July 20, 19*5*5
COXMUNICATIONS TO THE EDITOR
oxidation of cholesterol. None of the products administered as aqueous colloids have had a carcinogenic effect, although a number of them administered in sesame oil have produced positive results, indicating t h a t the presence of sesame oil is an essential link in the carcinogenic chain.6 As recently reported,' cholesterol a-oxide, A*-cholestene:I,O-dione, arid the 1: 1 complex5 of 6/3-hydroxy-A4cholestene-3-one and epicholesterol when injected into mice in sesame oil produced fibrosarcomas in 19-43y0 of the mice at the average age of 13-14 months. These compounds produced negative results when administered as aqueous colloids. We wish now t o report discovery of a still more potently carcinogenic oxidation product. AS-Cholestene-%one, an easily isomerized and highly reactive substance t h a t is oxidized by lead tetraacetate to the doc-acetoxy derivative,8 combines with molecular ovygen in hexane solution at 25' to give a compound t h a t crystallizes from methanol in needles, m.p. 180")XEtoH 236 mp ( E = 16,SSO). Anal. Calcd. for C27H4403 (416.62) : C, 77.83: H, 10.65. Found: C, 77.52: H, 10.72. The substance gives an immediate color with sodium iodideacetic acid (thiosulfate titer: mol. wt. 425) and is (111), reduced to B~-hydroxy-A4-cholestene-3-one
I
I1
I
dants may increase resistance to spontaneous carcinogenesis is under investigation. CHEMICAL LABORATORY OF HARVARD LOUISF. FIESER UNIVERSITY, CAMBRIDGE, MASS. THEODORA W. GREENE SANTABARBARA COTTAGE HOSPITAL FRITZ BISCHOFP GUILLERMO LOPEZ RESEARCH INSTITUTE, SANTA BARBARA, CALIFORNIA J. J. RUPP RECEIVED J U N E24, 1955 POLARIZATION OF T H E 1850
BAND OF AMIDES'
We have recently carried out a measurement of the direction of the transition moment vector for the first N,V transition in amides.2 Observations were made on sublimation flakes of myristamide3 (ClsH&ONH2) with light polarized along the a and b crystallographic axes, which are also the principal directions, in the region between 2300 and 1600 A. The orientation of samples in these experiments was obtained crystallographically with the aid of X-ray precession photographs. The ratio of the optical densities observed along the principal directions over the long-wave-length one-third of the main band was found to be constant a t D a l D b = 14.62. This gives for the orientation of the electric moment 0 = f arctan 0.262 where 0 is measured from the a axis. The two orientations referred to the amide group itself were computed from the known crystal structure of myristamide3 and are 9.1"and 26.7' from the line joining the nitrogen and oxygen (see Fig. 1).
/
I11
+
A.
Sir:
OH
OOH
3929
C\
m.p. 193", U D 27.2" Chf, mixed m.p. 193-194". Since the ultraviolet absorption corresponds to that of 111, and since reduction of hydroperoxides is known to cleave the oxygen-oxygen bond, the substance is identified as 6/3-hydroperoxy-A4-cholesFig. 1.-Possible orientations of the electric vector for the tene-3-one (11). 