November, 1964
STEROIDAL CARBAMIC ACIDa-LACTONES
solution. The crystalline product, which separated, was filtered with suction, washed well with water and cold ether and dried tn vacuo (12.4 g., 6270). Recrystallization from methanol yielded an analytical sample, m.p. 170-172'. Compound X ( R = R, = C6H5).-sodium borohydride (5.0 g., 0.335 mole) was added to a warm mixture of IX ( R = RI = C6&, 5.0i g., 0.0105 mole) in methanol (125 ml.). The mixture soon became homogeneous and a white crystalline solid separated. hfter 10 min., water (500 ml.) was added and the product was filtered with suction and dried in z'acuo (4.11 g., 815;). An analytical sample, m.p. 210-212.5", was obtained by recrystallization from a methanol-ethanol mixture. Compound XI ( R = RI = C6HS).-To a warm, st,irred solution of IX (It = Iil = CeH5, 5.0 g., 0.0105 mole) in 1-butanol (150 ml.), sodium (15.5 g., 0.67 g.-atom) was added in portions. The mixture was boiled a t reflux for 2 hr. Hydrochloric acid (l87C, 200 nil.) was added and the mixture was distilled until the distillate was clear and homogeneous. The residue was treated with additional HCl (IO?;, 200 nil.) and the acidic solution was extracted with three 70-ml. portions of ether. The aqueous layer was made alkaline with animonia and the resultant white precipitate was filtered, washed with water, and dried in vacuo (2.2 g., 65%). Recrystallization from methanol yielded an analytical sample, 1n.p. 210-212".
739
Compound X I (R = CeH5; R1 = CH20H).-A mixture of IX (R = C&, R1 = C02CzHs; 18.5 g., 0.039 mole), lithium aluminum hydride (25 g., 0.66 mole), and tetrahydrofuran (400 ml.) was stirred and boiled a t reflux under a nitrogen atmosphere for 4 days. Ether (1 1.) was added and the stirred mixture was hydrolyzed by dropwise addition of water (200 ml.), Inorganic salts were separated by filtration with Celite. The filter cake was washed with ether, and HC1(18%, 800 ml.) was added to the combined organic layers. The aqueous layer was separated, washed with four 250-nil. portions of ether and made alkaline Kith concentrated ammonia. The mixture was extracted with chloroform and the organic extract was washed with saturated aqueous XaC1 and dried (Na2S01). Removal of solvent in L ~ X O yiclded a viscous oil, which crystallized upon trituration with methylene chlorideether. Recrystallization from the same solvent pair yielded an analytical sample, m.p. 156-158".
Acknowledgment.-We wish to thank Dr. H. Graboyes, Smith, Kline and French Laboratories, for the preparation of several synthetic intermediates and N r . H. Farber for technical assistance.
Steroidal Carbamic Acid y-Lactones EUGE~XE FARKAS
AND
JOHX A. SWALLOW
T h e Lilly Research Laboratories, Indianapolis 6, Indiana Received July 27, 1964 The reaction of.steroida1 17a-ethynylcarbinols with alkyl isocyanates was used to prepare the 17-carbamoylcarbamate, carbamate, and carbamic acid ?-lactones in the aromatic A-ring, androstenone, and 19-norandrostenone series. The biological testing of these resultant compounds was performed and an interesting gradation in activity in the progestational area was observed.
Substitution of various functiona1 groups a t the 17position of the steroid nucleus produces interesting and useful alterations in the pharmacological activity of the resultant compounds. While the well-documented 17a-alkyl substitution in the 4-androstene molecule increases the androgenic-anabolic activity, the 17a-ethynyl-17fi-hydroxy substitution results in progestational activity.' The corresponding 17a-ethynyl-17p-hydroxy-19-nor steroids are especially important and useful progestational and anovulatory agents. The spirolactone grouping,* on the other hand, transforms these same androstene steroids into clinically useful antiniineralocorticoid agents. Utilizing the results of Easton, et al.,3 of this laboratory, the reaction of various steroidal 17a-ethynyl17P-carbinols with alkyl isocyanates was examined with the goal of introducing a C-17 spirocarbaniic acid y-lactone group. The reaction of 17a-ethynylestradiol-3-methyl and -ethyl isocyanate gave two products. The more polar product, obtained in the greater amount, was found to be the carbamate (IIA) as evidenced by the analysis and the infrared and 1i.ni.r. spectra. The infrared spectrum showed two peaks a t 2.9 and 3.04 p which corresponded to - T H and acetylenic hydrogen, respectively. The carbonyl region (1) 0. v S t Whitelock, E d , "New Steroid Compounds X i t h Progestational Actirity," Ann. X. Y . Acad S e a , 71, 479 (1958), C Djerassl, L. hIiramontes, G Rosenkranz, a n d F Sondheimer, J . Am. Chem SOC.,76, 4092 (1954). (2) J 4 Cella a n d C M Kagawa, %bid., 79, 4808 (1957). (3) N R. E a s t o n , D. R Cassady, a n d R. D Dlllard, J . Ow. C h e m , 27, 2927 (1962).
