Dec. 5, 1962
COMMUNICATIONS TO THE EDITOR
4507
dence supports the interpretation that the broadT~ in terms of p1and 0, the half width a t half height of the broadened line. ening is due to exchange. 1/71 = 8 - 81 (4) (8) Alfred P. Sloan Foundation Fellow, 1960-1964. Application.-Raman spectra of sodium trifluoro- DEPARTMENT OF CHEMISTRY acetate ion in the presence of excess sodium hy- UNIVERSITY OF MINKESOTA MAURICE M. KREEVOY~ c. ALDEN MEAD 14, MINNESOTA droxide show an intense peak a t 1433 cm.-l (prob- MINNEAPOLIS RECEIVED SEPTEMBER 4, 1962 ably the symmetrical C=O stretching f r e q ~ e n c y ) ~ with a width a t half height of 15.0 an.-'. In the presence of excess base the width is independent GLYCEROL AS A PRECURSOR OF RICININE' of trifluoroacetate concentration. Trifluoroacetate Sir: also has a fairly intense peak a t 843 cm.-', with By means of tracer experiments with young a width of 16.2 cm.-'. Trifluoroacetic acid in the presence of excess HC1 gives a strong peak a t 816 Ricinus communis L. we have recently established2s3 cm.-' with a width of 28.8 cm.-l. The latter two that succinic acid, or a closely related four-carbon are presumably the C-C stretching frequencies.j dicarboxylic acid found in the Krebs tricarboxylic I n solutions containing trifluoroacetic acid, its acid cycle, is a specific precursor of carbon atoms 2, conjugate base, and hydronium ion, the trifluoro- 3, and 7 in the biosynthesis of ricinine I. Since an origin outside of the Krebs cycle was acetate peak a t 1433 ern.-' is broadened by as much as 7.2 cm.-', and the two peaks around 830 indicated for carbon atoms 4,5, and 6, g l y ~ e r o l - l - ~ ~ C cm.-l both broaden and tend to merge. Both and glycer01-2-'~C were fed in separate experithe 1433 cm.-l peak and the 816 cm.-' peak also ments to young Ricinus plants. The percentages tend to shift toward higher frequency by 2-5 of the incorporated radioactivity located a t the 0cm. in mixed solutions. Spectra were measured methyl, N-methyl, and nitrile carbon atoms of the on a Cary model 81 photoelectric Raman spectro- isolated ricinine have been reported already.2 We now wish to describe degradative procedures photometer, equipped with two Toronto-type mercury arc lamps. Solution temperatures were to isolate the carbon atoms a t positions 4, 5, and around 40'. The lines are roughly Lorentzian in 6 of the ricinine molecule, and applications of these procedures to the ricinine obtained from both shape. C glycerol-2-14C, as well as, in The broadening of the 1433 cm.-l peak was g l y ~ e r o l - l - * ~and used to obtain k1 for trifluoroacetate ion by means part, to ricinine from the previously reported sucof eq. (4). These results are shown in Table I. cinic acid-2,3-I4C and sodium acetate-2-I4Cfeeding Solutions were made up simply by mixing tri- experiments. Ricinine I was converted to 4-methoxy- l-methylfluoroacetic acid and water. Concentrations were obtained by interpolation, using the degree of dis- 2-pyridone 11, m.p. 114-116°,4 which was reduced lithium aluminum hydride to l-methyl-4sociation results of Redlich and c o - ~ o r k e r s . ~ ~with ' Concentrations could also be inferred from the in- piperidone 111, b.p. 55-60' (12 mm.),5 identified tensity of the 1433 cm.-' line in the Raman spectra. by comparison with an authentic sample. ReacThe average difference between the two estimates tion of I11 with phenyllithium provided the known IV, m.p. of the trifluoroacetate concentration was 10%. 