) effects origi - ted by cholinergic or ant! cholinest erase compound e . WIN2299 had intravenous and subcutaneous LD,os for the mouse of 80 ugJkg and 460 mg/kg, respectively, according to information supplied by the Sterling-Winthrop Research Institute. Daily intramuscular doses of 29.3 ~gImouse, beginning on the day before irradiation was started and continuing unfit death, prolonged aligh~cly (and more than atroplne) survival of mice subjected to whole-body irradiation with 550 R of xradiation, according to They and Rhodes (277~. The mortality was still 1001. These authors attributed the prolongation of life to diminution in activity of intestinal smooth muscle by the antlcholinergic compound, with c onsequently slower leakage of inf ec tive and toxic entities into the body of the mouse f rom the intestinal Ibsen. Pennes and Hoch (2 78) administered tablets of WIN 2299 to seven patients. Two patients given doses of 2 mg exhibited principally sedative effects. In addition, one of these people described.vl.sual tIlusions and "hypereensitivity to light and sound." Four patients given 6 mg experienced illusions, delusions, and feelings of unreality. The single patient given 10 fig had full-blown delirium, with complete disorientation and visual and auditory hallucinations. No information was supplied on the time courses of these effects. Pink (279) supplied some information about the time course of effects of WIN 2299. Be gave 2.0-3.2 mg intravenously and found that the subjects- began to fee} restless and excited by about 10 min after the injection. These initial effects might develop into tenet fearfulness with visual illusions and de3-usior~. Complaints of dryness of the mouth were elicited by questioning, but were not made spontaneously. The heart rate did not change u~ess or until excitement and hyperactivity developed. The effects of these doses of WIl1 2299 lasted for 2-3 h. Grob et al. (147) reported the effects of oral doses of 0.5-2.0 mg of WIN 2299 administered on seven occasions to two volunteers who weighed 46 kg (volunteer I) ant 77 kg (volunteer 2~. A dose of 0.5 me to volunteer ~ produced feelings of fullness and heaviness of the head, of being far away from the immediate en~rtronment, and of walking unsteadily with subjective and objective impressions of mental depression, with spells of crying. line same dose given to volunteer 2 produced ~ 96

l only a sensation of heaviness of the head that began about 22 min after ingestion of the dose and that lasted for about an hour. A dose of ~ mg to volunteer ~ produced the came effects as the smaller dose, but to a greater degree. There was co effect on heart rate, pupil diameter, or muscular strength. Effects began about 45 man after ingestion and ended about 5.5 ~ after togestion. This dose had marked effects on volunteer 2, beginning 20~40 man after ingestion with feeli"e of siowed movement of the arms and the legs and of slowed speech a" thought. These effects were followed by seneations of swelling of the tongue and face, dryness of the mouth, wealcmese of the limbs, mild giddiness, and marked mental depression. ObJecti~rel~y, there was obvious mental depreselon, with listlessnese, apatly, asthenia, weeping, poor attention, slowed speech, thought, and response, impaired memory (particularity for events in the immediate paste, mental clouding and confusion, inability to perfone simple calculations, and drowsiness. Th. Qll~^t ~ ~ Art ~~ at ~d ~ ___ _~ ~ _ inability to perfone simple calculations, and The subJect's grip strength did cot change, but the ability to hold arms or hands raised from a position of rest decreased. There was alight objective uneteadicess tn walking and slight aster) otaxic ef feet . There were vl8U8 illusions and a slightly increased activity of tendon reflexes. The heart rate and pupil diameter did not change. A dose of ~ me of WIN 2299 to volunteer ~ during a period of taking daily oral doses of ~ me of TEPP produced the same effects as before, but time to onset was longer (90 min) and the effects were a little less severe than when TRAP was not being taken. On the basis of this experience, two additional volunteers (weights, 66 and 53 kg) who were receiving doses of 13-20 me Of TEEP durtrtg 5-10 h were given doses of ~ mg of WIN 2299. Ingestion of WIN 2299 had a slight inhibitory effect on mild 8ympt0~8 induced by the TEPP, beginning after 20-40 "n, but little or no effect on moderately severe 8"pt0~8. ~ second dose of WIN 2299 of the she size as the first talten 45 min after the fires dose, or the ingestion of a single dose of 2 fig, had slight to moderate inhibitory activity on moderately severe symptoms induced by TEPP. These changes began about 30 min after tagestlon of WIN 2299. The effects of WIN 2299 on the central nervous system were less severe in these subjects than in those who had not received TEPP. Atropine was a Scatter antagonist of the effects induced by TEPP than AN Z299. NONESTER ANTICROLINERGIC CO}£POU=S . Ta ble IN contains the inf ormation that could be obtained on the single~dose lethal activities of the two subetances in this group, benzetim'&e and mepiperphe~dol. Istfor~ation on the toxicities of these compounds was supplied by Janssen Phar~aceutica and Sharp and Dohee Institute for Medical Research, re specs ively . Benzeti~de is 2-( N~benzyI-4-piperi dyI)-2-pheny~glutarimide, and mepiperphenidoI is I-( 3-bydroxy-4-phenyI-S- methy~hexyI)-] methy~piperidinium bromide. I 97

BENZETIMIDE In addition to the information on 8in81 e-dose LD,os for benzetimide, JaD88eD Pharmaceutics supplied extensive information on subacute and chromic toxicities of this compound. Rata fed diets containing benzetimide or given daily subcutaneous doses 5 d/wk for 14 wk gained weight at a lower rate than control animals when they were fed a diet containing 10 mg/100 g, but not when they were fed one containing 1 mg/100 g or were given subcutaneous doses of 10 or 80 mg/kg. Daily subcutaneous injections of 1.25 mg/kg did not affect the rate of growth of the rats. In general, organ weights in these animals were below those in eontro1 rats. Brain weight was not reduced, however, so its relative weight increased. No pathemas or histopathologic changes were found consistently. Rats fed dicta contalaing benzetlmide at up to 10 mgtlOO g for 21 mo had no unusual gross mortality due to the compound. Among the rats that died in the control group, males accounted for 37. 5X of the deaths; among the animals fed diets containing benzetimide ~ males accounted for 52.2: of the deaths. Among those fed the diet containing the large&l concentration of benzetimide, males accounted for 62. 5% of the deaths. Pregnant rats f ed diets containing benzetimide at up to 20 mg/IOO g on days 6-15 of pregnancy for one or three generations gave birth to litters of the same adze as those borne by control rats and produced no abnormal offspring. Pregnant rats given daily subcutaneous doses of 0.63-160 mg/kg during the f {rat 21 ~ of pregnancy experienced no striking effects at doses below 10 mg/kg. At 10 mg/kg, but not at 5 mg/kg, there was a de crease in the incidence of implantations to 60Z of the females, compared with 90~92: i:` the controls. With tally doses of 40 mg/kg, one of 20 dams died before parturition, and the incidence of implantations decreased to 10: of the females. At the highest dose (160 mg/Icgy, three of 20 dams died, and no implantations were found. The weights of the offspring at birth were reduced slightly, but no abnormal offspri n8 were report ed . Pregnant rabbits given benzetimide by gavage at 0. 31 or 2.S mg/kg on days 6-~8 of pregnancy had increased numbers of dread and resorbed fetuses (28. 5X Ire. 7. 7Z in the control group), and one of 59 offspring born to the rabbits given the largest dose of benzetimide was deformed. These various experiments have demonstrated that benzetimide in the dose ~ used gave no evidence of mutagenic or carcinogenic activity and only questionable evidence of teratogenic activity. High dose. of the compound did interfere with reproductive processes. Janssen and Niemegeers (280), on the basis of testo with prophylactic administration of benzetimide to rats given pilocarpine hydrochiorlde intravenously at 80 mg/kg, concluded that benzetimide had sufficten~c action on the central nervous system to Justify its conaideration as a t}leraupeutlc agent in parkinsonism. They considered also that it was one of only three compounds, in a total of 23 anticholinergic subetances examined, that were able at submydria~ic doses to prevent lactlmation and salivation induced by pilocarpine. I 98

Soutidn et al. (281) examined the bloding of [38Jbenzetlmlde to subcellular fractions of the rat caudate nucleus and found both in vitro and in vivo that the (+) isomer was bound to the particles in a fraction composed of meo~branea and nerve endings to the extent of 4-7 times the (-) isomer. Fractions composed more nearly exclusively of membranes or of nerve endloga bound the labeled material less extensively than the fraction that contained both atructurea. 81ndiDg of benzetimide in vitro was reduces markedly by atropine and by carbachol, but only slightly, if at all, by d-tubocurarine. Beld and Arlene (282) measured the bindlag of benzetio~ide to microsomes from smooth muscle isolated from bovine tracheae ant to partlelea In homo~enatea of bovine cautate nuclei. Plotting the amount of ~ 38] benzetlmide bound to the microsomes as a function of the concactration of [3HJbenzeelmlte to contact filth them yielder a blphasic curve for (~)benzetimide, whereas thee for (-)benzeti2eide was rectlllnear. At a benzetloIlde concentration of about 6.3 ~M, 9 times as much of the (+) isomer was bound to ~crosomes fro. vascular tooth muscle as of the (-) isomer. The partleles In a homogenate of caudate nucleus bound about 5 times as much of the (+) isomer as of the (-) isomer. The saturable component of the material that bound (+~benzetlo~ide hat · higher affl~lty f or the isomer than the noneaturable -component, which bound (-)benzetlmlte about as well as it bound the (+) laomer. When the microsomes were exposed no a large concentration of unlabeled (+)benzetimlte before addition of {38~-labeled (+)benzetimite, binding of the labeled isomer aeemet to be only to the unsaturable component. Atropine wee found to bind to the same entities as benzetimide and to block binding of [3H] (+~benzetimide by the saturable component of the crosomal preparation. Karger (283) gave 17 volunteers benzetimide intravenously at 1.0~10.0 ~g/kg. He estimated that the MED,O for lowering the ac ore tn the Ember Facility Test Leo 75: of the baseline value for 50X of a group of subjects was 8.9 ~g/kg. M1 Den who met this criterion for a minimal central action had difficulty in focusing, which lasted in one case for 24 h. Xlapper et al. (162) re-examined one subject who had received benzetimite at some unstated time in the past. No evidence of long-term effects was found. MEP IPER~NIDOI. White et al. (152) gave six volunteers intramuscular lOjecti0~8 of 100-300 fig of mepiperphenidol (1.17-3.63 mg/~) and found that all six had blurred vision ant receding of the' near point of vision. These effects hat nearly disappeared by the following mornlug. Urinary urgency and an increase of 10.8 fig in glean diastolic pressure were particularly strlklag effects. Me mean Systolic pressure increased by 5.9 ~ fig, ant the ocean heart rate by 45.3 beats/mln. The mean breathing rate increased Slightly. All the subjects co~plalnet of dryness of the mouth, and all but one of becoming drowsy and of feeling unsteady in walking. The dose of mepiperphenido] I 99

increased the effectiveness of photic driving of the BEG in half the subjects and reduced the alerting reaction in one of the six subjects. Two subjects complained of eye {irritation. Nine subjects given ineramuscular inJectione of I.95~6.5 mg of atropine sulfate (0.031-0.064 mg/]cg) and 150-300 mg of mepiperphenidol (2.21-3.99 ag/kg) all had blurrtry' of vision and marked receding of the near point of Pistons which was still somewhat farther away than normal on the following morning Clean, 10.1 tat firs. 5~3 i~aeJo The mean diastolic and systolic blood pressures increased by 10~0 and 5~6 ~ Hg, respectively. The mean heart rate increased by 3769 beats/Bin. All nine subjects complained of dryness of the mouth, and eight of drowsiness and ataxia. Four of these sub jects experienced slight nausea. Three complained of eye irritation. According to the data supplied ~ the Sharp and Dohee Institute for Hedical Research, mepiperphenidol is one-tenth as active as atropine in antagonizing the actions of methacholine chloride on the dog. If this Prague t8 used to calculate an aeropine-equi~ralent dose for the subjects who received both atropSne and mepiperp-henidol, those who received sufficient doses of the two subetanees to yield atropine-equi~ralent doses of more than 0.320 mg~kg had a four-in-se~ren chance of developing nausea. The four persons who experienced nausea in this group were three men and one worn; the remainder of these subjects given atropine-equivalent doses above the threshold dose were two men and one wooers. In the group given atropine alone, the man who was the only subject who complained of nausea had received atropine sulfate at 0.037 mg/kg. Eight other subjects (ale men and two women) hat been given larger doses of atropine; two of these subjects complained of headache (accompanied by dysmerria in one), and a third complained of vertigo. In the group given mepiperphenidol alone, a woman who had reset an atropine-equivalent dose of 0.131 mg/kg complained of Blight nausea. Four subjects tn this group (three men and one woman) received larger doses of mepiperphenidol than the ~ssuseat.ed subject, without making any complaints . On comparing the changes in the mean values for the objectively measured properties induced by the three drug regimens used by White et al. (152), those for the subjects given both atropine and mepiperphenido] were below those for the subjects given either compound alone for systolic and diastolic blood pressures and were above those for the subjects given either compound alone for the rate of respiration and the near point of vision. The change in the mean pupil diameter for the subjects given both compounds was the same as that due to mepiperphenidol alone and greater than that due to atropine alone, whereas the change in mean heart rate for the subjects given both agents was almost precisely halfway between those due to the individual compounds. On the basis of tin analysis, aeropine and mepiperphenldo} appear to be mutualig. antagonistic in relation tO the regulation of blood pressure, to be somewhat antagonistic in their actions on heart rate, to be slightly mutually antagonistic with respect to breathing ~ 100