revealed a broad band centered a t 5.83 p. The n.m.r. spectrum4supported the structure IIA by dernonstrating the presence of a single N-ethyl grouping. The minor, less polar product assigned structure I A apparently resulted from the reaction of the ethynyl carbinol with 2 molecules of isocyanate. The infrared spectrum indicated no free -NH but a band for weakly hydrogen-bonded -KH a t 2.98 and also showed an acetylenic hydrogen a t 3.01 p. The carbonyl region contained an intense band a t 5.84 p for the -0CON- and medium intensity bands at 5.98 and 6.53 p for -SCOSH-. The n.1n.r. spectra supported structure IA showing two methyl triplets nearly coincident at 6 1.17 ( J = 7.0 c.P.s.) a =CH singlet a t 6 2.68, an 8-component multiplet a t 6 3.30 for XHCH2CH3 in which coupling with N H is 9.0 C.P.S.and with methyl 7.0 c.P.s., a quartet a t 6 3.78 for -CH,K+CO j,, and a broad signal for hydrogen bonded liH at 6 8.5. These spectra and the analysis favor the assigned structure, IX. As in the previous work, 3,6a,5b treatment of the carbamate (IIA) TTith sodium niethoxide yielded a new compound, IIIA, whose infrared spectrum had strong absorption a t 5.63 p in support of the carbamic acid y-lactone structure. I n the n.ni.r. the exocyclic methylene gave an AB pattern centered a t 6 4.18 for (4) T h e n.m.r. spectra were determined using a HR60 Varian i n s t r u m e n t with CDCla a s solvent a n d TblS a s a n internal standard. T h e 6-values are rpported as p.p.rn. ( 5 ) (a) R. Sisido, K. Hukuoka, M. T u d a , and H. Nozaki, J. Org. Chem.. 27, 2663 (1962); (b) P. J. Stoffel a n d A. J . Spezide, ibid., 28, 2814 (1963).
which Av is 12.1 C.P.S.and JAB = 3.3 C.P.S. The niethk'urther, in the progestational assay, as iiitmurd by ylene attached to the nitrogen shows a noneyuivalencc the AlcPhail procedure,' only t he Iac~loiiw (111(') slio\rcd good activity. By injection, compound HI(' corresponding to Av = 15.6 c.P.s., J h ~= 1.1 c.P.s.. (It = CH,) was found to be twice ab active as 17aand J c H P c H s = 7 C.P.S. Although the available data do not completely eliminate the oxygen-closed st rLi!3et liyiiyl-19-nortestosterone but \vas only oil?-half a': activc orally. Surprisingly, tlie spirolacton~I\ i t h R = ture, the nitrogen closure is favored since structurr ('?HZsho\wd only oiic-half the activity of Iia-('t liyitylIIIA is completely consistent with the physical dat a l!)-riortestostc~roii(,h y iiijectiori and no nctivit y orally. and because of the closely analogous work of EastoiiJ Siitrbc t lie precursor of t hc carbaiiiic acid lact oil(&, t licx where this closure was established cheniically. cart)aniat e, had no appreciable progestal ional activity. The exocyclic double bond in the spiro-4-met hylene~ h i hactivity appears t o hc unicluc~lyassociated with t I.\(, oxaaolidone is hindered since this olefinic bond resisted 1-ing-closed conipouiids. Thc st ructurca -act ivit y rvlnhydrogenation under a variety of conditions. Oiily t ionship.: of thcsc lattcr compounds indicai o t t i o t~ conditions which were vigorous rnough t o reduce t IN. i t ringeiii st iuctural requircnieiits operating 1 i ~ wiliaii aromatic A-ring, Pd-C catalyst, pressure, arid hcai 111 ihe parent et hyriylcarbinols \vlierc> t Iic corresponding would effect reduction. et hyiiyl-21 -iiic.thyl-Xand -21 -chloro'-lO-iior st ( w i d '"and tlic l~-tctrahydropyraii~-l1ic.r" 01 l l i i ~ compound proceeded with difficulty because of the rclaI ia-c~thyriyl-li3-liydrosyc*oiiipuuiids, t I\(' 1 i - c ; i r h tive insolubility of this compound. =Ilthough t 1iv iitatcl 111 m c h of tlic serici, A4,13. or ('. lvad- io ;I d i t 1 ~ i i l u lower isocyanates gave some reaction, best yields n-erc obtained using n-propyl isocyariatc leading t o t l i c ~ 1 1 0 1 1 111 biological activity. carbanioylcarbamate, the carbamate. and the carbainic acid spirolactone. In the 19-nor series the carbamoylExperimentalI2 carbamate lvas not obtained in any appreciable arnouiit . However., the carbamate and the carbamic acid lacton(. 