Tobacco Documents Online

Date. . Nameoflnvestigotbr(s)e(inciudeTitleandDegrees) Herbert McKennis, Jr., Ph.D., Professor of Pharmacology z Uutttutton & • Address: i`ledical College of Virginia, Richmond, Virginia 23219 Factors Controlling the Biological Disposition of Pyridine .;Coupounds of Tobacco Smoke {~. . . _ :• I. ProposedStartingDate: July 1, 1967' =` S. AnGcipoted Duration ofithis Specific Study: 1967-197D (3 years) 'it has been possible to•describe the broad outlines of many of the routes involved in the 6. Brief Descripton of Objectives or Specific Aims: From a series of experiments with support of the Couneid for Tobacco Researeh-U.S.A., .:~r -__c:;, i, ..c;.•oammalian metabolism of nicotine, nornicotine, and 3-acetylpyridine -- three of the many ; pyridine compounds that occur in tobacco and tobacco smoke. [The general nature of the 4igiRf. FI previous studies are given in the publications which are enumerated in an attached 1ist.) Through. isolation or total synthesis it has been possible to supply many of the mammalian .::pharmacologica3 properties of many nicotine metabolites. •' metabolites to biological investigators and. to assist them in a variety of work on the +=" , It is now desirable for us and interesting to us to expand our knowledge of the factors ~.='~'''which determine the disposition of pyrid3ne compounds arising from tobacco smoke and eaptore oiocnemicai1 inieraccions OeLWCeID Yml1AC WNpvucuw. rLC16 a~s aireauy in tne puo- lished literature some data that suggests that desensitization or tachyphylaxis to nicotine arises in part through a stimulation of the enzymes that are concerned with, the : aetabolism-of nicotine. It has been suggested that this stimulation of metabolism is ; produced by nicotine and:that other substances have a similar capacity to enhance the metabolism-of nicotine. Direct studies on nicotine metabolism-are however very limited ., in their scope. "determine the nature of stimulation or inhibition of metabolism at almost all- of the key C points that are involved in mammalian systems. Similarly, it should be possible to determine individual differences in the capacity to metabollize nicotine and related pyridine compounds. (continued on sheet la attached). 7. Give a Brief Statement of your,Working Hypothesis: Current data on the effect of various smoke components on the metabolism and transfer of nicotine and other alkaloids are limited or apparently contradictory. A study of these factors should be helpful to an- understanding of the.biological effects of tobacco smoking. have already described,, and through synthetic routes which make possible the labelLing of nicotine and related in almost any desired position, we believe that it is now possible to Through the current availability of synthetic routes to nicotine metabolites, which we [o-`mm a- CiAW 49MM ~ ~

Brief Description of Objectives or Specific Aims: (continued) ~.,_ . ~ . .. . . -~~. ..',. -.. ,. , . . .-. In the case of some of the simpler metabolites of nicotine, such as methylamine, 'it is "i relatively easy to obtain a broad picture of the nature of compounft which interfere with metabolic events. Work with 1`'C-methylamine in our own laboratory (The Pharmacologist, Fall 1967, in press) and others illustrates that interference with oxidation of rz ;}` T methylamine maY be produced by some types of monoamine-oxidase inhibitors. The data " suggest, however, that the inhibition may be referrable to-a so-called methylamine : oxidase which has not been fully characterized. Liver preparations obtained from rats after administration of monoamine-oxidase inhibitors have thus far been shown to be as fully capable as control livers in tests which, involve a spectrophotometric measurement of the disappearance of•nicotine. From another series of experiments, the data now suggests that the oxidation of nicotine can be impeded by the presence of large quantities of cotinine. It would, be desirable to study possible effects of other pyridine compounds 'T of smoke on the metabolism of nicotine and related substances. The.need for a biologically. , oriented study appears to be re-emphasized by apparently conflicting chemical data on the _ oxidation of nicotine. For instance, the pyrolidine-N-oxide of nicotine has been considered; (by J. C. Craig) to be the logical key intermediate in the biological.formation of cotinine, while Linnell has reported that the same oxide in vitro inhibits the oxidation ;..;: ~f nicotine to cotinine. 0

