COMMUNICATIONS TO THE EDITOR
Aug., 1948
out by the fact that a simple visual examination of iodine solutions of equal concentration in benzene, toluene, o-xylene, mesitylene and amethylnaphthalene shows that the color shifts stepwise in that order ending with a brown solution. Preliminary measurements of the absorption spectra of these iodine solutions show an absorption band in the ultraviolet region similar to that of benzene. This work is being continued and a complete report of the results will be given in a paper soon to be submitted for publication. I
2833
THE CRYSTALLINE TRIHYDROCHLORIDES OF STREPTOMYCIN AND MANNOSIDOSTREPTOMYCIN
Sir:
The preparation of the crystalline reineckate, sulfate,' helianthate,2 and the calcium chloride double salt2of streptomycin and the reineckate of mannosidostreptomycina has been reported. To date, there has been no published information on the crystallization of a simple mineral acid salt of either of these antibiotics. We now wish to report that, starting with relatively pure material, we have obtained the trihydrochlorides of streptoDEPARTMENT OF CHEMISTRY HANSA. BENESI mycin and mannosidostreptomycin in the crystalUNIVERSITY OF CALIFORNIA JOELH. HILDEBRAND line state from methanol solution. BERKELEY 4, CALIFORNIA The streptomycin trihydrachloride crystallizes RECEIVED JULY15, 1948 with two molecules of water of crystallization as monoclinic prisms showing birefringence. The crystalline material was shown to be a single sub8-PELTATIN, A NEW COMPONENT OF stance by ,a modification of the Craig counterPODOPHYLLIN current distribution technique' and thus to be free Sir : of mannosidostreptomycin. On heating on the The fractionation of the drug podophyllin by hot-stage, the dihydrate decomposes gradually chromatographic adsorption on alumina has without melting. When the trihydrochloride was yielded, beside podophyllotoxin and a-peltatin,l dried a t 55' in vacuo, it had the following analytia new crystalline substance in about 4% yield for cal composition: c , 34.86; H, 6.36; C1, 14.25 which the name P-peltatin is proposed. The new (Calcd. for CzlH39N7012.3HC1.2Ha0 : C, 34.54; compound possesses about the same high necro- H, 6.36; C1, 14.57). After drying a t 100' in tizing activity2 for mouse sarcoma 37 as a-pelta- vacuo, the anhydrous material showed [a tin. -236.1' (1.Oyoin water) and the following analyti&Peltatin crystallizes from alcohol in colorless, cal data were obtained: c, 36.27; H, 6.14; transparent prisms, m. p. 231.1-238.0' (shrinks a t N, 14.29; C1, 15.69 (Calcd. for G1H39N7012.3HC1; 225.5') cor.; [a]% -115' (c, 1.009, absolute al- C, 30.50; H, 6.13; N, 14.19; C1, 15.40). : 63.75; cohol). Anal.a Calcd. for C t ~ H ~ 0 8C, When assayed with K. pneumoniae in a brothH, 5.35. Found: C, 64.0; H, 5.0. Calcd. for dilution test,6the trihydrochloride dihydrate had a three methoxyl groups: 22.5; found, 22.2. Molec- potency of 820 units/mg. and on this basis the ular weight values (Rast) for derivatives of both anhydrous material would have an activity of 891 a- and @-peltatinagree with the formula CnZHnOs units/mg.6 and indicate that the peltatins are thus isomeric The trihydrochloride of mannosidostreptomycin with podophyll~toxin.~a-Peltatin has one less crystallizes in the form of hexagonal plates which methoxyl group than 8-peltatin and podophyllo- are isotropic. By means of the counter-current toxin. distribution m e t h ~ dthis , ~ material was also shown Beside the methoxyl content, a- and p-peltatin to be a single entity and t o be free of streptodiffer in their color reactions with sulfuric acid and mycin. in the properties of their derivatives. With conAfter drying a t 55' in vacuo, the trihydrochlocentrated sulfuric aad, both peltatins give an im- ride was found to have the following analysis: mediate yellow color, rapidly turning reddish C, 36.45; H, 6.26; C1, 12.14 (Calcd. for C27H49brown with a-peltatin and green with 8-peltatin; N701.1.3HC1.2HzO: C, 36.47; R,6.35; C1, 11.96). the final color with both peltatins is red. A series When dried a t 100' in vacuo, the anhydrous maof derivatives of the peltatins has been prepared terial showed [C~-]"*~D-54.1' (1.0% in water) and will be reported at a later date. (1) J. Fried and 0. Wintersteiner, Science, 104, 273 (1946). Structural and biological studies with @-peltatin (2) R. L. Peck, N . 0. Brink, F. A. Kuehi, Jr., E. H. Flynn, A. are in progress. Walti and K. Folkers, THIS JOURNAL, 67, 1866 (1945). J Z 6 s 6 ~
NATIONAL CANCER INSTITUTE NATIONAL INSTITUTE OF HEALTH U. S. PUBLIC HEALTH SERVICE JONATHAN L. HARTWELL BETHESDA, MARYLAND WENDELL E. DETTY RECEIVED JULY22, 1948 (1) J. L. Haftwell, TEIS JOURNAL, 69, 2918 (1947). (2) Unpublished results of Joseph Leiter and Faith Jouvenal.
(3) By Mrs. M. M. Ledyard and Mrs. Evelyn Peake National Institute of Health. (4) W. Bocsche and J . Niemann, Bcr., 61,1633 (1932); E.Spiith, F. Wessely and E. Nadler, ibid., 66, 125 (1933).
