164 Journal of Medicinal Chemistry, 1971, Vol. 14, N o . 2 general methods employed are illustrat,ed for symmetrically or unsymmet,rically substituted carbinolamines by the following 2 examples. l,l-Di-(4-fluorophenyl)-3-dimethylaminopropan-l-ol.--h s o h of 29.0 g (0.2 mole) of et,hyl 6-dimethylaminopropionate in 50 ml of anhyd EtzO was added gradually with stirring to a cooled (0') EtjzOs o h of a Grignard reagent prepd from 105 g (0.6 mole) of p-BrC6H4F and 14.6 g (0.6 mole) of Mg. After stirring in the cold for 1 hr and then refluxing for 2 hr, the cooled mixture was decompd with 200 ml of a 10% w/v soln of NH4Cl. Basification with N H 4 0 H and extraction with CHCla ( 3 X 250 ml) afforded a CHCb s o h which on drying and evapn gave 38.0 g of a crude oil. This crude product was dehydrated without further purification. 3-Dimethylamino-l-( 4-fluorophenyl)-l-phenylpropan-l-ol.p-Dimethylaminopropiophenone.HC1 (21.4 g, 0.1 mole) was added portionwise to a cooled (O'), stirred EtzO soln of the Grignard reagent prepared from 35 g (0.2 mole) of p-BrCsH4F and 4.8 g (0.2 mole) of Ng. After stirring in the cold for 1 hr and then refluxing for 2 hr, the cooled mixture was decompd and worked up as above. The crude colorless solid (I1 g j was dehydrated without further purification. l,l-Di-(4-fluorophenyI)-3-dimethylaminoprop-l-ene(25).-A soln of 41.0 g (0.14 mole) of the carbinolamine in 250 ml of AcOR and 80 ml of concd HC1 was refluxed for 30 min. The s o h was concd under reduced pressure, dild with HzO, and basified wit,h NHIOII. The base was extracted EtzO, and EtzO soln was dried
and evapd. Diatn under reduced pressure gave 20.5 g of ail oil. The base was converted into its hydrochloride in anhyd EtzO. Crystii of the hydrochloride from EtOAc-EtOH gave an anal. sample, mp 211-213'. 1,l-Di-( 4-fluorophenyl)-3-dimethylaminopropane (53).--A Soln of .5 g of the olefin.HC1 in 20 ml of EtOH was hydrogenated a t atm temp and pressure using 2.5 g of 57, Pd-C. After 1 mole of Hr had been absorbed, the catalyst was filtered, the soln was concd, and the product was pptd with EtzO. Filtration give 3.0 g of material. Crystn from EtOAc-EtOH gave an anal. sample, mp 188-180". 1,l-Diphenylprop-1-enylamine (I).-Diethyl phosphonoacelate (106 g) was added dropwise to a stirred suspension of X'aH (15 g) in dry dimethoxyethane (500 ml) at' 0". The mixture was stirred for 1 hr after the evoln of Hz ceased, and a soln of benzophenone (91 g ) iit dimethoxyethaiie (100 nil) was then added dropwise. The mixture was stirred at, 20" for 2 hr, then poured into HZO (2000 ml), and extracted (EtfO). Evapn and distn of the ext,ract gave @,p-diphenylacrylonitrile (63 g, mp 48-49'), This (60 g ) was added dropwise to a stirred soln of LAH (20 g j in dry Et20 (500 nil) at -20". The mixture was stirred for 2 hr at --2O0 and for 15 min at 0". Excess LAH was destroyed by addn of HzO, and the organic phase was aepd, dried (RIgS04), and evapd. 1,l-Diphenylprop-1-enylamine (1) was isolated from the residual oil as its hydrochloride (58 g, nip 206-208") by addn of ethereal HCl. The hydrochloride, recrystd from EtOH-EtzO, had mp 214-216".