1 8. Details offiExperimentat Design and Procedures: (Attach Separate Pages) With the aid of isotopically labelled.nicotine and its metabolites it is feasible to study by a number of different methods factors influencing the disposition of nicotine. It is anticipated that during the current studies both in vivo and!in vitro methods will be employed. _ Initial experiments to determine the suitability of the rat as an experimental animal for initial studies have already beem conducted. Liver microsomal preparations from both :young and old rats of the >:,'?Ystar strain have been examined for their capacity to metabolize . ;;;,y nicotine in vitro. The general picture currently presented is that equivalent fractions " by weight of liver that are obtained from older animals (250-550g body weight) do not ~: metabolize nicotine as effectively as do the respective fractions from younger animals (150-175 g body weight). It is not currently:known whether or not the observed deficiency •in the older animals can be attributed to a lesser amount of metabolizing cellular •'•.fractions per unit of liver weight, or an overa•11 lessening of metabolic ability that exists throughout the animal. It would-be desirable to settle this point and to determine ; whether or not the elimination of unchanged nicotine by the older animals suffers any ` impairment. The additional matter of elimination can.be settled without modification of the experimental design of the metabolic experiments. Questions of this type can be .•"answered by comparing the ability of groups of young and old'animals to'metabolize (continued--on sheet 2a attached) • . 9. • Physical Facilties Available (Where Other than Administering Organization Indicate Geographical Location) Laboratories and animal facilities are.located on the fourth and fifth floors of McGuire Hall at the Medical College of Virginia. Gas chromatography and isotope counting facilities are available in areas convenient to the working laboratories but separate from these. The equipment list includes a preparative ultracentrifuge, polarimeter, Wa bu a lratus, hydrogenation apparatus, and!the general equipment of chemical, and 1~ A~citiortequirements: biological laboratories. , Additional requirements, if any, would depend!upon the outcome of the research. J • 11. Biographical sketches of all:principal and professional personnel (append) 12. List of publications: (Five most recent as pertinent) (append) -~~ _..,...,.....- .._.KS,.r. ~ . . • ... .. ... ._ . ..- . _.._ .. .._ .~.._....._._,.. .__ .. ...._.,_..,...- .~.~.,.~,.._,..~ ......_

• 2a. 8. Details of Experimental Design,and Procedures: (continued) 1'+C-labelled nicotine and to eliminate unchanged alkaloid. For all of this isotopic material is now available.as a result of our synthetic work and previous nutriculture: nicotine-1`'C (random), nicotine-14C-methyl, and nicotine-2-14C. -"`'In additiom, six of the mammalian metabolites of (-)-nicotine are available in both• _aF . "' isotopic and'non-isotopic form so that quantitative data can be obtained. 0 .

R: REDACTED MATERIAL •- E. Consvmabie Supplies (iist by catepories) --Chemi.cal Reagents .Isotopes Glass ware : Animals and feed Year 2 Year 3 C. Other: Expenses Qtemize) Reprints and page charges •Travel (Presentation of reports and conferences) E.. Overhead (] 5% of A+ B+ C) EstimatedFuture Requirements: Salaries Consumable Suppl. It is understood thotthe applica in applying fona grant have re the Council's "Statement oi. Pot and?erms Under Which Project Sub Totai I OtherExpenses PermanentEquiR. 250 500 Ovencead Total S,S00 _ 1,000 1,000 6,675 52,175 6,000_ 1,000 1,000 7,050 55,050 // ~ Siqnature 1'..~- ~ t. / / /y~.( r t and institutionai officen 644-9851 xt. 8153 ~ ad and found accept661t - cy Containing Conditions Signature Telephone Grants Are Made-`° wu ~aoflk..ohhtlmGru6°n ~•1NJE. ~L [;~: ;S VICE Te4ephone n i ML VJ ~a );h W 'L•-•- "~ +~ at meetings Ar=

i , List financial support fea research from oPl sources, tndudinp own instituNon, for this and/or related research projects. . Quantitative Methods for the'Detesmination*of the Distribution of Nicotine and Its Congeners in Biological Systems Institution supplies salary of principal-investigator and all secretarial help currently obtained through other funds None other than this application for the Council for Tobacco Research - U.S.A. VD99PSC00t

R: REDACTED MATERIAL B'iographical'Inyormation on HERBERT I•icKEtJi+tll S , JR. '9-.. ~- _ _ ~ Attended elementary schools in Scarsdale, N.Y. and Loomis School, Windsor, Conn. , Harvard S~.II . , ~ and Cornel•-l. Fh..D-. Chemist at Nuodex Products Co., 1938-1939, and Ciba Pharmaceutical Products Co,, 1940-1942, Appointed Assistant in Biochemistry, Cornell University Medical College, 1942; Assistant Professor of Chemistry, Medical College of Virginia, 1945; Instructor in Physio- logical.Chemistry, the Johns H'opkins-University, 19:46; Associate Professor of-Biochemistry, Medical College of Virginia, 1948; fie•ad, Basic Sciences Res'earch Department, Naval C. E. Laboratory, 1949; Associate-Professor-, 1953, Professor of Pharmacology, Medical College-of Virginia, 1955. Biochemist; specialist in intermed!iary metabolism, anti- biotics, alkaloids, polymeric compoundis. U..S. and-foreign patents: soil stabilization and antispasmodic. Has done a wide variety of scientific-advisory and technical consulting work for industrial firms, government agencies,' scientific organizations, and universities. Visiting Professor•, University of Chile, 1960; Hono•ra•ry member-, Faculty of Medicine, University of Chile, Sociedad de Biologia, de Santiago. Sigma Xi. Phi Lambda Upsilon. American-Chemical Society, American Society of Biological Chemists, The Society for Experimental Biology and Medicine, International Oceanographic R'oundation, The New York Academy of Sciences, American Society for Pharmacology and Experi- mental Therapeutics, American Associ!_&ti:orn for the Advancement of Science, Society of American, Mil-itary Engineers, American Institute of Chemists, Virginia Academy of Science, Society of Toxicology.

