Synthesis of Potential Biodynamic Drugs

SYNTHESIS OF POTENTIAL BIODYNAMIC DRUGS

(SUMMARY)

A THESIS SUBMITTED FOR THE DEGREE OF Doctor of PhjIosophM IN Chenalstrq TO THE ALIGARH MUSLIM UNIVERSITY, ALIGARH

BY AKHIL Q. JHINQRAN

DIVISION OF MEDICINAL CHEMISTRY CENTRAL DRUG RESEARCH INSTITUTE LUCKNOW - 226001 JUNE, 1983 SUMMARY The work embodied in the thesLs is concerned with the synthesis of some potential post-ooital antifertiliiy agents, spermicides and antidepressants. This work is described in four chapters. In the first chapter the present status of seco-steroids is re-viewed as this topic has not so far been properly reviewed. The second chapter describes the synthesis of some 5»6-seco-estradiols and their antiimplantation activities. In the third chapter the synthesis and bioevaluation of N-substituted a-(2-aminoeth5'-l)acrylophenones and 3-amino-2-substituted met]iyleneindan-l-ones as possible spermicides has been described. The fourth chapter consists of the synthesis and pharmacological evaluation of some iminothiazolidine derivatives as potential antidepressants.

CMj£tGr__I

The sex steroid hormones, estrogens, progestogens and androgens play an important role in the regulation of mammalian reproductive system. Compounds which could interfere with the action of ihese hormones could in principal block conception. One approach to 1iie design of such agents is to build analogues of these hormones, yjtilch. lack one of the bonds in different rings of the tetracyclic stiucture of the steroid (seco-steroid); puch compounds having greater flexibility would thus be able to assume a larger number of discreet conformations, only one of which, would mimic the prototype and would Uius have altered affinity for "the receptors involved and hopefully a changed biological activity profile. In this approach a number of 5»6-seco-steroids have been synthesized and "their contracoptLve activity studies are described in Chapter II.

_Gha£;fcer_n

Syniai_esis of 5,,6-sec(>-estradiols 1~"^ In earlier work -^ carried out in tliis laboi'atory 2a, 63~diethyl"3g"(P"hydrcxyphenyl)"trans<"bi

was 23.85^ as compared to estradiol* In this chapter wts have described the ^nthesis of other tvro isomers of 1 vi z. 2a, 6p~diethyl-3a-(_p-hydro3:yphenyl)-trans-bicyclo[ 4» 3* 0] nonaii-7p-ol (_11 .cig-a^^ti-jteiaSlS) ^^'^ 2p» 6p~diethyl-3p(p- hydroxypheny 1 )-tran3-biqyclo[ 4« 3• 0]nonan-713-ol (I2, cis sy n>-trans) (Scheme 1). With ihe hope of increasing the oral absorption and consequently the activity 'uhe synthesis of Ta-ethynyl derivatives of IJL, and 1 lias also been carried out« Synthesis of the corresponding p-methoxyphenyl derivatives of 11 and _12 (i.e. 16 and 18, respeotively) has also been carried out.

Synthesis of 17a-ethynyl-13p~ethyl-5, 6-seco-estradiols

Hie ketoenol 6 was prepared "by the condensation of 2-ethylcyclopentane~l,3-cLione(.2) with propyl vinyl ketone (2), As a refinement over the method of Grupta et al. this condensation was carr5-ed out by simply stirring the two reactants in demineralized water f otr 5 days under nitrogen atmosphere, unlike the earlier method where the tt-ro were re fluxed in methanol in the presence of an alkali. Cyclization of the triketonc (4) thus obtained with _£-toluene sulfonic acid gave 2,6p-diethylbicyGlo[ 4» ^•OJi'ion-l- en-^, 7-dione (^). Reduction of 7-keto group of _5 with sodium borohydride in a mixture of isopropanol and ethanol afforded _6. Treatment of _6 with j>-methoxyphenyllithium in ether-THF at -40° furnished 2, 6p-.dLethyl-3-hydroxy-3-{^ me thcKy phenyl )bi cyclo[ 4» 3« 0] non-l-en-7p-ol (J), which without isolation was converted to the dienol 8 by the addition of one drop of HCl. Catalytic hydrogenation of 8 in ethanol over lOfo pd-C at R05?, till 2 mol of hydrogen was absorbed furnished a mixture of two isomers (_9 and 10). These two isomers were obtained in pure form in a ratio of r^ 0 0 a

^ O^Gf^

d^^

7 8

Gontd... OH C)J~ H H 4 \ \\^ H. H H 5 CH„0 5 10

g V N^

H0-^"^^ ^ 11 12

,o CsOH

Contd.. • 6

vvC=CH

9 > r<^^

vvCsCH

10 -

17 13

a, De mine rail zed water/Ngf b, PTSAi c, NaBH.} d, p-BrCgH^OGH /n-BuLi| e, HClj f, IC^ Pd-C/H2» 2 molj g, KOH, diethylene gLyoolj li, Jones reagentj i, LiG=OH/ethylene diamine

Scheme 1 40:60 by fractional crystallization from a mixture of ether and hexane. In the nmr spectrum the major isomer showed a highly shielded OH^ resonance (0.I5 6)» This isomer 10 was assigned cis-syn-trans stereochemistry on the basis of earlier vjork . The reason for this was -tiiat due to sterio interaction between S-CHpCH, and 2-CH2CH„ group the latter would be held in a plane away from the former. As a result of this steric compression the ring may undergo some c on for ma Honal chaiage which would bring 2-.ethyl chain T-dthin the shielding cone of the phenyl group. Formation of 1£| in major amount could be due to less steric hindrance for the a-face attach of hydrogen during reduction. Demethylation of 3 and 10 separately with potassium hydroxide in dieth^ylene glycol/hydrazine hydrate under nitrogen aljnosphere gave the corresponding phenols viz- 2a, 6p- and 2p, Sp-diethyl-3a/|3- (_2-Jwdroxyphenyl}-^trans-•bicyclo[ 4» 3* 0]nonan--7p-ol (12 and 12)-

To synthesize the correBpondj.ng 7a-ethynyl derivative (C-I7 of estradiol molecule), 3 \TQ,B oxidised to the 7-keto analogue (ij) by Jones reagent and subsequently ethynylated with lithium ace tylidej ethylene diamine under an atmosphere of acetylene by the method of Hlraga . Similar oxidation and ethyr^lation reaction of 3 and 1_0 afforded the corresponding 7a:-ethynyl-3p-(jg-.methoxyphenyl) derivatives 16 and 18. 8

¥ith a view to increase liie oral absorptivity of 1, the synthesis of ihe corresponding 7a-ethynyl compound ^ was carried out according to Scheme 2. Controlled catalytLo hydrogenation of 8 in ethanol over lOfo pd-0 at B.TP till 1 mol of hydrogen was absorbed gave 1^, which on Birch reduction gave 2a, 6p-diethyl-3j3-(p-methoxyphenyl)-ti3ns--bicyclo[4»3»0] nonan-7p-ol (20), Jones oxidation of 2j) and subsequent ethynylation furnished the corresponding 7a-ethynyl compound 22 (Scheme 2)»

Demethylation of 2jD, Jones oxidation and subsequent ethynylation of the product thus obtained (2j) produced the desired compound _2J..