Notes Synthesis of 3-Trifluoromethyl Steroids'" ABR.4HlM
F. P.\SCCAL'b
A N D I\I.kNFRED
E. JvOLFF'c*
Department of Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco, California 94122 Received July 28, 1970
The function of the carbonyl moiety of C-3 of steroid hormones in eliciting the biological response has been the subject of intensive study in this laboratory. Among C-2 and/or C-3 substituted steroids prepared to test the possibility that T bonding12high electron density, or H bonding2 might be important in terms of steroid-receptor interaction have been steroidal cyclopropanes2and nitro derivative^.^ In the present study, the preparation of steroidal C-3 substituted CF, derivatives was undertaken, inasmuch as CF3 represents a center which is both electron rich and capable of H bonding. The introduction of a CF3 group by photochemical addition of CF31across a double bond has been applied to 3p-ethoxy-17p-hydroxypregna-3,5-dien-20-oneace* Address correspondence t o this author. (1) (a) This investigation was supported in part by a Public Health Service Research Grant AM-05016 from the National Institute of Arthritis and Metabolic Diseases, U.S. Public Health Service. (b) Taken from a part of t h e Ph.D. Thesis of A. F. Pascual, University of California, San Francisco, Calif., 1969. (c) I t is a pleasure to acknowledge the contribution made t o this work by Barbara Rehermann Yicente who carried out preliminary studies on the photolysis reaction as part of her h I S . research. (2) hl. E. Wolff, W.Ho, and R . Kwok, J . M e d . Chem., 7 , 577 (1964). (3) 1.1.E. Wolff and R . C. Boguslaski, i b i d . , 11, 285 (1968).
tate4 but such reactions with unconjugated olefinic steroids have not been reported. In the present study, several methods were tried in preparing the 3-CF3 steroids. Attempted conversion of a 3-COzH substituent into a 3-CF3 group by reaction with SFd4failed. Likewise, reaction of a 3-keto steroid, dihydrotestosterone acetate, with F3CMg15 yielded only starting material. Finally, employing a modified method of Godfredsen and Vangedal,6 the 3-CF3 steroid derivative was obtained. A solution of 5a-pregn-3-en-20-one (1) in CCL CF31, and a small amount of pyridine was irradiated with uv light for 8 hr under ?rT2, Only one product, 3atrifluoromethyl-4~-iodo-5a-pregn-20-one (2), was isolated from the reaction mixture although S isomeric adducts (4 cis and 4 trans) could be formed from this reaction. The stereochemistry of 2 was established from further reactions as shown in Scheme I and from the nmr spectra of the adduct and its derivatives. The C-19 Me peak of 2 was shifted 0.36 ppm downfield compared with 1 and upon hydrogenolpsis of the iodo group with LAH, the C-19 ?\'le resonance shifted 0.33 ppm upfield. As it is well known that electronegative groups produce a deshielding effect (0.25 ppm) on the C-19 Me group when they are in a 1:3 diaxial
(4) (a) TV. R . Hasek, IV. C. Smith, a n d V. A. Engelhardt, J . Amer. Chem. D.G. Martin and F. Kagsn, J . Ore. Chem., 27,
Soc., 88, 543 (1960). (b)
3164 (1962). ( 5 ) (a) R . N. Hasaeldine, J . Chem. Soc., 1273 (1954). (b) E. T. McEee, R. D. Battershell, a n d H . P . Braedndlin, J . Ovg. Chem., 88, 1131 (1963). (6) W. Godfredsen and S. Vangedal, Acta Chim. Scand., 16, 1768 (1961).
