May, 1944
AN
ACYCLICSUGAR ORTHOACETATE
Pechmann condensation of ethyl acetoacetate and phenols does not always yield coumarins as previously reported. 2. Although sulfuric acid causes the condensation of 3,5-dimethylphenol and ethyl acetoacetate to a coumarin, this same reagent results in the formation of a chromone from 4-chlor0-3~5-dimethylphenol and ethyl acetoacetate. 3. The structure of the chromone was established by various hydrolytic reactions and by
lCONTRIBUTION FROM
THIE
805
condensation with benzaldehyde. 4. The coumarin expected in the initial condensation was synthesized b y nitration of 4,5,7trimethylcoumarin, followed bq' reduction and replacement of the amino group by chlorine. The proof that the product was a coumarin and that the chlorine was in the 6-position was determined by ozonolysis to 5-chloro-4,6-dimethyl-2-hydroxyacetophenone. URBANA, ILLINOIS
RECEIVED MARCH 2, 1944
CHEMICAL LABORATORY OF THEOHIO STATE UNIVERSITY 1
An Acyclic Sugar Orthoacetate BY M.L.WOLFROM AND D. I. WEISBLAT 1-Chloro-1-thioethoxy-uldehydo-D-galactose pentaacetate (11) has been prepared by the action of a dry ethereal solution of hydrogen chloride u on l-thioethoxy-aldehyh-D-gglactose hexaacetate On.repeating this procedure in larger quantities, a crude preparation of I1 was treated with ethanol and silver carbonate. The main reaction product was 1x1, designated D-galactose diethyl monothioacetal pentaacetate and previously synthesized2in a Similar manner from pure 11, prepared by the action of a mixture of acetyl chloride and phosphorus oxychloride upon D-galactose diethyl mercaptal pentaacetate. In addition there was obtained in low yield a product IV, to which is assigned an orthoacetate structure.
8)
an orthoacetate structure in an acyclic sugar derivative. Experimental 1-Thioethoxy-aMekydo-Pgalaetose Ethyl 1,2-0rthoace-
tate Tetraacetate.-Crude unrecrystallized l-chloro-lthioethoxy-aldchydo-D-galactosepentaacetate (m. p. 9698", 45 g,), prepared from l-thioethoxy-aldckydo-D-galactose hexaacetate as previously described' and thoroughly washed with an ether-petroleum ether (1 :1) mixture, was added to a suspension of 25 g. of Drierite (anhydrous calcium sulfate) and 60 g. of silver carbonate in 250 cc. of absolute ethanol. The reaction was then allowed to run at room temperature, under mechanical stirring and protection from atmospheric moisture, for approximately sixteen hours. The inorganic salts were removed by filtration through a bed of Super-cel (Johns-Manville)and the clear filtrate was made just opalescent with water. One crop of crystals was de?Et SEt posited on cooling to ice-box !Et ?Et I OEt temperature; a second and H&--oA~ Hk-Cl Hd-OEt H-C-0 third crop of crystals were I >&-CHI obtained by treating the I I HCOCOCI successive filtrates in a like HCOCOCHi manner. The first two crops HC1 EtOH I 1 -+ A c d H AcOCH (15-16 9.) were recrystallized AcOCH I I AQKO~ I from six parts of hot absoAcO H lute ethanol; m. p. 103-105°, ACOCH spec. rot. +Jo" (25", c 4, abs. I I I I HCOAc HCOAc CHCL, D line). This prodHCOAr HCOAc uct was therefore idenaed I I CHSOAC CH~OAC I C!HtOAc CH~OAC (mixed m. p. with an authentic sample unchanged) as D IV I 11 I11 galactose diethyl monothio The orthoacetate structure of IV, designated acetal pentaacetate for which the following constants recorded:' m. p. 104-105", spec. rot. +50" (20°, r I-thioethoxy-uldehydo-o-galactoseethyl 112-0rth0- are 3.5, abs. CHCL, D line). The third crop (6-8 9.) had acetate tetraacetate, is based upon the fact that a melting point of 85-95". Pure material was obtained one acetyl group in the compound was resistant after five recrystallizations from four parta of hot metlanol; yield 1.5-2.0 g., m. p. 126-126" (mixed m. p. with to alkaline hydrolysis but was removable by acid D-galactose diethyl monothioacetal pentaacetate of m. p. hydrolysis. This fact, among others, has been 104!