rate and activity of the ciliary bodies in the eye, and to be net ther antagonistic nor additive in action on the circular muscle of the iri s. Jager (94) found, in in vitro experimente, that the a~cropinase in rabbit serum was inhibited strongly by mepiperphenidol, whereas the atropinases in rabbit and guinea pig liver were comparatively resistant to inhibition by mepiperphenidol. n TAB a ~ XTt1 RE This ~xture contained 50 mg of atropine sulfate, 210 me of N-diethylaminoethyl benzilate (benaceyslne), and 2 g of ~ rime thylenebi ~ ~ 4-hydroxyioLinome thy~pyridinium)dibromide (TRIBE) in 100 m1 of solution. Wlles and Fort (284) summarized the results of experiments in which rats, rabbits, togs, ant monkeys were given intramuscular injections of graded single doses of TAB. Rate of the two sexes were studied separately. All the rabbits used were Bales. Dogs and monkery were unselected as to sex, but the groups of ads of these two species contained approximately equal n~bere of the two sexes. The first sign of toxic actions seen in the rat, the rabbit, and the dog was acacia; the first one seen in the monkey was ptosis of the eyelids. Table I-5 summarizes the information obtained from the studies summarized by Wiles and Ford (284) 884 by Lee et al. ~ 2 85) . The intramuscular doses that resulted in the first signs of an effect in 50: of intact animals of the various species and in death of 50% are given. TAB appeared to be more lethal (by a factor of 1.67) to female rats than to male ones, although the threshold toxic doses f or the two -sexes were identical. The rat apparently was more resistant to intramuscular TAB than the other species tested, the male rat having an intramuscular LD,o nearly 4 times the mean of those for mice, rabbits, dogs, ant monkeys. Dogs given-22.6 mg~kg had the acti~ritsr of their serum glutamic-oxaloacetic transaminase (SOOT) increased during S h after injection by a mean of nearly 85X. Thereafter, SOOT fell; at 7 d after injection, it was not significantly different from the control value. Monkeys that died 2-3 ~ after injection had liver necrosis and deposition of collagen in the renal gio~enlli. Monkeys had a series of alterations of the activity of SOOT similar to that reported for the dog, the mean activity 5 h after injection being increased by more than 118: above the baseline activity and decreasing slowly thereafter. Lee et al. reported that all mice that died did so within 2 d after a single injection of TAB, which these tn~restigators concluded was co more lethal than the IMB-4 in the mixture. Wiles and Ford (286) recorded the results of an experiment in which animals were given subcutaneous (rats) or intramuscular (dogs and monkeys) doses of TAB at t.2-41.2 mg/kg 5 t/wk for 4 wk. Rats were hyperactive for about ~ h after each injection, whereas the dogs (beagles) and monkeys were less active after each injection than they had been before it. The response of the iris to illumination of the retina in the I 101

dog and the monkey became sluggish to absent, depending on the dose of TAB. Three rats, all from a group given doses of 12.9 mg/k8, died during the experiment; a male died after the fifth dose, and two females died after the last (twentieth) dose, but before they were to be subjected to necropay. No definite cause for these deaths was found. No other animals used as subjects in the experiment died during the 4~wic study. Nolgate and Sidell (287) reported the results of an experiment in which three volunteers were given intramuscular injections of 2 m} of TAB and three other volunteers were given ones of 4 ml. By 9 win after injection, the mean performance in the Number Facility Test had barn reduced to SIN (2 mI) o; 59: (4 mI) of the baseline performance; by 5.5 h, performance was 98-9 9X of the baseline pert ordnance. Me heart rate increased slightly (by 6.6: after 2 I; by 44X after 4 mI) within the 0.5 h after injection; 6.5 h after either injection, it was below the original value. Systolic and diastolic blood pressures increased slightly after injection and then slowly fell to below the original values within 4.5-6.5 h after inJeetion. The pupil diameter increased after both doses, and both near and far visual acuities decreased. The ~O8t colon complaints voiced by the subjects of the experiment were of dryness of the mouth, muffled and indistinct speech, dizziness' weakness of the arms and legs, diminution of the sense of balance, and slowing of reflex responses. Other complaints made less frequently included those of thirstiness, sleepinese, disorganized activity, distraction by Douse, restIessness, uncoordinated mental activity, inability to focus the eyes, blurred vision, muscular twitching, weariness, discomfort in the throat durlog swallowing, malaise, inappropriate desire to laugh, pauses, weakness of the voice, and rubbery legs. Sided] (288) described an experiment similar to that of Holgate and Sidell (287) in which four volunteers were given intramuscular injections of 2 m} of TAB solution and another four were given 4 mI. The smaller dose resulted in a decrease in the mean score in the Number Facility Test of 40% during the 15 min after injection, whereas the larger dose resulted in a decrease of 78Z during the Awe Scone. The score returned to its preinjection value at a Beast of 2.2S h after injection of the smaller dose, whereas 2-5 h were required for a return to the preinJection Prague after the larger dose (no mean time for the four volunteers given this dose was stated). Three of the four Len given the larger dose experienced hallucinations. The smaller of the doses admir.isteret by Sidel1 resulted in a slight increase tn supine systolic blood pressure for l-1.5 h after Injection and a more marked and prolonged increase in supine diastolic blood pressure. Standing systolic blood pressure was 5-8 mm Ng below supine systolic blood pressure, whereas standing diastolic blood pressure was about the same as that in the supine position. Hean pupil diameter at 5 h after injection was about 0.5 mm greater than that bef ore injection. Af ter the larger dose of TAB ~ supine ant standing systolic pressures were increased for about 0~75 h af ter the injection, and supine diastolic pressure was . . ~ ~

increased for about 3.5 h. Standing diastolic pressure was about the same as that before injection. At 5 h after injection, the mean diameter of the pupil was 2 ~ greater than that before injection. Sidell reported that the sub jecto who received the smaller dose of TAB had xerostomia, blurred vision, "dizzluase" or light-headedness, drowainess, slightly impaired coordination, and some difficulty with concentration and memory. These appeared 5-10 min after injection, and the subjects generally were free of symptoms by about ~ h after injection. One subject was markedly nauseated, vomited, and complained of a severe headache. me subjects who received the larger dose were grossly uncoordinated, one being unable to stand without assistance, in addition to having xerostomia and blurred vision. Three of the four men had hallucinations, poor attention spans, impaired ability to concentrate, and poor memory. The effects started within 5 min after injection, peaked at about 30 man, and had largely subsided by 2 ~ after injection. Two of the subjects, one from each dosage group, who had been made quite uncomfortable by the effects of TAB were treated with physostigmine 18 min after receiving TAB. In both cases, the toxic symptoms were ameliorated within 10-15 min after administration of the physostigmine. Ilolgate and Sidell (289) summarized the effects of intramuscular injection of 2 m] of the TAB solution into three subjects who were exposed in a hot room at 35°C and a relet ive humid) ty of 90X f or ~ h bef ore the dose of TAB and f or 2 h after the injection. Merely increasing the temperature and the humidity was not fount to be sufficient to produce an ir~rease in heart rate, so long as the subjects remained at rest. Prom the dose-response data reported earlier, a single dose of TAB failed to have much effect on this rate. Yet the combination of TAB and increased temperature and humidity greatly increased heart rate. Cognitive function was not significantly altered under the conditions of the control tri als, although there was a tendency for the exerc ising subjects to have lower ac ore s in the Number Facility Test than the group at rest. Comparison of the effects of a~single dose of TAB in the lose-response study with those in this study indicates a synergistic relationship between TAB and thermal stres s. Holgate and Sidell (290) described an experiment in which eight volunteers were given intramuscular injections of 2 o'1 of the TAB solution after having trained in the performance of a lever-pressing tack that involved both estimation of time and vigilance; four other volunteers were given intramuscular injections of 4 ml of the TAB solution after training in the same task. The 2-m! and the 4 ml doses induced increases in the mean heart rate of 30X and 90:, respectively. The maxima change in heart rate occurred 10-20 win after injection and lasted for about 4 h after the larger dose. After the smaller dose, the subjects continued to be able to estimate time reasonably well and to respond effectively to visual signals in the vigilance port ion of the task. The larger tose of TAB resulted in failure by the subjects to initiate timing I 103

re sponges and to re spond to visual signals in the vigilance part of the task. Auditory signals of the erroneous estimation of Dime and of failure to re spond to the visual signs Is still evoked intermittent, but usually late, responses by these subjects. Holgate and Sidell concluded that the subjects could continue to function at a reduced level after the lower dose of TAB, but were virtually incapacitated after the larger (lose e It is apparent from the signs and" symptoms of intoxication by'rAB that the sublethal effects are due primarily to the anticholinergic components of the Cloture. Lois impression is strengthened by the reported beneficial effects of administration of physosti~ine to seriously intoxicated subjects. However, the data in Table 5 on the lethal effectiveness of the TAB mixture, in comparison with data in Tables ~ and 2, show that the LDso of TAB is closer to that of ~B-4 than to thee of ei the r of its anticholinergic constituents. Lee et al. (285) estimated that the intramuscular single-dose I~50 of TMB-4 for male mice was S3.5 mg/kg, whereas that of TAB was 64.5 mg/kg. The corresponding figures for benaceyzine hydrochloride and for atropine sulfate were 9201 and 604 mg/k~, respectively. Mice that died after doses of TAB, of IMBo4, or of benacryzice hydrochiorlte did so after bouts of clonic and tonic convulsions, whereas those which died after doses of atropine sulfate were severely depressed and ataxic. Mice given benacryzine hydrochloride were very sensitive to tactile or auditory stimuli, which often originated seizures. This triggers ng effect was not reported for the mice given either TAB or THB-4. The hindleg paralysis described as affecting only those mice given TAB may have resulted from a combination of the ataxic action of atropine and the blocking activity of TIdB~4 at the neuromuscular junction. DISCUSSION The initial inters st of the chemical defense establishment in anticholinergic compounds was related to their use in antagonlsing the toxic actions of the nerve gases. The early research on the biologic actions of this group of compounds performed by Edgewood Arsenal am its contractors was directed toward identification of the ii~t~ within which atropine might be used advantageously as a prophylactic and therapeutic agent a8ainet intoxication by inhibitors of cholinesterases arid toward determination of whether any synthetic anticholinergic compound, wNlch ~ghe be more readily avatiable than a~cropine during ~ convict, had safety and efficacy at least equal to those of atropine. For exile, the group of investigators at the Johns Hopkins Medical School (Abaer McGehee Harvey, David Grob, Joseph Lillienthal, Jr., Richard Johns, John Harvey, etc. studied several synthetic anticholinergic compounds that had seemed in preliminary test. with animale deco have some superiority over atropine in an;tagonlsin8 the toxic effects of inhibitors of cholinesterases. This was done in part by treating cholinergic crises, in patients with myasthenia gravis who were being given inhibitors of cholinesterase (DFP, TEPP, ~ 104

Sarin, and neostigmine) Lo increase their muscular strengths with the synthetic antlcholinergic compounds and comparing the results of such treatment with those of similar administrations of atropine. Some normal volunteers also were used in this sort of research. Unf ort unately, none of the anticholinergic compounds tested as outlined above was found to have any superiority over atropine. The research with WIN 2299 summarized in one of the reports (147) included in the group submitted for thle survey is an example of the work of the group at the Hopkins. Other groups doing research on anticholinergic compounds in the same era were those at Galesburg State Research Hospital (Harold Himwich and associates), Galesburg, Ill. (144,157), at the Montefiore Hospital (Yale Koskoff and associates), Pittsburgh, Pa. (158), and at the University of Illinois School of Medicine (Archer Gordon ant associates), Chicago, Ill. (27,151~. After a decision had been made to seek a new type of chemical agent. among compounds that had disruptive actions on the normal functions of the central nervous system, anticholinergic compounds because of interest again, because of the well-known activities of atropine and acopola~ne in producing a temporary toxic, psychotomimetic state. Itost of the reports in this survey belong no this later period, which starlet in about 1958. TAB, the last substance discussed in this survey, was intended to be a therapeutic agent in nerve-gas intoxication for self-administration by soldiers, as well as for use by medical personnel. It combined the pert pheral and central anticholinergic actions of atropine and benacryzine with the principally peripheral cholinesterase-reactivating activity of trimedoxl;oe (THB-4~. It was devised in approximately L975 after Russian chemical protective kits captured by the Israelis were f ount to contain somewhat similar preparations. Although the anticholinergic substances considered in this survey differ in chemical cons~ltution, all seem to produce about the same effects. There is some variation tn the balance between central and peripheral actions among these subetances, the compounds containing quaternized amino functions having almost exclusively peripheral actions when moderate doses are administered. The anticholinergic compounds said to have been adminlatered to volunteers, but not mentioned in the reports submitted for this survey, may reasonably be expected to have actions similar to those of the 24 that have been discussed, because of the similar! ty of actions among the 24. Single-dose Lassos for mice, rats, and rabbits have been found for 17 substances other than atropine that permit their lethal activities-to be related to that of atropine. On the basis of atropine - 100, the lethal activities of single doses of the 18 substances fall in the following order of increasing potency : methyIscopolammoniu~, scopolamine, d-hyoacyamine, atropine, meehylatropinium, WIN 2299, caramiphen, EA 3834, 302,668, benzetimite, mepiperphentdol, 3-quinuclidinyl benzilate, benacryzine, L-2-alpha-tropinyI-L- (pheny~cyclopentyl)-glycolate, TAB, tropiny} benzilate, Ditran, I 105

and I`-2-alp}~a-tropinyl benzilate. No information on the single-lose lethalities of EA 3443, EA 3580, 302,196' 302,212, 302, 282, 302, 53 7, 3-quinuclidinyI-L-( pheny~cyclopentyI)- glycolate, - or 2~ethyl-3-quinuclidinyI-L- (pheny~cyclopentyI)-glycolate were provided or could be found in the literature. Such information should exist; in its absence, no precise judgment on the safety of these compounds can be made. That is not to say that they are unsafe. The human studies carried out with these compounds indicated that none had persistent effects on the volunteers beyond several days at ter single doses. Among the 13 former volunteers who had been given at least one of the compounds with which this report is concerned and who were re-examined by Klapper et al. (161) in 1970~1971, only one' of two who hat received 3~quinuclitinyl benzilate, reported having experienced a flashback. The other abnormalities reported by or found in the nine former volunteers who had been given atroplne, scopolamine, or 3~quinuclidinyl benzilate probably were unrelated to their serving as volunteers. The four men who had been given EA 3834, 302, 668 9 L-2-alpha-tropiny! bensllate, or benzeelmide apparently had no abnormalities. Among the 16 compound for which doses effective in producing some definite alteration of function were stated, data are available for Il after latramuscular injection. The other f ive compounds were administered by intravenous injection. Because we have in no case an ludication of the relative effectiveness of intramuscular and intravenous injections of these five agents, it is impossible to rank am 16 in order of relative effectiveness. Among the compounds adminlotered to volunteers by intramuscular injection, the most active in inducing a degradation in normal function-~ost commonly assessed on the basis of the dose required to cause loss of at least 25X of the predose score in the Number Facility Test by 50X of a group of subJecte-~as EA 3443, followed fairly closely by EA 3580, EA 3834, and 3~quinuclidiny] benzilate. This ranklug agrees with that by Lavallee (50), except for BA 3834, which was not included in that author's rating. Less active than 3-quinuclidiny} benziJ-ate, in order of decreasing effectiveness, were mepiperphenidol, 3~quinuctidinyI-~-( pheny~cyclopentyl )- glycolate, 2~ethyl-3-quinuclidinyI-~-( pheny~cyc lopentyl) - glycolate, L-2-alpha-eropinyl benzilate, scopolamine, methyIscopola~onium, and atropine. Among the compounds sd=nistered by intravenous injection, the most active were 302,212 and 302,537. The intravenous doses of these two compounds required to induce a definite degradation in normal function in the volunteers were larger than the intramuscular dose of 2~e thy I-3-quinuc lidinyI-~-( pheny~cyclopentyI) -g lycolate required to satisfy the same criterion; thus, one surmises that these two compounds are probably weaker than the one or two agents that follow 2`ethyI-3~quinuclidinyI-L-(phenyI- cyclopentyI)-glycolate in the list of compounds injected intramuscularly. The other compounds injected intravenously, I 106

