COMM~JNICATIONS TO THE EDITOR
Nov., 1949 CH3 AcO
CHa
I
C=O
0
I
c=o
1. Na-Alcohol ______, 2. CrOa-HOAc ,
I1
1 2 ( 4-Acetoxyprogesterone
allo-Pregnan3,12,20-trione
CH3 I
HO
C=O
0
12(a)-Hydroxypregnan3,20-dione
CH3 I
)yYc=o ) 0
Pregnan3,12,20-trione
3857
powerful analgesic surpassing morphine in clinical tests2. 1,2,7-Trimethoxynaphthalene,m.p. 38.5-39.5 ', b.p. 133' a t 1 mm. (picrate3, m.p. 113')) gave by reduction* with sodium and alcohol, the crystalline ketone, m.p. 76' (anal. calcd. for CloHgO(OCH&: OCH,, 30.1. Found: OCH3, 29.5, 29.3) , characterized as the semicarbazone, m.p. 191-191.5', and the 2,4-dinitrophenylhydrazonej m.p. 167' dec. (anal. calcd. for C18H1808N4: C, 56.0; H, 4.7; N, 14.5. Found: C, 55.7; H, 4.6; N, 15.0, 14.8). The structure of the ketone was shown by oxidation, with alkaline permanganate, to hemipinic acid, identified by its m.p.6 (177179') and by the m.p.6 (166-167') and characteristic fluorescences of the pure anhydride. 2,7-Dimethoxynaphthalenesimilarly4 gave on reduction 7-methoxy-2-tetralone, m.p. 27-28', b.p. 124-126' (1.5 mm.); semicarbazone, m.p. 174-176' (anal. calcd. for C12H1,O2N3: C, 61.8; H, 6.5. Found: C, 62.1, 62.1; H, 6.4, 6.4); 2,4-dinitrophenylhydrazonem.p. 177-181' (anal. calcd. for C17H16N406: C, 57.3; H, 4.5. Found: C, 57.2, 57.5; H, 4.4, 4.6).
to lead t o the allo-configuration a t C-5. Nevertheless, we (2) Schnider and Grussner, Helw. Chim. Acta, 81, 821 (1939). have prepared the corresponding isomer, pregnan-3,12,20(3) Chakravarti and Pasupati, J . Chem. SOL.,1859 (1937). trione (IV), by the mild oxidation of an authentic sample of (4) Cornforth, Cornforth and Robinson, ibid., 689 (1942). 12(c~)-hydroxypregnan-3,20-dione (111). It had the follow( 5 ) Perkin, ibid., 109, 922 (1916). ingproperties: rn.p.204-206', [ ~ ] S +181", D [cY]~ +225O ~u~~ (6) Dobbie and Lauder, ibid., 6'7, 19 (1895). (chloroform). AnaE. Calcd. for CllH3003: C, 76.3; H. OF CHEMISTRY MILTOND. SOFFER 9.2. Found: C, 76.0; H, 9.1. Reichstein and von ArxS DEPARTMENT COLLEGE report for pregnan-3,12,20-trione: m.p. 201-202'; [ ( Y ] ~ ~SMITH D J. CHARLES CAVAGNOL MASS. HILDAE. GELLERSON +182 * 7, [ C Y ] ~ ~ M B I+219 * 8 (ethanol). A mixture of IV NORTHAMPTON, The with I1 showed a melting point depression of 36'. RECEIVEDOCTOBER 19, 1949 melting point of each of these compounds was depressed 10-20" by the ~ i o n from e hecogenin. Since the properties of elZo-pregnan-3,12,2O-trione(11) DEGRADATION OF GLYCOGEN TO ISOMALTOSE are different from those of the samples derived from hecogenin and botogenin, some doubt must be entertained as to Sir: the structures of the degradation products from both of Methylation studies' have indicated that the these sapogenins. glycogen molecule has a highly ramified structure We thank Parke, Davis and Company for their help.
composed of a-D-glucopyranosyl units joined 1,4 with branching a t C6 on one out of twelve units. As additional evidence in support of this structure THEWHITMORE LABORATORIES R. B. WAGNER we report the isolation of crystalline 6-a-~-glucoSCHOOL OF CHEMISTRY A N D PHYSICS THEPENNSYLVANIA STATE COLLEGE JAMBS A. MOORE pyranosyl-P-D-glucopyranose octaacetate (P-D-iS0F. FORKER STATECOLLEGE, PBNNSYLVANIAROBERT maltose octaacetate) from an acetylated acid RECEIVED OCTOBER 10, 1949 hydrolysate of glycogen. Animal (rabbit liver) glycogen (5.00 g., [aylz6~ SYNTHESES IN THE DIRECTION OF MORPHINE. I. +200', c 0.92, water) in 2% concentration was 7-METHOXY- AND 7,8-DIMETHOXY-2-TETRALONE. hydrolyzed a t 100' in 0.05 N sulfuric acid for nine hours (degree of hydrolysis ca. 75%). After acid Sir : We wish to report the synthesis of 7,s-dimeth- neutralization with barium carbonate and ion reoxy-2-tetralone, which may serve as a useful moval with exchange resins (Amberlite IR-100 and intermediate for elaboration in the direction of IR-4))the amorphous solid obtained on solvent remorphine and certain of its degradation products,' moval was acetylated with hot acetic anhydride and may open a way for the preparation of physi- and sodium acetate. The resultant sugar acetate ologically active substances oxygenated a t points mixture (6.08 g.) was chromatographed2 on Magcorresponding to the 3 and 4 positions in mor- nesol-Celite under such developmental conditions phine. 7-Methoxy-2-tetralone may serve in the that monosaccharides were removed from the syntheses of substances similarly substituted in column. P-D-Glucose pentaacetate was identified, (1) W. N. Haworth and E. G. W. Percival, J . Chem. SOC.,2277 the 3 position; and is of particular interest in view W. N. Haworth, E. L. Hirst and F. Smith, ibid., 1914 of the recent report that 3-hydroxymorphinane is a (1931); (1939). (5) Reichstein and von Arx, Xclu. Chim. Acto, 98, 747 (1940).