s- Edward R. Bowman Born: West Virginia 1927 Medical C.>llege of Virginia Citizen U.S.A. Richmond, Virginia Education: -Concord College B.S. 1952 (Biology - Chemistry). West Virginia University 1953 (Physi:ology). Duke University 1955-56• (Graduate Student-in Physiology). %=. Medical College of Virginia PheD. 1963 (Pharmacology). Experience: 1961 - present • 19'56 - 1958 1955 - 1956 1954 - 1955 ~ 1952 - 1953 1952 195 0 - 1951 •.19'47 - 1950 •1944 - 1946 Research Associate Department of Pharmacology Medical College of Virginia Richmond, Virginia Graduate Student, Major - Pharmacology Minor - Physiology & Biochemistry Medical College of Virginia, Richmond., Virginia Research Assistant Department of Pharmacology Medical College of Virginia Richmond, Virginia Graduate Student, Major - Physiology Minor - Anatomy Duke University Durham, North Carolina Bacteriologist State Department of Health Richmond, Virginia Graduate Student, Physiology West Virginia University Morgantown, West Virginia Student, Biology & Chemistry Concord College N Athens West Vir inia O , g O U.S. Army ~JI EA Student, Biology & Chemistry Concord College ~ Athens , West Vir inia g Q? U.S. Arm.y

R: REDACTED MATERIAL 0

PUBLICATIONS OF PROJECT WORK SUPPORTED BY THE COUNCIL FOR TOBACCO RESEARCH - U.S.A. 1. Synthesis and properties of pyridylalanines. Herbert McKennis, Jr., and Edward R. Bowman. Virginia Journal of Science, 8, 314 (1957). 2. Metabolism of y-(3-pyridyl)-Y-oxobutyric acid. Lennox B. Turnbull, Edward R. Bowman, and Herbert McKennis, Jr. Federation Proceedings, 17, 325 (1958). 3. Y-(3-Pyrid)rl)-Y-methylaminobutyric acid as a urinary metabolite "of nicotine. Herbert McKennis, Jr., Leiinox B. Turnbull, and Edward R. Bowman. Journal of the American Chemical Society, 79, 6342 (1957). 4. Metabolites of nicotine and a synthesis of nornicotine. Herbert McKennis, Jr., Lennox B. Turnbull, Harvey N. Wingfield, Jr., and Lovell J. Dewey. Journal of the American Chemical Society, 80, 1634 (1958). 5. The role of cotinine in nicotine metabolism Herbert McKennis, Jr., Lennox B•. Turnbull, and Edward R. Bowman. Abstracts of Communications, IV. International Congress of Biochemistry, Vienna, Sept. 1-6, 1958. 6. 7. 8. A constant rate infusion apparatus. Quentin S. McKennis, Edward R. Bowman, and Herbert McKennis, J Toxicology and Applied Pharmacology, 1, 61 (1959). Metabolism of nicotine to (+)-Y-(3-pyridyl)-y-methylaminobutyric acid. Herbert McKennis, Jr., Lennox B. Turnbull,*and Edward R. Bowman. Journal of the American Chemical Society, 80, 6597 (1958). Metabolism of nicotine in the human and excretion of pyridine compounds by smokers. Edward R. Bowman, Lennox B. Turnbull, and Herbert McKennis, Jr. The Journal of Pharmacology and Experimental Therapeutics, 127, 92 (1959). Demethylation of cotinine in vivo.. N O ~ Herbert McKennis, Jr., Lennox B. Turnbull, Edward R. Bowman, and Einosuke P+ada. Journal of the American Chemical Society, 81, 3951 (1959). GJ CA ~,. Oxidation of cotinine in vivo. ~ Edward R. Bowman, Lennox B. Turnbull, and Herbert McKennis, Jr. ~ ~ Federation Proceedings, 18, 371 (1959).