Journal of Medicinal Chemistry, 1971, Vol. 14, N o . 2 165
NOTES
&
SCHEMEI1
SCHEME I
-
&--R1
B k
-
H
P
H
9, R- OAc; R1= H 10, R OH; R, = CH,
5
1
=i
I
CHj R 1. alumina
MeOH
F3C’* 13,R=OAc;R,=H 14, R = OH; R, = CH3
11,R = OAc; Rl = H 12, R = OH;R1= CH3
I‘d-C-Hz
($p ‘\H,
F3C’”
15, R = OAC;R, = H 16, R = OH; R,= CH,
H /I01
CH, I
”
F,C”* 4a
8
relationship with the C-19 Me and a small deshielding effect (0.0-0.3 ppm) when they are in the equatorial or a-axial position,’ the iodo group in 2 must be 4P, and the product is either the trans adduct 2 or the corresponding cis isomer, 3P-trifluoromethyl-4p-iodo-5apregnan-20-one. Dehydrogenation of the last compound with base via a trans elimination mechanism should give a A3-olefin considering that the 3 proton geminal to 3-CF3 is more acidic than the 5 proton. On the other hand, a similar treatment of 2 should give a A4-olefin since only the (7) N. S. Bhacoa and D. H. Williams, ”Applications of N M R Spectrosin Organic Chemistry,” Holden-Day, Ino., San Francisco, Calif., 1964, pp 13-41.
COPY
F,,C”‘
I1
17, R = OAc; R, = H 18, R = O H R1= CH3
7
IPd-C-Hz
+
R
- -R1
H
3
1. LAH, AcZO 2. LAH
5 proton is on a trans-diaxial relationship with the iodo group. Upon refluxing the adduct in methanolic KOH,* olefin 3 was obtained as indicated in the nmr by the presence of a doublet ( J = 4 Ha) centered at 5.2 ppm. The large downfield shift (0.23 ppm) of the (3-19 Rile peak of 3 compared with that of 1 strongly suggested a A4-olefin.9 Moreover, the 19Fnmr spectrumlo showed a doublet ( J = 10 Hz) centered at 603 Hz on the high-field side of external CeH6CF3 reference. This indicates the splitting of the F atoms by the 3-H and therefore, confirms the assignment of the double bond to the 4 position. Catalytic hydrogenation of 3 in MeOH with Pd-C gave a mixture of 3a-trifluoromethyl-5P-pregnan-20one (4a) and 3a-trifluoromethyl-5~-pregnan-2O-methyl ether (4b).” The 5P configuration of 4a and 4b is evident from the nmr spectra. In both compounds, the C-19 Me peak is shifted 0.15 ppm downfield compared to the 5a isomer. This large difference of the 19-Me of 5a- and 5P-steroids is well established.’ The 5a isomer, 3a-trifluoromethylda-pregnan-20-one (8)) was prepared by hydrogenolysis of 3a-trifluoromethyl4P-iodo-5a-pregnan-200-01 acetate (6) with LAH followed by CrOs oxidation of the resulting 20P-hydroxy derivative 7. The absence of any 5a isomer from the catalytic hydrogenation reaction mixture indirectly confirms the ( 8 ) The same product mas obtained by passage of the solution through neutral alumina (grade I). (9) A double bond a t the 4 position is known t o shift the 19-angular methyl peak downfield by 0.25 ppm.7 (10) The 19F nmr spectrum \vas obtained by Dr. William Budde a t Midwest Research Institute, Kansas City, Mo. (11) The reduction of ketones t o Me ethers under similar conditions has been reported by S. Nishimura, T. Itaya, and M. Shiota, Chem. Commun., 422 (1967).