in order of decreasing activity, are 302,668, 302,282, ant benzetimide . Regarding the 16 compounds for which incapacitating doses--usually calculated as the doses required to degrade the score in the [lumber Faclilty Test to not more than 10: of the predose acore--were given, three routes of administration were used: intramuscular in Section, intravenous injection, and ingestion. lye most effective of the compounds injected intramuscularly was 3-quinuclidinyl benzilate, followed in order by EA 3443, EA 3580, 3~quinuclidinyI-L-(phenyI- cyclopenty! )-glycolate, I.-2-alpha-- ropinyI-L- ~ pheny~cyclopentyl)-glycolate, I~-2-alpha-tropin,,! benzilate, scopolamine, 302,196, atroplne, and Ditran. The most actively incapacitating of the intravenously injected compounds was 302, 212; itS i~apacitati~ dose was larger than that of 3~quinuclidinyI-L-( pheny~cyclopentyI)- glycolate, so it probably fits into the series of compounds given by intramuscular injection two or more compounds below 3-quinuclidinyl-L-<pheny~cyclopentyi)-g, ycolate. Of the two compounds administered orally, EA 3834 was slightly more than twice as effective as WIN 2299. Its effective oral dose was about 64: greater than the ID,o for Intramuscular L-2-alpha-troplny} benzilate. EA 3834 apparently ranks fairly high in comparative ability to produce incapacitation, and WIN 2299 probably falls near the middle of the hierarchy of compounds with this type of activity. Among the seven compounds injected intramuscularly for which times to onset of effect were stated, the most rapidly acting was 302,196, followed closely by scopolamine. Then followed in order L-2-alpha-~ropinyl benzilate, 3-quinuclidinyl benzilate, 3~quinuclidinyI-L-< phenylcyclopentyI)-glycolate, 5\ 3580, and 2 "ethyl-3-quinuclitinyI-L-(pheny~cyclopeneyl)-- glycolate. Among the intravenously injected compounds, the one with the fastest onset of action was 302,66B, with a time to onset of 30 min. whereas intramuscularly injected scopolamine ant L-2-alpha-troplny} benzilate had times to onset of 15 and 60 ~in, respectively. WIN 2299 hat a time to onset of 40 min after ingestion, whereas EA 3834, by the same route of administration, had one of 168 min. The two compounds other than 302,668 injected intravenously, 302,537 and 302,212, had times to onset of 90 and 240 min. respectively, so they probably fall in the middle and lower parts of the entire hierarc}~. Two of the 12 compounds for which durations of effects were stated are outstanding in thle regard: 3-quinuclidinyl-1~-(phenylc~rclopentyl)-glycolate and 3-quinuclidinyl benzilate, with mean durations of incapacitation of 69 h. L-2-alpha-'rropinyl-L-(phenyl- cyclopentyl)-glycolate came in a creak third, with an estloIatet duration of incapacitation of 27 he The other nine compounds for which this measure is available had durations of action between 90 min ant 6 h. In order of increasing duration of effect they are EA 3580, 302,196, 2~e thy 1-3-quinuc lidinyl-L-( phenylcyclopentyl)-glyc orate, L-2-alpha-tropinyl benzilate, scopolamine, EA 3834, WIN 2299, 302, 668, and 302, 212. I 107

Because of the particularly long durations of their effects, 3-quinuclitinyl benzilate and 3-qulnuclidinyl-L-(pheny~cyclopentyI)-glycolate (EA 3167) might be more hazardous to both corporeal and mental health than the other compounds on which reasonably complete information to available. However, 3-quinuclidiny} benzilate has one of the most favorable ratios of LDso for small rodents to IDso for humans. The corresponding ratio for EA 3167 cannot be stated, because no data on its LD50 for experimental animals have been provided. Of the five qutnuclidiny} compounds on which information derived flora experiments with h~an subjects was provided, we have measurements of lethality for experimental animals only for 3~quinuclidinyl benzilate. The only basis on which we can compare this group of compounds is effect on the volunteers. Of these five compounds, 3-quinuclidiny} benzilate is the most potent as an incapacitating agent, and it and EA 3167 (the next 08t potent of the five as an incapacitatlag agent) have part Ocularly long durations of action, requiring 3-5 d f or complete recovery. The other three compounds in the group have durations of incapacitation of about 2-6 h. As ~ group, these compounds are not particularly rapid inducers of whatever changes follow their administration; even after intravenous injections 302,212 and 302,S37 required 4 and I.5 h, respectively, to induce incapacitation. After intramuscular injection, 3-quinuclidinyl benzilate, EA 3167, and 301,060 required i.25, 2, and 5 h, respectively, to induce incapacitation. These long times contrast with 12 min for 3023196--N-methyI-4-piperidly~cyclopentyl-~-propynyl)- glycolate. They probably reflec t comparatively slow penetration into the brain and adsorption onto nerve cells there. The dais for the differences is not readily apparent. In both animals and man, the earliest symptoms and signs that follow administration of any of the acticholinergic c ompounds are restlessness, a f eelir~g, of dryness of the mouth, dryness and flushing of the skin, and decreased motility of the gastrointestinal tract. The effects produced by these compounds can be categorized as effects on or through the ~ central nervous system, effects on peripheral effectors, and effects on or through nucleotides. The effects mediated through anticholinergic actions on the central nervous ejetem include restiesenese, shortened attentiveness and decreased ability to concentrate on a topic, sensation of dlzzinese or faintness, tiredness, lassitude, drowsiness, sensations of heaviness and of altered shape of the limbs, apprehension, flattening of the EEG and decrease in the predominant frequency, weake~i~8 or abolition of the visual alerting reaction in the EEG, increase in photic driving of the EEG, ataxia, nausea, mental slowing, underestimation of elapses time, decreased sensitivity to pain, obstinate progression, vomitir~g,, incoherent and extravagant ideation, xanthopoia and other chromatopsias, disturbed sleep, hypers~efiexia with development of Babinaki' ~ sign, illusions, disorientation, hallucinatione (predominantly visual), somnambulism, delirium (possibly violent), and coma with occasional convulsions. I 108

Effects of anticholinergic compounds that may depend primarily on actions at neuroeffector Junctions include trynese of mouth, difficulty in swallowing, hoarseness and dry cough, dryness of nasal mucosa and nasal stuffiness, dryness of con junctivae, relaxation of smooth muscles of the intesti - } tract with decreased peristalsis, ~rasodilatation in conjuncti~rae and selerae, brightness and dryness of the eyes, mydriasis and cycloplegia, flushing of face and upper chest, increased body temperature during exercise or in a hot environment, temporary bradyeardia (rarely, but possibly, with cardiac standatill), tachyeardia, slightly decreased systolic blood pressure, increased diastolic blood pressure, headache or eyeache, difficult urination, dyspnea, atrial fibriliatlon, a trial ventricular dissociation, and ventricular fibrillation. The only compound in this group whose fate in the body has been studied to a moderately satisfactory extent ts atropine. Some information on the metabolic fates of 3-quinuclidiny] benzilate and of tiethylaminoethy} benzilate is a~ratiable ( 193, 219), but it does not account completely for all parts of the molecules. The binding of 3-quinuclidinyl benzilate to nerve cells (193,194) and to organelles from such cells (195) has been studied, but no detailed studies of its detoxification, pharmacol~netice, and molecular pharmacology have been fount for use in this review. Little or no information on the biochemical aspects of the activities of the anticholinergic compounds surveyed here has been found. Whether such information exists in literature that has been withheld from consideration for this surrey is unknown. So far as the general impact of antichollnergic compound on human health is concerned, the principal ill effect that has been attributed to long-ter~ administration of these compounds ts the induction or exacerbation of the dyakinesta that may occur after long~continued use of neuroleptlc compounds. This condition is characterized by fnter~l~tent protrusion and rotation of the tongue and by chewing or biting movements of the Jaws and" mouth, in many cases with athetold or choreifom movements of the limbs (292-296~. The condition has been reproduced in anthers, at lease in part, by administration of neuroleptlc drug e (297, 298~. Scopolamine was found to increase the intensity of the experimental syndrome induced in mice by such bugs as methy~phent~ate and teflutlxol (29B,299~. By analysts of electromyographic records from patients with tardive dyakinesia during administratione of various drugs and procedures, Ma no and associates (300,301) concluded that the dyokinesta is induced by relative hyperfunction of dopaminergic neurons in the striates established by excessive inactivation of cholinergic neurons by anticholinergic drugs. There is abundant evidence that medication with anetcholinergic drugs is contraindicated in human tardi~re dysicinesia (302-309~. Burnett et al. (308) found that the improvement in tsrdive dyokinesia after disconti'2uance of the anticholinergic drug, when both an antlcholinergic compound and a neuroleptic drug had been used to treat schizophrenia, was not due to any change in the patients' serum concentration of I 109

the neuroleptic compound, but muat have been due to withdrawal of an effect by the parasympatholytic compound. Gerlach et al. (310) had advanced essentially the acme idea in suggesting that centrally active anticholine Tic drugs administered to patients with eardive dyakinesia increased the predominance of dopamine in the brain seem (eatabliahed there by the induction by neuroleptic drugs of supersensitivity to that effecter substance) by decreasing the effect of acetylcholine on striatal receptors and thereby disturbing the balance between chollnergic and dopaminergic effects. Martys (311) investigated the drugs responsible for "certain' and "probable" reactions in 335 patients in a total of 817 patients treated during a parlor of years, findlog that 51X of all the reactions were canaet by drug a that hat actions on the central nervous system. The most common of these reactions were droweiness, nausea, dizziness. diarrhea, headache, and dry mouth. When the symptoms collected by tnvestigatore (311-314) who had used a variety of antichollnergic drugs for various purposes are arranged in order of decreasing frequency of specific complaints, the f ollo~ring fiat results: incoherence, disorientation, f rushed faciea, hallucination, mydriasia and cyclopedia, restlesaness, hyperactivity, picking or plucking olotiona with the fingers, ataxia, motor incoordi~tion'~ coma, tachycardia' co~uslon9 dryness of the mouth. hyperreflexia, apprehenston, fear. somnolence, fever (above 37.~°Cy, retrograde amnesia, toxic delirium, nausea, diarrhea, headache, noc curia, urinary retention, impotence, painful ejaculation, tremor, geatralgia, rash, itch, and vomiting. Pfeiffer et al. (312) found that children generally were somewhat more sensitive to antimuscarinic drugs than adults; the root mean square of the difference in order numbers based on the frequencies of signs and symptoms of intoxication by 3-quinuclidinyl benzilate between adults and children was 2.66. Children had higher frequencies of occurrence than adults for 10 of the 15 effects counted. Craig (315) analyzed the nature of death of 48 patients in a mental hospital whose deaths were attributed to asphyxia and found that 14 of the patients had choked to death. Because therapeutic regimens for psychotic patients commonly include combinations of neuroleptic, antimuscarinic, and tricycItc antltepresasut drugs and because all these categories of drugs have the ability to induce senaitization of neuronal effectora to dopamine or to prevent access of acety~choline to cholinergic neuron 1 effectora, or both, Craig and associates (316) undertook a further study of the asaociation among alteration of the gag reflex, possible drug-induced necrologic syndromes' and the adminlstratlon of drugs to chronically hoapitallsed psychiatric patients. Of the 58 patients in the study (316), 34 were conaltered to have normal gag reflexes, mine to have variable responses to touching the posterior pharugeal wall, and 15 to have definitely impaired gag reflexes. The last two groups were combined for the analysis of drug use. All 58 patients had been receiving neuroleptics and antiparkinsonism drugs; 12 I 110

(35Z) of the 34 patients with co~1 gag reflexes had received antlcholinergic bugs, whereas 15 (62~) of the 24 patlente with variable or impaired gag reflexes had received anticholinergic drugs. In the group with normal gag reflexes, only 14 (413) had facial-oral dyakinesia, and only 19 (Shy had dyokinesia of some part of the body. In contrast, 19 ~ 79X ~ of the patients with variable or impaired gag reflexes had facial-oral dyalcinesia, and 20 (83X) had dyakineala of some part of the body. Craig_ al. concluded that patients with tardive dyakinesis, and pare Ocularly those receiving anticholinergic drugs, are at high risk of having impaired gag reflexes and, consequently, of ha~rin`8 swallowing impairments that increase their risk of death by asphyxia associated with entrance of food or other foreign material into the larynx. InJections of atropine and hyoacine into chicken eggs on the fourth day of incubation hare resulted (291) in increased rates of death of the embryos (e.g., 33: tiering on the twelfth day of incubation vs. 9IZ alive in eggs into which saline was injected) and of malformation (e.g., 33: of those alive on the twelfth day of incubation vet 03: occurrence in the control eggs). Gastroachisis was the most common malfo~atlon, accounting for about 7IX of the abnormalities found in the embryos. Despite this finding in the closed environment of the egg, there have been few findings suggestive of a teratogenic effect of anticholinerglc compounds in mammals. One report (317) described an acranial and anencephalic fetus with complete atelectasis of the lungs, hemorrhage into the lungs, and bypoplasia of the adrenals born to a woman who had been treated during pregnancy for a duodenal ulcer with 10 mg of oxyphencyclamine two or three times a day. (As the saying is, one swallow does not make a assumer; or, to corrupt Virgil's imperative, ab uno non tisce omnes.) The survey by the General Practitioner Research Group (136) fount only one among 57 instances of reproductive accidents in the group of 661 women who had taken drugs of some sort during the first trimester of pregnancy that could possibly be related to an anticholinergic drug. It should be noted, however, that two-thirds of the accidents of reproduction found in this survey were related to the use of antihistaminic bugs, many of which have some anticholinergic activity . Although anticholinergic drugs are useful in ameliorating the results of lack of inhibition of contractions of the detrusor muscles of the bladder, the danger of causing the development of hydronephrosis by using such drugs in cases with obstruction of urinary outflow from the bladder has been noted ~318). With respect specifically to 3-quinuclidinyl benzilate, several drugs containing the quinuclidine ring, including quloidine, are in more or less regular use. The closest relative to 3~quinuclidiny} benzilate is clidinium bromide, which is a component of Librax. This preparation consists of one-third clidinium bromide and two-thirds chiordiazepoxide and is administered usually in unit doses of 7.5 ma, one or two such doses being prescribed three or four times a day. Several papers (319-323) have reported side effects experienced by I 111