-2- 11. Depressor activity from nicotine metabolites. Edward R. Bowman, H. B. Kennedy, Jr., Einosuke Wada, and Herbert McKennis, Jr. - Abstracts of Communications, XXI. International Congress of Physiological Sciences., 41, Buenos Aires, August 9-15, 1959. The isolation andistructure of a ketoamide formed in the metabolism of nicotine. Herbert McKennis, Jr., Edward R. Bowman, and Lennox B. Turnbull. Journal of the American Chemical Society, 82, 3974 (1960). 13. Methylation-in the.metabolism of (-)-nicotine. •Lennox B. Turnbull, Edward R. Bowman, and Herber•t'McKennis, Jr. Federation Proceedings, 19, 268 (1960). 14. The excretion and metabolism of nicotine. Herbert MeKennis, Jr. Annals of the New York Academy of Sciences, 90, 36 (1960). 15. Norcotinine (desmethylcotinine) as a urinary metabolite of nornicotine. Einosuke Wada, Edward R. Bowman, Lennox B. Turnbull, and Herbert McKennis, Jr. Journal of Medicinal and:Pharmaceutical Chemistry, 4, 21 (1961). 16. Depressor effects arising from (-)-cotinine. Joseph F. Borzelleca, Edward R. Bowman, and Herbert McKennis, Jr. The Pharmacologist, 2, 72 (1960). 17. Oxidation of nicotine-C-14 and nicotine-methyl-C-14 in vivo. Lennox B. Turnbull, Einosuke Wada, Edward R. Bowman, and Herbert McKennis, Jr. Federation Proceedings, 20, 172 (1961). 18. Metabolism,of nicotine in the human. Edward!R. Bowman, Lennox B. Turnbull, and-Herbert McKennis, Jr. Virginia Journal of Science, 9, 438 (1948). 19. Degradation of (-)-cotinine in the human. Herbert McKennis, Jr., and Edward R. Bowman. Abstracts of Communications, V. International Congress of Biochemistry, Moscow, August 10-16, 1961, p. 393. 20. The mammalian degradation of (-)-nicotine to 3-pyridylacetic acid and other compounds. Herbert McKennis, Jr., Edward R. Bowman, and Lennox B. Turnbull. Proceedings of the Society for Experimental Biology and Medicine, 107, 145 (1961). 21. Metabolism of (-)-cotinine in the rat. Is Sorell L. Schwartz, Edward R. Bowman, and Herbert McKennis, Jr. Virginia Journal of Science, 12, 196 (1961).

-3- 22. Demethylation in the metabolism of (-)-nicotine. Herbert McKennis, Jr., Lennox B. Turnbull., Sorell L. Schwartz, Einosuke Tamaki. and Edward R. Bowman. The Journal of Biological Chemistry, 237, 541 (1962). .23. Metabolism of (-)-cotinine to a keto acid. . Sorell L. Schwartz, and Herbert McKennis, Jr. -Federation Proceedings, 21, 183 (1962). 24. .Studies on the respiratory and cardiovascular effects of (-)-cotinine. Joseph F. Borzelleca, Edward R. Bowman., and Herbert McKennis, Jr. The Journal of Pharmacology and Experimental Therapeutics, 137, 313 (1962). 25. Studies on the metabolism of (-)-cotinine in the human. Edward R. Bowman, and Herbert McKennis, Jr. The Journal of Pharmacology and Experimental Therapeutics, 135, 306 (1962). _ 26. Routes in the mammalian metabolism of (-)-nicotine to 3-pyridylacetic acid. Sorell L. Schwartz, Edward R. Bowman, and,Herbert McKennis, Jr. Abstracts of Papers, 16th Tobacco Chemists' Research Conference, Richmond, Virginia, Sept. 26-28, 1962, p. 7. 27. Studies on the metabolic fate of 3-acetylpyridine. Lennox B. Turnbull, Edward R'. Bowman, and Herbert McKennis, Jr. Abstracts of Papers, 16th Tobacco Chemists' Research Conference, Richmond, Virginia, Sept. 26-28, 1962, p. 6. 28. The corrected structure of a ketoamide arising from the metabolism of (-)-nicotine. Herbert McKennis, Jr., Lennox B. Turnbull, Edward R. Bowman, and Sorell L. Schwartz. Journal of the American Chemical Society, 84, 4598 (1962). 29. N-Methylation of nicotine and cotinine in vivo.' Herbert McKennis, Jr.,•Lennox B. Turnbull, and Edward R. Bowman. The Journal of Biological Chemistry, 238, 719 (1963). 30. Studies on-the degradation of the pyrrolidine ring of (-)-nicotine in vivo. Formation of Y-(3-pyri,dyl)-Y,-oxobutyric acid. Sorell L. Schwartz, and Herbert A•fcf:ennis, Jr. The Journal of Biological Chemistry, 238, 1807 (1963). 31. The synthesis of hydroxycotinine and studies on its structure. Herbert McKennis, Jr., Lennox B. Turnbull, Edward R. Bowman, and Einosuke Tamaki. Journal of Organic Chemistry, 28, 383 (1963). 32. (-)-Cotinine. Edward R. Bowman, and Herbert McKennis, Jr. Biochemical Preparations, 10, 36 (1963).