Synthesis of 2a,6p-dietliyl-7a-ethynyl-3p-(cyclohex-2-.en-4-

one )-toaiis-.bicyclo[ 4« 3» 0] nonan-7p'-ol

Reduction of the aromatic ring in _2^ was carried out with lithium in liquid ammonia in the presence of a protic solvent such as ethanol to give ^ (Scheme 5). Oxidation of 2 3 could not be carried out with Jones reagent as this 5 results in the rearomatization of ring A • Opponauer oxidation of 23 gave the 7-keto analogue 2_6. Ethyaylation of _26 and treatment of the product 27 thus obtained gave 28, the desired 5>6-seco analogue of norgestrol. 9

a, l.(3fo Pd-G/Hg, 1 moll b, K-NH„j o, Jones reagentj d, Li^C=CH/ethylene diaminei e, KOH, d3.ethy3.ene gl^odl.

Scheme 2 10

OH N a 20 > H

H H GH„0' 5

25 26

.svCaOH

(5^^^"^^

28 27

a, Li-NH /G2H^0H| b, Al [t-BuO]^ /benzenoj c, LiCsOH/ethyIcnediaminei d, HGl/methanol

Scheme 5 11

Synthesis of 2, 6p-diethyl-5-(2-«ie-thyl-4-.hydrc)ix:yphQnyl) bicyclo[ 4r 3» 0] non-l-en-7p-ol

Condensation of ^ with 2-methyl-4—methoxyphenyllithii^m gave 2, 6p-diethyl-3-hydroxy-3-.(2-.methyl-4-meihoxj'-phenyl) lDicyclo[4-i-5«0]non-.l-en~.7p-ol (29). Unlike 7 this hydroxy intermediate (_29) was quite stable and ihe dehydrati-:jn could he carried out only after refLuxing 2J in benzene with _£--toluenesulfonic acid. Catalytic reduction ^ in ethanol over lOfo Pd-C at R!CP till 1 mol of Hp was absorbed resulted in the formation of _31» I*urther reduction of 2, 5-double bond was tried with 10^ Pd-C, a mixture of dfo wet and lofs pd-C, Raney-nickel and PtOp at pressures ranging from RIP to 1500 p.s.i., but without success. Steric hindrance could be a reason for the inertness of the double bond towards reduction. The migration of this double bond to 5,4-positLon was then attempted but this also could not be carried out successfully. iEriethylsilane in GP„GOOH, a hydiride donor could also not reduce ttiis productj the reaction with this reagent went only to the intermediate stage J4» Forcing conditions for the forward reaction gave back Jl, the starting material. 12

29 12

^ ii

y Reduction

a, 2-Methyl-4-tromoanisolG/n-BuIij b, pTSAj c, 1C^» Pd-G/H2» 1 inol» d, Jones reagenti e, KOH, diethylene glycol.

Schemo_ 4 13

P„COOO H

H aH,o 0 14

Jones oxidation of yL gave the 7-keto analogue J^ whereas demethylation of ^1 led to the oori<5sponding hydroxyphenyl compound y^ (Scheme 4).

£ha£ter_III

One of the oldest and simplest method for fertility regulation is the use of spermicides for vagina. Though the failure rates encountered with these agents are as high as five percent, they find their use mainly out of the disadvantages which users find or fear with other methods. In the begining of this chapter a brief review about the known spermicides has been presented. Mongst tiie currently'- used spermicides nonylphenoxypolyethoxyethanol (nonyl^9 or nonoxynol-9j ^) and p-methaxyphenylpolyoxyethylene ether (menfegol or TS-88, _^) are the most important representative

^^;^^X^^O-(CH2CH2 0^ (0CH2CH2)gOH Me

55 14

Some of the most common spermicides available today suffer from the drawback of rather low order of aolivity and their action is pH dependent* This emphasizes the need for developing more active spermicides which would also be effeo-uive in a broad pH range. 6 In earlier work from tliis laboratory it was reported that N-substituted cc-aminomethylacrylophenones (J7a) «,«- diaminomethylacrylophenones (,^.Tb) and "their quaternary salts exhibit strong spermicidal ac'iivity, which are a novel class of spermicides. In a study of structure-function and

R-^ R- ^^

tfiuH nwii^

structure-activity relationship of this novel class of spermicidesir the synthesis of N-substituted a-(2-aminoethyl- acr3?"lophenones {^) and 2-substituted methylsneindan-lr-ones

^^^^ 58

(12) wa^ undertaken eiid their activity is described in this part. 15

Synthesis of N-substLtuted a-(2-aminoethyl)acrylophenones

To carry out the synthesis of Jg, various substituted Y-chlorobutyrophenones _40 were used as starti3:ig material. The corresponding Y-s-2ii.J^o"but3n:ophenones (jp.) used as intermediates were obtained by refluxing _£0 with an appro priate amine in the presence of potassium carbonate (Scheme 5) The condensation of JJ. >d.th paraformaldehyde was carried out by refluxing the two in hexanoic acid, instead of acetic acid ^fliich was used for the synthesis of 27a. In the present case a higher boiling acid had to be used since acetic acid could not fetch successful results. Thus the reaction of _41 with paraformaldehyde in hexanoic acid gave the desired compounds _^_. n /

R=^,Br,OH S or H / IT =Pi peri dine, Morpholine, \ Diethylamine or N~ P heny Ipi pe ra z± ne / a, N J b, HCOpH/hexanoic acid

Scheme 5 16

Synthesis of 3-aEiin(>-2-substLtuted methyleneindan-1-ones

The synthesis of target moleoules J_9 was achieved starting from various indaii-l-ones (J2). Benzylic "bromination of ^ vrith N-bromosuocini!ia.de in CGI. in presence of henzoj^'l peroxide yle3.ded aryl substituted 5-bromoindan-l-ones(4^5) (Scheme 6), Reaction of _43 witla different amines resulted in the formation of 1iie corresponding ^-a^ino deriva'kives J4» iO-dol condensation of _44 and various substituted aromatic aldehydes in # alcoholic KOH afforded the desired compounds _2?»