(3) J. Fried and E. Titus, J . B i d . Chem., 168, 391 (1947). (4) A modification of the counter-current distribution described by Titus and Fried ( J . B i d . Chem., 174, 57 (1948)) has been deveioped by our colleagues Drs. Flaut and McCormack which eliminates the appearance of the tautomers of the two streptomycins in the Craig diagram. (5) R. Donovick, D. Hamrc, F. Kavanagh and G.Rake,J . Boct., 10, 623 (1945). (6) Based 09 the F. D. A. working standard. Spectrophotochemical asgays based on a maltol method, similar to that published by G. F. hfueller (THIS JOWRNAL, SO, 195 (1947)), have confirmed these microbiological results.
COMMUNICATIONS TO THE EDITOR
2834
Vol. 70
of such a compound would lead to compounds of the structure (111), which type of structure has recently been proposed for ketoyobyrine, on the basis of an exhaustive study of its chemistry% and likewise on the basis of a comparison of its absorption speetnun with that of rutaearpine.2b Pending more complete presentation of our various syntheses of the type of structure represented by (111),we wish to record our synthetic confirmation of this proposed structure for ketoyobyrine. 6-Methylhomophthalic acid, m. p. 1W0, was prepared from 0-tolylacetic acid* by conversion Via the Arndt-Eistert reaction into o-tolylpropiDIVISION OF Cmaaxw DEVELOPMENT LEONJ. HEUSER onic acid, which was then treated according to the method of Mercer and Robertson.' CondensaE. R. &UIBB AND SONS MORRISA. DOLLN. J. ERICT. STILLERtion with tryptamine yielded N-(@-indolrlethyl)NEWBRUNSWICK, 6-methylhomophtfialimide, m. p. 228 ConRECEIVED JULY19, 1948 version of the latter into the corresponding homophthalamic acid,' m. p. of picrate 147O, THE SYNTHESIS OF KBTOYOBYRINE methylation of the acid with diazomethane, m. p. sir: of methyl ester 222O, dec., followed by ring For several years we have been investigating closure with phosphorus oxychloride, yielded the possibility of synthesizing the basic ring struc- ketoyobyrine, m. p. 3 1 6 - 3 1 8 O , dec. ture of yohimbine by ring closure of isoquinolylA d . Calcd. for C ~ ~ H ~ ( I OC,N ~79.98; : H, ethyl oxindoles of the type represented by formula 5.37; N, 9.32. Found: C, 79.43; H, 5.55; N, (I).l The introduction of the double bond in ring 9.24. Comparisons of the ultraviolet absorption D presented numerous f i c u l t i e s because of the spectrum of synthetic ketoyobyrine with that of ease with which compounds of the type (I) (as the product of natural origin showed the two to well as the yohimbine molecule itself) suffer be identical. Absorption maxima for synthetic cleavage at the nitrogen atom of ring D. More- material: at 385,366 and 340 mp, log z 4.40,4.51 over, we have pointed out* that 1,2-dihydroiso- and 4.52, respectively. quinolines like (I) are virtually unknown. (2) (a) Woodward and Witlcop, T a ~ JOURNAL, s TO, 2409 (1948); Accordingly, our efforts were later directed (b) Raymond-Hamet, Compt. r e d . , 116, 137 (1848). toward the preparation and ring closure of com(3) Julian, Karpel, Magnani and Meyer, Tms JOURNAL, TO, 180 pounds of the type represented by formula (11) (1948). (4) M u m and Robertson, J . Ckm. Soc., 288 (1938). H H
and gave analytical data: C, 38.25; H, 6.22; N, 11.31; C1, 12.52 (Calcd. for CorH&017*3HCl: C, 38.01; H, 6.14; N, 11.49; C1, 12.47). When assayed with K. preumoniae in a broth dilution test,' the anhydrous mannosidostreptomycin had a potency of circa 210 units/mg.* Additional information on the properties and activities of these crystalline hydrochlorides will be published a t a later date. We wish to express our appreciation to Dr. R. Donovick, Mr. R. Blue, and Mr. D. Lapedes for the bio-assays, Mr. F. Rum-Alesi for the countercurrent distributions, and Mr. J. Alicino for the micro-analysis.
.
(5)
Cf.Haworth, Perkin and Pink, J. Ckm. Soc., 1709 (1926).
'MEGLIDDEN COMPANY PERCY L. JULIAN RESEARCH LABORATORIES WILW J. ICABPBL SOYA PRODUCTS DIVISION ARTHUR MAGNANI CHICAGO,ILLINOIS EDWINW. MEYBR RECEIVED JULY 24,1948
YJ
R/' (I)
(11) (R = CHs or H)
(R = CHI or H)
N
R
(111) Ketoyobyrine (R
-
CHI)
where the appropriate double bond of ring D could be introduced without difficulty. Ring closure (1) Julian, Magnani, PiLl and Korpel, THISJOURNAL, TO, 174 (1948).
THE OZONIZATION OF THE MALEIC ANHYDRIDE ADDUCT OF DEHYDROERGOSTERYL ACETATE
sir: A recent publication of Bergmann and Stevens' describes the preparation of the maleic anhydride adduct of 3(~)-acetoxy-9,11-oxidobisnor-5,7choladienic acid (11) by the ozonization of 9,lloxidoergosteryl acetate-maleic anhydride adduct. For some time previous a study of analogous reactions has been under way in our laboratories and we now wish to report the preparation of the maleic anhydride adduct of 3(/3)-acetoxybisnor5,7,9-cholatrienic acid (I) by the selective ozonization of 9,ll-dehydroergosteryl acetate-maleic anhydride adduct (111). A solution of the dehydroadduct (111) in methylene chloride was treated with two equiva(1) Bergmann and Stevens, J . Org. C k m . , 18, 10 (1948).