patients advised to take Librax, usually for ulcers of the gsatrointestinal tract. The most colon side effects of use of thle mixture were dry mouth ant constipation. Other fairly colon effects were droweinese, headache, gastritis, nauseas and vomiting. ~L180 mentioned in these papers were blurred vision, ataxia, tachyeardia, and difficult mic~curition. Feo~alea were reported (319) to develop high r heart r tea and greater tendencies to co-=tipatton than males; msles had a greater incidence of difficult alcturition than females. There was one report (321) of petechiae and thrombocytopenia in a woman who developed these conditions while salting one unit dose of Librax three times a day. After-~acontlnusoce of Librax, her platelet count, which had fallen to 12,000/mm3, became normal (200,000-500,000/mm3) within 5 A. Although N-benzylcortropiny1 benzilate was fount (324) to be more potently antimusca~ic than 3-quinuclidinyl benzilate, the latter compound produced greater disruption of the behavior of Lice and rats than any of eight other antimuscarinic compounds teated, includlog N-benzylcortropinyl benzilate. Although one of the two former aubJecta who had been given 3-quinuclltinyl benzilate re-exa~ned by Klapper et al. (162) was the only former subject to have had an apparent flashback. a conclusion that 502 of these subjects experienced flashbacks probably would be highly erroneous. The available evidence from studies with both animals and man is that recovery from intoxication with this bensllate, although it occurs slowly, is complete. Mice and guinea pigs given five subcutaneoua doses per week of 3-quinuclidinyl benzilate at 150 ug/kg for 3 wk (total maximal dose, 2.25 mg/kg) were reported (~84) to have developed no lasting signs of toxicity. The experience in experiments with both laboratory animals and man has been that generally the mydrlatlc and cycloplegic effects on the intrinsic smooth muscles of the eye are the ones that persist longest after doses of 3-quinuclldiny1 benzilate. The cytotoxic actions of this benzilate (206,207) do not seem to have been reflected in definite heritable mutagenic effects. The general conclualon of this survey ts that there is co final evidence that any compound surveyed cartlea a direct hazard of persistent dlminutlon of human health and normal function tn the doses used by Edgewood Aree"l and its contractors or an indirect hazard of abnormality of structure or function in offspring of the former volunteers. However, the cryptic activities of these compounds Deed to be studied much more intensively than they have been. I Il2

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TABLE I-3 Single-Dose LD,o. of Phenylisopropylglycolic Acid Esters Route of LDsos~, mg/k~ Animal Administration EA 3834 302,668 Mouse I.Y. 63.0 81.1 I.P. P.O. Rat I.~1. I.P. 120.0 383.0 63.5 153.4 I.M. ~ 262 -- Rabbit I.V. 25.1 30.0 Cat ~ V __ 4 9 . 0 Dog I.V. 49.5 65.0 Monkey I.V. 29.4 100.0 I 115

TABLE I-4 Single-Dose LD,os of Nonester Anticholinergic Compounds Route of istration Hepiperphenidol Benzetimid House . I.Y. 20.0 46.0 SOCO -- 640 P.O. 970.0 305 Rae Icy. -- 37.6 O_ ~0 __ __ S O C O P.OO Guinea Pig S.Ce P.O. ~ 116 160 404 153 825

TABLE I-5 Single-Dose Minimal Effective and Lethal Intramuscular Doses of TAB teal Hale mice Male rats Female rats Male rabbits Dogs Monkeys 36 (27, 48) 36 (10, 134) 45 21 21 LD c^. ms/k~a 61 + 4.2 251 ( 210, 301 ) 154 (131, 179) 66 80 50 Figures in parentheses are 95X confidence Itoaits. The standard deviation of the mean is given for mice. Single values were derives from experiments whose results were not suitable for calculation of appropriate statistics. I Il7

REFERENCES 1. Herbert , J. J., Durant , N. N., Reynolds , L. S ., ttolle , Henderson, E.G. 19~: Characterization of End-Plate Modif ication by 62-69. Conductance in Transected Frog ^ecle: Drugs. J. Pharmacol Exp. Therap. 216: Ringer, S. 1875: ~ Handbook of Therapeutics, Fourth Ed., New York, Wm. Wood ~ Co., pp 494-516. 30 Dale, N.ll., Feldberg, W., Vogt, M. 1936: Release of Acetylcholine at Voluntary Motor Nerve Endings. J. Phystol. 86: 353-380. Hodgkin, A.L., Huxley, A.F. 1952: Currents Carried by Sodium and Potassium lone Through the Membrane of the Giant Axon of Loligo. J. Physiol. Il6: 449~4720 50 Hodgkin, A.L., Burley' A.F. 1952 A Quantitative Deacription of Hembrane Current and its Application to Conduction and Excitation in Nerve. J. PhysiolO 117: 500~544. 6. CaStillo, JO tel. Katz' B. 1954: Me Membrane fame Produced by the Neuromuscular - Transmitter. J. Phyelol. 125: 546-565. 7. KrnJevic, K., Mileti, a. 1958: Acetylchollne in Mammalian Neuromuscular Transmission. Nature 182: 805-806. S. Evans, D.~.L., Schild, H.0., Thealeff, S. 1958: Effects of Drugs on Depolarized Plain Muscle. J. Physiol. 163: 4 74-485. Takauchi, A., Takauchi, N. 1960: On the Pemeability of End-Plate Embrace During the Action of Transfer. J. Phy siol. 154: 52-6 7 10. Durbin, R.~., Jenkinson, D.~. 1961: Me Effect of Carbachol on the Permeability of Depolarized Smooth ^sele to Inorgante Ione. J. Physiol. 157: 74-89. 11. Poseur, W.~. 1966: Fundamente1 Aspects of Neuromuscular Transmiselon. Ann. N.Y. Aced. Sci. 135: 110-135. 12. Feldberg, W., Gadd~, Jig. 1934: The Chemical Transmitter at Synapses tn a Sympathetic Ganglion. J. Phyalol. 81: 305-319. Perry, W..., Talesnik' J. 1953: The Role of Acetylcholine in Synaptic Transmission at Parasympathetic Ganglia. J. Physiol. 119: 455-469. ~ 118

14. Longo, Vie. 1955: Acetylcholine, Cholinergic Drugs, ant Cortical Elec trical Ac tiYity . Experientia Il: 76-78 . Eccles, JeCo ~ Eccles, Rim., Fatt, P. 1956: Pharmacologic Inves~eigatione on a Central Synapse Operated By Acety~choline. J. Physiol. 131: 154-169. 16. Hagiwara, S., Tasaki, I. 195~8: ~ Study on the Hechanism of Impuse Transmission Across the Giant Synapse of the Squid. J. Physiol. 143: Il4-137. 17. KrnJevic, K., Pumain, R., Renaud, L. 1971: The Mechanism of Excitation by Acety~chollne in the Cerebral Cortex. J. Physiol. 215: 247-268. 18. Forrer, G. R. 1950: Atropine Toxicity in the Treatment of Schizophrenia. J. Hich. State }2~. Soc. 49: I84-~85. 19. Goldner, R.D . 1956: Experience of Use ~ of atropine toxicity therapy] in Private Practice. J. Nerv. Ment. Dis. 124: 276-280. ~ 20. Miller, J.J.; Schwarz, H., Forrer, G.R. 1957: Atropine Coma Therapy. Oral Presentation to Michigan State Medical Society. 21. Forrer, G.~. 1956: History [of Atropine toxicity therapy! and Future Research. J. Nerv. Hent. Dis. 124: 256-259. 22. Wada, T., Horigome, S., Sakurada, T. 1960: Clinical Experience of the So-Called "Atropine Toxicity Therapy [Forrer]". Tohoku J. Exp. Med. 72: 398-404. 23. Schwarz, H. 1956: Statistical Evaluation [of a tropine ~ oxicity therapy] . J. Nerv. Ment . Dis. 124: 281-286. 24. Ketchum, J.S., Sidell, F.R., Crowell, E.B., Jr., Agha jantan, G.R., Hayes, A.H., Jr. 1973: Atropine, Scopolamine, and Di ban: Comparative Pharmacology "d Antagonists In Man. Edgewood Arsenal Technical Report 4761. 25. Life Sciences Department, Space and Information Sy8tem8 Division, North American Aviation, Inc. 1962: Effects of BZ on Pilot Performance. Contract No. DA18-108-CML-6644, Report No . SID 62 - 71. 26. Stoll, H.C. 1948: Phar~acodys~amic Considerations of Atropine and Related Compounds. Am. J. Med. Sci. 215: 5 77-5 92 . 27. Gordon, A.S., Frye, C.W. 1955: Large Doses of Atropine. Low Toxicity and Effectiveness in Anticholinesterase Intoxlcation. J. An. Hed. Assoc. 159: Il81-~84. ~ 119

2 ~ . Bachrach, W . H. 1958: As~ticholinergic Drugs. A.. J. Digest. Dis. 3: 743-799. 29. Eger, Eel., IT, 1962: Atropine, Scopolamine, and Related Compounds. Anesthesiol. 23: 365-383. 30. Andrews, I.C., Belonal~y, BeIoo ~ 1973: Parasympatholytics. Chin. Aces the s . 10: Il-29. 31 . Mirakhur, R.K., Clarke , R. S As., Bundee, J. W., McDonald , J.R.9 1978: Anticholinergic Drug e in Anesthesia. A Survey of thelr Pre sent Position. "aesthe s. 33: 133-138 . 32. Shunt, L.E., Bowes, J.B. 1979 : Atropine and Hyoscine . Anaesthes. 34: 476-490. 330 Molitor, H. 1936 The use of Bulbocapaine in Pre-Anesthetic Hedication. J. Pharmacol. Exp. Therap. 56 85-96e 34e Graham, JODOP., Lazarus, S. 1940: The Actions of He thyI -Atropin-e Ni bate [E~ydrin ] . JO Phaneacol . Exp . The rap. 70 165-1700 35e Cahen, R.L., ~rede, K. [952 lIomatropine Methy~bromide: A Phar~acological Reevaluation. J. Pharmacol. Exp. Therap. 105: 166-177. 36. Becker, B.A., Swift, JoG. 1959 Effective Reduction of the A~ute Toxicity of Certain Pharmacologic Agents by us of Synthetic lon ~change Resins. Toxicol. Appl. Phamacol. I: 42-54. 37. Boyd, C.E., Boyd, E.1~. 1961: The Acute Toxicity of Atropine Sulfate. Canad. Hied. Assoc. J. 85: 1241-12440 38. Buckett, W.R. ~ Haining, C.Go 1965: Some Pharmacological Studies on the Optically Active Iso~ere of Ilyoacine and Hyoacy~ine. Brit . ~J. Pharmacol. 24: 138-146. 39. Rosen, H., Blumenthal, A., Panasevich, R., McColl~, ~J. 1965: Dimethy} Sulfoxide [DMSO] as a Solvent in Acute Toxicity Deter~inaelons. Proc. Soc. Exp. Biol. I2ed. 120: 511-514. 40. Stockhaus, K., Wick, H. 1969: Toxizltltsunterschiede von pharmaka beI subcutaner, intragastraler und intraduodenaler applikation bei ratter. Arch. Internat. Phaneacodyn. Therap. I8t): 155-161. 41. Friedman, A.H., Walker, C.A. 1972: The Acute Toxicity of Drugs Acting at Cholinocepti~re Sites and 24-Hour Rhythms in Brain Acety~choline. Arch. Toxikol. 29: 39-490 I 120

' 45. 42. Albanus, L. 1970: Central And Peripheral Effects of Anticholinergic Compounds. Ac ta phar~acol. Toxicol. 28: 305-326. 43. Boyd, C.E., 80yd, E.M. 1962: The Chronic Toxicity of Atropine Administered Intramuscularly to Rabbits. Toxicol. Appl. Phamacol. 4: 457-467. 44. Ishidate, M., Jr., Hayashi ~ M., Sawada, M., Matsuolta, A., Yoshikawa, K., Ono, M., Nakatate, fit. 1978: Cytotoxicity Tests on Medical Druge--Chromosome Aberration Tests with Chinese Hamster Cells in Vitro. Bull. Nat. Inst. Eye. Sci. 96: S5--61. Butros, J. 1972: Melon of Some Cardiovascular and Neurobumeral Agents on the Formation of the Chick Embryo Heart . Teratol. 6: 16 7-~80. 46. Grunfelt , Y.F., Balaze , T. 1979: Receptor Supersensitivity in the Dam and Adrenergic Preponderance in the Progeny after Treatment with Atroplne in Pregnant Ra t s. Toxicol. App] . Phar~acol. 48: A104. 47. Waskell, L. 1978: A Study of the Mutagenicity of Anesthetics and their ltetabolites. Hut. Res. 57: 141-153. 48. Vrba, M. 1967: Chromosomeaberrationen hervorgerufen durch Scopolaminum Hydrobromatum. Humangenet . 4: 371-373. 49. Upbeat, M.E., Ramirez, L. 1980: - Persistent Effects of Prenatally Administered Scopolamine. Paycholog. Rep. 46: 633-636. 50. I`avallee, R.J. 1971: An Examination of the Adequacy of the Secondary Animal Screen as a Predictor of Human Incapacitation with a Selec ~ Group of ~ Anticholinc~gic Compounds. Edgewood Arse - l Technical Hemorandum 100-~. 51. Ariens, E.J. 1966: Receptor Theory and Structure-~tion Relationships. Mv. Drug Res. 3: 235-285. 52. Schallek, W. ~ Smith, T.H.F. 1952: Electroencephalographic Analysis of Side Effects of Spasmolytic Drugs. J. Pharmacol. Exp. Therap . 104: 292-298. 53. Wescoe, W.C., Green, R.C. ~ McNamara, B.P., Prop, S. 1948: 'the Inf luence of Atropine and Scopolamine on the Central Ef feces of DFP. 3. Phar~acol. Exp. Therap. 92: 63-72. 54. Funderburk, W.H., Case, T.J. 1951 : The Effect of Atropine on Cortical Potentlala. BEG Clin. Neurophysiol. 3: 213-223. 55. Macht, D.~. 1923: APharmacodynamic Analysis of the Cerebral Effect ~ of Atroplne, Romatropine, Scopolamine, ant Related Drugs. J. Pharmacol. Exp. Therap. 22: 35-48. ~ 121