Condensation of paraformaldehyde with jA was carried out by reHusing the tvro in acetic acid for 4 b.. 'Hbls gave a mixture of two isomers ^ and A5 which could not be, however, separated. !Ehe nmr spectrum shewed one to be having exocyolic double bond {^) and -tie o'liier with endocyclic double bond ^5«

Chapter IT

At the start of this chapter a brief account of the known tricyclic antidepressant agents has been ^ven. Most of these compounds have an associated anticholinergic activity which is a serious limitation mth them. Ihese drugs suffer from some other drawbacks such as hepatotoxicity and a very late onset of action- Hence, there is a need for 17

lA^ ^r 42 43

^ •N-

R = H or CH„0 M = Pi peri dine, morpholine, diethylamine, t-butylamine; N-methylpiperazine or ]:l-plienylpipera2d.ne.

R' = OgH^CHO, o-FGgH^GHO, ^(CH^O)CgH^CHO, p-(lT02)CgH.CH0 or H a, NBS, CCl./Benzoylperoxidej b, m , Ace tone/K^^O^j c. Mo KOH alcoholic.

Scheme 6 18

non--fericyclio antidepressants.

During a course of broad primary screening of compounds in this Institute, ^te;an£^-2-.[N-(2-hydroxy-l,2,3,4- tetrahydro-l-napli'ttiyl)]iminothiazolidine was found to possess marked anld.depressant activity , oanparable to that of imipramine, an important member of the tricyclic anti depressants. Eiis novel lead proved worthy of further exploitation and prompted the synthesis of some iminothiazolidine derivatives (4^ and 52h

Synthesis c£ iraino-chiazolidine derivatives

To achieve the sjmthesis of various iminothiazolidine derivatives (J^ and _5,9) different benzocycloalkanones (_£7) were used as starting mate^-lal. (treatment of _47 with hydroxylamine hydrocthloride in ethanol in the presence of an alkali under refluxing conditions srielded the corresponding oxime JS, which on catalytic hydrogenation over IC^ pd~C furnished the amino derivative ^^9 (Scheme ?)• Condensation of j|9 id.th benzoylisothiacyanate in anhydrous acetone afforded the benzoyl thiourea derivative 30, Lebenzoylation of ^0 with lOfo NaOH solution gave the thiourea derivative _§!. Oyclization of "the thiourea ohain in ^ with bromoethylaniine b.ydxobromide was carried out by allowing the mixture of these t^ro in ethanol to reflux for 24 h. Ihis resulted in the desired compound _46. 19

.^^^^^-^^ a (CH^)^ R-

b v

Z ^ R --

.^^^"^V^^ R -p'

NH-0~im.

a, NH2OH.HCI I b, HgfPd/G] | c, CgH^CONCS \

d, 1(^0 NaOH I e, Br(CH2)2NH2«HBr

R = 5-0CH-, 6~0CH- or H, 5 5 X = GH2 or 0, n = 1 or 2

Scheme 7 20

The synthesis of trans-2-[N-(6-hydroxy-6,7 »8,9- tetrahydro~5H-benzocyoloheptene)]iMnotMazolicline (_59) was caiTied out according to Scheme 8. Hhe starting mate-^ial •benzocyclohepten-5-one (benzsuberone, J7» X=CH2» R=H and n=2) was treated with sodium borohydride so as to provide the alcohol (,§2)» Dehydration of. 52 with potassium bisulfate afforded 52.* ^^ bromohydrin SJ was obtained by the reactiou of 52 ^Jii^h N-bromosuccinimide in dimethyl sulfoxide^water under nitprogen atmosphere. _54, when allowed to stir vdth ammonia solution gave the amino aloohol ^ via an epoxide as intermediate. ^|5^ans opening of this epoxide gave the 5- amino-6-hydrox.7-6,7 >8,9-tetrahj/-dro-5H-benzocycloheptene (_55 ) • The nmr spec-fcrum of this product _55, showed it to be a mixture of two isomers which could not bo separated at this stage. Condensation of ^ irith benzoylisothiacyanate gave a mixture

°'^ J£§J1S ^'^^ .ci-? benzoyl thiourea deriva'tives (56_ and 37 respectively). These two isomers X'fere separated by fract ional cr3''stallization from acetone-water mixture. The stereochemistry of 56 and 52 was confirmed by NOE experiment- Hydrolysis of _5^ and subsequent cyclization of the thiourea derivative _58 with bromoethylai'iine hydrobromide afforded the desi.red compound 59»

BIG £i^-isomer 5Xt however, behaved in an entirely different fashion. The hydrolysis of 57 with 2fa ethanolic 21

NH-C-NH-C-Ph It W 56 + S 0 55

'OH NH-O-ITH-O-Ph II M S 0 57

g 16 ^ ->

OH I! ^ S

a, UaBH^i b, KHSO., A t c, IBS, IMSO/Waterj d, WH^OHi f, IQffo NaOHj g, Br(CH2)2M2.HBr

Scheme 8 22

NaOH solution resulted in the formation of two products 60 a.nd Gl» 60 was characterized as iiie ^csL^s-amino alcohol and _61 as a urea derivative. Elemental analysis of 61 showed the absence of sulfur. 'Ihe identity of these two products was confirmed on the basis of nmr, ir and mass spectra and elemental analysis.

Hydrolysis 57 J..

