~~~-CYPT,C)PR STEROIDS OPYL
Septembrr 1967
793
combined extracts were washed (dilute NaHCO3, HzO). After drying, t,he solvent was removed under reduced pressure to give 5.18 g (55%) of crude prodiict. l i e .tallizatioii from etherpetroleum ether furnished 3.6 g (40 of xvhite crystals, mp 192-195". I n a pilot experiment, the product was recrystallized from ether-petroleum ether to give white crystals: m p 195-197'; [a]n -11': no significant absorpt,ion in the ult,raviolet; Xmnx 2.94, hyl ether (eliited n i t h 30YG ether). Further elution, with 40-605 ether, gave T.85 g of crude 2. Two recrystallizations from acetonitrile provided 4.90 g of pure 2: nip 120.5-122.0'; [aI25ju +44.3'; infrared, vmax 2.87, 3.25, 3.42, 3.50, 6.23, 6.32, 6.66, 6.84, 8.09 p ; Iimr (external TMS standard) signals, 435-415 (aromatic ring), 249 (OCHI), E3 (angular CHI), and 50lopropyl) [lit.2mp 116-116.5", [ a ] " D +42.0" (c O . i 9 ) ] . Anal. Calcd for C22H3002: C, 80.92; €1, 0.26. Bound: C, 80.72; H, 9.2%. In an alteriiative purification procedure, the crude product from the reaction of 920 mmoles of cyclopropyllithium with 460 mmoles of estrone methyl ether (1) was added to 250 ml of ethanol, 30 ml of AcOH, and 28 g (150 mmoles) of Girard "P" reagent. The solution was refluxed for I hr, cooled, diluted with 2 1. of cold water, :tiid extracted with 3 1. of ethei,. l h e separated i l f ) L. G . ifersliherrer, E. G. Shipley, a n d 11. K. Ifeyer, $'roc. S o c . IizptZ. Bzol. .Wed., 63, 175 (1953). 113) H . D . Lawson. C. G . Eeller, J. I3. Golden, a n d E . I. Severinghuas, Endominology,24, 35 (193Y). (14) C. Clauberg, Z e n t r . Gynoekol., 54, 2 i 5 7 (1YSO). (15) +Melting points were determined in open capillaries in an oil bath witli a calibrated thermometer, and are accurate to + l o . Optical rotations were measured using 1 % solutions of the comrounds in CHCla; ultraviolet spectra were measured using solutions in Y;ic/, EtOH. T h e infrared spectra were determined with K B r pellets. and the nmr spectra were recorded for CDCk solutions. T h a n k s a r e exprrssetl t o l l r , IedNaCl solution (100 ml). After having been dried (Na2SOk),the solution was evaporated and the residue was crystallized from hexane to give 0 . i 0 g of 3, mp 125-128". The iiifrared spectrum showed characteristic dihydroanisole absorption a t 5.92 and 6.02 M , and a sharp hydroxyl band a t 2.9 p . Only end absorption was seen in the ultraviolet region. iZ second crop of crystals was also obtained: 0.07 g, mp 122-126". The analytical sample (from ether-hexane) had mp 123-129" (129-131' in an evacuated capillary), [ a ]25uf94.0". Anal. Calcd for C22H3202: C, 80.44; H, 9.82. Found: C, 80.6T; H, 10.05. 17a-CycIopropyI-17-hydroxyestr-5( lO)-en-3-one (4).-A solution of 0.197 g (0.61 mmole) of 3 i n 20 ml of methanol was mixed with a solution of 0.23 g of oxalic acid dihydrate iii 3 ml of Eater. After being kept for 40 min at room temperat.ure, the solution was transferred to a separatory funnel with 125 ml of ether. The ether phase was washed (YaHCO,, H20, saturated NaC1). Ilrying arid evaporation of the ether solution left a residue which was ci.yatalliaed from ether-hexane t o give 0.125 g of 4, mp 145-150'. The iiifrared spectrum showed C=O (5.86 p ) and OH (2.85 p j absorption. 