NOTES
July 1966
The ketone I11 is converted into 17a-ethynyl-1,4dimethylestra-l,3,5(10)-trien-l7/i?-ol (IVa) with either potassium acetylide or the lithium acetylide-ethylenediamine complex. When heated with acetic anhydride and pyridine, IVa gives the acetate IVb, while successive carboxylation and hydrogenationla transform IVa into the spirolactone, 3-(17P-hydroxy-l,4-dimethylest ra-l,3,5(10)-trien-17 a-yl)propionic acid lactone (V). Experimental Section6
1,4-Dimethylestra-l,3,5(lO)-trien-17-one (III).4-To a stirred solution of 20 ml of 3 J!f methylmagnesium bromide in diethyl ether and 200 ml of anhydrous ether was added a mixture of 10 g of androsta-1,4-diene-3,1i-dione17-ethylene ketal7 in 700 ml of anhydrous ether. The reaction mixture was stirred and heated under reflux for 2.5 hr. Then it was cooled in an ice bath and decomposed with a saturated solution of NH'CI. The ether phase was separated, washed successively with water and a saturated solution of XaC1, dried (Na2S04), and distilled to dryness under reduced presslire to afford a viscous oil. The oil was treated with a solution of 9 ml of 6 S HC1 in 90 ml of 957, ethanol. The mixture was stirred a t room temperature for 0.5 hr. Then it was diluted wit,h water and extracted with ether. The ether extract was separated and worked up as previously described. T h e resriltant viscous oil was chromatographed on TO0 g of silica gel. Elution of the column with benzene gave 3.3 g of a colorless crystalline product, which melted a t 126-127' after crystallization from ether-pentane (lit.' 126-128'). It proved identical with an authentic sample of IT' prepared by the Cr03 oxidation' of l,Pdimethylestra-1,3,8( lO)-trien-17p-o1 (111)" obtained from either 17p-hydroxyandrosta-l,4dien-3-one( I a ) or its acetate (Ib). 17~-Ethynyl-1,4-dimethylestra-1,3,5(lO)-trien-l7~-ol (IVa). A,-Acetylene was passed for 1 hr into a solution of 15 g of potassium t-butoxide in 200 ml of t-butyl alcohol and 100 ml of toluene, stirred, and maintained in a n ice bath. A solution of 1.6 g of I11 in 40 nil of toluene was then added as acetylene was still being passed into the solution. After 4 hr, passage of acetylene was stopped. The reaction mixture was stirred for an additional 12 hr during which time i t was permitted to warm to room temperature. The reaction mixture was then diluted with a large volume of a saturated solution of NH4Cl. The organic phase was separated, washed with water, and distilled to dryness under reduced pressure. The residue was chromatographed on 80 g of silica gel. Elution with 27, ethyl acetate in benzene gave Va, which was crystallized from ether-pentane, mp 170.5-172', yield 0.31 g. T h e melting point was raised to 174-174.5' on recrystallization from ether-pentane; XKBr 3425, 3311, 3289, 801 em-'. A n a l . Calcd for C22H280: C, 85.66; H, 9.15. Found: C, 85.87; H, 9.22. B.-A mixture of 1.36 g of 111, 2.7 g of lithium acetylideethylenediamine,E and 100 ml of tetrahydrofuran (freshly distilled from methylmagnesium bromide) was stirred a t room temperature for 2 hr. The reaction mixt,ure was decomposed with a dilute solution of NH&l and then extracted with ether. T h e ether extract was washed successively with water and a saturated solution of SaC1, dried (Sa&04), and distilled to dryness under reduced pressure to afford a viscous orange-red oil. T h e oil was chromatographed on 180 g of silica gel. Elution with benzene, followed by crystallization of the solid product from ether-pentane, afforded 0.54 g of IVa, mp 171-171.5', which was identical with that obtained by procedure A. From the mother liquor an additional 0.12 g of IVa, mp 164.8-167.5O, was obtained. 17~~-Ethynyl-1,4-dimethylestra-1,3,5( lO)-trien-17p-o1 Acetate (IVb).-A solution of 0.78 g of IVa, 6 ml of pyridine, and 8 ml of acetic anhydride was maintained at 95' for 18 hr after which time it was concentrated to about 2 ml by distillation under reduced pressure. The residue was diluted with ice water and extracted with ether. The ether extract was washed successively ( 6 ) Melting points were taken on a Fisher-Johns melting block and are corrected. ( 7 ) 11.J. Gentles, J. B. Moss, H. L. Herzog, and E. B. Hershberg, J . A m . Chem. Soc., 80, 3702 (1958). (8) 0 . F. Beumel, .Jr., and R. F. Harris, J . Org. Chem., 28, 2775 (1963); ibid., 29, 1872 (1964).