OH Wdr 60 8 0^9

Synthesis of pyrimidinone deriva-cives

A large number of pyrimidinone derivatives synthesized in this Institute were found to possess stimulant or depressant activity. This prompted to utj.lize the thiourea deriva-lives _51 for the synthesis of some pyrimidinone derivatives (H)*

S-Me-thylation of some thiourea derivatives (57» X=0, R=H, n=l and X=CH2> E=H, n=2) was carried out by refluixinc it in absolute methanol with methyl iodide (Scheme 9), so as to produce 6^» The I-substituted isotMouranium salt 62 was formed as an intermediate which was converted to 6^ by bicarbonate treatment. OyclizatLon of 63 with ethj^-l 23

propi da-be in neiiianol afforded 6J- [ 5-(4-oJ'Omar^3fl)-2.- methylmGroapto-.4( 5H)-pyrimidinone aiid 3-(6,7f8,9-tetrahydro- 5H-bGnzocycloIieptcne)-2-methylinercapto-4( 3H)-P3nriinidinone].

,^^x^^^ , (™2'„

NH-C-ITH2 NH-C=NH2 I s S-CH. 51 62

ra-c=:NH S-GIL 63

a, MeI/methanol| b, NaHCO_j c, CH=C-COOG^Hc ^ 25

Scheme 9 24

Synthesis of 5~[ (N-substLtuted 5"amin{>-2-l-iydroxypropjl) n.---m-m«r^ »l* n t^jm^ ^» oximino] -6,7»8,9-te tralnydr o-5H-'benzoq^'- oloheptene a

A reoent publication has shown "the importanoG of arauatiQ ODcime ethers as effective p-adroner^c blockers. Kiis prompted to synthesize th.e title compoi^nds (^6).

Reduciion of the cximino derivative (JS) of benzsuberone with sodium methoKide/methanol and subsequent reaction \d.th epichlorohydrin in anhyd^rous IMF gave the epoxide ^. Opening up of this epoxide 65 ''.•d.th various amines gave the title compounds 6§.

/ N-OU-CH2CHCH2IT \ 66 OH + - a, 1. NaO He/methanol, 2, epichlorohydrin/ljviF, b, M^ (morpholine, t-butylamine and H-phenylpiperazine

Scheme 10 25

Bi ologioal Actlvl^ty

Most of the compounds have been evaluated for their •biolo^cal ao'oivity. In the antifertili"ty tes'ling £4 was found to be ilie most promising molecule of ihese studLesj it showed lOCffi antiimplantation activity in female rats at a dose of 1 mg/kg when adsiin-.stored orally. testing at further lcn«rer doses is in progress._2_4 liad 25«4-9^ receptor binding affinity as compared to estradiol. 11 and 12 have also ehajn lOOjS antiimplantation activity at 10 mg/kg when. admini.stered Grails'- to rats. Tvro other interestingly active molecules were JO and JjL. Both these compounds showed lOQp/b antiimplantation activity in rats at an oral dose of 2 mg/kg. Screening of 11, 12., XQ ^^^ ^ ^"^ lower doses is under process.

None of the compounds showed any noteworthy activity in spermicidal and antidepressant screening. 28

EBFIHEINJOES

!• J.S.Bindra, A* T.Nejryarapally, R.C.Gupta, V.PtKamboj and F.Aiiand, J. Med. Ohem., 18, (1975), 921-925. 2. S.Durani, A.K* Agan-ral, R.Sa^cena, B.S.Setiy, R.C.Gupta, P.Jj.Kole, S.Ray and IT.Anand, J. Steroid Biocheni., 11, (1979), 67-77. 3. R.O.Gupta, S.Durani, A.IC. Agarwal, V.P.Kamboj and H.inand, Ind. J. Cliem., 19B, (1^0), 866-890, 4. K.IIiraga, Cheu. pharm. Bull., l^, (1965), 1289-1294- 5. H.J.Ringold, G.Rosenkranz and P.Sondheiirier, J. Am. Chem. 3oc., 78, (1956), 2477. 6. P.FautLyal, Ph.D. Dlssei-tation, Lucknow University, luckno"W, (1981). 7. U.K.Sliukla, Ph.D* Eissertation, Kanpur University, • Kanpur, (1979)• 8. E. A* Gupta, Ph.D. DLsserta-fcion, Meerut University, Meerut, (1981).

9. G.Leclerc, A.Mann, C. G.¥ermuth, N.Bieth and J.Schwartz,

J. Tied. Cham., 20, (1977)y 1657. SYNTHESIS OF POTENTIAL BIODYNAMIC DRUGS

A THESIS SUBMITTED FOR THE DEGREE OF Doctor of Ph/losophM IN ChemistPLi TO THE ALIGARH MUSLIM UNIVERSITY, ALIGARH

BY AKHIL G. JHIWQRAN

DIVISION OF MEDICINAL CHEMISTRY CENTRAL DRUG RESEARCH INSTITUTE LUCKNOW - 226001 JUNE, 1983 T2690

•-^*fim'

THESIS SECTION

CHE ^ To ^'

'f *«* ^ MY PiRMTS * Telex : 0535-286 Telegram : CENDRUG Phone : 32411-18 PABX ^J'^^rS 2260O1 (TRcT) CENTRAL DRUG RESEARCH INSTITUTE Chatter Manzil, Post Box No 173 LUCKNOW—226001 (INDIA)

No. Date June 6,1985

CERTIFICAIB

This is to ceriift'- that the work embodied in this thesis has been caridLed out by Mr.Akhrll G.Jhingran under our supervision. He has fulfilled the requirements for the degree of Doctor of Philosophy of the .Aligarh Muslim University regarding the nature and period of investigational work. Ihe work included in this thesis has not been submitted for any other degree and, unless otherwise stated, is all original.

I |\|vXly:)( KV>aA\4 (WASim EAHM.AN) (NITTA MUBD) Head FNA Department of Chemistry Director A»M-U#, Aligarh. Central Drug Research Institute Lucknow. I wish to record my deep sense of gratitude and indebteajiess to Dr.Ni-fcya Anand, PITA* Director, OentrsJ- Drug Research InstLtute, Lucknow who has been my mentor and guide in more than one sense of the vrords and for his sagacious and indespensable guidance throughout the course of these studies and making it worthy of presentation.

I -^-Jish to convey my sincere thanks to Dr.¥asiur Rahman, Head, Department of Chemistry, Aligsirh Muslim University, ALigarh for his abiding interest and invaluable help rendered during the tenure of this work.