166 Journal of Medicinal Chemistry, 1971,
Vol. 14, S o . 2
orientation of the 3-CF3. Catalytic hydrogenation of A*-steroids in a neutral solvent is known to give a mixture of 3a and $3 isomers n-ith the 3p isomer prcdominating.'? Introduction of R bulky group nt the 38equatorial position increases steric hindrance oii the p side. Thus, hydrogenation of 3p-methoxy-4-cholestene occurs most15 from the a side to give 60ycof the 3a isomer.13 Conversely, a 3a-axial substituent buch as CF, should increase steric hindrance on the N d e and give the $3 iqomrr. From this evidence, the addition product of cp31 m d 3a-pregn-3-en-2O-one is 3a-trifluoromethyl-4Piodo-3a-preg1ian-20-orie (2). .I similar sequence of reactions was carried out in the androstane series as shon-n in Scheme 11. Biological Te~ting.~~--Compounds 2, 3, and 8 I\ ere tested for progestational activity 111 a Clauberg-typc tchtl' at L: level of 3.0 mg rabbit arid nere inactive at t h i i level. Conipi)unds 11, 12, 13. 14, 15, and 16 ITerc a t levels of tested for myotrophic androgenic 3.0 mg/rnt arid were inactive at this level. It TI :is concluded that n trifluoromethyl carinot assume the d e of thc 3-oxo group in progesterone and testosteroric. CY
Experimental Section" Photolytic Additions of Trifluoroiodomethane to A3-Steroid Olefins.-A sohi of 1 g of t8hesteroid olefin in 150 ml of CC1, and 2 ml of pyridine was placed in a vessel equipped with a Dry Ice roiidenser, a water-cooled lamp well, and a gas inlet tube at the bot,tom. Enough Hg was added to cover the bottom of the vessel. The s o h was chilled in an ice-salt. bat,h and treated with ail exce>s (20-30 g ) of CFJ. The system was irradiated with LIV light for 8 hr while N, was bubbled slowly t,hrough the solii. The uv light was generated by a 2OO-W, high-pressure mercury lamp and contained in a water-cooled quarts immersion well equipped with a borosilicate filter. The irradiation was interrupted every 2 hr to clean the lamp well. ilt the end of the reaction, the Dry Ice condenser was removed to allow the excess reagent to evap. The yellow s o h was washed with HyO, dried (Na,So,), and evapd to yield an oil which slowly crystd upon cooling. The product was recrystd several times from XeOH. 3~-Trifluoromethyl-4p-iodo-501-pregnan-2O-one ( 2).-3 01Pregri-3-en-2O-one (1) I * (1 g) was allowed to react with excess CF31 under the conditions described above. The product was worked up in the iisiial manner. I?ecryatn from XeOH afforded 0.66 g (4Ocl) of 2, mp 145-150". Further recrystns from MeOH gave the anal. sample: mp 156-138°; [elz2D ,5Y0 i c 1, CHC1,); iiinr (CL)Cl,) 6 4.61 (m, 4a-€1)) 2.1 (s, 2 l - H ~ ) ,1.14 (s, 19-H), and 0.6 ppm (s, 18-Ha). Anal. ( G ~ H ~ Z F J C, O )H, I. 3a-Trifluoromethylpregn-4-en-20-one(3).-Compd 2 (230