4-hydroxy- 1-methyl-4-phen~lpiperidine~ This strongly suggests that the intensity of the 115-116', which was oxidized with potassium per1433 cm.-l band is proportional to the trifluoro- manganate to benzoic acid V, m.p. 121-122', with acetate concentration and otherwise independent the carboxyl carbon atom originating from carbon of the composition of the solution. Table I also 4 of ricinine. Nitration of ricinine I afforded the 5-nitro derivgives the rate constant k-l for the conversion of trifluoroacetic acid, as determined from the re- ative VI, m.p. 163-164' (anal. Calcd. for CsH704N3: C, 45.94; H, 3.37; N, 20.09. Found: C, quirement that equilibrium be maintained. 45.94; H, 3.39; N, 20.19) which on treatment TABLE I with calcium hypobromite yielded tribromonitroAVERAGE LIFETIME OF TRIFLUOROACETATE IONIN VARIOUS methane (bromopicrin) VII, with the carbon atom SOLUTIONS originating from position 5 of ricinine. The bromoMolar 10-11 ki 10-11 k-1 picrin was identified by comparison with an CFaCOzCFsC02H Hf sec.-1 sec. -1 authentic sample prepared from picric acid.7 1.8 3.5 1.8 6.9 3.5 Catalytic reduction of the pyridone I1 yielded 1.5 1.5 1.5 4.7 4.7 1-methyl-2-piperidone VIII, b.p. 110-105° (12 mm.) 0.81 0.24 0.81 1.6 5.4 (mercuric chloride derivative, m.p. 117-120°8) which was hydrolyzed with hydrochloric acid, The rate constants cited above were used to (1) N.R.C. No. 7107. calculate the shape of the doublet centering around (2) P. F. Juby and Leo Marion, Biochem. Bioghys. Res. Comm.. 6, 830 cm.-l. The general shape of the band is 461 (1961). moderately well reproduced. The band shape ( 3 ) P . F. Juby and Leo Marion, Can. J . Chem., in the press. (4) E . Winterstein. J. Keller and A. B. Weinhagen, Arch. Pharm., calculated by adding the unbroadened lines is 513 (1917). worse, but the difference is not great. This evi- ass, (5) R . E. Lyle, R . E. Adel and G. G. Lyle, J . Org. Chcm., 14, 342 ~
~~~~
7
(5) R . E . Robinson and R . C. Taylor, Sgectrochimico A d a , 18, 1093 (1962). (6) 0. Redlich and G . C. Hood, Discussions Faraday Soc., No. 2 4 , 8 7 (1957). (7) G. C. Hood, 0. Redlich and C. A. Reilly, J . Chem. P h y s . , as, 2229 (19.5.5).
(1959). (6) Aktieselskabet "Ferrosan," Danish Patent 60,592 (1943) : Chem. A b s . , 40, 408@ (1946). (7) J. Baddilep, G. Ehrensvlrd, E. Klein, L. Reio and E . Salriste, J . Biol Chcm., 183, 777 (1950). (8) C . R l t h , A n n . , 489, 107 (1931).
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COMMUNICATIONS TO T H E E D I T O R CH.0
0
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feeding acetate-2-14C and succinic acid-2,3-I4C. From results reported previously2 it is clear that a t least part of the glycerol is broken down to siinplcr units prior to incorporation. However, the fact that carbon 5 of ricinine from the glycer01-2-'~C experiment contains an appreciable percentage of the incorporated activity while carbon 4 and carbon 6 contain little or no activity suggests that part of the glycerol was incorporated intact into ricinine a t positions 4, 5 , and 6. The high activity a t carbon 4 and carbon 6, and low activity a t carbon 5 after the feeding of g l y ~ e r o l - l - ~lends ~ C support to this suggestion. (12) National Research Council of Canada Postdoctorate Fellow (13) American Chemical Society Petroleum Research Fellow.