My vocabulary fails me to express adequately my humble gratefulness to Dr.S.Ray, Scientist, Medicinal Chemistry Division for his incessant encouragement, valuable discussions and affectionate behaviour which brought my research efforts to the success of thLs dissertation. It gives me immense pleasure to thank Dr.R.C.Gupta, Scientist, for his interest and guidance at all stages of this work. I convey my sincere thanlcs to Drs. A»P« Bhaduri and S.Sharma, Scientists for their valuable suggestions and stimulating criticism frcm time to time. I am highly grateful to Dr.S.P.Popli, Depuiy Director and Scientist-in-charge, Medicdnal Chemistry Division for providing the laboratory faciliiiGs.

I am also iiiankfUl to Dr.B.W-DhaTfan, Deputy Director and his colleagues in the DL"vision of Pharmacology and Dr.V.P.Zamboj, Deputy Director, Dr.M.M.Singh and Dr. 1*K. igarwal of Endocrinology Division for providing the •biological screening results.

Words would be inadequate in expressing my sincere thankfulness to I>rs.(Miss) P.Nautiyal and S. Abuzar for their most I'dlling cooperation and stimulating companionship during the course of these studies. Hie ready help and cheerful oonpany of Drs.S.IC. Bhatia, R»B.Sharma, R. D-Bindal, SMv Kumar, S. Gupta and E>K.Jain and Mr.S.N.Bindal is gratefully acknowledged.

Blanks are also die to the technical staff of Regional Sophisticated Instrumentation Centre, ODRI, for providing spectroscopic and analytical data and Mr«S.D«K« Chopra and his colleagues of Glass Blowing Section.

Lastly, I acknowledge the financial assistance by the Council of Scientific and Industrial Research, Nevr Delhi.

(AKHH G. JHINffiM) CONTENTS Page

LIST CF ABBREVIATIONS i • « PREP AGE • • « GHAPTBR I 1.1 SBCO-STBROIDS AB SEX HQRlldTES 1 1. 2 Re f erence s ... 52 OHAPTSR II 5,6-3ECO-BSQr.ADIOLS MD RELATED CCSVIPOWDS 2.1 Introduction and Basis of work 55 2.2 Synthesis of 2a, 6p- and 2p,6B- di e thy 1-3a/p (p-by dr oxy phen^rl)- trans-bicyclof^. 5* 0] nonan-7p-ol anSlâ, 6j3-die thyl-7a-e thynyl- 3a~( ^-hydroxy phenyl )-:s:S2;S- bicy olo[ 4. 3. 0] nonan-Tp-ol 62 2.3 Synthesis of 2a, 6p-diethyL-7a- e thynyl-3p-(_p-m8 thoxy/hy droxyphenyl)-trans.-bicyclo[ 4« 3» 0] nonan-7p^^^ôI . • • 69 2.4 Synthesis of 2a, 6p-diethyl~7a- e thynyl-3p-( qy cl ohex-2-en-4-one). trans-hiqy clo[ 4* 3» 0] nonan-7p-ol 72 2.5 Synthesis of 2, 6p-diethyl-3-.(2- me thyl-4~hy droxyphenyl )bi cycl o 74 [ 4* 3* 0] non-l-en-7p-ol 78 2.6 Experimental ••• 98 2.7 Biological Activity.. 104 2.8 References ... 0H1P!2ER III EiEVBLOPlvfflNT CF SPERIÎCIDAL AGENTS 3.1 Spermicidal agents - AJI Overview • • • 107 3.2 Introduction and Basis of work • •. • • • 122 3»5 Syntliesis of IT-substituted a-(2- aniinoethyl)acrylophenones 124 3.4 Synthesis of 3-amino-2-su'bst3.tuted methyleneindan-l~ones 126

3.5 Experimental ... I50 3.6 Biological Activity. • 151 3«7 References ... 155 CHiPlEF IV JiUTIDBPRESSMT AGMTS 4»1 A short review on Antidepressants ••.• 161 4.2 Basis of work ... 18 5 4.3 Synthesis of iminothiazolidine derivative s »• • IS 6 4.4 Synthesis of 3-(4-C!raTianyl)-2- methylmercapto-4(3H)~pyrimidinone and 3-(6,7,8,9-tetrahydro-5H-ben20cycloheptene )-2- methylmerGapto-4(3H)- pyrimidinone ••« 192 4.5 Synthesis of 5-[ (N-substituted 3-amino-2-hy droxy propyl) o:cimino]- 6? 7 j8,9-te trahydr o--5H-ben20- cycloheptene «.. 194

4.6 Experimental ••• 196 4.7 Biological Activity.. 219 4.8 References ... 220 :L;

LIST _(^,^ _£iB]XBYlJ^Tipi!^B

AG : ace tyl hh ! broad liump hs : broad singlet bp ! boiling point bs r broad singlet d { double t MSO dLmGthyl siL].f aside IMP ! dimethyl formamide Ether ; diethyl ether Hz , hsrtz IE. ! infra red M"^ ! molecular ion m ! multiplet m ' HS.'^S m.p. : melting point IJJffi s nuclear magnetic resonance N'OE ! nuclear overhauser effect _o : ortho

^ : J^T& Ph J phenyl p«s.i. !, per square inch q ! quartet s ; singlet S»0» ! subcutaneous t : triplet TPA ! trifluoroace-feio acid PREPACS ii)

intie single biggest problem facing the country today is our inability to control the increase in population. Ihis is in large due to the inadequacy of the available methods of fertility regulation as applicable to the socio-cultural milieu of the Indian setup. !Qie development of contraception in the early I960's has been a milestone in the field of fertility regulation and has been a major factor in the control of population in the developed countries and some of the developing countries as well. As a 'pill' requires continuous daily intake for at least 21 days of every menstrual cycle, this requires a strong motivation and a more highly developed social system where women have the availability to store the drugs - conditions which are not available in largely rural setting in India. Tlierefore, this contraceptive ' pill' has not been able to make any big headway in the family planning programme in India, hence the urgent need for an alternative method for contraception. A more suitable approach in the Indian setup would be drugs •vtiich could be taken much less frequently or only when they are needed after the coital act. Ihis could be achieved by having compounds which could interfere -^dth hormonal support that is needed for conception. The first t\fO iii) chapters of the present -work are related to the development of some seco-steroids which could possibly interfere with the action of estrogens or progestogens for successful implantation of the ovum.

The possible side effects of systemically used contraceptives has fooussed attention on agents that could be used externally such as spermicides. Chapter III of the thesis deals with the synthesis and the biological evaluation of rather a new class of spermicides.

