Tobacco Documents Online

TIII: Cut•rcu.VbR Tox:ACCO RrS n.AR('}I-U.S.A., INC. July 31, 1s7~ WQ-1 0t}IIO 1l1 MEMORANDUM TO: Drs. Ford, Hockett, Meier, Sommers and Stone FROM: W. U. Gardner SUBJECf: Application 1161R2. A. M. Tometsko, Ph.D., Litron Laboratories Ltd., Rochester, N.Y. "Probing Nicotine Receptor Sites." Progress. Photosensitive and conventional probes have been synthesized and used to study the functionlstructure relationships of nicotine receptors. One of the cationic photosensitive substances synthesized was 4-azido-2-nitro- nitrophenoyl nornicotine (NAP-N1C). It competes with nicotine for receptor sites. Rodent malaria (Plasmodium vinckei) has been used to find sensitive means of identifying vital sites of drug action. A manuscript reporting the effect of photosensitive chemicals on the rodent malaria was submitted with the progress report. Work done with Dr. Abood with (-)-nicotine and with a number of nicotine and pipPridine analogues has been reported in the current grant year although the work was done previously. (-)-nicotine was more than 100 times more active than (+)-nicotine in producing the prostration-immobilizing syndrome. Projection. During the coming year two prokaryotes (E. coli and P. pudita) and two eukaryotes S. cervisiae and human blood cells will be used to (1) determine the mechanism of transport intci the cells, (2) evaluate the ability of synthetic derivatives to compete with nicotine for transport sites, (3) use photosensitive derivatives as probes to check on inhibition of nicotine transport, (4) synthesize derivatives with high affinity for the transport site, and (5) synthesize macromolecular nicotine derivatives to study further nicotine receptors on cell membranes. The compounds to be synthetized are listed and procedures to be used described. Finally, the limitations of studies of this type are listed: (1) possible non-specificity of transport proteins, (2) toxicity to the cells, (3) possible carcinogenicity of some of the chemicals, and (4) possibility of a pseudoaffinity labeling. Budget. Salaries of two professional (20% of time) and one technical person are requested. The remainder is for consumables and services. The indirect costs are 36% of TDC and a profit of 6% of TDC. To EJ. tg. U. Gardner WUG:ek