mg) was dissolved in Et&, adsorbed onto neutral A1203, aiid left standing for 2 days. ilfter elution with Et20 and recrystn from aq IIeOH, there wa.5 obtained 45 mg of 3, nip 110-14:3°. ther recrystn from the same >olveiit teni gave the mi:~l.dample: mp 143-14.-1"; [ a ] 2D2 lv54" !c 0.*5, CHCls); iimr (CUC13: 6 ,j.28 (111, &H), 2 . i (3, 21-113), 1.01 ( c , 19-HaJj and 0.ti5 ppm ( s , 18-H3): 19F iiinr (CHCl,) 603 Ha on the high-field . d e of eslernal I'hCF, (d, J = 10 Hz, 3-CFa). Ana/. (CP?H,IF30)C, II. 3a-Trifluoromethyl-5/3-pregnan-20-one (4a).---To a s o h of 100 mg of 3 in 50 ml of 3leOH was added 300 mg of Pd-C. The nrixtiire \vas hydrogenated at 0.2 kg/cm2 for 2 hr. After ; i i i add1 100 mg of catalyst8was added hydrogenation 15-as coiitd for another 2 hr to coriiplete the reactioii. After filtration and evapii of ?he m h e n t , :t mixture ( 2 spot;; o n tlcl was obtained. It w:k iepd tiy prepsrative tlr i i h g pet ether-Et20 !9: I ; colveiit syhtern. The lower halid yielded 11.6 nig of 4a: nip 104-10>o; [ a 1 2 2 u +Wi" ( ( 2 (I,:, CHC1.3); nmr (CIjC13) 6 2.1 is! %l-EI.3), 0.06 (., 1F)-H3),and 0.6ppni is; 18-H.3). A n d . (Cy2H,,F3C)j C. H. The upper halid ww recrystd from sq NeOH arid afforded 1i.6 iiig of :~~-trifluoro1iiethyl-2ll~-metho~y-.~~-pregiiaiie (4b), m p 164-165'. Fiirther recryr;tii from ncl LIeOH gave the aiinl. sample: mp 167-168": [ a I z 2+ 1~2 " i c 1, CHC1,); nnir (CIX1,) 6 3 2 4 i s ) 'LO-OCHa), 1.05 (d>J = (i IIz, 21-H4),0.96 (q, 19-H3\, and 0.67 ppni lii-l13i. A n a l . (C:,H3;F30) C, II. 3~Triflnoror.-.ethyl-4~-iod0-5a-pregnan-20/3-01 Acetate ( 6 ) . -~ 2OB-IIydroxy-T,cu-pregti-:3-eiie acetate (,;)ls (750 mg) was allowed to re:ict xith escesi CF31iinder the same miiditions described ill the general procedure. The mixture was worked lip iii t,he iisiial manner, The critde producmt w a s rerrystd twice from 1IeOII to yield 300 mg (4.";) of 6 : m p 171-172': ; a : ? ?1) +11" ! C 1. CHCl?): nmr (Cl)Cl,) 6 4.82 ini, 20a-H), 4.65 (m, 4a-H): 2.01 i-, z ~ - I I ) ,4.6.5 iII1, 4 a - ~ 11, 2.01 i., ~ O ~ - O Ai,i: ~ I (d, , J = ri m, 21-Ha), 1.1.5 is, lB-H?), :tiid 0.63 ppm (q, lX-H3). Anal. (C:?rH.,,FJO) c , 11, I. 3a-Trifluoromethyl-501-pregnan-2OJ-ol (7).--A s o h of 250 nig of 6 i t ] 50 in1 of anhyd E120 was t,reated with 300 m g of LrlH and 5tirretl for 1 day at, room temp. After destroyirig excess LAH wiih EtOAc, the mixture was treat,ed n-ith a satd iolii of Sa-I< tartarate. The Et,?O layer wit2 septl atid !he acj layer w:i~estract,ed twice with Et,0. The c~;mhined ethereal est,raci was wnahed witli H,O. dried iXia2SO;),:iiid evqld. The product x a i piirified by prepnrntive tlc aiid recrya1.d from nq LIeuCO t o give 101 mg of 7, nip 16-169'. Further recry-tn afforded the niial. ple: mi) 167-169': 11 + S o i c 1> CHC1,); n m r iCIIC1,j 6 (in, 20a-I%),1.1%(ti, J = 6 1 1 21-11?)> ~ ~ 0.S' '5, 1'3-tI.3) : l r i r l O.7,j ppm (s, 18-113). d n u l . I C22€13iF&))C, IT. 3~-Trifluoromethyl-5a-pregnan-20-one (8).-&1 suit! of .ICrO, reagent was added dropwise at room temp t o a stirred soln it1 20 ml of LIe2C0 iiiitil the oraiige color of the bed. 2-PrOII was added to destroy the ewe.-; reagent. -kfter coitcg the mixture riiicler redLiced pres>iire, F i r 0 was ndded. The ppt \vas filtered and xvnshed with IipO. ReFut u from aq MeOIL nfl'orded 6 J nig of 8, mp 113-144".