GO3
with a 5% solut,ion of N a H C 0 3 and water, dried (Sa,SO,), and dist'illed to dryness under reduced pressure. The residual viscous oil was chromatographed on 80 g of silica gel. Elution with beuzene gave a colorless viscous oil. This was evaporatively distilled a t 180' (0.1 mm) to afford I V b as a colorless amorphous product, XKBr 3311, 2119, 1761, 1892, 1250, 1241, 1229, 806 cm-'. Thin layer chromatography indicated that the product was homogeneous. A n a l . Calcd for C.XH&: C, 82.24; H, 8.63. Found: C, 82.17; H, 8.65. 3 4 17p-Hydroxy-l,4-dimethylestra-l,3,5( lO)-trien-l7a-yl )propionic Acid Lactone (V).-To a stirred solution of 15 nil of 3 J I methylmagnesium bromide in diethyl ether and 25 ml of tetrahydrofuran ( T H F ) was added over a period of 10 min a solution of 2.4 g of IVa in 30 ml of T H F . The reaction mixture was distilled to remove the diethyl ether. Then it was stirred and heated under reflux under S2for 15 hr. The cooled reaction mixture was stirred for an additional 22 hr while CO, was cont,inuously passed into it. A11 ice-cold, dilute solution of H2S04 was added. The resultant solid was collected, washed well with water, and dried, mp 105-110". A solut'ion of the solid (2.7 g ) and 0.83 g of triethylamine in 100 ml of 95% ethanol was hydrogenated over 0.3 g of 5% Pd-C at room temperature and atmospheric pressure. After the calculated amount of hydrogen was absorbed, hydrogenation was stopped. The filtered solntion was distilled to dryness under reduced pressure. The residue was triturated with dilute HC1 to afford a colorless crystalline product. The product was collected, washed well with water, and dried. Crystallization from methanol-water gave 1.65 g of 5'; mp 200-202.5"; kKBr1792, 159.5, 804 cni-1; [ o ~ ] ~ O D $8.9" ( C 1, CHC13). Anal. Calcd for C43H3002: C, 81.61; H, 8.93. Forind: C, 81.74; H, 8.85.
Synthesis and Pharmacological Evaluation of a- Substituted 1-Naphthylacetic Acids GIANFRAKCO PALA, TIBERIO BRUZZESE, >IARIIZZI-UBERTI, AXD GERMAXO COPPI
ERNEST.4
Research Laboratories, Istituto De Angeli S . p . A . , Milan, Italy Received J a n u a r y 4, 1966
Continuing our investigation on the pharmacological propert,ies of a,a-disubstituted 1-napht,hylacetonitriles and 1-naphthylacetamides,' we have prepared 21 a-substituted 1-naphthylacet'ic acids for pharmacological screening, including studies of acute t,oxicity and ant,iinflammatory, antipyretic, analgesic, diuretic, choleretic, and hypoglycemic action, as well as the in uitro ant,ibacterial and ant,ifungal activities. The monosubstituted acids were prepared by hydrolysis of t'he nitriles with dilute sulfuric acid (see Experimental Section, methods A and B). This procedure failed to give t'he disubstit'ut'ed acids, only the corresponding amides being obtained as previously reported. l'' However, these acids were easily prepared by reaction of the amides with isoamyl nit,rite in glacial acetic acid, in the presence of HC1 (method C), as described for aalkyl-substituted 1-naphthylacetic acids.2 ,411 the acids were isolated and identified as the hydrochlorides (Table I ) . The results of the pharmacological screening are reported in Table 11. Compared with the corresponding (1) (a) S. Casadio, G . Pala, E. Crescenzi, T. Druzeese. E. MarazziUberti, and G . Coppi, J . .\fed. Chem., 8 , 589 ( 1 9 6 5 ) ; (b) S. Casadio, G . Pala, T. Bruzzese, E. Crescenzi. E. hlarazzi-Cberti, and G . Coppi, i b i d . , 8 , 594 (1965); ( c ) G . Pala, S. Casadio, T. Bruzzese, E. Crescenzi, a n d E. Maraazi-Uberti, i b i d . , 8 , 698 (1965). (2) S. Casadio, G . Pala, T. Rruzzese, and E. Maraezi-Uberti, F Q ~ ~ Q C O (Pavia), Ed. Sei., 17, 797 (1962).