(1) F i c m and Holmes, THIOJOURNAL, W, 2549 (1088); 60, 2a19
~ i ~ a oc.L~, ) ; J . cim. SOC., aaw (ioaa.
(2) M.L. Wolfrom, L. W. George8 and I, L. Miller, THIO Joumhr. IO. 47s (iom; T I , la6 (1949~
3858
COMMUNICATIONS TO THE EDITOR
Vol. 71
by melting point and rotation, in the effluent. The material from the lowest zone consisted of p-D-maltose octaacetate (m. p. 15&160°, unchanged on admixture with an authentic specimen; [ c Y ] ~+62.5", ~D c 1.1, chloroform). The material from the next higher zone was rechromatographed in the same manner, and the eluent from the lower zone which crystallized from ethanol was identified as p-D-isomaltose octaacetate (m. p. 144-145", unchanged on admixture with an authentic specimen; [ c Y ] ~4-98", ~ D c 1.0, chloroform); yield 92 mg.
made for therapeutic use have been found to be rich in a principle which appears on the basis of chemical and biological properties to be analogous to the apoerythein in gastric juice. Other biological materials tested, including commercial pepsins, contain very little or none of the active substance. Less than 2000 parts by weight of a concentrate prepared from hog gastric mucosa completely counteracted consistently the microbiological growth stimulation of one part of erythrotin. For preparative purposes hog gastric mucosa has been used, and the principle can be preDEPARTMENT OF CHEMISTRY THEOHIOSTATE UNIVERSITY M. L. WOLFROM cipitated from an aqueous extract by alcohol, COLUMBUS 10, OHIO A . N. O'NEILL3 acetone or ammonium sulfate (80% saturation). RECEIVED OCTOBER 8, 1949 The principle is highly selective in its action and inactivates erythrotin but does not diminish the (3) Corn Industries Research Foundation Fellow in the Departbiological action of the end-products of erythrotinment of Chemistry. catalyzed processes which can substitute for this vitamin in microbiological assays-methionine (Escherichia coli test) and desoxyribosides (LactoERYTHEIN AND APOERYTHEIN AND THEIR RELATION TO THE ANTIPERNICIOUS ANEMIA bacilli tests) .4 PRINCIPLE The complex formed when erythrotin combines Sir : with apoerythein decomposes upon heating (120' Normal gastric juice has been found to contain a fifteen minutes) into erythrotin (or a compound non-dialyzable, heat labile substance which com- which cannot be distinguished from it biologically bines, apparently stoichiometrically, with erythro- or chromatographically), and a residue no longer tin,' (vitamin B u ) ~to form a complex (erythein) possessing the ability to bind erythrotin. In comwhich is non-dialyzable and not dissociated by bined form erythrotin is not as susceptible to dedialysis. Erythrotin in this combination is not struction by alkaline or oxidative treatments available to microorganisms (Escherichia coli, which inactivate the unbound vitamin, since heat Lactobacillus lactis Dorner, Lactobacillus leich- liberation following such treatment of the complex mannii), but is released by heat, much as biotin is yields the original erythrotin activity. These experiments point to the probability that released from avidin, whereupon it is again microbiologically active. Heated gastric juice contains apoerythein is the intrinsic factor of Castle or an no principle capable of combining with erythrotin. important component thereof. Clinical trials are Quantitative determination of heat labile now in progress to test this conclusion. principle (apoerythein) is readily performed by We are deeply indebted to Dr. William Shive measuring in an erythrotin assay (Escherichia for generous supplies of erythrotin before vitamin coli)' the inhibition of growth resulting when Bl2 was available and for prepublication disaliquots of the juice are added (unheated) to closures concerning erythrotin tests, and to Dr. cultures containing just sufficient erythrotin to Edward Campbell, Eli Lilly and Company, who elicit a maximum response. The erythrotin com- furnished biological preparations and gastric bining capacities (millimicrograms of erythrotin samples. per ml. of secretion) of samples of gastric juice THEBIOLOGICAL INSTITUTE AND from normal and anemic subjects were found to be THE DEPARTMENT OF CHEMISTRY AND respectively, 20, GO3, 60, 15; and 5,