560 Z~irblis, P., Kondritzer, A.A. 1966: Adsorption Of lI3-BZ and Ci4-Atropine to the Mitochondrial Fraction of the Rae Brain. Edgewood Arse - ] Technical Report 6~042. 57. Farrow, J.P., O'Brien, R.D. 1973: Binding of ~4tropine and Escarole to Rat Brain Fractions and its Relation to the Ac ety~choline Receptor. Moi. Pharmacol. 9: 33-40. 58. Hiley, CeRo ~ 8urgen, A.S.1t. 1974: The Distribution of Muscarinic Receptor Sites in the Nervous System of the Dog. J. Neurochem. 22: 159-162. 557. Burgen, A.S.V., Riley, C.R., Young, J.M. 1974: The Properties of ~acarinic Receptors in Mammalian Cerebral Co rt ex . Bri ~ . J. Pharmacol. 51: 2 79~2 85. 60. Yamamura, H.I., Kuhar, M.J., S~yder, S.~. 1974: In Vivo Identificatlon of Huscarinic Cholinergic Receptor Binding in Rat Brain. Brain Re~. 80: 170-176. 61. Ya;eao~ura, H.I., Snyder, S.H. 1976: Muacarinic Cholinergic Blnding in Rat Brain. Proc . Nat ~ Acad . Sci ~ 130 S 0 71: 1725-1 729. 620 Snyder, S.H-., Chang, K.J., Kuhar, lI.J., Yamamura, H.I. 1975: Biochemical Identlfication of the Mammalian Muscarinic Cholinergic Receptor. Fed. Proc. 34: 1915-1921. 63. Yamamura, H.~., Chang, K.J., Kuhar, M.J., Snyder, S.~. 1975: Cholinergic Muscarinic Receptor: Bioebemical and Light Autoradiographic Localization in the 3zain. Croat. Chem. Acta. 47: 475-488. 64. Krieger, D.~. 1967: Central-Nervous-System Adrenal Interrelations. Third Quarterly Progress Report, DAAAol5-6 7-C-0189 . 6S. Paton, W.D.M., Rang, lI.P. 1965: The Uptake of Atropine and Related Drugs by Intestinal Smooth Muscle of the Guinea Pig in Relation to Acety~choline Receptore. Proc. Roy. Soc. I.ondon Bl63: I-44. 66. Gaddum, J.H. 1937: The Quantitative Effects of Antagonistic Oruga. J. Physiol. 89: 7P-9P. 67. Stephenson, R.P. 1956: A Modificatlon of Receptor Theory. Brit. J. Pharmacol. Il: 379-393. 68. Aruniakshama, O., Schild, 0. 1959: Some Quantitative uses of Drug Antagonists. Brie. J. Pharmacol. 14: 48-58. 69. Paton, W.D.M. 1961: A Theory of Drug Ac~cion Based on ehe Rate of l~rug-Receptor Combination. Proc. Roy. Soc. London Bl54: 21-69. . 1~

Yamamura , H. ~ ., Snyder , S . H. 1974: Muscarinic Chol~inergic Receptor Binding in the Longitudinal Muscle of the Guinea Pig Ilium with [3~] Quinuciltiny} Benzi3-ate. Clot. Pharmacol. 10: 861-867. 71 . Sayers , A. C ., Bllrl~i , Il. R. 19 76 : Ant iacetycholine Activities of Psychoactive Drugs: A Comparison of the [3H]-Quinuclidiny} Benzilate Binding Assay with Conventional Me shots. J. Pharm. Pharmacol. 28: 252-253. 72. Witter, A., Slan~en, J.L., Terpatra, G.K. 1973: Distribution of 3H - ethylatropine in Rae Brain. Neuropharmacol. 12: 835~41. 73. Michaelis, }£., Finesin8er, J.E., Feaster, F. de B., Erickson, R.W. 1952: Effect of DFP and Atropine on the Free Acety~choline in Rabbit Brain. J. Pharmacol. Exp. Therap. 106 407. 74. Berry, J.F., Store, E. 1956: Acetaltehyde and Acetoln in Brain During Acetaldehyde Intoxication. Quart. J. Stud. Tic. 17: 190~194. 75. Giar~an, N.J., Pepeu, G. 1962: Drug-Induced Changes in Brain Acetylcholine. Belt. J. Phamsco}. 19: 226-234. 76. Giarman, Id., Pepeu, G. 1964: The influence of Centrally-Act_ng Cholinolytic Drugs on Brain Acety~chol~lne Levels. Bri t . J. Pharmacol. 23: 123-130. Aquilonius , So-., Lundholm, B., Winbladh, B . 1972 : Effects of Some Anticholinergic Drugs on Cortical Acety~choline Release and Motor Acti~rlty in Rats. Europ. J. Pharmacol. 20: 224-230. 78. Szerb, J.C. 1964: The Effect of Tertiary and Qua ternary Atropine on Cortical Acety~choline Output and on the Electroencephalogram in Cats. Canad. J. Physiol. Phar~acol. 42: 303-314. 79. Rommelapacher, H., Kuhar, H.J. 197S: Effects of Drugs and Axotomy on Ace ty~choline Level e in Central Cholinergic Neurons. Arch. Pharmacol. 291: 17-21. 80. Berger, F.~., Kletzltin, M., Margolin, S. 1966: The Action of Certain Tranquilizers, Antidepressants, and Anticholinergic Drugs on Hippocampal Afterdischarges. Proc . 1st Internat . Symp. Antidepressant Drugs, Excerpts Hed. Internat. Congress Series No. 122: 241-246. 81. Frances, H., Jacob, To 1971: Comparalson des Effete de Substances Cholinergiques et Anticholinergiques sur lea Taux Cerebraux d'Acety~choline et sur lea Mobilize chez la Souri ~ . Psychophamacol. 21: 338-352 .

82. Herink, J., Golda, V., Brdina, i., Nemecek' S. 1973: The Ef feet of Three Anticholinergics on the Evoked ggraSSivlty in Noneal and Septal Rat.. Active Nerv. Sup. 15: 131. 83. Usui, S., Iwahara, S. 1977: Effects of Atropine Upon the Hippocampal Electrical Activity in Rats with Special Reference to Paradoxical Sleep. EE(: Clin. Neurophysiol. 42: 510-517. 84. Takeyasu, K., Uchida, S., Noguchi, Y., Fudita, t1., Saito, K., Hats, F. j Yoshida, H. 1979: Changes in Brain Huscarinic Acetyl~choline Receptors and Behavorial Responses to Atropine and "omorphine in Chronic Atropine-Treated Rats. Life Sc. 25: 585-592. 85. Bell, C., Gershon, S., Carroll, B., lIolan, G. 1964: Beha~rioural Antagonism to a New Psychotomimetic: JB-329. Arch. Internat. Phar~acodyn. Therap. 147. 9-25. 86. Bell, C., Gershon, S. 19640 Experimental k`ticholinergic Psychotomimetics: Antagonism of Yohimbine And Tacrine {THA]. Med. exp.. 10: 15-21. 87. Evert~busch, V., Gelling, E.M.K. 19S3: Distribution and Excretion of Radloactlvity in Mice Following the Administration of C14-Labeled Atropine. Fed. Proc. 12: 319. 88. Pulewka, P. 1932: Das Auge der weissen Haus aln pha~kologisches Testob jekt . I. ~~teilung: eine Nethode our quantitativen Bestimmung kleinster blengen Atropin und enterer Mydriatika. Arch. exp. Path. Phamakol. 166: 307-318. 89. Tonnesen, H. 1950: The Excretion of Atropine and Allied Alkaloids in Urine. Acta pha~acol. topical. 6: 147-164. 90. Evertabusch, V., Gelling, E.M.K. 1956: Studies With Radioactive Atropine. I. Distribution and Excretion Patterns in the Ilouse. Arch. Interest. Pharmacodyn. ~erap . 105: 175-192. 91. Gosselin, R.E., Gabourel, J.~., Kalser, S.C., Wills, J.H. 1955: The Metabolism of Ci4-Labeled Atropine and Tropic Acit in ~ce. Chemical Corps Medical Laboratories Re search Repore No . 339. 92. Gosselin, R.E., Gabourel, J.D., Kalser, S.C., Wills, J.~. 1955: Tne Metabolism of Ci4-Labeled Atropine and Tropic Acid in Mice. J. Pharmacol. Exp. Therap. Il5: 217-229. 93. Bernheim, F., Bernheim, tt. 1938: The Hydrolysts of Homa tropine and Atropine by tJarious Tissues. J. Pharmacol. Exp. 'rherap . 64: 209-216.

94. Jager, B.V. 1956: Progress Report, April 1956 Lo October l9S6, Contract flo. DA-18-108-CML-5421. 95. Ammon, R. 1977: The Enzymatic Cleavage of seYera] Phamaceutical Esters. Arzuetm. Forech. 27: 944-949. 96. Kal~er, S.C., W111s, 3.~1. ~ Gabourel, J.~., Gosselin, R. E., Epps , C.F. 1957: Further Studies of the Excretion of Atropine-Alpha-Ci4. J. Phar~acol. Exp. Therap. 121: 449-6~56. 97. Gabourel, J.D., Gosselin, R.E. lU58: The Kechanism of Atropine Detoxication in Mice and Rats. Arch. Inters~at. Pharmacodyn. Therap. Il5: 416-432. 98. Phillipson, J.D., Hanta, S.S., Gorrod, J.W. 1976: Metabolic N~Oxidation of Atropine, Hyoscine, and The Corresponding Nor-Alkaloids by Guinea-Pig Liver ~crosoma1 Preparations. J. Pharm. Pharmacol. 28: 687-691. 99. ~erner, G., Sch~idt, H.L. 1963: Inteneediary Metabolia of C14-Atropine in Several Ha~alian Systems. Abhand1. Deut. Akad. Wiss. Berlin, Klin. Chem., Geol. ~ Biol. 4: 207-216. 100. Kalser, S.C., Kelvington, E.J., Randolph, b2.M., Santomenna, 13.~. 1965a: Drug Iletabolism iQ Hypothermia. I. Biliary Excretisn of C14-Atroplne Hetaboli~e~ in the Intac~ and Nephrectomized Rat. J. Phar~acol. Exp. Therap . 14 7: 252-259. 101. Ka1ser, S.C., Ke1,rin~gton, E.J., Randolph, M.~., Santo}eenna, D.M. I965b: Drug Metabolism In }Iypothemia. II. C14-Atropine Uptake, Metabollam, and Excretion By Tne Isolated, Perfused Rat Liver. J. Pharmacol. Exp. Therap. 147: 260~269. 102. Winbladh, B. 1973: l~e Fate of Atropine in the Puppy. Acta pharmacol . toxicol. 32: 46-64. 103. Harrison, S.~., Jr., Bosin, T.R., Halekel, R.P. 1974: Ph~rsiological DIstribution of Atropine in the Rat. Pharmacol. Biochem. Behav. 2: 843-845. 104. Goseelin, R.E., Gabourel, 3.13., Wills, J.~. 1960: The Fate of Atropine in Han. C1in. Phamacol. Therap. 1: 59 7-603. 105. Ka1~er, S.~., McLain, P.L. 1970: Atropine Hetabollam in Han. Clin. Pharmacol. Therap. 11: 214-227. 106. Kalser, S.C. 1971: rhe Fate of Atropine in Han. Ann. N.Y. Acad. Sci . 179: 667-683. T t ~ ~

107. Ga~zner, P., Past, I., Juhasz, P., Javor, T. 1976: Gyogysserkinetilmi Vi zagalotok Nagydosisu [ 10~920 ma] Atropin Adasa Soran, 3H Atropin Izotop Heressel [1. a ma] depo-ozerepe ~ . Orv. Hetil. 117: 1988-1991. 108. Hayden, P.W., Larson, S.M., Lakshminarayanen, S. 1979: Atropine Clearance frosa Humar~ Plasma O J. Nucl O Hed . 20: 366-367. 109. Virtanen, R., Kant.o, J., Iisalo, E. 1980: Radiolmmunoa~say for Atropine and L-Hyoseyamine. Acta pharmacol. toxicol. 47: 208-212. lLO. 5;DRR~P. ~ ~ 1~Q. ~ __ _ 111. _r ~ _ ~ ~ · ~ · ^~ · A ~8e 0t suspected ~lladonna Poisoning. Irish J. Med. Sci. l: 27-29. Mexander, E., Morris, D.P., Ealick, R.~. 1946: Atropine Poisoning: Report of a Case wit}` Reco~rery After Ingestion of one Gram. New EngI. J. Hed. 234: 258-259. Mackenzie, A.L., Piggott, JeFeGo 1971' Atropine Overdose in ~ree Children. Brlt. J. Anaesthes. 43: 1088-3~090. 113. Sims, S.R. 1954: Poisor~ng Due to Bellatonna Plasters. Brit. Hed. J. lI: IS31. 116. Beach, G.O., Fitzgerald, R.P., Holmes, R., Phibbe, B., Stucicenhoff, H. 1964: Scopolamlne PoisoMng. New EngI. J . Med . 2 70: 13 54-13 55. 115. Cullumbine, H., McKee, W.lI.E., Creasey, N.H. 1955: The Effects of Atropine Sulphate Upon Healthy Male Sub jects. Quart. J. Exp. Physiol. 40:. 309-319. 116. Wyant, G.M., Dobkin, A.B. 1957: Antiaialogogue Drugs in Man. Comparison of Atropine9 Scopolamine [~-byoscine] and L-Hyoscyamine [Bellafollne ~ . Anaesthes . 12: 203-222. 117. Bachrach, W.H. 1958: Anticholinergic Druge. ". J. Digest . Di s. 3: 743-799 . 118. Herxheimer, A. 1958: A Comparison of Some Atropine-L~ke Orugs in ~n with Particular Beference To Their End—)rgan Specificity. Brit. J. Phaneacol. 13: IB4-192. 119. Hirakhur, R.K. 1978: Comparatlve Study of the Effects of Oral and Intramuscular Atropine and Hyoscine in Volunteers. Brit . ~J. Anaesthes. 50: 591-598 . 120. Schweitzer, P., [lark, H. 1980: The Effect of Atropine on Ca rdiac Arrhy thmias and Conduction . ~ . Am . Heart J. 100: Il9-127. 121. Schweitzer, P., Mark, H. 1980: The Effece of Atropine on Cardiac Arrhyt~ias and Conductlon. lI. Diagnostic Use of Atropine. Am. Heart J. 100: 255-261. I 126