33. Conjugate formation in the metabolisni of 3-acetylpyridine..' Lennox B. Turnbull, Edward R. Bowman, and Herbert McKennis, Jr. Abstracts of Papers, 17th-Tobacco Chemists' Research Conference, Montreal, Quebec, Canada, Sept. 22-25, 1963, p. 11. A comparative study of the metabolic release of methyl groups from a series of N-methylpyridinium compounds. Herbert McKennis, Jr., Edward'R. Bowman, Antonio Horvath, and John P. Bederka, Jr. ~ Nature, 202, 699 (1964). i • 35. Mammalian degradation of (-)-demethylcotinine. Sorell L. Schwartz, and Herbert McKennis, Jr. Nature, 202, 594 (1964). 36. Disposition and fate of (-)-cotinine-H3 in the mouse. Edward R: Bowman, Eskil Hansson, Lennox B. Turnbull, Herbert McKennis, Jr., and Carl G_ Schmiterlt5w. The Journal of Pharmacology and!Experimental Therapeutics, 143, 301 (1964). 40. Alternate routes in the metabolic degradation of the pyrrolidine ring of nicotine. Herbert McKennis, Jr., Sorell L. Schwartz, and-Edward R. Bowman. The Journal of Biological Chemistry,, 239, 3990 (1964). Herbert MeF:ennis, Jr., Sorell L. Schwartz, Lennox B. Turnbull, Einosuke Tamaki, and Edward R. Bowman. The Journal of Biological Chemistry, 239, 3981 (1964). 37. Additional routes in the metabolism:of 3-acetylpyridine. Herbert McKennis, Jr., Lennox B. Turnbull, and Edward R. Bowman. The Journal of Biological Chemistry, 239, 121S (1964). 38. Effect of cotinine and other nicotine metabolites in-yitro on duod'enum-and;ileum segments. K. S. Kim, Joseph F. Borzelleca, Edward R. Bowman, and Herbert McKennis, Jr. Federation Proceedings, 23, 330. (1964). 39. The metabolic formation of y-(3-pyridyl)-y-hydroxybutyric acid and its possible intermediary role in the mammalian metabolism of nicotine. 41. Metabolism of 3-acetylpyridine to an analog of mandelic acid. Lennox B. Turnbull, C. N1. Lukhard,, and Herbert McKennis, Jr. Toxicology and Applied Pharmacology, 6, 362 (1964). 42. Disposition and fate of nicotine in animals. Herbert McKennis, Jr. Tobacco Alkaloids and Related Compounds, U. S. Von Euler, editor, Pergamon Press, Oxford, 1965, p. 53.

43. Urinary excretion of conjugate forms.of 1-(3-pyridyl)ethanol aftei administration of 3-acetylpyridine. Herbert McKennis, Jr., Lennox B. Turnbull, Edward R. Bowman., and C. Norman Lukhard. The Journal of Biological Chemistry, 241, 1878 (1966). 44. The structure of dibromoticonine, a bromination product of nicotine. • New York, N. Y., Sept. 12-16, 1966. Abstracts of Papers, 152nd Meeting, American Chemical Society, Herbert McKennis, Jr., Edward R. Bowman, L. D. Quin, and R. C. Denney: 45. Studies on the synthesis and metabolism*of 4-(3-pyridyl)-4- methylaminobutyric acid-4-14C. Paolo L. Morselli, Edward R. Bowman, Helen H. 0ng, and! Herbert McKennis, Jr. Virginia Journal of Science, 17, 345 (1966). 46. The fate and distribution of 1-(3-pyridyl)ethanol methiodide in relation to the toxicity.of 1-(3-pyridyl)ethanol and 3-acetylpyridine. John P. Bederka, Jr., Eskil Hansson, Edward R. Bowman, and Herbert McKennis, Jr. Biochemical Pharmacology, 16, 1 (1967). 47. Studies on the separation of acidic metabolites of nicotine by gas chromatography. Herbert McKennis, Jr., Edward R. Bowman, and Mohammad Saeed Dar. Virginia Journal of Science, 18, 13 (1967). 48. Pharmacological action and intermediary role of 5-(3- -pyridyl)tetrahydrofuranone-2. Edward R. Bowman, Pavol lirdina and; Herbert McKennis, Jr. Federation Proceedings, 26, 616 (1967). 49. Metabolism of (')-Cotinine-2-Z 4C in the rat. Paolo L. Morselli, Helen H. Ong, Edward R. Bowman, and Herbert McKennis, Jr. Journal of Medicinal Chemistry, in press. 50. 1`1ethylamine metabolism in normal and, mao-inhibitor-treated Paolo L. biorselli, Edward R. Bowman,, and Herbert McKennis, The Pllarmacologist, in press. rats. Jr.