Chapter IV of the thesis is concerned with the synthesis and bioevaluatlon of some potential antidepressants. In the pharmacological screening programme carried out earlier in this Institute, 2-[N-C2-hydro2cy-.l,2,3,4- tetrahydro-l-naphthyl)]iminothiazolidine showed comparable antidepressant activity to imipramine, a clinically used tricyclic antidepressant. Ihls provided an impetus to synthesize some related iminothiazolidine deriva'fcives and study their pharmacological behaviour, which is described in this chapter. SEGO-SOEROIDS i3 SEX HCejviOTES 1.1 SSCO-STEROIDS AS SEX HCHMQITES

Sex hormones, namely estrogens, progestogens, and androgens, play a major role in cell biology, The mammalian reproductive system is regulated by these hormones. They also control the growth of certain benign and malignant tumours. An effort to use these hormones juidiciously to regulate fertility and for the treatment of cancer has lead to a number of drugs but has also thrown much light on the mechanism of hormone action.

Estradiol (I), progesterone (II) and testosterone (III) are the most important endogenously produced sez steroid hoxmones, Ihese steroidal compounds have in common a cyclopentanoperhydrophenanthrene ring system called 'gonane' and vaiy in having different subs tit uents and degree of unsaturation at certain planes.

II Ill

\fh.en one or more bonds anre removed from the gonane structure, the molecule is called a seco-steroid. This class of compound though described as intermediates for the preparation of steyoids, have been less studied from biological activiiy point of view. In this review an effort has been made to present what is knovm about tiie biological3y active seco-steroids in reference to the activLiy of the parent prototype. Since chemical naming of such compounds is often not based on seco-steroids their search in the literatu?:^ presents a difficult task. In our discussion of -seco-steroids, steroid numbering has been retained. • Wherever there is any deviation in the stereochemistiy from the parent steroid molecule it has been brought out clearly. The numerical value before the word 'seco' denotes the missing carbon-carbon bond. The terms nor-seco , dinor- seco and homo-seco have been used to denote secosteroids with two or three of the bonds missing and a seco ring with an additional bond respectively. In our discussion we have also included compounds which have more than one seco ring. Ohemistjy

A-Sec o-s te roids

In the group of A-seco~s-beroids that would come under sex-hormones some A-nor-3,5-seco-»androsten-3, 17p-diol derivatives and 4,5-seco-testosterones were reported by A, Kasal ,

A-nor"5-seco"androsten~3»17p-diol (4) (Scheme 1) was S3rnthesi25ed starting from testosterone which was converted to (5R)-3a-"acetoxy-5-methane-sulfonate (2) and treated with LiAlH. to give ^ as the major product (34'^) , Catalytic hydrogenation of ^ gave ihe corresponding androstane-diol (1), Preferential oxidation of ^ was carried out using pyridinium chlorochromate to afford 5-hydroxy-A-nor"3,5~ seGo-5*"androsten-17-one (_^),

For the preparation of the 4f5-seco-analogues, testosterone pivalate was converted to 5-oxo-17p"Pivalaxy- A-nor-3>5-secoandrostan-3-oic acid 6_, esterified and treated with benzyl mercaptan to give the ben2ylthioenol ether .7 (Scheme 2). Desulfurization with Raney Ni led to methyl 17p-pivaloxy-A"nor-5>5'-secoandrostan-5-oate (8) which was hydrolysed and converted to its lithium salt. Addition of Meli on to the acid salt gave 4,5-seco-testosterone (£), Jones oxidation of ^ gave the corresponding 17-oxo-steroid 10. QPiv

AcO -k>

HCK^ 3

HO' 5 Piv = (CH^)^CGO a, LiAlH. I b, Cat./[H2]» ^* Py:ridinium chlorochromate

Scheme 1 iv QPiv

HOOG 0:>-N^ Me02G

a y OH QPiv

ROOC 0 8

10 Contd».. .sMe 4-1^

Hv^- ^ 11

a. Eaney~Ni| b.l. Hydi-olysis, 2. MeLi» c. Jones reagent» d. MeLi

Scheme 2

Preparation of the 17-methyl analogue 12 was carried out from 4 through its Eyridinium chloitjchrornate oxidation to 11 and 2 molar addition of MeLi«

Biolgc^ical Activity

The A-nor-3,5-secoandrostenes did not exhibit ary noteworthy activiiy. In the 4,5-secoandrogens, 12. had relatively high affinity for SHBG. She testoseterone analogue ^^ was weakly androgenic. These 4,5-seco-steroids showed high RBA for androgen receptors and were found to be distinctly antiandrogenic, particularly 1.2.

B-Se_co-;Gteroids

B-Seco-steroids of the estrone series possessing hormonal activity were prepared as follows which led to the required ^^.TiS^-^a^itl-tTa^^ compounds along vdth cj.s-ajij;i^~1grans. and cis-syn-traas stereoisomers. la-, andlp, Ya-acetoxy-S-ethyl-ep-methyl-^-C^-methoxypheny•tl ) bicyclo[4«3.0]non'-2-enes

The synthesis of Bindra et al, involved the construction of a thia-B-homo steroid utilizing Torgov •5 6 approach-^' which on desulfurization and subsequent reducxion gave B-seco-compounds (Scheme 3),

1. a 2. b MeO 12 14

1* d 2.e Me O-^^^^ix-^ MeO 15 16

1. f 2. g Contd... ^3- h 8

QAc OAc

Me O'^'^^^ 17 OB

V QA.C OAc

+ 160-"^=^^=^ y

20 19 a. Y-Butyrolactone, lTa» b. PPAj c. Ref.5,6j d. (nH2)2CSj AcOH» e. 2-methylcyclopentane-l,3-dione ,

KOH} f. LiAlH^ (OBu'*^)^Hi g. AC20,Pyi h» TsOHj i. Raney-Nii j. Pd/C, H^ (1 mol)

Scheme 5 9

2-.AL}5yl-6^methyl-3-aryl-biGyclo[ 4. 3. 0]nonan-7-ols •»-m •^>Wfc -ll-^i-i'l • »1..