-105°, 93'1, spec. rot. +64" (ao, c 4, abs. CHClo, employed in establishing the orthoacetate struc- D lime)'. The substaiice crystallized in transparent, elongated ture* in the cyclic sugar derivatives. In formula and exhibited properties and solubilities similar . to ._ IV, the placement of the sulfur atom and the size prisms the isomeric D-galactose diethyl monothioscetal pentaof the ring are probable but arbitrary. To our acetate, except that it was more sensitive to aciditv. knowledge, this is the first recorded Occurrenceof A w l . Calcd. for CIOHI?OOS(CH&O)S: C, 497;)9; H, (1) M. L. Wolfrom, D. I. Weisblat and A. R. Hanze, THIS JOUR6.71; S, 6.67; CHiCO (4 equiv.), 8.33 CC. 0.1 N NaOH NAL, 64, 3246 (1940). per 100 mg.; CH&O (6 equiv.): 10.41 cc. Found: C, (2) M. L. Wolfrom and D. I. Weisblat, ibid., 0,878 (1940). 49.92; H, 6.73; S, 6.86; CHICO (alkaline hydrolysis, (3) K. Freudenberg and E. Braun, Nelurtuisscxscheftm, 16, 393 Kunz ____and Hudson' method), 8.39 CC.; CHnCO (arid
H-ro + Acoc
__f
~~
(1930); E. Braun, Bm.,68B, 1972 (1930); H. G. Bott. Haworth nnd R. I.. Hirrt, J . Ckcm. Snc. 1396 (1930).
W. N
806
W. H. PEARLMAN
hydrolysis, Freudenberg and HarderKmethod), UI.52 cc.
summary 1. There is reported the synthesis, in crystal(5)
K.Preudenberg and M. Rarder. Ann., 4Bs,230 (1923).
Vol. 66
line form, of the first acyclic sugar orthoacetate (IV), designated l - t h i o e t h O x y - a - D - g ~ a c tose 1,2-ofioacetate tetraacetate. COLtAfBUS,OHIO
RECBNED FEBRUAXY 21.194
[CONTRIBUTION FROM THE PHYSIOLOGICAL LABORATORIES, CLARK UNIVERSITY]
A Study of the Neutral, Non-Saponifiable Fraction of Ox Bile' BY W, H. PEARL MAN^ A theoryBpostulating a biological conversion of TABLEI cholesterol to the bile acids has recently received COMPOUNDS ( O T H E R THAN CHOLESTEROL) ISOLATED FROM experimental support in an investigationa carried NON-SAPONIFIABLE FXACTION OF Ox BILE out with the aid of deuterium. One might expect THE NEIJTFLU,, Tentative -Derivatives-to find metabolic products intermediate between Mbp.," empirical M ;E:,!' cholesterol and the bile acids in bile since it con- Compd. C. formula T m tains these steroids in large amount. Studies of Monoacetate 216-217 A 300 CnH& the comparative biochemistry of the bile have alMonobenzoate 155-157 B 192-193 GiH1601 Dibenzoate 234-235 ready furnished clues as to the nature of the hypothetical intermediates. The bile of elasmobranchs2 C 255-257 %.