1-he ultraviolet spectrum indicated the presence of a small amount of conjugated ketone as an impurity: Aman 240 mp ( e 350). S o vinyl protons could be seen iii the nmr spectrum, and the cyclopropyl and angular methyl group protons were apparent. I n a scaled-up preparation (7.0 g of steroid, 21.3 mnioles), a 93% yield of comparable material was obtaiiied. Recrystallizatioii from ethyl acetate-hexane afforded the analytical sample, mp 150.5-153' (in an evacuated capillary 152.5-154"), [ a ]2 5 +156.4'. Anal. Calcd for C21H3002: C, 80.21; H, 9.62. Found: C, 80.34: 11, 9.66. 17~u-Cyclo~ropyl-17-tgdroxyestr-4-en-3-one (5).-Oiie milliliter of 2 AV YaOH solution was added to a solution of 2.0 g (6.36 mmoles) of 4 in 75 ml of methanol. After 30 min a t room temperature, the isomerization was complete, as indicated by thin layer chromatography (tlc). The base was neutralized by the addition of a few drops of AcOH, and the solution was then evaporated to dryness a t room temperature under reduced pressure. The residue was taken up in ether and the solution was washed, dried, and evaporated. Crystallization of this residue from ether-hexane gave 1.i g of 5 , mp 132-133", [ O ~ ] * ~+13.i0, D ,,,,A, 241 nip ( E 17,600). ilnal. Calcd for C21H3002: C, 80.21; H, 9.62. Found: C, 79.99; H, 9.39. A second crop of crystals, 0.21 g, nip 129-132.5", was also iholated. 111 later preparations of this compound, a higher melting polymorphic form of 5 was always isolated, mp 144.5-146" [lit.2 mp 139.5-140.5', [ C Y ] ~ ~ +11.1", 'O 240 mp ( e 16,900)]. 17~-CycIopropyl-17-hydraxyestra-4,9( lO)-dien-3-one (6).--A solutioil of 12.6 g (40 mnioles) of 4 iii 250 nil of d1.y pyt,i.iiile was stirred arid held at 3 ?C 2" with an ic.e bath, while 14 g (44 mmoles) of pyridinium bromide perbromide was added in small portions during 30 min. The pale yellow mixture, now containing some precipitate, was stirred a t room temperature for 5 hr and then was diliited with 3 1. of cold water. The precipitation product was collected by filtration, washed with water, dried, and recrystallized from ether. Three crops of crystals, totalling 9.70 g arid melting in the range 147-151.5' were obtaiued. The analytical sample of 6 (from ether) had nip 151-122'; [a]"D -205.8';
~
September 1967
[l'icu,ltia-t/]Oxazo~~~o STEROIDS
water arid evaporated to dryness under reduced pressure. To the vellow solid residue there was added 200 ml of methanol, 16 g ;)f NaOAc.3H20, 40 ml of water, and 16 ml of AcOH; the resultiiig solut,ion was refluxed for 4 hr. The methanol was evaporated arid 200 ml of 4 S HCI was added to the residual mixture of oil aiid water, which was then extracted (CH2C1,). The extract was washed with dilute base and water and was then dried and
799
evaporated. Chromatography of the residual oil over silica gel (elution with 1: 1 ether-pentane) followed by two crystallizatioria from acetonitrile gave 4.25 g of pure 15, mp 162.5-163.5", 242 mp (E 16,700), [ c u ] ~ ~ $70.2" D [lit.? mp 158-159", [CUI% +62.2" (c 0.98), Xma, 241 mp (E 16,500)]. Anal. Calcd for CnHJIz: C, 80.44; H, 9.82. Fouiid: C, 80.27; H, 9.93.
Steroids Possessing Nitrogen Atoms. 111. Synthesis of New Highly Active Corticoids. [17a,3-6a-d]OxazolinoSteroids' G I A S G I A C O ~ I OXATHASSOHS, G ~ O R GWINTERS, IO AXD
EMILIO TESTA
Sleroid Group of Research Laboratorzes, Lepetzt S.p.*i.,Jlilan, Italy
Recezved Maich 13, 1967 The preparatioit of [17a, Ifj,-cl]-2'-n~ethylosazolirlo analogs of prediiisotie, prednisolotie, a d !)oi-fluorc)prediiisolone, from lin-a~ido-5a-pregiiaiie-3P,160i-diol-l1,20-dione s,lB-diacetate, is described. Preliminary pharmacological data show that t,he osazolino analogs of prednisolone aiid 9,-fluoropredriisoloiie are significantly active when tested for neoglycogenetic and antiinflammatory activity in the rat.