Experimental SectionJ Chemistry.-~-Thc~ii1ieriiiedi:tte iiirrileq :illti :iiiiiil(i> were p~ep:iretI as iw.etitly reportecL1:1~', T h e title c-orlipouiids n w e ol)taiiied by three differelit method; \r.hich ;ire well illusirateti l)y the followiiig iiiethocls h, 13, :itid C,'. They :ire listed i i i T:ik)le I, nl(iirg wit.h yield,*, p h y s i c d c.onstarlth, :iird ai1alytirnl ciala. Method A. ~-(2-Morpholinoethyl)-l-naphthyIaceticAcid Hydrochloride (XVI). -.-CY-( 2-~1orpholiiioethyl)-l-naphthylacdonitrile (50 g, 0.176, nii)lc) IWY hydrolyzed by refluxing for 2 h r with a 1: 1 : L mixture of ('oiicenttxted 1-12804,glacial acetic. :icitl, atid water (19.5 1111). The renctiotl mixture w w cooled to roorii temperature, dilutetl \Tit11 water, :iiiti w ~ q u e o u1:ryer ~ W B S made :ilkalirie with 30 with ether a i i d ac:itiified i o p I T 1 wit1 solutioti TVBS pvapor:itcd t o dryiiesy utider redured pressure, n ~ i d the residue w i s extrac>ted four times with boiling eth:tnol (800 rirl ). The c-oinhiried extrticts werr twiporai ed to dryness : ~ n d llizetl frciiri ! ) O r ; ctIi:iiiol, giving rolorltw Method B. a-(2-Piperidinoethyl)-l-naphthylaceticAcid Hydrochloride [ X ) . ---u-( 2 - Piperidiiioel hyl)-l-iiaphthyl:i~~e~~it~it rile ( 4 0 g, 0.144 t m l e j was f i r 4 hydrolyzed :is deswibed iir inethod A . The reartion rnixture its theii cwoled, diluted with water, w-:ishetI with ether, aiid riiticle alkalitie n.ith 30c; NaOH. h i oil sep:irated which w a s w:r-hctl with ether, a i i d then coiiceiitrnted €I('] w:ih added t ( J pH 1 . The oil>- layer was agaiii separatecl a i r t i di.solved i i i 2-pr(ip>LlllJl,the ~ i ~ l i i t iwas ~ i i dried (CaCly), :tnd e t h w w:is 1hen added. Thr tarry precipitate obtniiied wv:w i q lized from 2-pi~op:iii1~l, giviiig :I r.olorlcss ( 107-.199".
Method C. ~-lsopropyl-a-(2-dimethylaminoethyl)-l-naphthylacetic Acid Hydrochloride (IVL-Hydrogel1 chloride v-:I~ &)wIy bubbled for 1.;ihi., :it roorii teiiiperature, through B c~)oletl ~~rliitiotii ) f ~ r - i ~ r 1 ~ ~ 1 ~ 1 1 ~ ~ ~ I - n - ~ ~ - d i r ~ 1 e t h y l a m i 1 1 o e t h ~ l ) - l - r ~ a p h
G 05 T.\BI,EI1 P H I R ~ I I C O L O G I C A LSCREENING
Compd I I1 I11 IV V vI VI1 VI11 IX X XI XI1 XI11 XIV
XV SVI XVII XVIII
XIX XX XXI 3Iorphine'HCl Phenylbutazone Hydroc hlorothiazide Dehydrocholic acid Chlorprops mide
Approx LDao (mouse), mg /ka ip
1120-1 230 560-630 1090-1 2 10 >so0 540-610 580-620 510-590 290-320 550-620 45-70 1100-1260 5T0-640 580-820 590-650 280-310 1140-1220 410-450 :390-410 1170-1250 590-630 > I600
Analgesic act. (mouse) Increase of reaction time, mg/kg
'lo"
1P
14 21 37 59 .X 8 38 58 10 28 12 23 34 50 14 16 34 36 13 28 9 6i 61
200 100 100 200 200 100 200 200 200 25 100 200 200 200 50 200 100 30 100 100 100 5 100
Hot plate test, 1 hr after treatment. hr after treatment. e Equimolar doses. Q
Antipyretic act. (rat) Antiinflammatory Max activity (rat) temp Inhibition deof edema, mg/kg crease, ma/ke: Ip OCC 5% ip
22 47 3T Inactive 26 50 Inactive 37 30 Inactive 45 Inactive 38 Inactive 18 Inactive
200 100 100 200 200 100 200 200 200 25 100 200 200 200 50 200 100 50 100 100 100
2 2 1 4 2 2 2 2 2 4 1 4 3 2 3 1 3 4 0 8 2 9 2 4 2 7 3 3 0 9 2 1 1 3 0 i 1 6 2 7 1 7
200 100 100 200 200 100 200 200 200 25 100 200 200 200 50 200 30 100 100 100 100
