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 crystalOF CALIFORNIA UNIVERSITY 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 MARYLAND WENDELL E. DETTY BETHESDA, 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.