&lo-mOc Monoacetate 187 D 232-233 GJ&OS~~ Monoacetate 111 contains a polyhydric alcohol, scymnol (GH460,) which is related to cholesterol; it can be E 202 C&n04 Diacetate 142.5 readily degraded in the laboratory to cholic a ~ i d . ~ , ~ Other alcohols of an alleged steroid nature have diol-3(/3),20(a)was suspected to be the isomer in been isolated from the bile of frogseand toad^^^^; question. This steroid had been prepared by of these, pentahydroxybufostane (Cd-b~Ob)can Butenandt and Miiller14and they reported a meltbe oxidized i n vitro to cholic acid.' A mechanism ing point of 189-190.5' uncor. Marker, et aZ.,I6 for the biological oxidation of the side-chain of had previously prepared the isomer from pregsteroids with 27 and 28 carbon atoms has been nandiol but their preparationmelted a t 182'. This discrepancy in melting points was noted by Butsuggested by Kazuno.' An exploratory examination of the neutral enandt and Miiller; they raised a doubt regarding fraction of ox bile failed to reveal the presence of the nature of their own preparation, stating that scymnol, etc. On the other hand, five compounds isomerization of the C-17 side-chain might have (other than cholesterolg) were isolated in small occurred since the reduction of the 20-keto group amount. Although the empirical formulas (see of the intermediary steroid had been carried out Table I) point to a steroid structure, these com- in an alkaline medium. The writer prepared pounds could not be identified with any known pregnandiol-3(@),20(a), following the procedure steroids. Structural proof must wait until more of Marker, et aZ.,l6in all respects except that the intermediate 20-acetate of pregnandiol-3(a),20material is available. Compound B, m. p. 192-193', has an empirical (a),was isolated. The final product melted at formula identical with that of pregnandiol. Since 182' ; purification by chromatographic analysis Compound B is digitonin-precipitable, pregnan- did not raise the melting point. The dibenzoate (not previously described) melts at 167-168'. (1) Aided by a grant from G. D. Searle and Company. Since the dibenzoate of Compound B melts at (la) Present Address: Department of Biology, Princeton f n i v e r 234-235', it is clear that Compound B cannot be S t y , Princeton, N. J. (2) L. F. Fieser. "The Chemistry of Natural Compounds Related identical with pregnandiol-3(B),20(cu). Comto Phenanthrene," Reinhold Publishing Cotp.. New York, N. Y., pound B thus remains unidentified for the time 1937. being; it is probably an (allo) pregnane derivative (3) K. Bloch, B. N. Berg and D. Rittenberg, J , B i d . Chcm., 14% containing a 3(@)-hydroxylgroup. 511 (1943). (4) R. Asiari, J . Biochcm. (Japan), 9% 319 (1939). Compound C contains either 25 or 26 carbon (5) W. Bergmann and Wm. T. Pace, T m s JOURNAL, 61, 477 atoms. This is an unusual number for naturally (1943). occurring steroids but it might be mentioned that (6) Y . Kurauti and T. Kazuno, 2. Dhysiol. Chcm., 269, 53 (1939). a C-26 acid has been obtained from bile.O (7) T. Kazuno, $bid., 966, 11 (1940). ( 8 ) H. Makino, ibid., 240, 49 (1833). The ever-recurrent question of artifact forma(9) No attempt waa made to isolate dihydrocholesterol which tion deserves mention in this report but, since the Schomhdmer, rl al.,lO have shown to be present to the extent of
{
1-2% in cholesterol from,v&ious organs; Pertzbornl' reports 3.6% dihydrocholesterol in ox bile. (10) R. Schoenheimer, H. V. Behring and R. Hummel. ibid., lS9, 93 (1930). (11) R . Pertzborn. Z. Ees. e x M . M c d . . 101, 350 (1937).
(12) All K.. p.'s in thia paper are corrected. (13) Empirical formula deduced from analytical data on aftate. (14) A. Butenandt and G. Milller, Bar., T i , 191 (1938). (15) R . E. Marker, 0. K a m a , E. L. Wittittle, T. S. Oakwood, E. J . Lawson and J. F. Laucius, TEIS JOURNAL, 6S, 2291 (1937).
