Table 11. 112 Vitro Antimicrobial Activity of ~,5-Dihalo~i\iir(ile-2-carboxylic Acid Derivatives
could be conclusively identified by comparing the nmr absorptions of the ethylidenedioxy groups. In 90 compounds ( c . g . , 7a) the absorptions appear a t M I C , ' fig ~~1 higher field than in their 93 isomers ((>.;I., 7b) as a result Stuph. P. E. P. of 3 downfield shift in the latter c;ise, which i:. to be asComyd uureus uerug" co: i ' uulg,i cribed to a diamagnetic effect of t h e aromatic, .A ring in 8 7.8 250 15.6 125 the ( b e n t )93 31.2 62.5 10 3 .Y 250 After protection of' the 20-keto and the ;i-hydroxyl groups 12 1.95 500 31.2 250 hy conversion to t,he corresponding ethylenedioxy and 13 3.9 > 125 31.2 125 methoxy groups the products were subjected to a Birch 18 0.62 125 125 >125 19 0.31 7.8 250 62.5 reduction6 and hydrolyzed to l h , t;ct-alkylidenediosy7 8 250 20 0.12 > 125 19-norprogesterone derivatives. In t w o cases thew 23 2.5 > 125 >125 >125 products were converted to 3-enol ethers. T h e whole rea(.24 > 100 > 100 > > 100 tion sequence is outlined in Scheme I and described in d e . 25 > 100 > 100 >loo tail for the ethylidenediox?- compound in the I5xperinic.n36 >125 31.2 >250 tal Section. T h e end products arc' listed in Table I with 38 15.6 >62.5 > 12.5 >125 39 3 .9 >12%5 7.8 > 125 some pharmacological d a t a . 41 7 .S >62 5 >250 >12.5 LVe assigned I he Y.?.l o c i configuration t o the Y l j - 1'3-ntir42 0 .Y7 >62.6 >250 >125 progesterone rierivati\.es on t h e following considerations. 46 15.6 > 125 12: >125 From Dreiding models ir is obvious that 14-:i-ketosteroids 47 .9 165 > 125 >125 with a 9d.103 configuration are only possible with ring €3 48 0 ,075 >62 . 5 >I26 >l2>j in the boat form which is thermodynamically unj'avorahle. 52 0 .8 >1Xi >250 >125 ~~___ _ _ _ _ ~ It is known that such steroids rearrange under ac,idic ('om Minimum inhibitory concentration. '' Pseudomorms ditions to the 93.10tk configuration i,ia a 1 5 ( lO)-:i-ketosy.-. c7cv-uginoso. Escherichici coli. Proteus uuigaris. tem.7 Since our 93 derivatives were fhrmed under strong11 acidic conditions from 1 0 ) steroids their roni'igurat ion> must be 93.IO(?. The crude material was crystallized from EtOH-H20 to give 14.2 On one occasion there was a little complication in t lit' g (78% yield) of white powder. m p 227--228". reaction sequence. After Birch reduction of the benzylideneAcknowledgment. We are grateful to Drs. W. A . Goss, dioxy compound it appeared that the benzylidenedioxy :\n.. ticins or suspensions in maize oil (50 mg/ml f o r rat:: h .?I, 100 (19711. mgiinl f'or rabbits). The rats used in most of these experi( 3 ) W. A. Goss and E. B. Cimijotti. Appl. Microhioi.. 16, 1414 j 19681. ments were the \Vistar-derived C p h strain f'roni TNO ( 4 ) A . A . Larson. C . hloore, .J. Sprague. B. Cioke, .J. hloss, and .J. (Zeist, Holland); i n some experiments (indicated in Table 0. Hi~ppe.,J.A m c r . ('hem. Soc , 78, 3210 (1956). 1) the strain "Orga (-'ph" from the same institute ~ i i h used. The rabbit s used were the "Bastard" strain bred b y TSO. 'The procedures used for testing the anticoncept ive and progestational activities were essentially t h e same as rt,l k t , 17tr-Alkylidenedioxy Steroids. 2. 19-Nor-9n,10~- ported ear1ier.l The resulth are summarized in Table 1. and Ig-Nor-gP, loa-progesterone Analogs .Just as the 19-methyl derivatives.I the li)-noralkyiidenedioxyprogesterones show 2111 optimum o i activith- in Arthur F. Xlarx, Herman J . Kooreman. Kapil D. Jaitly. the region of the propylidcnedioxy compounds (I< = Et: and Dirk van der Sijde* 12a and 12b). The very long pregnancy delay prodiiced by (:i.\t-Hrocades A'. L'., Research and Det,elopment Dit,ision, I 6 iR = Phi i y striking since t h e 19-methyl analog' did l ~ e l / t ,7'hc NethPrlnnds. Rpceiced Februar? 2.In%? not possess such acti\-ity. The activities of the % t and HwO(
17
1 . 4 (0.1) 0 . 1 (0.1) 2 . 2 (0.1)
3 . 5 (0.1) 2.8 (0.01) 4 . 0 (10) 3 . 8 (10) 3 . 5 (10)
(lo), 18
0 . 1 (1)
1 2 (10) 18 (lo), 24
15
3 . 0 (0.1) 0 . 7 (0.1)
“ A n a l y t i c a l results w e r e w i t h i n 0.4% of t h e t h e o r e t i c a l values. ( I n all c a s e s C and H w e r e d e t e r m i n e d . ) * T h e p r e g n a n c y d e l a y is g i v e n in d a y s following a single sc i n j e c t i o n (or oral a d m i n i s t r a t i o n ) of t h e d o s e s m e n t i o n e d in p a r e n t h e s e s in m g l r a t . ? T h e M c P h a i l score w a s d e t e r m i n e d a f t e r d a i l y sc i n j e c t i o n (or oral a d m i n i s t r a t i o n ) , for 5 d a y s , of t h e doses m e n t i o n e d i n p a r e n t h e s e s in m g / r a b b i t . “‘Orga rats” were u s e d i n s t e a d of “ W i s t a r rats.”