I n our previous paper2 we described the preparation and demonstrated the st'ructure of 5cu-pregnan-3p01-11,20-dione[ l i a ,16a-d]-2'-methyloxazoline (IIa) . Due to t8he chemical stability of t'he oxazoline ring, this compound represented an excellent material for continuing our studies aimed a t obtaining 17-nitrogen derivatives of steroid hormones. The present paper describes in detail the synt'hesis of the oxazolirie analogs of prednisone, prednisolone, and Sa-fluoroprednisolone. Starting from I I a which was prepared from the azide I by an improved method compared t o that previously described,2 we obtained excellent yields of pregnanetrione I I I a by oxidat,ion with chromic acid in acetone. The introduction of a bromine at'om followed by dehydrohromiiiatiori led to t'he 1J-dehydro derivative IV. Subsequent treatment with SeOs in t-butyl al(who1 gave the diene V. This compound was more easily obt'ained from the 2,4-dibromo derivat'ive IIIc by heating in dimethylformamide (DAIF) with Li salts. Attempts to introduce a bromine atom a t C21in the oxazoline 11, under various experimental conditions, have failed. This absence of reactivity is not unusual in 16.17-disubstituted pregnane derivat'ives and is reported, for instance, for 17a-bromo-3/3,16p-diacet~oxy-5a-pregiian-20-orie3 and 16a,l7a-dihydroxypregn-4-ene-3,20-diorle 16,17-ac~tonide.~Allen and Weisda reported further that lBcu,lia-isopropylidenedioxypregnaii-20-one derivat,ives failed to give both 20-semicarbazones arid 21-iodo compounds. By cwiitrast, good results n-ere obtained in our 21iodi1i:itioii of V wrording to the Ringold-St'ork method modified by Iiothman, et a1.j The 21-iodo derivative reacted regularly with triethylammonium acetate to give the oxazoline analog of prednisone acetate (VI). Conversion of VI int'o the 1I@-hydroxy derivative ( 1 ) 'rliid paper \ \ a s part of a communication presented a t t h e 2nd Intrriiational Congress of Steroid Hormones, Milan, I t a l y , M a y 23-28, 1966. ( 2 ) G. G . Nathansohn, Q. \Vintera, and A. Vigevani, Gnzz. Chi,,&. I t a / . , 96, 1x38 (1963). (:3) S . G. L e r i n e and 31. E. IVaIl, .I. A m . C h e m . Soc., 81, 2829 (1959). (4) (a) G. R. .\lien, J r . , a n d A l . J. Weiss, ibid., 81, 4968 (1959); (b) iiiid.,
82, 2840 (1960). (j)
E. S, Rotliinan, T. Perlstein, a n d 11. E. TVall, J . Ory. C h e m . , 26, 1Y66
(1960).
via the 3,20-bis(semicarbazone), reduction with cwmplex metal hydrides, and hydrolysis of the biesemicarbazone did not give appreciable yields of the required compound XII. I t was found t o be much more convenient t o reduce the 20-semicarbazone VII, obtained from IIa, with SaBH4. The facile conversion of I I a t o the semicarbazone VII, when compared with the lack of reactivity of 20-ketopregnaiie-l6a,l7adihydroxyacetonides, suggests that other factors, and not merely steric hindrance, must intervene in order t o explain the noteworthy different chemical behavior betlveen the D-ring-fused oxazoline- arid dioxolidinepregnanes. The 1lp-hydroxy derivative VI11 was hydrolyzed to IX by merely boiling with HC1 in aqueous methanol. This process has given better yields than other methods6which necessitate the use of pyruvic or nitrous acid. Selective oxidation of the 3-hydroxyl of IX, according t o Oppenauer, led t o the %keto derivative X, and the diene XI was obtained from this, by dibromination in dioxane and dehydrobroniiiiation with Li salts and DMF. 21-Acetoxylation performed as described for 1'1 furnished the oxazoline analog of prednisolone (XII). The synthesis of the Sa-fluoro derivative XV was performed in a routine manner, dehydrating7 the l l p hydroxy steroid XI1 to A1~4~9(11)-trierie XI11 ; on adding HOBr t o the 9,11 double bond and treating the bromohydrin with alkali the Sp,llp-epoxide XIV was obtained. Reaction of XIV with anhydrous HF supplied the required [licu,l6a-d]-2'-methyloxazoline of 9cu-fluoroprednisolone (XV) Biological Results.lO--The conil)ouiids have been examined for neoglycogeneticil and antigraiiulomiltous .8s9
(6) I n t e r d i n : (a) H. L. Herzop, C. C. P a y n e , 11. .\. Je\-nik, D . Gould, E. L. Shapiro, E. P. Oliveto, a n d E. B. Hershberg, J . Am. Ckem. Sor.. 77, 4781 (195.5); 01) E. P. Oliveto, E. Rausser, I,. Kelier, E. Sliapiro, D. Gould, a n d E. H. Heruliberg, i b i d . , 78, 1736 (1956); (c) N . 1). \Vendler, H u a n r 3Iinlon. a n d R l . Tishler, ibid., 73, 3818 (1951). (7) G. G. Haaen a n d 13. if-. Rosenburg, J . O w . Cliem.. 29, 1930 (1964). (8) J . Fried a n d E. F. Sabo. J . A m Ckrrn. Yoc., 79, 1130 (1957). (9) R . F. H i r x h m a n n , R . RIiller. .J. \Toad, a n d R . E. .Jones, ibid., 78, 1956 ( I 956). (10) T h e authors a r e indebted t o D r . P. Schiatti a n d Professor G. M a E i of t h e Lepetit Department of Pharmacology for kindly supplying these d a t a . (11) R. E. Olson, F..i. Jacobs, D. Richert. J. .i, Tliayer, K . ,J. Kopp, and N . J . \Vade, Bridocrzrioloyy, 36, 430 (1944).