+
(5;
+
Compd 9'9 (1.35g ' xy:ii allowed t o reart Tyith excess CF3I under the conditioii- described wni isolated i t i the yield 0.6 g of 11. nip l.i4-l.i8°. 'The :tiiul. sample had mp 160D l o (C 1, CHClj~t;l!nir !CIIC13) 6 4.6 (111, 2 , 4a-H 162': and 17cu-Hj, 1.16 (F, 17-C)Ac), 1.16 is, 19-IId, a i i d 0.78 pprn (h, 18-IIa). A ~ c z[ ~C .~ ~ H ~ P FC,~H, I OI.~ ) 3~-Trifluoromethylandrost-4-en-l7~-ol Acetate [ 13).-(>onipd 11 (400mg) was dil;solved in 20 nil of EtrO and adsorbed onto 30 g of neiitral A1203(grade I ) . hft,erit was left standing for 6 hr, the column was eluted with Et20. Evapii of t,he solvent gave 300 mg of m y s t solid whirh \vas recrystd from aq MeOH to afford 148 mg of 13, nip 87-90". Further recrystii afforded the anal. sample: mp 03-95'; [ a ] 2 D2 +88" (e 1, CIIC1,): nmr (CDC1,) 6 5.28 (m, 4-€11, 4.6 (m, 17~-i-I),2.03 (8, 17-OAe), 1.03(s, 19-H3), aiid 0.82 !. C, H. ppnl ( 5 , 1X-Ha). ~ T L C G (C22H31Fd(]?) 3a-Trifluoromethyl-5~-androstan-l7~-0l Acetate (15).-To a ~ l i ofi 96 mg of 13 i l l 100 ml of MeOH was added 300 mg of Pd-C. The mixture wa, hydrogenated :it 0.2 kg 'emz for 3 hr. After fiitratioii aiid elnpii of the solvent, a solid residue w w obtd.
+
(12) L. 17. Fieser and RI.Fieser, "Steroids." Reinhold, iYew ITork, 3.. >.., 1059, [>]J 271-273. (13) hl. C. Dart and H . 13. Henhest. J . Chem. S o c . , 3563 (1960). (14) Pharmacological tests irere performed a t the Endocrine Lahoratoriep, Madison, \Vis. (15) T. hliyake, .Vethods Hormone Kea., 2 , 135 (1962). (16) L. G . Hersliherger, E. G . Shipley and R . K . hleyer, f r o c . Sac. C z p . I h o l . . U d 83, I i 5 (1953). (17) Melting points were taken u i t h a Thomas-Hoover apparatus equipped
u i t h a corrected thermometer. I r spectra were obtained n i t h a PerkinElmer 337 instrument. Microanalyses were carried out by the Microanalstical Lab., Chemistry Department, University of California, Berkeley, Calif. Optical rotations were obtained in a 0.5-dm tube with a Rudolpli photoelectric polarimeter. X m r spectra vere obtained from samples in CDCls on Varian A\-60 .I and Jeolco J S h I 4H-100 instruments, respectively (ThIS). The 1QFnmr spectrum v a s obtained from a sample in CHCla on a Varian FI.\-100 instrument using PliCF3 as external atandard b y D r . Killiam I ~ u d d ea t Midwest Research Institute, Kansas City, h l o . (18) P. Longevialle and R . Goutarel, Bull. S a c . Chim. P r . , 11, : 2 2 3 (lSG5).
.
.
~~
19) .i, J l e d . C h e m . . 6, 1-50 (1983). I
Carpio, 11. Calzada. and E .