122. Wills, J.~., McNamara, B.P., Fine, A.E. 1950: Ventricular Fibrillation in Delayed Treatment of SEEP Poisoning. Fed. Proc. 9: 136. 123. Ludemann, H.ll., Cornblath, M., Gold, .4.J., Filbert, M.G., Udal-] , J., Haricabus , R., Cugell , D ., Freeman, G. 1955 : Ventricular Fibrillation Following Atropine in Acute Sarin Poisoning in Dogs. Medical I"aboratorles Research Re port 402. 124. Hardy, I.K., Wakely, D. 1962: The Amnesic Properties of F~roscine and Atropine in Preanaesthetic Medication. Anaesthes. 17: 331-336. 125. Dervent, B.,, Karacan, I. 1979: Effects of Anticholinergic Drugs on Human REM Sleep and Nocturnal Penile hmeacence. tang Sleeping 3: 81. Ostfeld, A.M., Machne' X., Unna, K.R. 1960: The Effects of Atropine on the Electroencephalogram and Behavior in ~ n. J. Phamacol. Exp. Therap . 128: 265-272 . 12 7. Ostfeld , A.~., Aruguete , A. 1962. Central Nervous System Effects of Hyoscine in Man. J. Pharmacol. Exp. Therap. 137: 133-139. 128. Commin, P., Bismuth, C., Gaultier, M., Mellerio, F. 1973: Apport de la Correlation Electroencepalographie-Clinique a 1 tActivite Anti cholinergique Centrale de la Physostig~ine . Agreso! . 19: 287-292. 129. Mellerio, F., Commin, P., Bismuth, C., Gaultier, M. 1979: L'electroencepalographie au Coure du Traitment par Eseri ne des Intoxications Anticholinergiques. Re~r. EEG Neurophysiol. 9: 5-~. 130. Hollender, M.~., Jamieson, R.C., McKee, E.A., Roback, H.B. 1978: Anticholinergic Delirium in a Case of Munchausen Syndrome. A - . J. Paychiat. 135: 1407-1409. 131. Aucamp, A.K., Heyer, C.`J. 1979: Epileptic Seizures Exacerbated by Anticholinergic Drugs . S . Af r. ]2ed . 3. 56: 423. 132. tfoskovitz, C., Moses, }~., Klawans, lI.L. 1978: Levadopa-Induced Psychosis: A Kindling Phenomenon. As'. J. Paychiat. 135: 669~675. 133. Holland, P., t~ite, R. G., Collirtge, 8. 1971: Atropine Sulphate Absorption in Humans Following Intra Muscular Injection of a Mixture of The Oxime, P2S, Ar~d Atropine. echnical Note No. 68, Chemical Defence Establishment., Porton l~own, Salisbury, Wilts. I 127

134. Hollant, P., Whlte, R.G. 1971: Atroplne Sulphate Absorption in Humane Following Intramuscular InJection of a Hixture of the Oxime, P2S-, and Atropine. Brit. J. Pharmacol. 42: 645P. 135. Hartin, H. de V. 1973: Atropine Sulfate Aboorption From an Intramuscular InJection of a Mixture of the Oxime, P2S, and Atropine in Exercising Humans. Brite J. Pharmacol.-67: 619P o 136. General Practitioner Reaearch Group 1963 Drug a in Pre8nancy Survey. The Practitioner 191: 775-780. 137. Hell31an, L.M., Morton, G.W., Wallach, E.D., Tollea, W.E., Fillisti, L.P. 1963 A~ Analysis of the Atroplne Test for Placental Transfer in 28 Normal Gravidas. Am. J. Obs . Gyn. 87: 650~659. 138. Mellin, G. W. 19640 Orugs In The Firot Trimester of Pregnancy and the Fetal Life of Nomo Sapiens. ". JO Obs . Gyn. 90: Il6 901180. 139. Hellman, L.~., Fillisti, L.PO 1965: "alysis of the Atropine Test for Placental Transfer in Gravidas with Toxemia and Diabetes. ". J. Obe. Gyn. 91: 797-B05. 140. Qullligan, E.J. 1965: Dlacusslon [of paper of Hellman ant Fillisti ], 1965. Am. J. Obe. Gyn. 91: 806-807. 141. ICivalo, I., Saarikooki, S. 1977: Placental Transmission of Atropine ae Full-Term Pregnancy. Brit. J. Anaesthes. 4 9: 101 7-1021 . 142. Onnen, I., Barrier, G., t'Athis, P., Sureau, C., Olive, G. 1979: Placental Transfer at the end of Pregnaccy. Europ. J. ClinO Pharmacol. 15: 443-446. 143. Horowitz, J.D., Goble, A.J. 1979: Drugs ant the Impaired Male Sexual Function. Drugs 18: 206-217. 144. Knuutila, S., K3lhk5nen, M. ~ Wilaka, M. 1976: tiotable Chromosomal Anomalies in a Woman Receiving Drugs During the Period of Viremia Af ter Rubella ttaccication. Hered. 84: 120~1230 145. Minich, V., Smith, H.E., Thompeon, D., Kornfeld, S. 1976: Detection of ~tagenic &CtiYit, iD 8~n Urice Using Mut.ant Strains of Salmonella typhimurI~. CancO 313 1253-125l3o 146. Aeara, M., Rennick, B. 1972: Renal Tubular Tranaport of A~etylcholine and Atropine: Enhancement and Inhibition. J. Pharmacol. Exp. Therap. IS2: 14-26. ~ 128

147. Grob, D., Harvey, J.C., Hartley, A. llcG. 1950: Quarterly Progress Report, Contract No. DA-18-108-C=-6~16. 148. I2arzulli, F.N., Cope, O.B. 1950: Subjective and ObJective Study of Healthy }tales $nJec ted Intramuscularly With 1, 2, Or 3 mg of Atropine Sulfate. Hedical Division Research Report Ilo. 24. 149. Himwlch, H.E. 1954: Effect of Large Doses of Atropine Sulfate on BEG ant Personality Structure. Medical La boratori es Contract Report No . 49. 150. Wechsler, R., Koskoff, Y.D. 1955: Effect on Beans of Moderate Doses of Atropine. Medical Laboratories Contrac ~ Report No. 56. 151. Andrews, P., Miller, R.D., Gordon, A.S. 1955: Evaluation of Atropinization by Various Route. in Rumans.Hedical La boratories Contract Report No. 5g. 152. White, R.P., Rinaldi, F. , }limwich, 11.E. 1955: Central ant Peripheral Nervous Effects of Atropine Sulfate and Darstine. Medical Laboratories Contract Report ~lo. 66. 153. Elkin, E.H., Freedle, R.O., Cott, lI.P. van, Fleishman, E.A. 1965: Effects of Drugs on HIl=An Performance. The Effects of Scopolamine on Representative lIuman Performance Tests. Technical Report I, Contract No. DA-18-035-A`YC-282[A] . 154. Kitzes, D.~., Keechum, J.S., Crowell, E.B., Jr., Balter, L. 1966: Factors Contributing to Individual Differences in Response to Incapacitating Agents. 1. The Contribution of Difference tn Body Weight and Performance Ability to Response Variance. Edgewood Arsenal Technical .`lemorandum Il4-3. 155. Cummings, E.G., Craig, F.N. 1967: Influence of The Rate of Sweating on the Inhibitory Dose of Atropine. Edgewood Arsenal Technical Report 4063. 156. Kligman, A.ll., Cbpelan, H.W. 1967: Annual Report, Contract No . DA-18-035-AMC-126 [A] . 157. Crowell, E.B., Jr., Ket chum, J.S. 1967: The Treatment of Scopolamine-Induced Delirium With Physostig~ine. Edgewood Arsenal Technical Report 4115. 158. Baker, W.J., Cott, H.P. van, Fleishean, E.~. 1967: Effects of Drugs on Human Performance: Scary of Results. Final Report, Contract No. DA-18-035~AMC-282[A] . 159. Safer, D.J. 1969: The Concomitant Effects of Mild Sleep Lose and Antichollnergic Drug. Edgewoot Arsenal Te chnical Report 4268. ~ ~ q ~

160. Hayes, A.H., fir., Copeland, H., Ket chum, J.S. 1969. The Effect of High Intramuscular Doses of Atropine Sulfate on the Human Electrocardiogram. Edgewood Arterial Technical Report 4220. 161 . Hayes, A. H., Jr., Copeland , H.~. , Ketchum, J. S. 1971 : Effects of Large Intramuscular Doses of Atropine on Cardiac Rhythm. Clin. Pharmacol. Therap. 12: 482-4860 162. Klapper, J.A., McCarroll, J.E., McColloch, M-.A. 1972: Long-Term Followup of Medical Volunteers. Edgewood Arsenal Te chnical Report 4 611. 163. Klapper, J.A., McColloch, H.A., Kysor, K.P., Sim, V.M. 1972: Personality Correlates of Response to Atropine, Scopolamine, and Glycolate Compounds. Edgewood Arsenal Te chnical Report 4 616. 164. Sidell, F.R., AghaJanian, G.K., Groff 9 W.A. 1973: The Re~rersal of Anticholinerglc Intoxication in Man with the Cholinesterase Inhibitor VX. Proc. Soc. Expo Biol. }2ed. 144: 72 5-730 . 165. Ketchum, J.S., Sidell, F.R., Crowell, E.B., AghaJanian, G.K., Hayes, A.H., Jr. 1969: Atropine, Scopolamine, and Ditran: Comparative Phar~acology and An~eagonists in Man. Proc. 1969 ~nit. Comm. Sci. Conf.: 64-83. 166. Ketchum, J. S., Sidell, F.R., Crowell, E.B., Jr., Agha jas~lan, G.K., Hayas, A.H., Jr. 1973: Atropine, Scopolam~ne, and Ditran: Compara~caive Pharmacology and An~agonists in Han. Psychophamacol. 28: 1~21-145. 167. Side?l, F.R. 1974: Modification by Diluento of Effects of Intramuscular A~cropine on Heart Rate in Man. Clin. Pharmacol. Therap. 16: 711-715. 168. Sidell, F.R. 1976: Use of Physostig~ine by the Intravenous, Intramuscular, and Oral Routes in the Therapy of Anticholinergic Drug Intoxlcation. Edgewood Arsenal Technical Report 76012. 169. Horenstein, H., P~hlicke , }I. 1938: Dber eine neue Umiagerungsreaktion und ihre Anwendung zur l~arstellung von Estern der Aminoalkohole. Ber. Deutech. Chem. Gesell~ch. 71: 1644-1657. 170. Hromatka, O., Csoiclich, C., Hofbauer, I. 1952: ~ber den Tropin-benziladureester. t£onatshefte Chem. 83: 1321-1325. 171. Sternbach, I..H. ' Kalser, S. 1952: As~ti~pasmotics. I. Bicyclic Basic ^\lcohols. J. Am. Chem. Soc . 74: 2215-2218. I 130

172. S~ernhach, L.H., Kaiser, S. 1952: Antlepasmotica. II. Esters of Basic Blcycli-c Alcohols. J. Am. Chem. Soc. 74: 2 219-22 21. 173. Randall, L.O., Benson, W.M., Stefko, P.L. 195;2: Spasmolytic Action of Blcyclic Basic Alcoho] Esters. Phamacol. Exp. Therap . 104: 284-290. 174. Archer, S., Bell, M.R. 1961: 2-Alpha-tropanyl Estere. U.S. Patent Application. 175. Atkinson, E.R., McRitchie, D.D., 1971: 'rotal Synthesis of the Four Isomeric 2-Tropanols. J. Org. Chem. 36: 3240~3241 . 176. Kimura, K.K. 3~960: Curren~ Information From Hoffman-MRoche, Inc., on Ro2-3308 as of February 1960. Chem. Warfare I.aboratories Speclal Publication 2-29. 177. Schallek, W. ~ Smith, 'r.H.F. 1952: Elec troencephalographlc Analysis of Side Effects of Spasmolytic Orugs. J. Pharmacol. Exp. Therap. 104: 291-298. 178. Denisenko, P.P. 1963: Ekaperimentalnoe i Klinicheskoe ~zuchenie Tsentralaich Cholinolitikov. Far~ako] Neirotropnykh Srevst~r, Akad. Med. Nauk. SSSR' Sb. 1963: 67-85. 179. Halpern, B.N. 193B: Recherches sur des Esters d'Aminoalcools doues de Proprietes Spasmolitiques. Arch. Internat. Pharmacodyn. Therap. 59: 149-194. 180. Lehmann, G., Knoefel, K. 1942: The Spasmolytic And Local Anesthetic Action of Some Derivatives of Fluorene-Carboxylic Acid and Related Compounds. J. Pharmacol. Exp. Therap. 74: 274-283. IB1. Larsen, V. 1955: The General Phar~acology of Benzilic Acid Diethylaminoethy] Ester lIydrochioride [Benaceyzine NFN, Suavitil, Parasan] . Acta phamaco}. toxicol. 11: 405-420. 182. Friedman, A.~1., Smith, C.M. 1957: Comparison of the Effects of Tropy! Benzilate and its N-aliyl Derivatlve with those of Atropine. J. Pharmacol. Exp. The rap. 119: 147. IB3. Friedman, A.11., S~ith, C.M. 1959: Pharmacological Properties of Tropyl Benzilate lIydrochioride ~ BAT] and i ts N-ally] Derivative {N1'B] . Arch. Internat . Phar~acodyn. Therap . 120: 160-~85. I

184. McN=nara, B.P. 1960: Research in Toxicology Division OR Effects of CS 4030 in Animals. Chemical Warfare Laboratorles Special Publication 2-28. 185. Fournier, E., Petit, L. 1962: Esters d'Acides Alpha Alcoole et d'Acides encombres en Position Alpha. Considerations sur leur Toxicite et leur Phar~Acodynamie. Therap. 170 1245-1249. 186. Wal, A. van de, Jr. 1962: A Compilation of Preliminary Toxicologlc-Pharmacologic Ir~ormatlon on Suggested Incapacitating Agents. Chemical Research and Development Laboratories Technical Report 3147. IB7. Lisunkin, Y. ~ I. 1962: Sensibiliziriuschee i Desensibiliziriuschee" t~llyanie Atropizu, BETE i Amlzlla na ProtivosudoroJolu Alcti~nost Pentafena i Tropataina. Byul. Ekep. Biol. my. 54~9~: 69-73. 188. Arthur D . Li etle Inc . and Sterling Winthrop Re a . Inat O 1966: Preelinical Pharmacology and Toxicology of Candidate Agent 2260860 Quart. Rept., ~ Mar. 1966~l Sept . 1966, Contrae t No . EA. IS-IO8-AlfC-103(A) . 189. Ketchum, J.S., Tharp, B.R., Crowell, E.B., Jr., Sawhill, CAL., Vancil' M.E. 1967: The Human Assessment of BE Disseminated Onder Field Conditions. Edgewood Arsenal Technical Report 4140. 190. Bagton, R.E., Garner, E. 1966: One Year Chronic Toxicity Studies of CIidinium Bromide in Rats. Report from Department of Pharmacology, Hoffman-laRoche , Inc ., to management . i91. CarSon, s. i965: Chronic fi-year] Oral Dosage Studies with Tarzan in Dogs. Report from Food and Drug Research Laboratories, Inc., to Hoffman-LaRoche, Inc. 192. Larsaon, Z.., Wallenateen, M., Wallerberg, G., Oatman, B., 1974: The lIydrogen Bond Condition in Some Anticholinergic Esters of GI,colic Acids. I. Acea phase. Suecica Il: 304-308. 193. Becker, M.J. 1964: Biochemical Studies of BZ and EA 3443. Final Report, Contract No. DA-~-104-C~-6631. 194. Yamamura, H. I., Kuhar, M.J., Greenberg, O., Snyder, S.H. 1974: ~acarinic Cholinergic Receptor 81nding: Regional Distribution in Monkey Brain. Brain Res. 66: 541-546. 195. Yamamura , B. I., Snyder , S . H. 1974 : Postsynaptic Localization of Huscarinic Cholinergic Receptor Binding in Rat Hippocampus. Brain Res. 78: 320-326. I 132