1850 A. electronic absorption band in amides. The hydroperoxide I1 was given in three spaced subcutaneous injections of 5 mg. each in sesame oil Keither of these values agrees with conclusions to each of 32 Marsh-Buffalo mice.g At the age of from recent experiment4 in which absorption a t 12 months, fibrosarcomas have appeared at the site 2100 A. was found to occur essentially along one of of injection in 13 of the mice treated (average tu- the principal directions (also here the c crystallomor age 9.6 mo.), and 17 of the remaining mice were graphic axis) of crystalline N-acetylglycine. I n still alive. hTo fibrosarcomas were observed in lit- that work i t was inferred that absorption must be ter-mates given the same amount of I1 in aqueous nearly along the carbon-nitrogen line of the amide colloidal solution, nor in a series of controls (up to group, a line which is only four degrees from one of 18 months of age) which received only sesame the principal directions. Possibly what was actuoil. ally measured was mainly a single allowed crystal Indirect evidence t h a t the precursor I can be transition occupying a small region at the extreme formed from cholesterol in the body is afforded by long-wave-length edge of the main absorption, Such isolation of probable transformation products : absorption would have to be along one or the other A4-cholestene-3-one,10 A4~6-cholestadiene-3-one11principal direction. Indeed we have found absorp(swine spleen, arteriosclerotic aorta), and coprosta- tion in the region from 2120 to 2250 A. a t the longnonelZ (ambergris). The possibility that antioxi- wave-length edge of the band for myristamide ( 6 ) F. Bischoff, G. Lopez and J . J . R u p p , Abst. Am. Chem. Sac., which is, within experimental error, exactly along March 3 - C (1954). the a axis, a principal direction. I n addition, we ( 7 ) F Bischoff, G . Lopez, J , J . R u p p and C L . Gray, Federation
X A
0
Proc., 14, 183 (1955). (8) L. F . Fieser and R. Stevenson, THISJ O U R N A L , 76, 1728 (19.54). (9) Supported in part by grant C 1586 C from t h e National Cancer Institute, National Institutes of Health, PHS. (10) V. Prelog, et al., Helu. Chim. Acta, SO, 1080 (1947). (11) E. Hardegger, L. Ruzicka and E. Tagmann, ibid., 26, 2205 (1943). (12) E.Lederer, e l al., ibid., 29, 1354 (1946).
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(1) Supported in part b y t h e Air Research and Development Command, Contract No. 4 F lS(600)-375. (2) J. S. H a m and J . R. P l a t t , J. Chcm. P h y s . , 20, 335 (1952): and H . D . H u n t and W. T. Simpson, THISJOURNAL, 7 S , 4540 (1953). (3) T h e crystal structure of myristamide was determined by J . D. Turner, Ph.D. Thesis, University of Washington, 19.53, and refined by R . F. Adamsky, private communication. (4) J, C. Ward, Proc. Roy. Soc. (London), A938,205 (1955).
COMMUNICATIONS TO THE EDITOR
3930
Vol. 77
have made theoretical calculations of the l / R 3 force modifications of the spectrum to be expected for myristamide (using both of the angles given it1 Fig. 1). These calculations support the interpretation of the 2120-2250 A. absorption as a crystal transition, in that the long-wave-length allowed component is predicted to occur in the a axis direction, split off by ca. 100 to 150 A., just as observed (and regardless of which angle is chosen). We have also made quantum-theoretical calculations of the direction of the transition moment for an amide model. I n the model an amide is regarded as a perturbed allyl anion5 with the nitrogen represented by a deepening of the coulomb potential on one of the end carbon atoms and the oxygen considered as being the same as carbon. The result is that the direction should be close to the nitrogen-oxygen line, but inclined toward the carbon nitrogen line. Of the two values fouiid in our experiments the one with an angle of 9.1' from the nitrogen-oxygen line is thus favored. Both the experimental and theoretical aspects of this investigation will be reported in detail in a forthcoming paper.