Journal uf Medicinal Chemistry, 1971, Vol. 14, N o . 2
NOTES
167
The desired product was purified by preparative tlc and recrystd Studies on the 4-Hydroxycoumarins. from MeOH to afford 45 mg of 15, mp 117-118". Further reSynthesis of the Metabolites and Some crystn gave the anal. sample: mp 117-118"; [ a ] 2 2D +9" (e, 1, CHCl,); nmr (CDCl,) 6 4.6 (m, 17a-H), 2.03 (s, 17-OAc), 0.97 Other Derivatives of Warfarin' (s, I%&), and 0.78 ppm (s, 18-133). Anal. (Ct~H33F302)C, H. 3a-Trifluoromethyl-5~-androstan-17p-ol Acetate (17).-A soln fir. A. HERXODSON, W. M. BARKER,* AND K. P. LINK of 200 mg of 11 in 50 ml of anhyd Et20 was treated with 1 g of LAH and stirred for 1 day a t room temp. After destroying the Department of Biochemistry, Uniiiersity of Wisconsin, excess hydride by careful addition of EtOAc, the mixture was Madison, Wisconsin 53706 treated with a satd soln of Sa-K tartarate. The Et20 layer was sepd and the aq layer was extracted with Et2O. The combined Received June 22, 1970 EtsO extract was washed with HtO, dried (Na~S04),and evapd to give a gummy residue. The crude product (1 spot on tlc), 3atrifluoromethyl-5a-androstaii-17@-ol, was dissolved in 10 ml of Studies on the metabolism of warfarin [3-(a-acetonylpyridine and 2 ml of A c t 0 and left standing for 18 hr a t room benzyl)-4-hydroxycoumarin J (I) in the rat have been temp. The mixture was dild with H20 and was extracted with described elsewhere. The high physiological activity EtzO. The ethereal extract was ivashed with 5yoHC1 and HzO, dried (Na2SOa), and evapd. The product was isolated by preparative tlc and recrystd from aq MeOH to give 50 mg of 17, mp ?H ~ H 2 C o C H J 99-100". Further recrystn afforded the anal. sample: mp 100101'; [ a ] * D2 $7" (c, 1, CHCl,); nmr (CDC13) 6 4.6 (m, 17a-H), 2.03 (s, 17-OAc), 0.82 (s, 19-H6),and 0.78 ppm (s, 18-H3). Anal. (C22Hd'LIz) C. H. . ~... 17a-Methyl:5a-androst-3-en-17p-ol(lo).-To a soln of 1.1 g of I Sa-androst-3-en-17-one*o in 100 ml of Et20-THF (4: 1) was added 25 ml of 3 X 3IeSlgBr in EttO. The reaction mixture was reof the anticoagulant precludes the isolation of excreted fluxed overnight, poured into ice, and acidified with 10% HC1. metabolic products in quantities large enough for clasThe Et20 layer was sepd aiid the aq layer was extracted sical chemical identification. Therefore, metabolic fate with EttO. The combined extracts were washed with 5% studies on warfarin were conducted using [ 4-I4C]warfaNa2CO3and H20, dried (Na2S04),and evapd to give a gummy residue. The crude product was dissolved in C& and adsorbed rin sodium. The excreted radioactive metabolites from onto 50 g of neutral alumina. The column was eluted with rats treated with the labeled compound were compared c ~ H , - E t t o ( 1 : l and 1:4). Recrystn from aq XeOH afwith known compounds, using chromatographic and forded 0.6 g of 10, mp 140-141" (lit.21 mp 139.5-140") from a isotope dilution techniques. This paper describes the similar method). 3cu-Trifluoromethyl-4@-iodo-17a-methyl-5~-androstan-17~-olsyntheses of 5 compounds shown to be metabolites of (12).-A quant'it'y of 800 mg of 10 was allowed to react with excess warfarin, namely, 6-, 7-, 8-, and 4'-hydroxywarfarin and CF3I under the coiiditioiis described above. The product, was 2,3-dihydro-2-meth~d-4-phenyl-5-oxo-y-pyrano [3,2-c]isolated in the usual manner and recrystd from 3leOH to afford [ 1Ibenzopyran. The syntheses of several hydroxylated 440 mg of 12, mp 104-115". Several recrystns from hIeOH gave warfarin derivatives which were not metabolites are the anal. sample: mp 117-120"; [ a ] z D2 -7" ( c 1, CHC13); nmr and (CDC13) 6 4.6 (m, 4a-H), 1.20 (s, 17-CH3), 1.17 (s, also described. 