-6- 51. On•the congeners of whiskey. Herbert McKennis, Jr.,:-and;Harvey B. Haag. Journal of the American Geriatrics Society, 7, 848 (1959). 52. ,Depressor effects arising from (-)-cotinine. Joseph F. Borzelleca, Edwar.d R. Bowman, and Herbert McKennis, Jr. •The Pharmacologist, 2, 72 (1960). 53. The excretion and metabolism of nicotine. Herbert McKennis, Jr. Annals of the New York Academy of Sciences, 90, 36 (1960). 54. The isolation and structure of a ketoamide formed in the metabolism of nicotine. Herbert McKennis, Jr., Edward R. Bowman, and: Lennox B. Turnbull. . Journal of the American Chemical Society, 82, 3974 (1960). 5S. Methylation in the metabolism of (-)-nicotine. Lennox B. Turnbull, Edward R. Bowman, and Herbert McKennis, Jr. Federation Proceedings, 19, 268' (1960). 56. L-y-Glutamylhydrazine and the metabolism of hydrazine. Herbert McKennis, Jr., Allan S. Yard, Elizabeth J. Adair, and J. H. Weatherby. The Journal of Pharmacology and Experimental Therapeutics, 131, 152 (1961). ` 57. Demethylation in the metabolism,of (-)-nicotine in vivo. Herbert McKennis, Jr., Einosuke Wada, Edward R. Bowman, and Lennox B. Turnbull. Nature, 190, 910 (1961). 58. Norcotinine (Desmethylcotinine) as a urinary metabolite of nornicotine. Einosuke R'ada, Edward R. Bowman, Lennox B. Turnbull, and Herbert McKennis, Jr. Journal of Medicinal and Pharmaceutical Chemistry, 4, 21 (1961). 59. The isolation of 3-pyridylacetic acidi, a urinary metabolite of (-)-cotinine. Edward R. Bowman,, Lennox B. Turnbull, and Herbert McKennis, Jr. Abstracts of Papers, 14th Tobacco Chemists'- Research Conference, Oct. 13-14, 1960, Winston-Salem, North-Carolina. 60. Oxidation,of nicotine-C14 and nicotine-methyl-C14 in viyo. Lennox B. Turnbull, Einosuke iiada-, Edward R. Bowman, and Herbert McKennis, Jr. " Federation Proceedings, 20, 172 (1961). 61. Inhibition by thyroxine of y-aminobutyrate-a-ketoglutarate transaminasc. Antonio Horva:th, Fernando Orrego, and Herbert McKennis, Jr. Federation Proceedings, 20, S (1961).

62. Factors controlling the metabolism of Y-aminobutyric acid. Antonio Horvath, Fernando Orrego, and Herbert McKennis, Jr. The Journal of Pharmacologyy and Experimental Therapeutics, 134, ,222 (1961). 63. Mammalian degradation of (-)-nicotine to 3-pyridylacetic acid and other compounds. . Herbert McKennis, Jr., Edward R. Bowman, and Lennox B: Turnbull. Proceedings of the Society for Experimental Biology and Medicine, 107, 145 (1961). 64. Selective toxicity. (Book review). Herbert McKennis, Jr. Revista Medica de Chile, 88, 864 (1960). 65. Los oxoesteroides, el uso de-hidrazidos fen6licos para su- detecci6n, caracterizaci6n y medici6n,. (Book review). Herbert McKennis, Jr. Revista Medica de Chile, 88, 626 (1960). 66. Metabolism of nicotine in the human. Edward R. Bowman, Lennox B. Turnbull, and Herbert McKennis, Jr. Virginia Journal of Science, 9, 438 (1958). 67. Metabolism of (-)-cotinine in the rat. Sorell L. Schwartz, Edward:R. Bowman, andiHerbert McKennis, Jr. Virginia Journal of Science, 12, 196 (1961). 68. Degradation of (-)!-cotinine in the human.. Herbert McKennis, Jr., and Edward,R. Bowman. Abstracts of Communications, V. International Congress of Biochemistry, Moscow, August 10-16, 1961, p. 393. 69. Aspects of the metabolism of isoniazid and acetylisoniazid in the human and the dog. Allan S. Yard, and Herbert McKennis, Jr. Journal of Medicinal and Pharmaceutical Chemistry, 5, 196 (1962). 70. Demethylation in the metabolism of (-)-nicotine. , Herbert McKennis, Jr., Lennox B. Turnbull, Sorell L. Schwartz, Einosuke Tamaki, and Edward R. Bowman.. The Journal of Biol~ogical Chemistry, 237, 541 (1962). Inhibition of the catabolism of aspartic-4-C14 acid by thyroxine in vivo. Antonio Horvath, and•Herbert McKennis, Jr. Enzymologia 24 91 (1962) N 0 , , . Studies on-the metabolism of (-)-cotinine in the human. G.? C11 .~ Edward R. Bowman, and Herbert McKennis, Jr. ~ The Journal of Pharmacology and Experimental Therapeutics, ~ 135, 306 (1962). ~