196. Boldt, R.E., Brendalen, E., Gillelan, W. 19-66 : Studies of Brain Microsomal ATP-ase Activity and lon Transpose in Tissues Treated with BZ. Edgewood Arsenal Technical Report 4043. 197. Jovic, R.C., Zupanc, S. 1973: Inhibition of Stimulated Cerebral Resplration In Vitro and Oxygen CoD8~ptioD In live in Rats Treated by ~olinolytic Drugs. Biochem. Pharmaco!. 22: IlB9-~194. 198. MeyerhUffer, A. 1972: Absolute Configuration of 3-QuinuclidinyI-benzilate and the Behavioral Effect in the Dog of the Optical Isomers. J. Hed. Chem. 15: 994-995. 199. Kabes, J., Fusek, J., Fink, Z. 1972: Effects of 3-QuinuctidinyI-benzilate on Spontaneous Motor Activity in Rats and Management of the Anticholinergic Syndrome by Some Psychotropic Drugs. Achier. Nerv. Sup. 14: 161-162. 200. Kabes, J. 1972: Ef facts of 3-QuinuclidinyI-benzilate on CAR Acquialtion and Performance of Rats in a Shuttle-Box. Ac tiv. Nerv. Sup . I4: 163-164. 201. Kabes, J. 1972: Rating Scale for Autonomic, Motor, and Central Effects in Dogs. Effects of Anticholinergic and Cholinergic Drugs. Actlv. Nerv. Sup. 14: 164-165. 202. Funk, J., Kabes, J., Fink, Z. 1972: Anticholinergic Effect of 3-Quinuclidinyl-benzilate. Activ. Nerv. Sup. 14: 165-166. 203. Poel, A.~. van Her 1974: The Influence of 3-Quinuclidiny~bencliate on the Behaviour of Rats in a Ci rcular Runway. Paychophar~acol. 36: 151-162. 204. Holmberg, G., Gershon, S. 1961: Autonomic and Psychic Ef fects of Yohimbine Hydrochloride. Psychopharmacol. 2: 93-106. 205. hlsek, J., PatocIca, i., Ba Gary J., IJrban, R., Kolesar, J., Ilerink. J., BrdIna, V. 1975: Antidotal Effect of I, 2, 3, 4-tetrahydro-9-aminoacridine [Tacrine] Against Poisoning with 3-QuinuclidyI-henzilate. Activ. Nerv. Sup . ~ 7: 252. 206. Sram, R.J., Kucherova, M. ~ Bardode], Z. 1975: Mutagenic Ac timidity of 3-Quinuclitylbenzilate . Artier. Nero. Sup . 17: 252-253. 207. Gram, R.J. 1975: Cytogenetic Analysis of 3-Quinuclidy~benzilate, 3-Quinuclidinol, and Benacryzine in Bone Marrow of Chinese Hammer. Attic. Nerv. Sup. 17: 253-254. I 133

208. Hughes, H.C., Bostrom, R.E., Goldstein, R.S., Ferrell9 JOFO 1969: The Effects of BZ on Spermatogenesis in the Dog. ~gewood Arsenal Technical Memorandum Il6-So 209. Jacobsen, E. ~ Skearup, Y. 1955: Experimental Induction of Conflict-Beha~riour in Cats: The Effect of Some Ar~ticholinergic Compounds. Ac ta pharmacol. toxicol. ll: 125-134. 210. Jacobsen, E., Sonne, E. 1955: The Effect of Benzilic Acid Diet}~riaminoethylester, ECl [8enaceyzine] on Stress-Induced Behaviour in the Rat. Acts pharmacol. ~oxicol. Il :- 135-147. 211. Jacobsen, E., Kehiet, H., Larsen, V., Mun~vad, I., Skinhod, K. 1955: The Autonomic Reaction of Psycho-neurotles to a New Sedative: Benaceyzin NFN, Suavitil [Benzilic Acid Diethylaminoethylester ECL]. Acta psychiat. neurol. scand. 30: 627-642. 212. Berger, FoMo ~ Hendley, COD., [ynes, TeEo 19S6 Pharmacology of the Psychotherapeutic Drug Benaceyzine [beta-diethylaminoethyI-benzilate hydrochloride]. Proc. Soc. Exp. Biol. Med. 92: 563-566. 213. McColl, J.~., Rice, W.B. 1962: Antagonism of Tremorine by Benaceyzine and Dioxolane Analogs. Toxicol. Appl. PhanmacolO 4: 263-268. 2140 Jacobsen, E., 1964: Benaceyzine. Chapter ~ in Gordon, tel., ed . 1964: Psychophar~acological Agents, tJol . I., Academic Press, New York and Lonton, pp. 287-300. 215. Shitov, E.E., 1965: itItyanie sulfate magnlya ~J kombinaschit ~ trank~ilizatorami na bioelecktricheskuyu i cholinesterasnuyu akti~rnost golovnogo mozga. Famakol . Tokaikol. 28: 13-17. 216. Banshchikov, V.M., Stoliarov, (;.~. 1966 : Patchotomimeticheskie Sredst~ra ~ Aneicholinergichessim l~eis tviem. Zh . Ite~rropaeol . Pai~hiat . Koroakeva 66:464-468. 217. Vojeechovsky, M. 1958: ~ Psychosis Caused by Benaceyzine [ntoxication. Acta paychiat. neurol. scand. 33: 514-518. 218. Vo~techov~ky, M., Vitek, V., Ryeanek, K., Bultasova, a. 1958: Psychotogenic and Hallucinogenic Properttes of Large Dosea of Benaceyzine. Experientia 14: 422-423. 219. Edel~on, J., Schiosser, A., Douglas, J.F. 1970: Benaceyzine Hetabolism in the Rat. Arch. Internat. Phanmacodyn, Therap. 187: 139-143. I 134

220. Davies, E.B. 1956: A New Drug to Relieve Anxiety. Brit. Med . J. I: 48 0-484 . 2~ . ~ymond, H.J., Lucas, C.J. 1956: Benaceyzine in Psychoneurools, with a Note on the EEG Changes in Noneal SubJecto. Brit. Hed. J. I: 952-954. 222. Kinross-Wrlght, V., Moyer, J.~. l9S7 : Obeer~rations Upon the Therapeutic Use of Benactyzine Suavetil. Am. J. Psychiat. 114: 73-74. 223. Vo ~techovoly, Il., tJitek, V., Ry~anek, K. 1966: Experimentelle Psychose nach ~Jerabrelchung von Benacryzine. Arzniem. Forech. 16: 240~242. 224. Rickels, K., Gordon, P.E., Jenkins, B.tl., Sablosky, L., ~llachos, V.A., Iteise , C. C., Whalen, E . M., Wilson, I) .A. 1971: Combination of ldeprobamate ant Benaceyzine (Deprol) and Con~ei~uents in Neurotic Depressed Patiento. Dis. Nerv. Syst. 32: 457-467. 225. Sidell, F.R. undated: A Summary of the Investigatione in Man wi~h BZ Conducted by the U.S. Army, 1960-1969. CSt. 000~13 7. 226. Freedman, T. 1962: Effec-~s of ~Z on Pilot Performance. Final Report, Contract No. DA-~-108-C.YL-6644. 227. Authur unkno~ 1962: Posalble Buman Estimates for BZ Baset on Relationship Between BZ and A~ropine. C.C. I£ed. Labs. Miscellaneous Paper. 228. Ketchum, J. 1963: rhe human Assessoaent of BZ. Chemical Re search and Development Laboratori es Techulcal ~femorandum 20~29. 229. Directorate of Medical Research, U.S. Army Edgewood Areenal I;965: Guide to the Management of BZ Cssualties. 230. Kitzes, D.~., Uancil, M.E. 1965: Estimate of Hinimal Effective Doses of BZ by the Intramuscular Route in Man. Chemical Research and Development Laboratories Technical Hemorandum 2-30. 231. Crowell, E.8., Jr. 1966: CS27,349: Estimate of the Incapacitating Dose in Man. Edgewood Arsenal Tech~cal Memorandum Il4-2. 232. Craig, F.N., t~cttichael, P.D., Robinson, P.F., Sldell, F.R. 1969: Effects of BZ on Temperature Regulation in Han. Edgewood Arsenal Technical Hemorandum 112-~. 233. Brown, H., 1965: A Review of the Phar~acology and Toxicology of C" 301, 060. Contract No. DA-~-108-AMC-lO 8( A) . I 135

2340 Mennear, J., Samuel, G.K. 1963: Pre~clinical Pharmacology Study - Monkeys. EA 3443. Contract No. I'A-l8-1013-A~C-7 8(A) . 235. Mer~near, 3., Crews, L.M. 1963: Pre-clinical Pharmacology Study - Dogse EA 3443. Contract No. DA-18-108-"C-78(A)o 236. Mennear, Jew. 1962: Pre-clinical Pharmacology Study - Dogs. EA 2545 and EA 3167. Contract No. DA-13-108-405-QfL-826. 237. Kitzes, }AL., Ketchum, J.S. 1965: Estimate of Minimal Effective Dose of EA 3443 in ttan. Chemical Research and Development Laboratories Technical }memorandum 2-29. 238. Havoc, J.~., Salter, 1~. 1966: A Study of Possible Residual Effects of EA 3443 and EA 2S8O on Cognitive Ability. Edgewood Arsenal Technical Report 4044. 239. Armstrong, R.D., Meeker, L.WO, Somere' L., O'[eary, J.FO, 1975: Behavioral Dose~Response Studles of EA 3643 and EA 3580 ia Rat8 Trained in the Sequential Response Test. Edgewood Arsenal Te chnical Hemorandum 75001. 240. Abood, L.G., Ostfeld, A.}l., Biel, J. 1958: A New Group of Psychotomimetic Agents. Proc. Soc. Exp. Biol. Hed. 9 7: 483-486. 241. Ostfeld, A.N., Abood, L.G., Harcus, O.A. 1958: Studies with Ceruloplasmin and a New Hallucinogen. Arch. Neurol. Psychiat. 79: 317-322. 242. Abood, L.G., Ostfeld, A., Biel, J.~, 1959: S true ture-Ac tiv! ty Re lationships of 3-Pi peridyl Benzila~es with Paychotogenic Properties. Arch. Internat . Pharmacodyn. Therap 0 120: ~ ~ 6-2 00. 243. Usdin, E., Efron, 13.11. 1972: Paychotropic Drugs ant Related Compounts, Second Editlon, Washington, O.C. U.S. Goverament Printing Office, p. 224. 244. Gershon, S., Olartu, J. 1960: JB 329- A New Psychotomimetic. Its Antagonlam by Tetrahydroaminacrine and Its Comparison with LS0, Hescaline' and Sernyl. J. Neuropaychiat. 1:283-292. 245. Biel, J.~., Nuhfer, P.A., Hoya, W.K., Leiser H.A., Abood, L.G. 1962: Cholinergic Blockade as an Approach to the Development of New Paychotropic Agents. Ann. N. YO Acad. Sci. 96:251-262. 246. Brown, M.L., Gershon, S., Lang, W.J., Korol, B. 1966: The Effects of Paychoactive Druge on the Behavioral Response to Ditran in Dogs. Arch. Internat. Pharmacodyn. Therap. 160:407~423. 1 136

247. Gershon, S., Angrist, B.M. 1973: Effects of Alterations of Cholinergic Function on Behavior. Chapter 2 in Proc. Of the 62nd Ann. Meet. Am. Paychopathol. Assoc., Cole, J.O., Freedman, A.M. , Friedhoff, A.J. , cd's, Baltimore, Johns Hopkins Press, pp 15-36. 248. Ketchum, J.S., Kitzes, D., Copelan, H. 1973: EA 3167: Effects in Han. Edgewood Arsenal Technical Report 6713. 249. Arthur D. Little, Inc., and Sterlin8-Winthrop Research Institute 1968: Supplement S. Additional Toxicity and Tolerance Tests with 226,086 and 226,169. Quarterly Report ~ Contract No. DA-~-lO8-AMC-103 (A) . 250. Hayes, A.H., Jr. 1967: CAR 301,060: Estimate of Minimal Effective Dose in Man. Edgewood Arsenal Technical Hen~orandum Il4-12. 251. Copelan, H.~. 1968: tfi050 of Agent 834. Flual Report, Contrac ~ No . DAAAl5~68-C~0627. 252. Averill }I.P., ldcHamara, B.P., Callahan, J.F., Oberet, F.W., Farrant, R.L. 1970: Toxicology of EA3834A in Animals. Edgewood Arsenal Technical Memorandum 106. 253. Sim, V.M. 19 71: Compound 302, 668: S~'nmary Report . Edgewood Arsenal Special Publication 100~95. 254. Sidell, F.R., Braun, B.G. 1972: EA 3834A: Effects in Man Af ter a Single Oral Dose. Edgewood Arsenal Technical Report 459 7. 255. McCarroll, J.E., Markis, J.E., Ketchum, J.S., Houff, C.W., Sim, V.~. 1972: EA 3834: Effects on Performance of a Small Inf entry Rif le Element ~Jnter Simulated Combat Conditions. Edgewood Arsenal Technical Report 4633. 256. Cucinell, S.A., Cummings, E.G., Holgate, S.H. 1975: Sweat Inhibition and Performance Decrements in Han Following Percutaneous Exposure to EA3834. Edgewood Arsenal Technical Report EB-TR-76004. Copelan, Hip. 1967: HE:DSO of Agent 668. Final Report, Contract No. DA-18-035-AMC-126 (A). 2S8. Karger, S. 1968: Incapacitating Dose of CAR 302, 668 in Man, and Efficacy of Physostigmine as an Antidote. Edgewood Arsenal Technical Hemorandum 116-20. 259. Sidell, F.R., Karger, S. ~ Simons, C.J., Weiner, J.T. 1970: Compound 302, 668: Aerosol Administration to Man. Edgewood Arsenal Te chnical Report 4395. 260. Copelan, H.W. 1967: MED50 of Agent CAR 302,212. Report No . I\t, Contrac ~ No . DA-18~035-A.MC-126 (A) . ~ 137

