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TO THE EDITOR COMMUNICATIONS
ZYGADENUS ALKALOIDS. II. THE OCCURRENCE OF HYPOTENSIVE GERMINE ESTERS IN ZYGADENUS VENENOSUS
Vol. 74
acid determination, 17.52 mg. of neogermidine was equivalent to 5.77 ml. of 0.009126 N sodium thiosulfate; calcd. for germine monoacetate mono-aSir : methylbutyrate, 6.05 ml. Alkaline hydrolysis of A recent study' of the alkaloidal constituents of neogermidine afforded germine, acetic acid and aZygadenus venenosus revealed the occurrence in the methylbutyric acid. The acids were identified by plant of the esters veratroylzygadenine and vanil- conversion to their p-phenylphenacyl esters which loylzygadenine as well as the alkamines zygadenine were characterized after chromatographic separaand germine. We wish to report now the isolation tion. Methanolysis of neogermidine afforded profrom a batch of this plant collected in June, 1951 of toveratridine. The large change in rotation atthe germine esters neogermitrineJ2ge~midine,~ pro- tending the methanolysis of neogermidine to prot~veratridine,~ and a new diester isomeric with toveratridine suggests that the site of attachment of germidine, for which we propose the name neoger- the acetyl group on the alkamine germine is the midine. same as that of the labile acetyl group in both neoFractionation of the chloroform-extractable al- gerrnitrine and germitrine. kaloids of Zygadenus venenosus (WATS.)5 by 8Acetylation of neogermidine with acetic anhyplate countercurrent distribution using benzene and dride and pyridine yielded acetylneogermitrine, phosphate buffer at pH 7.1 yielded, in addition to 1n.p. 248-249' dec., identical with a sample prethe alkaloids obtained previously, the germine pared by acetylation of germidine.2 Acetylation monoester protoveratridine (from the 0-plate frac- of protoveratridine under the same conditions also tion), Protoveratridine crystallized as rectangular gave acetylneogermitrine. These facts show that plates upon reprecipitation from alcoholic acetic the site of attachment of the a-methylbutyryl acid solution with aqueous ammonia; m.p. 272- group is the same in each of the four germine esters 273' dec.; [a]2 2 -9' ~ (c 0.76, pyr.). Anal. Calcd. isolated from Zygadenus venenosus. C32H510BN: C, 64.73; H, 8.66; N, 2.36, Found: Pharmacological experiments carried out with C, 64.79; H, 8.62; N, 2.62. neogermidine a t the laboratory of Professor 0. The filtrates after removal of the crystalline al- Krayer a t Harvard Medical School indicate that kaloids from the plate-8 fraction were combined the circulatory action in the cat and the veratrinic with the material from plates 4 to 7, and the amor- effect on the frog muscle are similar to those of phous mixture obtained upon lyophilization was germidine. designated the "organophilic" fraction. The filThis work was supported (in part) by grants trates after removal of the crystalline alkaloids from the National Institutes of Health and Eli from the plate-0 fraction were combined with the Lilly and Company. The assistance of Eli Lilly material from plates 1 to 3, and the residue obtained and Company in gathering and extracting Zygadeupon lyophilization was designated the "hydro- nus venenosus'is gratefully 'acknowledged. philic" fraction. OF CHEMISTRY Neogermitrine and germidine were obtained by DEPARTMENT HARVARD UNIVERSITY S. MORRIS KUPCHAN 24-plate countercurrent distribution of the organo- CAMRRIDCE 38, MASSACHUSETTS C. V. DELIWALA philic fraction using benzene and 2M acetate bufRECEIVED MAY22, 1952 fer a t pH 5.5.2 The identity of these substances was confirmed by mixed melting point and infrared spectral comparisons with authentic specimens A NEW DISACCHARIDE PRODUCED BY from Veratrum viride kindly provided by Dr. J. LEUCONOSTOC MESENTEROIDES Fried. Sir : The hydrophilic fraction was subjected to 8-plate In studies on the enzymatic synthesis of dextran countercurrent distribution using chloroform and 2M acetate buffer a t pH 5.5. Neogermidine was from sucrose by Leuconostoc mesenteroides we have obtained by chromatography on alumina of the ma- noted that under certain conditions as much as 370 terial recovered from plates 5 to 8. Neogermidine of the sucrose is diverted to the production of a new which has the following properties: crystallized from benzene as heavy prisms; m.p. disaccharide Crystallizes in the form of bars from methanol221-223' dec.; [ C ~ ] ~-60' ~ D (C 2.00, PYr.); [ L Y ] ~ ~ D ethyl acetate; m.p. 161-163'; [ C Y ] * ~-8.8' D after -25' (c 2.00, chf.). Anal. Calcd. CS~HBOION:3 minutes, -6.8' after 24 hours (c, 4; H20); analC, (34.22; H, 8.38. Found: C, 64.15; H, 5.70. yzes for Cl~HzzOll; reducing power by Somogyi Neogermidine thiocyanate crystallized from ace- method 46y0 of that of fructose; low order of reactone as needles, m.p. 247-249' dec. Anal. Calcd. tion with hypoiodite (9% by Willsttitter-Schudel C34H530loN.HNCS: C, 60.50; H, 7.82; S,4.61. method; turanose gave 11%); positive Seliwanoff Found: C, 60.16; H, 7.87; S, 4.65. In a volatile test for fructose; hydrolyzes to glucose and fructose as shown by paper chromatography; phenyl(1) S. M. Kupchan and C. V. Deliwala, THISJ O U R N A L , 74, 2382 iI!l.52). osazone (analyzing for &HsNdOe) as needles from ( 2 ) J. Fried, P. Numerof and N . M. C o y , ibid., 74, 3041 (1952). We wet ethyl acetate (m.p. 186-188') ; yields an amorwish t o thank Dr. J. Fried for private communication of these results phous phenylosotriazole which hydrolyzes to Dprior to publication. glucose ( [ c Y ] ~ ~+52') D and D-glucose phenylosotri( 3 ) J Pried. H. L. Whitcand 0. Wiuterrteiner, ;bid.,?%, 4621 (1950). ( 4 ) G. Salzberger, Arch. Phorm., 298, 462 (1890); W. Poethke. {bid., azole (m.p. 197'; [ a I z 5 D -81'). 275, 571 (1937). From this preliminary work it would appear (5) Plant gathered in northeastern Oregon in Juuc, 1951. We are that the sugar is a D-glUCOSyl-D-frUCtOSe with the grateful to Dr. Reed Kollius, Gray Herbarium. IInrvard University, for glucosyl unit probably in the pyranose form since confirming the identity o f the planl.