DIVISION OF PURECHEMISTRY NATIONAL RESEARCHCOUNCIL OTTAWA, CANADA
J. E S S E R Y ~ ~ P. F. JUBY'* LBO MARION E. TRUMBULL'~
OCTOBER20, 1962 RECEIVED THE CORRECTED STRUCTURE OF A KETOAMIDE ARISING FROM THE METABOLISM OF (-)-NICOTINE
Sir:
(CH,),N(CH,),C0,CH3
Ix
1,
2,
CH,=CH(CH,),CO,CH,
HLOL HEAT
X OSO,/NaIO,
HCHO
XI methylated with formic acid and formaldehyde and esterified with diazomethane to methyl 5-dimethylaminovalerate IX, b.p. 90' (20 m ~ n . ) character,~ ized as its methiodide (anal. Calcd. for CgH2o02NI : C, 35.88; H, 6.69; N, 4.65; I, 42.13. Found: C, 35.89; HI 6.81; N, 4.77; I, 42.02). The N-oxide of IX when pyrolyzed yielded the methyl ester of allylacetic acid X , identified by comparison of its infrared spectrum with that of an authentic synthetic sample, b.p. 127-128' ( i G 0 mm.) (anal. Calcd. for CsHloOz: C, 63.13; H, 8.83. Found: C, 63.3; H, 8.86). The olefin was cleaved using sodium periodate and osmium tetroxide'O and the liberated formaldehyde XI, containing carbon 6 of ricinine, isolated as the dimedon derivative, m.p. 191-192'.11 The results obtained with the use of the procedures outlined above are recorded in Table I. TABLEI LOCATION OF RADIOACTIVITY IN RICININE of activity of ricinine
Peecursor
c-4
Acetate-2-I4C Succinic acid-2,3-I4C Glycerol-l-~4C G l y c ~ o l - 2"-C
0.0 0.0 25.8 2.2
C-5
0.3 0.0
2.2 38.8
C-6
Not datd. Not detd. 19.7 0.0
As expected3 there is virtually no activity associated with carbons 4 and 5 of ricinine after (9) R. Willstatttr and W. Kahn, B o . , 31, 1853 (1904). (lo) R. Pappo, D. S. Allen, R. U. Lemieux and W. S. Johnsun, J . OTE. Chem., 21, 478 (1956). (11) R. E. Reeves, J. A m . Chem. Soc., 63, 1476 (1941).
During the course of studies' on the metabolism of (-)-nicotine, a ketonic metabolite CloHl&TsOa was isolated from the urine of dogs, and later from r a t s 2 This compound, which has amphoteric properties, as exhibited in its behavior toward ion exchange resins, was ostensibly obtained in pure form and then hydrolyzed to a keto acid C9H9N03 and methylamine. The oxime of the keto acid then mas subjected to a Beckmann rearrangement. The isolation of 3-pyridylacetic acid from the reaction products of this rearrangement was a crucial point in the conclusion' that the ketoamide had the structure of y- (3-pyridyl)-Poxo-N-methylbutyramide. On the basis of current data, which include both a degradation and a synthesis, the foregoing structure which arose through technical errors of a yet undetermined nature is incorrect. Since the ketoamide, now shown to be r-(3-pyridyl)-y-oxo-N-methylbutyramide, appears to occupy a key role as an intermediate in the mammalian degradation of the pyrrolidine ring of (-)-nicotine to y-(3-pyridyl)-yoxobutyric acid,a y-(3-pyridyl)-y-hydroxybutyric acid,4 3-pyridylacetic acid,Kand other substances, a summary of data necessary to the immediate correction of the erroneous formula is presented. By a procedure patterned after the original,' the ketoamide m-as isolated from the urine of dogs after oral administration of (-)-cotinine. The colorless crystals, R, 0 74 upon paper chromatography in the ammonia were obtained (1) H. McKennis, Jr., E. R . Bowman and L. B. Turnbul1,J. A m . Chem. SOC.,82, 3974 (1960). (2) H. McKennis, Jr., L. B. Turnbull, S. I,. Schwartz. E . Tamilki and E. R . Bowman, J . B i d . Chcm., 237, 5 4 1 (1962). (3) S. I*. Schwartz and H. McKennis, Jr., Fedcralioa Proc.. 21, lS:? (1962) (4) L. B. Turnbull, E. R . Bowman and 11. McKennis, Jr., ihid., 17, 325 (1958). (5) H. McKennis, Jr., E. R. Bowman and I,. U. Tnruhull, Proc. SOC.Ex#. B i d . Med., 101, 145 (1961). (A) H. McKennis, Jr., L. B. Turnbull, H. N. Wingfield, Jr., ilud L. J. Dewey, J . A n i . L I M N I . SOC.,80, 1634 (1958).