73. Metabolism.of (-)-cotinine to a keto acid. Sorell L. Schwartz, and Herbert McKennis, Jr. Federation Proceedings, 21, 183 (1962). 74. The_excretion and metabolism of triethylene glycol. Herbert McKennis, Jr., Robert A. Turner, Lennox B: Trunbull, .and Edward R. Bowman. W. W. Muelder, M. P. Neidhardt, and"Carl L. Hake. - Richard Henderson, Herbert G. Nadaeu, and Samuel Spencer. Toxicology and Applied Pharmacology, 4, 411 (1962)1. 75. Studies on the respiratory and cardiovascular effects of (-)-cotinine. Joseph F. Borzelleca, Edward R. Bowman, and Herbert McKennis, Jr. The Journal of Pharmacology and-Experimental Therapeutics, 137, 313 (1962). 76. Routes in the mammalian metabolism of (-)-nicotine to 3-pyridylacetic acid. Sorell L. Schwartz, Edward R. Bowman, and Herbert hlcKennis, Jr. Abstracts of Papers, 16th. Tobacco Chemists' Research Conference, Sept. 26-28, 1962, Richmond, Virginia, p. 7. 77. Studies on the metabolic fate of 3-acetylpyridine. Lennox B. Turnbull, Edward R. Bowman, and Herbert McKennis, Jr. Abstracts of Papers, 16th Tobacco Chemists' Research Conference, Sept. 26-28, 1962, Richmond, Virginia, p. 6. 78. Acetylhydrazine as an intermediate in•the metabolism of aroylhydrazines. Lennox B. Turnbull, Allan S. Yard, and Herbert McKennis, Jr. Journal of Medicinal and Pharmaceutical Chemistry, 5, 1327 (1962). 79. N-Methylation of nicotine and cotinine in vivo. Herbert McKennis, Jr., Lennox B. Turnbull~, aE-d Edward R. Bowman. The Journal of Biological Chemistry, 238, 719.(1963). > 80. The synthesis of hydroxycotinine and:studies on its structure. Herbert McKennis, Jr., Lennox B. Turnbull, Edward R. Bowman, and Einosuke Tamaki. Journal of Organic Chemistry, 28, 383 (1963). 81. The corrected structure of a ketoamide arising from the metabolism of (-)-nicotine. Herbert McKennis, Jr., Lennox B. Turnbull, Edward R. Bowman, 82. and Sorell L. Schwartz. Journal of the American Chemical Society, 84~, 4598 (1962). (-)-Cotinine. Edward R. Bowman, and Herbert McKennis, Jr. Biochemical Preparations, 10., 36 (1963). ti

83.. Studies on the degradation of the pyrrolidine ring of (-)-nicotine in vivo. Formationm of.7-(3-pyri•dyl)-Y-oxobutyric acid. Sorell L., Schwartz, and Hkerbert McKennis, Jr. The Journal of Biological Chemistry, 238, 1807 (1963). 84. Conjugate formation in the metabolism of 3-acetylpyridine. Lennox R. Turnbull, Edward R. Bowman, and Herbert McKennis, Jr. Abstracts of Papers, 17th Tobacco Chemists', Research Conference, Sept. 22-25, 1963, Montreal, Quebec, p. 11. 85. Metabolic release of methyl groups from a series of N-methylpyridinium compounds. Herbert McKennis, Jr., Edward R. Bowman, Antonio Horvath, and John P. Bederka, Jr. Nature, 202, 699 (1964). 86. Mammalian degradation•bf (-)-demethylcotinine. Sorell L. Schwartz, and Herbert McKennis,, Jr. Nature, 202, 594 (1964). 87. Disposition and fate of (-)-cotinine-H3'in the mouse. Edward R. Bowman, Eskil Hansson, Lennox B. Turnbull, Herbert McKennis, Jr.; and Carl G. Schmiterlbw. The Journal of Pharmacology and:Experimental Therapeutics, 143, 301 (1964). 88. Additional routes in the metabolism of 3-acetylpyridine. Herbert McKennis, Jr., Lennox B. Turnbull, and Edtaard R. Bowman. The Journal of Biological Chemistry, 239, 1215 (1964). 89. The metabolic formation of Y-(3-pyridyl)-y-hydroxybutyric acid and its possible intermediary role in the mammalian metabolism of nicotine. Herbert McKennis, Jr., Sorell L. Schwartz, Lennox B. Turnbull, Einosuke Tamaki, and Edward R. Bowman. The Journal of Biological Chemistry, 239, 3981 (1964). 90. Alternate routes in the metabolic degradation of the pyrrolidine ring of nicotine. Herbert McKennis, Jr., Sorell L. Schwartz, and Edward R. Bowman. The Journal of Biological Chemistry, 239, 3990 (1964). 91. Effect of cotinine and other nicotine nmetabolites in vitro on duodenum and ileum segments. K. S. Kim, Joseph F. Borzelleca, Edward R. Bowman, and Herbert McKennis, Jr. Federation Proceedings, 23, 330 (1964). 92. Metabolism of 3-acetylpyridine to an, analog of mandelic acid. Lennox B. Turnbull, C. N_ Lukhard, and Herbert McKennis, Jr. Toxicology and Applied°Pharmacology, 6, 362 (1964). ..~,.~._.;~::....,..d.~~...~.».-.: ~~w...