261. Sidell, F.R., Ketchum, J.S., Harkis, J.E., Rysor, K.PO 1972: Compound 302,196: Intramuscular Administratlon to Han. Edgewood Arsenal Technical Report 4634. 262. Pylkko, 0., Samuel G. 1964: Preclinical Pharmacology Study - Dogs. EA3392 and EA 3580A. Contract No. DA-~-IO8-~HC-7 ~ O 263. Samuel, G., Crews, L. 1964: Preclinical Pharmacology Study - Rats. EA3580A. Contract No. DA-~-lO8-A~C-~(A). 264. MacFariant, H., Goldberg, M., Crews, L., Greendyke, R., Samuel, G. 1964: Preclinical Pharmacology Study - Rats. EA3 580B . Contract No . DA-18-035-AMC-120( A) . 265. Agha janian, (:.K., Kitzes, D.L., Harper, D.G., Bottiglieri, N. 1965: EA 3580A: Estimate of Hinimal Effective Dose in Man. Chemical Research and Development Laboratories Technical Memorandum 2-12 . 2660 Institute for Behavioral Research 1965: The Effects of Two Dose Level. of a CR1'L Compound Upon a Delayed M at chi ng Perf ormance wi t h Human Voluntee r SubJec sac a . Contract No. DA-~-lO8-AMC-26<A)-CP3-40. 2670 Crowell, E.B., JR. 1965: EA35804: Estimate of Incapacitating Dose in Man. Chemical Research and Development Laborat vies Technical Memorandum 2-20. 268. Baker, W.J. ~ 1966: The Effects of Drugs on Human Perf ormance: rhe Analysis of Responses Over Time in Weight-Adjusted vs. Fixed-Dose Studies of Psychoactive Chemicals. Technical Note No. 2, Contract No. 1)A-~-035~AMC-282( A) . 269. Kitzes' D.L., Ketchum, JeSo ~ Weimer, 3.T., Farrand, RoLe 1967. Human Assessment of EA 3580 by the Aerosol Route. Edgewood Aresenal Technical Report 4051. 2700 Baker, W.J. 1967: Effects of Drugs on Han Performance. Relation of Performance Deerements Under 3580A to SubJect-Related Variables. Technical Report No. 5, Contrac ~ No . DA-18-035-AllC-282( A) . 271. Allen, R.P., Safer, D.J. 1968: Exercise and Incapacitating Effects of EA3580A in man. Edgewood Arsenal Technical Memorandum Il4-15. Copelan, H.~1., 1967: ~050 of Agent 282. Final Report, Contrac ~ No. DA-~035-AMC-126~) . 273. Safer, D.J. 1968: Estimate of Incapacitating Dose of Compound 302,282 in Han. Edgewood Arsenal Technical Memorandum 114-21. I 138

274. Brown, H. 1966: A Review of the Pharmacology and Toxicology of CAR 302, 53 7 ~ Report ~ Contrac t No . DA-l8-108-AMC-lO8( A) . 275. Leib, G. 1969: Minimal Effective Dose of 302'537 in Man. Edgewood Arsenal Technical Memorandum 100-~. 276. Fine, E.A., Wills, J.H., McNamara, B.P. 19SO: Compound WIN-2299 as an Antidote in Poisoning Produced by Anticholines~cerase Agents. Medical Division Research Report 7. 277. Haley, T.J., Rhodes, B.Mo 1953 Influence of Peripheral Cholinergic Blocking Drugs on Survival Time in X-ray Irradiated Hice. Science Ill 139~140e 27~3o Pennes, H.H., Hoch, P.H. 1957 Psychotomimetice, C1 inical and Theore tical Considerations: Hamine, WIN-229 9, and Nalline . AH. J. Psychiat . ~13: 887-892.233 Brown, H., 1965: ~ Review of the Pharmacology and Toxicology of CAR 301, 060. Contract No. DA-18-108-h~C-108( A) . 279. Fink, H. 1960: Effect of Anticholinergic Compounds on Post-Convulst~e Electroencephalogram and Behavior of Psychiatric Patients. BEG Clin. Neurophysiol. 12: 359-369. 280. Janssen, P.A. J., Niemegeers, C.J.E. 1967: The Peripheral and Central Anticholinergic Properties of Benzeti~ide (R-4929) and Other Atropir~e-I~ke Drugs as Measured in a New Anti-Pilocarpine Test in Rats. Psychophamacol. 11:231-254. 281. Soudidn, W., Widagearden, I. van, Ariens, E.J. 1973: Dexetimide, a Useful Tool in Acetylcholine-Receptor Localization. Europ. J. Pharmacol. 24: 43-48. 282. Beld, A.J., Ariens, E.J. 1974: Stereospecific Binding as a Too] in Attempts to Focalize and Isolate Muscarinic Receptors. Part II. Binding of (+~-Benzeti~ de, ~-)-Benzetimide and Atropine to a Fraction from Bovine Tracheal Smooth Muscle and to Bovine Caudate Nucleus. Europ. J. Pharmacol. 25: 203-209. 283. Karger, S . 1969-: Effects of 8enzeeimide Given Intravenously to Humans. Edgewood Arsenal Technical Memorandum MO. Il4-24. 284. Wiles, J. S., Ford , O.F. 1976 : Acute Intramuscular Toxicology of TAB in Several Animal Species. Part I. of Investigational New Drug Application, pp. 153-205. ~ 139

285. Lee, C.-C., Castles, T.R., Bridges, R. 1976: Fourteen Day Acute Toxicities of TAB Mixture and Each of Its Constituents Following a Single Intramuscular Dose in Hale Mice. Part M of Investigational New Drug ADD1ICatiOn PP . 209 - 21 7. 2436. Wiles, J.S., Ford, Dip. 1976: Subacute Toxicity of TA.B in Several Animal Species. Part N of Investigational New Drug Application, pp. 220~264. 287. Holgate, S.H., Sidell' F.R. 1976: A Dose-Response Study on TAB ant Benactyzine. Part Cl of Supplement ~ to Investigational New Drug Application, pp. 42-90. 288. Sidell, F.R. 1976: TAB. Safety and Tolerance Study. Part D1 of Supplement ~ to Investigational New Drug Application, pp . 92-~10. 289. Holgate, S.H., Sitell, F.R. 1976: Me Effects of TAB in Stres sful Environment . Part El of Supplement ~ to Investigational New Drug Applications pp. ll8-15~)o 290. Holgate, SeHo, Sidell, F.R. 1976: The Effect of TAB on Operant Pe rf ordnance . Part Fl of Supplement I to Investigational flew Drug Application, pp. 152-206. 291. McBride, W.G. 1980: The Effects of Antlcholinergic Drugs on the Development of the Chick Embryo. IRCS Hed. Sci. ~ :537. 292. Sigwaid, J., Bouttier, D., Courvolsier, S. 1959. Les accidents neurologiques des medications neuroleptiques. Rev. Neu~ole 100: S53-5 95. 293. Sigwald, J., Bouttier, D., Rdymondeaud, C., Plot, C. 1959: Quatre cas de dyokinesie facto-bucco~11nguo~masticatrice a evolution prolonSee secontaire a un traitement par lea neuroleptIques. Rev. Neurol. 100. 751-7S5. 294. Uhrbrand, L., Faurbye, A. 1960: Reveralble and Irreversible l~yokinesia After Treatment with Perphenazine, Chlorpromazine, Reserpine, and Elec troconvulsive Therapy. Psychopharmacol. I: 408-418. 295. Hunter , R ., Earl, C. J., Thornicrof ~ , S . 1 964 : An Apparently Irreversi ble Syndrome of Abnormal Movement Following Phenothiazlne Medication. Proc. Roy. Soc. Hed. 57:758-762. 296. Faurbye, A., Rasch, P.J., Petersen, P.B., Brandborg, G., Pakkenberg, H. 1964: Neurological Symptoms in Pharmacotiserapy of Psychoses. Ac ta psychiat . scand . 40 :10-27. I 140

297. PJalland, 8., Nielsen, I.M. 1974: Enhancement of Methy~phent~ate-Induced Stereotypies by Repeated Admlaistration of Neuroleptics. Paychopha~col. 34 :IOS-lO9. 298. Chrlatensen, A.~., Neilson, I.M. 1979: Dopan~tnergic Super Sensitivity: I=fI:uence of Dopamine Agonists, Cholinergice, Anti-Cholingerics, and Gouge Used for the Treatment of Tardive Dyakinesta. Psychophamacol. 62:~-116. 299. Christenesen, A.~., Nielsen, I.M. 1979: On the Supersensitivity of Dop~'n~ne Receptors After Single and Repeated Atministration of Neuroleptics in: Tardive Dy~kinesia - Research and Treatment, Smith R.C., Fann W.E., Davis S.llo, Domino, E.F., ed.'s, New Yoric, Spectrum. 300. Hano, T., Sobue I., Hirose K., Iakag! S., Watanabe lI., Okamoto S., 1973: Dyalcinesias Induced by Anticholinergic Drugs. An Electro Physiological Analysis. Excerpta Med. Internat. Congr. Ser. (IOth Internat. Con~gr. Neurol. 296 :44. 301. Mano, T. 1974: [Dyakinesia Induced by Anticholinergic Orugs - An Electrophysiological Amalysis]. Rinsho Shi nkeigaku 14: 255-2 63. 302. Fahn S., David, E. 1972: Oral-Eacial-Lingual Dyakinesia Due to Anti cholinerglc Medication. Trans . Am. Neurol . Assn. 97: 277-279. 303. Fahn, S., Da~rid, E. 1973. Oral Dyskinesia Secondary to Anti cholinergic Drugs . Excerpt . Hed . Internat . Congr. Ser. (lOth Internat. Congr. Neurol. ~ 296:43-44. 304. Birket-Smith, E. 1974. Abnormal tovoluntary Movements Intuced by Anticholiner~ic Therapy. Ac ta neurol. scand . 50:801-~. 305. Birket-Smith, E. 1975: Abnoneal In~roluntar~r Movements in Relation to Anticholinergics and Levadopa Therapy. Acta neurol. scand. 52:158-160. 306. Chouinard, G., Annable, L., Ross-Chouinard, A., Nestoroa, J.N. 1979. Factors Related to Tardive Byskinesia. Am. J. Psychiat . 136: 79~83. 307. Chouinard , J., Annable , L., Ros - Chouinard , A., Kropoky, M. I. 1979: Ethopropezine and Benz:ropine in tieuroleptic-Induced Parkinsocism. J. Clin. Psychiat. 40 :147-152. ~ 141

3080 Burnett, G.B., Prange, A.J., Jr., Wilson, I.CO Jolliff 9 L.A., Creese , I. C., Snyder' S.H. 1980: Adverse Ef fects of Aneicholinergic Antiparkinsonian Drugs in Tardive Dyakinesia. An In~restigation of Hechanism .,Jeuropsychobio]. 6 :109-120. 309. Tanner, C., Goetz, CoGo ~ Weiner, W.J. 1980. Anti chollnergics and Tardive Dyakinesia. Am. J. Psychiat . 137:1470. 310. Geriach 9 J., Reisby , N., Randrup , A. 1974: Dopamf nergic Hypersensitivity and Cholinergic Hypofunction in the Pa~hophysiology of Tardive l~ysicinesia. Psychophar~acol. 34: 2lo35. , Martys, C.R. 1979: Ad~rerse Reactions to Drugs in General Practice. Brit . Med. J. II :1194-~197. 312. Pfeiffer, C.C., Murphee, H.B., Jenney, E.~., Robertson, HoGo ~ Randal~l, A.ll., Bryan, L. 19590 Hallucinatory Effect in Man of Acety~choline Inhibitors. Neurol. 9 :249-2500 313 . Wulf f, H. R., Soltof t, J., Gudmand-Hoyer, E., Molmann, K.~., Bonnevie, O., Petersen, H.E.~., Overo' K.F. 1977: Behandling af ulcus duoden] med en ny bicyklisk f orbindelse: prindamin. Ugeskr . Laeg . 139: 2062-2064 . 314. Dry, J., Pradalier, A., Palma, H. di, Lanoue, R. 1980: Essai clinique en double aveugle contre placebo de la mequitezine chez des patients non hospltalises. Therap. 35 :189-195. 315. Craig> T.J. 1980: .Medication Use and Deaths Attributed to Asphyxia Among Psychiattlc Patlents. Am. J. Psychiat. 137 :1366-1373. 316. Craig, T.J., Richardson, H.A., Bark, N.~., Klebanov, R. In press: The Association of Tardive Byskinesia, Impairet Gag Refiex, and Anticholinergic l3rug Use. 317. Piotrowski, J., Zahorsici, A. 1967: Aencephalia plod~u matict leczonej z powodu wrzodu durmastnicy. Prz. Lek. Z3: 799-801. 318. Novicici, D.E., Willacher, M.K. 1979: Anticholinergic-Induced lIydronephrosis. Urol. 13: 324-325. 319. Cromwell, H.A. 1968. Double-Blind S~udy of a Psychotropic Anticholinergic Grug in Gastric Disorders. Med . Times 96: 933-9 38. 320. Taylor, W.J.~., 1970. A Comparati`.re Determinatlon of Side Effects Associated with the Oral Use of Three Anticholinergic Psychotropic Drugs. Internat. Ztschr. klin. Pharmakol. Ther. Toxikol. 3:1-13. I L42

321. Reallne, S. Hofatetter, J.-~., 1973. Traitement du colon Irtltable par une nouvelle asaociatlon o~etlca~eneeuse (chlordiazepoxyde, bromure he clldlatum, propypheDazone). Praxis 62: 1S19-1525. 322. Celada , A., Berreros, V., Rudolph, 11. 1977 : Thrombocytopenia Purpura During Treatment with Libras. Brit, Med. J. I:2S8. 323. 81eger, G. 1980: Lea traltementa medlcaux au long coura de la maladie ulcereuse en tehora dea pouseeea. Med. Q~ir. Dlgestiv. 9: 4S3-456. 324. Gabel, N.W., Abood, L.G. 1965: Stereochemlcal Factore Related to the Potency of k~tlcholinerglc Psychotomlmetlc Drugs. J. Hed ~ Chem. 8: 616-619 ~ 143

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