_
~
~
rated in L ' U C U O . The residue (450 g ) was crystallized from 1.25 I. ol MeOH to yield ,316 g i77.570) of 5: m p 230-235": uv 264 nm ( ( , 1605, 1675, 1490 cm !C-=-C arom); 5 (1. s. 4 HI. 6.6-7.5 ppm ( 2 . A - H , I and 2 H). :%,I . i l k , 1 Sa-Trihydroxy-19-norpregna-1,3,3(10)-trien-20-one (6a). A mixture of 714 g o f t h e tetraene 5 , 14 1. of CH2C12. 7 I. 01 MeOH. and 180 g ot' 107~Pd on C was hydrogenated at atmospheric pressure. After 3 hr at room temperature the steroid was conipletely hydrogenated to a n approximately 2: 1 mixture of the isomers 6a and 6b. After removal of the catalyst by filtration the snlution was evaporated in ~'ucuo.The residue was crystallized from :iI . of MeOH. The bulk of the 9d isomer 6b was obtained in practically pure crystalline form (136.5 gi. The mother liquor was evaporated in L ' U C U O and the crystalline residue was treated with :X:l I . ol boiling CeHe. After cooling the bulk of the 9n isomer 6a was obtained in nearly pure form (307 g ) . The latter product was recrystallized from CcH6 and from MezCO to yield 234 g (42.6%i i i 1 pure 6a (From mother liquors a second crop of 14.5% was ~ 1 1 tained. I : m p 214--248": u v 216 nni ( f 87.50). 281 (23001. 2% 12100!. :i,1 kt, 17n-Trihydroxy-19-nor-9d-pregna1,3,3(10)-trien-20one (6b). The tirst crop from the preceding experiment (ls36,5g~ was recrystallized from CH2C12-MeOH T O yield 12'. g 117.87~) of pure fib (From the mother liquor, combined with tnose from the preceding experiment. ii second crop of :1.87~was obtained.!: m p I67 ItiY0; uv 217 nm I I 6700). 221 (60001. 281 (1800),2-55 (1600). :]-Hydroxy-1&,, I7tu-ethglidenedioxy-19-norpregna-1,3,5(10)trien-20-one(ia).A mixture of 38.5 g ((1.18 mol) of the trihydroxy compound ria, 360 nil of dioxane. 360 ml ( 2 . 7 moll of paraldehyde. anci :1 in! of 70% HC104 was stirred for 15 rnin at room temperatrire. The reaction wai stopped by the addition ~f 100 ml of saturated XaHC'O3 solution and the produ vas crystallized h y the addition of 2 I. of water to yield 5 9 . j p (9 % 1 of ia: m p 200-201': nmr I . : I R (3, d . J = 5 cps. Me acetali. 5 i ppm ( 1 . q . ./ = 5 cps;. H acetal i. :%-Hydroxyl h , I Sa-ethylidenedioxy-19-nor-9d-pregna1,:1,5(10)-trien-20-one (7b). A mixture of 42 g (0.097 moll of 6b, 200 mi of' dioxane. 200 ml 11.5 mol) of paraldehyde. and 1.7 ml of 70% HC104 was stirred for 15 niin at room temperature. After treatment of the mixture with N a H C 0 3 solution the product was crystallized by the addition of 2 I. of water to yield 28.9g of crude Sb. f'ry>tallization from a 1:l mixture of hleOH and water yieldc a d pure 7b (24.7 g. 71%): mp 177-180": nmr 0.88 (:{. d , ./ = ,5 cps. >le acetal!. 4.93 ppm (1. q , .J = 5 cps. H acetal,. 3-Methoxy-1 Jn, 17a-ethylidenedioxy-19-norpregna1,4,d( 10)trien-20-one (Xa). T o a stirred mixture of 50,68 g (0.14 moll ot the :i-hydroxy compound 7a, 900 mi of CHC13. and 250 ml of K O H solution was added at 20"in 5 min 50 ml (0.53 mol) of I., and the mixture was stirred at c.a 15'. After 40 min a further 80 ml of the KOH solution and 16 ml of MezS04 were added and the stirring was continued for l..5 hr. The organic layer \vab separated, washed with water. and evaporated i t 1 cncuo The residue \vas rrystallized from 800 mi of 50% SleOH t o yield 50.9g (96.6%I i ) f Sa. An analytical sample \\as obtained by recrystallizatiiln frum EtOH: m p 16:3-164', :I-Methoxy-lkr,1 Sci-ethylidenedioxy-20,20-ethylenedioxyI+norpregna-l,3,,5(lO)-triene(9a). A mixture of 1.5 1. of CsH6. 