sive confirmation obtained through an 1I-methoxyalloyohimbane (IIa, R = R' = H ) (reserpane) synthesis which is stereochemically unambiguous insofar as the CI6-Cznasymmetric centers are con~erned.~ 6-Methoxytryptaminee (obtained through the route : 6-methoxyindole -t 6-methoxygramine methosulfate -t 6-methoxyindoleacetonitrile -+ 6-methoxytryptamine) was alkylated in boiling dimethylformamide by ethyl dl-cis-2-bromomethylcyclohexaneacetate,'~~ affording the lactam of dZ-cis-N-(P-3'indolylethyl) -2-aminomethylcyclohexaneacetic acid (111) [benzene solvate), m.p. 72.5-74.0" (Calcd. for CnoHa6Sz02.C6H6: C, 77.19; H, 7.97. Found: C, 77.02; H, 7.93). Heating I11 with phosphorus oxychloride in benzene, followed by platinum-catalyzed reduction of the unisolated A"ring-closed product, yielded the desired til-all0 base IIa, which melted a t 209-210" after crystallization from methanol. Infrared spectral comparison of chloroforni solutions of IIa and a reserpane derived by reduction from reserpone (IIb, IC, R ' = 0)9 showed the two substances to be, apart from the racemic nature of the former, identical.'" DEPARTMEKT OF CHEMISTRY D. L. PE-I.ERSOX~ Publication of our views on the nature of the remaining asymmetric centers in reserpine and deOF WASHISGTOS TI., T. Srhwsos UNIVERSITY SEATTLE, WASHINGTOS serpidine, including evidence relating thereto, is RECEIVED J U T E 1.3, 1955 :i 11 ticipatcd. 15) Calculation waF facilitated by use of full conlixuration interacAcknowledgment.--The authors wish to express tion A.S.IVf.0. calculations on allyl anion (H. D H u n t , D L Peterson their gratitude t o the Department of Health, Weland Tir. T. Simpson, t o be published) fare and Education for financial support (Grant NO. (6) National Science Foundation Predoctoral Fellow 19.54-195.5. G-3S92'! and t o S. R.Penick and Co. for a gift of reserpine. ( 5 ) T h e matter of t h e stereochemistry a t C Ain o m synthetic prodiict
T H E D/E CIS RING JUNCTURE O F RESERPINE
Sir: Reserpine, the potent hypotensive agent obtained from various species of Rauwolfia, has been Because of its proxiassigned the structure I
is deferred for t h e present. (0) S.Akabori and K. Saito, B e y . , 6SB, 2245 (1930).
( 7 ) G . Stork and R . Hill, THISJOURNAL, 76, 949 (1954). ( 8 ) Unpiiblished results obtained in this Laboratory. (9) C . P. Iluehner, H . B. hIacPhillamy, A . F. S t . Snrlr6 and E. Schlittler, Tins J O ~ J R N A I . ,7 7 , 473 (195.5). ( I O ) n ' c should like t u thank Dr. Schlittler and Dr. St. Andr6 for their n;siitance in establishing t h e identity of t h e two specimens. Tn addition, o u r base Ita was shown t o be identical with material obtained b y them via a different synthetic route.
DEPARTMEXT OF CHEMISTRY EUGENE E. VAS TAMELEN USIVERSITYOF WISCOXSIS PAULD. HANCE MADISON, WISCOSSIS KENNETH V. SIEBRASSE PAULE. ALDRICH RECEIVED MAY3, 1955 OCH3
I
R
R'
TIa, b
POLY-GLUTAMYL PTERIDINE COENZYMES
.%: It was recently demonstrated that the conversion mate relation to deserpidine (formula I with the 11methoxyl replaced by - H ) , Schlittler, et al., sur- of serine to glycine by a bacterial extract is demised that reserpine too possesses a D/E cis (allo) pendent upon DPX, Mn++, pyridoxal phosphate, ring juncture3; and Wintersteiner, et u Z . , ~ on the orthophosphate and catalytic levels of a new coenbasis of the intramolecular N-4 quaternization of zyme, Co C.1,2 Co C is isolated from Clostridium methyl reserpate tosylate, inferred the same rela- cylindrosporum, and substitutes for b u t is not identionship. We wish to report for this view conclu- tical with known folic acid derivatives. By means of fractional acetone precipitation, chromatography (1) L. Dorfman, A . Furlenmeier, C. F. Huebner, R . Lncas, H . B. on cellulose columns, and repeated paper chromaMacPhillamy, J. hl. Muller, E. Schlittler, R. Schwyzer and A. F. tography in various solvent systems,3five groups of St. Andre, H d v . Chim. Acta, 37,59 (1954). (2) N. Neuss, H . E. Boaz and J. W. Forbes, THISJOURNAL,76, pteridine derivatives with Co C activity have been 2463 (1954). separated in relatively pure form from extracts of
(3) H . B. MacPhillamy, L. Dorfman, C. F. Huebner, E. Schlittler and A. F. St. Andre, ibid.,77,1071 (1955). (4) P. A. Diassi, F. I.. Weisenborn, C . M. Dylion and 0. Wintersteiner, ibid., 77, 2028 (1955).
(1) B. E. Wright, Biochini el Biophys. Acta, 16, 165 (1955) ( 2 ) B. E . Wright, F e d . PTOC., 14, 308 (1955). (3) B. E ITright and E. R. Stadtman, unpublished data.