0.83 ppm, (s, 18-H3). Anal. (C21H32F3IO)C, HI I. Chemistry.-Warfarin may be synthesized by the 3a-Trifluoromethyl-l7~-methylandrost-4-en-I7p-o1 (14).-A Michael addition of 4-hydroxycoumarin to benzalacquant,ity of 366 mg of 12 was dissolved in 50 ml of 10% H20 in etone under a wide range of acidic or basic condition^.^ hIeOH containing 2 g of KOH. The mixt,ure was refluxed for 2 The 4-hydroxycoumarin used in this reaction can be hr. After cooling, HzO was added to ppt the product. This was collected, washed with HzO, aiid dried under vacuum. Recrgstn easily prepared by the method of DickensonJ4using from hexane afforded 124 mg of 14, mp 149-131'. Further reo-hydroxyacetophenone and diethyl carbonate. Rowcrystn from hexane afforded the anal. sample: mp 149-150"; ever, this method did not prove successful for the synthe[alZ2 D +77" ( c 1, CHCl,); nmr (CDCI,) 6 5.28 (m, 4-H, 1.20
sis of any of the desired dihydroxycoumarins from appropriate dihydroxyacetophenones, possibly due to the 3~-Trifluoromethyl-17a-methyl-5p-androstan-l7~-01(16).-To extreme insolubility of the disodium salts in the reaction a sohi of 100 mg of 14 in 75 ml of MeOH was added 300 mg of solvent, benzene. Blocking the nonortho OH group Pd-C. The mixture was hydrogenated at 0.2 kg/cm2 for 4.5 hr. by formation of a benzyl ether provided an intermediate I t was filtered and the filtrate was evapd to yield a solid residue. The residue was recrystd from hexane twice to afford 38.6 mg of which would undergo the desired reaction with 16, mp 189-190". Further recrystn from hexane afforded the (EtO)2CO. anal. sample. mp 189-190", D -4" ( c 1, CHC13); nmr Therefore 6-, 7-, and 8-hydroxywarfarin were synthe(CDCl,) 6 1.22 (s, 17-C&), 0.98 (s, 19-&), and 0.83 ppm (s, sized as shown in Scheme I. 18-H3). Anal. (C21Ha3F3O) C, H. The synthesis of 5-hydroxywarfarin, kvhich could not 3~-Trifluoromethyl-17a-methyl-5a-androstan-l7p-o1 ( 18).-A soln of 100 mg of 12 in 30 ml of anhyd Et20 was treated with 400 be achieved by Scheme I, proceeded by the route shown mg of LAH and stirred overnight a t room tern,?. After destroying in Scheme 11. the excess hydride with EtOAc, the mixture was treated with a It was found later that this method of synthesis also satd soln of Ka-K tartarate. The E t & layer was sepd, and the aq gave 6-, 7-, and 8-hydroxytvarfarin in considerably betlayer was extracted twice with EttO. The combined extracts were wabhed with HtO, dried (Na2S04),and evapd. The product was isolated by preparative tlc and recrystd from sq MeOH to give 31 mg of 18, mp 131-134". Further recrystn from 3leOH gave the anal. sample: mp 134'; l 0 1 1 ~D~ +2" ( c 1. CHCI?):nmr TCDCl,) 6 1.21 (s,-17-CHi). 0.85 (s, 19-H3), and 0.82 ppm (s, 18-H3). A n d . (C21H33F,0) C, H. (20) 5ol-Androst-3-en-17-one was obtd from 5,-androst-3-en-178-01 acet a t e (9) by hydrolysis of t h e 17-hcO group with 2% MeOH-KOH followed by COa oxidn of t h e corresponding 17-OH group. (21) P. D . Klimstra, U. S. Patent 3,166,578 (1965); Chem. Abstr., 63, 9307c (1865).
*
T o whom inquiries concerning this work should be addressed a t the Pharmacology Department, International Minerals a n d Chemical Corp.. Libertyville, Ill. 60048. (1) T h e work described herein was submitted in partial fulfillment of the requirements for t h e Ph.D. degrees of M . A . Hermodson and W. M . Barker, University of Wisconsin, Madison, Wis. (2) W. hl. Barker, M . A . Hermodson, a n d K . P. Link, J . Pharm. E z p . Ther., 171, 307 (1970). (3) C . Schroeder, P h . D . Thesis, University of Wisconsin, Madison, Wis. (1955). (4) H. G. Dickenson (to Ward, Blenkinsop a n d Co., Ltd.), cyclic lactones, U. S. Patent 2,449,162, Sept 14, 1948 [Chem. Abstr., 48, 694 (1949)l.