a Me o-^^^^ MeO-"'^^^ 21

(f^^^^ Me MeO 25 24

Oontd»«. 10

+ R

R = H, CH„ or C2H5> R'=H or Aci R =H or CH^

a. Ref. 5,6i b. {NH2)2GS, AcOH; c. 2-me thy ley elope ntane-1, 3-dione, KOHj d. TsOH; e. NaBH^j f. lOfo Pd/C,H2 (1 raol)i g. K-NH,| h. l(^c Pd/C, H2 (2 mol)

Scheme 4

Method 2

A more convenient synthesis based on the procedure of Boyce and White hurst was reported by G-upta et al ' . Starting from 2, 6p -dialkyl-7p -hydroxy -bicyclo[ 4. 3.0] non~l- en-3-one (j^), by reaction with ^-methoxyphenyl-lithium followed by dehydration was obtained the dienol ^1., which afforded three of the possible four isomeric compounds under different reduction conditions (Scheme 5), 11

2.b

MeO^

Me 0-^ ^2

d V \k

..^^^-^^

H0^^ ^ 54 15

Contd«.. 12

, OH

Me 0^^ 31

R' OH

HO^-^^^ Myiecre ^

^ H

R'^GHj or 02H^ , R=CH^, C2H5 or O^H^

a, j-CMeCgH^li , b, TsOH , c, IC^. Pd/C, Hg (2 mol), d, KOH, diethylene glycol , e, 1(^ pd/C, H2 (l mol), f, K-NH,

Scheme 5 13

B-Se c 0j3r o ^e ste rone

5,6-Se CO progesterone and testosterone compounds were prepared by Grossley and Dowell from 3p ,20p--diacetoxy~ pregn-5-en-7-one (j^J (Scheme 6), which was epoxidised to 40, which on treatment with _g-tolylsu.lfonylhydrazine and hydrogenation of the 5p ,20p~diacetoxy~5,6-secopregn-6"yn-5- one (Jl), thus obtained furnished 5p ,20p-diaGetoxy-5,6-seco- pregnan-5-one (J;2). Reaction of j;2 with sodium dihydro bis-(2-methoxyethoxy) alumina te gave 5p,5(a &p)» 20p~ trihyd3X)Xy-5,6-seco-pregnane (4;^) which on treatment with silver carbonate on celite, hydrolysis and subsequent oxidation gave 5,6-secopregn-4-ene-3,20"dione (44). DDQ dehydration of 44 gave 1,2-dehydro derivative ^^^, 6,7- Dehydro~5»6--secoprogesterone (46) was obtained through partial liydrogenation of the acetylene compound 41,

For the preparation of the 5»6-secotestesterone acetate (48) the enamine 4J generated from 4j4 was reacted with peracid and subsequently hydrolysed,

Biol^Qg^cal_^_Acjtijl^

In the seco-estranes of type A, it was observed that l2;§^.''§liM.'"i£aSS isomers were more active than the other two isomers, in ethyl substituent at G-15 (R^Et) was found to be superior for biological activiiy as compared to the 14

AcO^

l.a 2.b

AcO AcO' 19

1. C 2.d fAcO. ^ k^ <- (^^1

AcO^

^ AcOs^^

g -T'

Oontd../. • 15

h 44 ->

OAc

H<:>-^ ^ \ V 47

V-

a. NaBH. f "b. m-ohloroperbenzoio acid j-c. p-Tolylsul- phoi]ylli;^'(i-razinei d. KpCO- i e. Hg/PtOp ) f. Sodium dihydrobis-(2-metlioxyethoxy)-aluminate i g« Silver carbonate on celite, hydrolysis and oxidation i h. DDQ Schema 6 16

corresponding methyl compounds. Compounds -witliout an alkyl substituent at 0-8 (Rn=H) were inactive at a 20 mg/kg dose (rat). No activiiy lias been reported for compounds having R-,=Me, Biological activiiy profile of compounds having IL=Et or Pr is showi in Table 1,

In the 5,6-secoprog9sterone and testosterone series, s,c, administration of 1 mg/kg (rat) of 5,6-secotestosterone acetate produced anabolic and androgenic activity equivalent to 0,25 mg/kg of testosterone propionate, A1.1 other compounds of this series were devoid of biological activity,

C~Seco-steroids

A series of C-seco-estradiol derivatives were prepared in our laboratory a,1:5-15 ^^^ elsewhere starting from estradiol through a sequence of reactions in which the stereochemistry of the parent compound was retained. 17

Table 1

Antiimp- Uterotrophic SI, Compound lantat- activity RBA ion acti-'" Dose " Uterine No. viiy MED mg/kg wt(mg) IOO5I (5 days) (mg/kg)

1. Estradiol 100

2, Estrone 9.27+0.05 0,01 0.01 65

7.95+2.15 0.33 77

5»06+1.65 10 1.0 50

10 1.0 73

6.38+0.84 10 0.75 49.5

23.85+4.75 0.02 51.2 18

The title compound was prepared"] 2a starting from IT-acetyl-estradiol-^-methyl ether (43.) (Scheme 7), its oxidation and hydrogenolysis led to the seco-compound J\^, The 17H)H was protected as THP ether and the carbonyl funotion was reduced to alcohol ^ and deoxygenated through its mesylate ^ by LiAlH. treatment. The THP group was subsequently removed to give 9,ll-seco-estradiol-3-fflethyl ether (^). Demethylation of ^ under alkaline condition gave the required seco-steroid (^7).

49 50

1 .b 2.C \ ' OR OIHP ^^

Contd... 19

OOHP

MsOHoC^"""

r^^^^^f

MeO MeO-

^ 54

\v OOHP

MeO 55

a. OrO ('''Ref.l2b); b. Hg/Pd-O, MeOH-HGl»

G. 1(^* HaOH (ethanolic); d. DHP/PTSJLj e. LiAlH^l

f. Metliyl sulfonyl clxLoride/Pyriclinei g. HCl,

Die thy lens glycol, KOH

Scheme 7 20

11,12-Seco-

^R OAo

a ^

MeO 56 (R=Ac)

MeO 60 5_9

a. CrO^j b. MeMgli c. Fa/im„

Scheme 8 21

9 «ll-Seco-G-liomoestradiol

(a) Treatment of ^l with MeLi gave"^'14^ tlie ketone 6£ V7hich on l/olff-Kishnor reduction furnished the title compound

6^.