3 : i n 1 1 ~ 1t C H 2 0 H ) 2 . and :3 g of p-toluenesulfonic acid was reflusrd with energetic stirring for 1 hr under N2 ciu a water-absorbinp trap il'illed with C:aC12 and a molecular sieve,: 59.9 g of the 2O-keto compound 8a was added and the mixture was rel'luxerl i n the hame way t'or 28 h r . After cooling the layers bvere separated. the glycol laver was extracted with 250 ml of C&. and the ccimhined extracts tvere washed with TaHC0:j solution and water. 'Yhe solution was concentrated ;n t'actcij (with a drop 01' pyridine) I O 250 ml and the product was crystallized by addit i o n ot heptane: yield 31.30 g (47.0%). For analysis a sample was prt>paredhy crystallization from hIezCO: m p 182-18-1". ,'bMethoxy-140,litr -ethylidenedioxy-20,2O-ethylenedioxy19-norpregna-2,5(lO)-diene (loa). Into a reaction vessel with gas inlet. stirrer. dropping fiinnel, and reflux condensor (with C o n acetone). closed with drying tubes. NH:%gas. dried over CaO. was introduced, until t ' n . 850 ml was condensed in the vessel; 200 ml 01 d r y T H F and 4.6 g ( 0 . 2 mli of Na cuttings were added. After seine niinittrs stirring of the intense hlue solution. a solution of 5 g ((1.012 mol! uf the aromatic compound 9a in 4 ml 10.068 mol) of absolute EtOH and 150 ml of dry THF wab added cia the dropping funnr.1 in 1 hr. The mixture was stirred for 0.5 hr and then a mixture ( i f ' X ml (0.135 moli of EtOH and 20 ml of T H F was added in 15 m i n The blue color disappeared and the mixture was niaintaiiied at the distillation temperature of S H 3 for I hr. Then
11 g ( t u . 0 . 2 mol, of powdered NH4CI was added and the SH3 was allowed to distil off. The mixture was diluted with 7 5 n i l of water and the product was filtered and washed with ~ a t e r yield 5 . 0 g (10070) of loa. From uv measurement at P 7 nm 1 ' c l l : peared that this product contained cn 15% of the aromatic c i i n i pound 9a: ir 1694. 1668 c Mel. l..'ll i:i.C . 21 hlel. 1 s , Ohlei. 3.95 r-l. rn, CH = .-) qx,H acetal. together n i t h small absorptioni due t o !]ai. 1 In,17~t-Ethylidenedioxy-l9-norpregn-J-ene-:l,20-dione ( I 1 a). Product 10a \vas hydrolyzed by refluxing 5 g nf' the s t m i i d in I I mixture of 200 m i of hleOH and 60 ml ot 6 .\ HC'I d u r i n g 15 min. The solutirin \vas cooled and neutralized n i t h g ( i t NaOAi. and water. The MeOH was removed distillation in ti0 m i the concentrate extracted ivvith isohu 1 methyl ketcinr. Tht, P Y tract was washed with nater and eva irated in c'ai'ur,. The w h i due was chromatographed ( i n si1ii.a gel ivlth C',;HI; \lc'z('O ~
1711 ( 2 0 c = - 0 1 . 16ti9 20-one (17a).A mixture of 20 ml of dioxane. 2 ml iO.00125 m i i l i ot triethyl orthoformate. and 100 mp I J I p-toluenesulfnnic acid \'i a s stirred tor 1 hr at rooni temperature: 2.5 g 10.0067 inoh ot l l w I;n-propylidenedii~xy-l9-norpregn-4-ene.:~,2~ 1-diiinr. I 2 a 1 an(I 1 ml r ~ t triethyl orthoformate w r e added anti the iiiixtuic- \va. stirred for 1.3 h r at room temperature. T h e mixture uai n e u t r a ired a i t h 11.1 nil of I,>ridinr and the product \vas crystallized ti> addition