1 7 ~ E t h y n y l Carbamoylcarbamate (IA) and 1 7 ~ E t h y n y l were readily prepared using both inethyl and ethyl Carbamate (IIA).-A solution of 2.0 g. of liol-ethyn!.lestrrtdi[il isocyanate. The infrared and 1i.iii.r. spectra ~ v ( w 3-methyl ether, 23 nil. of ethyl isocayanate, aiid 0.15 g. of I)abc~11~ was heated a t reflux for 62 hr. xit,h an additional 5 nil. of ethyl completely analogous to those discussed in series A. (at
I
I1
0
C
Pharmacological Activity.-The biological act ivit i of these new compounds, particularly the carbamic acid lactones, mirrored the activity of the parent ethynyl carbinols. Surprisingly, none of the various spiro compounds showed any appreciable antideoxycorticobterone activity. I n the -4 or aromatic series, compounds IIA arid IIIA had about the estrogenic activity of estradiol while the carbaiiioylcarbaniate IA was almost devoid of estrogenic properties at these same doses. The substituted androstenorie compounds, series B, possessed only a low order of activity in all assays studied. The 19-norandrostenone or series C compounds proved to be of the most interest. I n the antiestrogen assay,6 although the carbamate IIC (R = CHE) showed no activity, the lactone IIIC had onehalf the activity of 17wet hynyl-19-nort est osteronc~. (6) R. A. Edgren a n d D. W. Calhoun, I'roc 589 (1956).
S O L L z p i l Bid
lled
92,
isoq-anate being added after. an initial elapse of 24 hr. The solvent was renioved under vacuuni, and tlie residue was clironi:rtographed on 75 g. of neutral alumina using benzene as tlic initial solvent. The carbanioylcarbamate was eluted with 10' 2 ether-benzene arid recrystallized from methanol to give 0.11 g., m.p. 214-215". .tncil. Calcd. for Cz;H,,X:04: C:! 71.65; 13, S.0"; S . (?.IS. Found: C, 71.27: €I, 7.99; S, 6.12. Further elution with 25-50yc ether-benzene gave tlie carbamate (IIA) which 'ivas recrystallized from methanol to give 0.62 g., n1.p. 89-91". .-lnal. Calcd. for &Ha1N@a: C, i 5 . 3 5 ; H, 8.19; S , :3.67. b'ou~ld:C, 75.28; H,Y.14; N,3.57. Carbamic Acid Lactone (IIIA).--A solution of 0.2 g. of ITA, 5 rnl. of sodium methoside solution (0.1 g. of sodiuni in 10 nil. o f methanol), and 17 ml. of reagent rnet'lianol was heated at reflux overnight. Some of the solvent was evaporated, and the solut,ion was poured into excess water. The mixture was extracted with ethyl et,her, and the combined ethereal solutions were washed with sodium chloride solution, dried ( Na$304), and concentrated in zsacuo. The residue was recrystallized from inethanol to give 0.12 g., m.p. 148-149". .lnu/. Calcd. for C2aH31?;@s:C, 75.55: H, 8.19: 9 , 3.67. Found: C,75.32; H,8.38; S , 3.18. Hydrogenation Studies.---Since no reduction had ocmrretl under milder condit,ions, 0.2 g. of IIIA was reduced using 0.2 g. of 5.; Pd-C i n 70 ml. of ethanol with 63.72 kg./cni.' (900 p.s.i.) o f hydrogen at 70' for 5 hr. After filtration the solvent, was evaporated, and since the residue did not readily crystallize, it was exanlined spectrally. The ultraviolet spectrum revealed rrrily %r; of rnethos\- aroniatic remaining, and the infrared
(7) 11. IC. L l ~ ~ P h i t iJl ., T ' / ~ ~ j s ~ o(London), Z. 83, 145 (1935). 18) .\. David, T. Hrtrtley. I ) . K . RIellson, a n d 1'. Petroa., J . Pliarm. I'h