.-10- + 93. Disposition and fate of nicotine irn animals. Herbert McKennis, Jr. ' Tobacco Alkaloids and Related Compounds, U. S. Von Euler, editor, Perganon Press, Oxford, 1965, p.'53. 94. Viewpoints of the study of drug toxicity. (Book review). Herbert McKennis, Jr. American Journal of Pharmaceutical Education, 28,469 (1964). 95. Urinary excretion of conjugate forms of 1-(3-pyridyl)ethanol after administration of 3-acetylpyrid2ne. Herbert McKennis, Jr., Lennox B. Turnbull, Edward R. Bowman, and C. Norman Lukhard. The Journal of Biological Chemistry, 241,.1878 (1966). 96. The structure of dibromoticonine, a bromination product of nicotine. Herbert McKennis, Jr., Edward R. Bowman, L. D. Quin, and R. C. Denney.

Papers Published During the Period of This Report i 1. Urinary excretion of conjugate forms of 1•-(3-pyridyl)ethanol after administration of 3-acetylpyridine. Herbert McKennis, Jr., Lennox B. Turnbull, Edward R. Bowman,, and C. Norman Lukhard. The Journal of Biological Chemistry, 241, 1878 (1966). 2. The structure of dibromoticonine, a bromination.product of nicotine. Herbert McKennis, Jr., Edward R. Bowman, L. D. Quin, and R. C. Denney. Abstracts of Papers, 152nd Meeting, American Chemical Society, New York, N. Y., Sept. 12-16, 1966. 3. Studies on the synthesis and metabolism of 4-(3-pyridy.l)-4- methylam%nobutyric acid-4~I`'C. Paolo L. Morselli, Edward R. Bowman, Helen H. Ong, and Herbert McKennis, Jr. , Virginia Journal of Science, 17, 345 (1966). 4. The fate and distribution, of I-(3-pyridyl)ethanol methiodide im relation to the toxicity of 1-(3-pyridyl)ethanol and 3-acetylpyridine. John P. Bederka, Jr., Eskil Hansson, Edward R. Bowman, and Herbert McKennis, Jr. Biochemical Pharmacology, 16, 1 (1967). 5. Studies on the separation of acidic metabolites of nicotine by gas chromatography. ` Herbert bicKemtis, Jr., Edward R. Bowman, and Mohammad Saeed Dar. Virginia Journal of Science., 18, 13 (1967). 6. Pharmacological action and intermediary role of 5-(3- pyrid'y1)tetrahydrofuranone-2. - Edward R. Bowman, Pavol lirdina and Herbert McKennis, Jr. Federation Proceedings, 26, 616 (1967). 7. b1etabolism• of (=)-cotinine-2-14C in the rat. Paolo L. Morselli, Helen H. Ong, Edward R. Bowman, and Herbert McKennis, Jr. Journal of Medicinal Chemistry, in press. 8. Methylamine metabolism in normal and mao-inhibitor-treated rats. F.1 Paolo L. Morselli, Edi,rard~ R. Bowman, and Herbert McKennis, Jr. O The Pharmacologist, in press. CO CA 4 0? M ~ ~ . . o~

Talks Given During the Period of this Report 1. Studies on the synthesis and metabolism,o.f 4-(3-pyridyl)-4- methyiaminobutyric acid-4-14C. Paolo L. Morselli, Edward R. Bowman,•'tlelen`II: Ong, and Herbert McKennis, Jr. Annual Meeting 'of the Virginia Academy of Science, Harrisonburg, Virginia, May 4-6, 1966.- 2. The structure of dYbromoticonine, a bromination produce of nicotine. Herbert McKennis, Jr., Edward R. Bowman, L. D. Quin, and R. C. Denne'y:--i52nd-Meeting, American Chemical Society, New York, N. Y., Sept: 12-16, 1966. 3. Some new-synrthetic routes to metabolites of nicotine. IIerbert.. McKennis, Jr., Biological Seminar, Medical College of Virginia, Richmond, Virginia, Sept. 28', 1966. 4. Quantitative methods for the determination of the distribution of nicotine and its congeners in biological systems. Iierbert McKennis, Jr., American Medical Association 14orkshop,, Colorado Springs, Colorado, Nov. 1-3, 1966. 5. Studies on 1''C-labell-ed nicotine metabolites. Edward R. Bowman and Herbert McKennis, Jr., American Medical Association Workshop, Colorado,Springs, Colorado, Nov. 1-3, 1966. ~ a... r. n.. . Pharmacological action and intermediary role of 5-(3-pyridyl)tetrahydrofuranone- Edward R. Bowman and Pavel Itrdina. S1st Annual Meeting of the Federation of American Societies for Experimental Biology, ;-Chicago, Illinois, April 16-21, 1967. . Pharmacological effects of some nicotine metabolites and,related r-compounds. --K.--S. Kim and J. F. Borzelldca, S1st Annual Meeting of the Fed'eration of American Societies for Experimental Biology, Chicago, Illinois, April 16-21, 1967. 8._ PentafluropropionyLation in•the determination of nicotine. "`llerbert McKennis, Jr., S. C. Srivastava, and Edward R. Bowman. Annual Meeting of the Virginia Academy of Science, Roanoke, Virginia, May 6, 1967.