18
100
1 6
100
30 13 13 25
17
RESVLTR
Iliuretic activity (rat) Test volt,/ control mdkp vol PO Inactive 1.15 Inactive 1 29 1 19 1.1.5 1.12 Inactive Inactive 1.30 Inactive 1.27 1.13 1.14 Inactive Inactive 1.18 Inactive Inactive Inactire Inactive
1.56
50 50 50 50 50
.5 0 50 50 50 50 50 50 50 50 50 50 50 50 50 50 30
Choleretic activity (rat) Increase of bile flow, 7cd my'kge 1 hr 2 hr id 32
18
Hypoglycemic activity (rat) Blood sugar decrease, m g / k g
'Zf Inactive
i3
14
Inactive
Inactive
80
Inactive
36
83
9i 134 92
39 131 78
28
43
120
92
42
100
83 87
Inactive 13 :3 5
100 100 50
38 Inactive
50 100
Inactive Inactive Inactive 11
50 100
12 Inactive
100
(4) P. Iheresidue, giving a solid product which, on crystallization from ethanolligroin ( b p 75-120°), gave colorless crystals, mp 2277228". Pharmacology.-The acute toxicity, and antiinflammatory, analgesic, and diuretic activit,ies were investigated according to the techniques previously described.'& The antipyretic action was studied in rats made pyretic by brewer's yeast, according to Smith and Hamb0urger.l The activity on the choleresis was investigated in rats, using the biliary fistula technique of 3larazziUberti and Turba.j The hypoglycemic action was measured in rats, according to the procedure described by CeriotL6 The antibacterial and antifungal activities were measured against Micrococcus pyogenes z,ar. aureus ATCC 6538 P, Bacillus subtilis ATCC 6633, Escherichia coli McLeod ATCC 10,536, Salmonella typhi T 30 Roma 31 507, and Candida albicans ATCC 10,231, using the serial dilution technique.' All compounds were administered as the hydrochlorides in aqueous solution. Morphine, phenylbutazone, hydrochlorothiazide, dehydrocholic acid, and chlorpropamide were used as standards for comparing the analgesic, antiinflammatory-antipyretic, diuretic, choleretic, and hypoglycemic activities, respectively .
PO
100
d
50
S-aliie+ a t 1 and 2
Effect of Organic Compounds on Reproductive Processes. 11. Alkylating Agents Derived from Various a,w-Alkplenediols \T. A. SKIWER, J. H4Y-FORD, T . E. SHELLESBERGER, A\D
IT. T . COLWELI.
Life Scaences Research, Stanford Research Znstzticte, .lienlo P a r k , Calzfornia Receaved January 10, 1966
A program of synthesis of alkylating agents having related carrier moieties but with a variety of alkylating functions has been under way in our laboratories. These compounds are being evaluated for their effect on reproduction in the housefly (Musca doiizesfia L.), mice, and ,Japanese quail. Previous studies have shown that ~~,~'-bis(aziridiriylacetyl)-l,8-octaniethyle1iediamine' inhibits reproduction in the housefly at 1 arid 0.1% concentration when added to their feed. I t was of interest to see whether similar conipounds derived from a,@-alkanediols would also affect their reproduction. The compounds synthesized (2-21) are shown in Table I. The bisbronioacetyl esters (2, 6, 10, and 14) were best prepared by the addition of diol to chilled bronioacetyl bromide. The iodo derivatives (3, 7, 11, and 15) mere prepared from the bromo compounds using S a 1 in acetone. The aziridinyl derivatives (4, 8, arid 12) were derived froin the bronio compounds by the addi(1) IT-. A. skinner, H. C. Tone, T E. Shellenberger, and G. \V Nevell, J . ,\fed. Chvm , 8, 647 (1965).