51 (R=Ac)

a. MeLi i b. Diethylene glycol, KOH

(b) Chinn et al, ^ prepared seco-steroids Jl ^^^ JA starting from _trans~2-(6-methoxy-2-naphthyl)-5~oxocyclopentane acetic acid (^) following the Scheme 9, When R was H, a mixture of trans-anti and trans-syn products 66 a, and §6b were obtained while only trans -anti product resulted with R=CH^ in 6^, 22

65, a. R=H b. R=CH

MeO HeO 68 a, R=H 67

b, R=CH, \

N1/ OH R I^^O^CSCH OAc

HO-n

iS a, R=H 71 b, R=GH, Oontd* 23

Me 0 MeO 70 72

l.e 2.f

C=OH

MQO MeO J2(R=H» R=Ac) 74

a, E-lMgBr, GH=CH| b, Diazomeiiianej c, MeMgBri d, AC20/Py I e, BPJBt20, HgOj f, Methanolic HCl

Scheme 9 24

OAc

MeO

58 15

77 (R=H) 76

78 (R=Fx)

a. pb(OAc),, triiyl chloride

Scheme 10 25

9,ll-Seco-C~norestradiol 16 ^nthesis of the title compound was best achieved' through a modified halodecarboxylation 17 of ^, Thus treatment of ^8 with lead tetraacetate in the presence of triiyl chloride gave the halo compound 7^ (Scheme 10) which vras subjected to hydrogenolysis and subsequently treated with la/NE, to give 9,ll-seco-0-norestradiol-3-methyl ether (XL) and could be demethylated under alkaline condition to J8 (R=H).

11.12-Soco-^stradiol

Pyridinium chlorochromate oxidation of XL (R=Ac) afforded the 9"keto compound ^ (Scheme 11), Grignard reaction -^cith MeMgl and its subsequent Na/NH^ reduction furnished an isomeric mixture of trans~syn-trans and trans -an ti -trans products 81 and 82,, This mixture could be demethylated under alkaline condition to the corresponding hydroxy isomers 8J and 84,

MeO 77 (R=Ac) 79

Contd..• 26

MeO 80

82 81

a, Pyridinium chlorocliromatei b. MeMgli c, Na-NH^j d. KOH, diethylene glycol

Scheme 11 27

8,14-Seco-estronG

A total synthesis of 8,14-S8co-estrone w£is achievod'1 8 utilizing the dione 8^ obtained as an intermediate in the Torgov synthesis-^* of estrone. Catalytic reduction of the Sjll-double bond gave _86 which was converted to the mono- thio ketal 82 (Scheme 12) atid subjocted to dosulfurization to give the methyl ether 88, Demethylation of 88 gave the required compound Sg, The diol ^ was obtained by sodium borohydride reduction.

MeO' 85 86

38 87

i/ Contd. -. 28

§.? 90

a, Bthanedithiol, TsOHj b, Raney-Nij c, Pyridine hydrochloride; d, HaBH.

Scheme 12

9 * 11-Seco-progesterone

The title compound was prepared "by Orossley et al. 19 starting from the readily available 9-dehydro progesterone (2k)» Oonjugated. double bond V7as reduced by Li/N"H^ and the 5a-oompound thus obtained was subjected to ozonolysis and then treated with sodium borobydride to get the C-seco compound ^ (Scheme 13), The hydroxy group from C-11 >ras removed through mesylation and the 4,5-

0^^

91 H 92 (R=Ac)

^V^

H H 94 12 (Ri=OH)

^v^

95 96 a, LiAlH-i b, 1. 0™, 2. NaBH^j c, DDQ, pTSAi d, [Hg], soluble Rhodium catalyst

Solieme I3 —fc'^^**"!*- ' 30

Biolo^:^ical ActivHy

The RBA, antiimplantation activliy and uterotropliic activiV results of some of the ahove G-seco-estradiol derivatives is listed in Table 21 7 ,

It was observed that JO and X4 show estrogenic actLvi-ty at 1 mg dose level. Only 6^ displayed some antifertility activity at a dose of 4 mg.

The best compound of this group was Jd (R=H) wiiich exhibited 100'/^ inhibition of pregnancy in rats at 2,5 mg/ kg when administered s,c, J8 had a RBA of 78.90?^. None other compound possessed significant activi-ty 8,14-seco- estrone derivatives failed to interfere with implantation in rats at 10 mg/kg dose. 31

Table 2

Compd. RBA A.I.actiYity

Oral S.G. Oral S.C.

X8(R=H) 78.90 5.0 2,5 60 100 £0+61 7.20 5.0 40 84+8^ 9.10 2,0 14 65 9.80 5.0 0 51 25.20 10.0 100 5.0 61.0 5.0 0 10.0 5.0 100 100 5.0 65.0

D-Seco-g-beroids

INfo of 'otie most potent and early compounds in seoo- steroids were the D-seco analogues of estrone and equilenin, 2 0 called doisynolic acid (2J7) and bisdehydrodoisynolic acid (S8), Further structural modifications resulted in dl-cis- bisdehydrodoisynolic acid (2_g_) and its methyl ether (lOO).

/\V'^ "OH

97 96 32

v^^^'^OH

99, R=H 100, R=CH-

(97) and (_28^) could be prepared by alkali fUsion of estrone (IQI) and equilenin (102) respectively.

A A ^(96) KOH

101 102

2T Racemic bisdehydrodoisynolic acid was obtained ~ by reacting methyl l-keto-2-m0thyl-7-methoxy-l,2,3,4- tetrahydro~2-phenantbrene-carboxylatG (lO;^) with Eil^IgBr.

OQMe BtMgBr COCMe "^ Raoemic (98)

MeO 102 104 33

15 ^16-«Sooo-progesterone

.22 The -fcitle compound was prepared starting from I6a» 17a-di hydroxy pregnenolone acetate (10^),'its conversion to 16,17-dihydroxy-D-homo-androstane derivative 110 and subsequent cleavage of the D-ring (Scheme 14-).

..»^ OH ,XNOH

ACCK' 105 AoO 106

\J' ,QR X^

H(r"^- 107(R=Na) 106

110 (R=H,R'=Me) 109 (R=H)

Contd-.. 34

111 (R=Me, R'=OHO) 112

112 115

114 a, HOOOH, mnO.i b, Na/CgH^OHj c, OH,I, IMP| d, HaBH^i e, aqueous HCl-OHCl i f, CH_Lij g, NalO.j h, (Rhodiumtrichloride trihydrate, ph,p)+ CH_0N| i, K^ t-BuOV^BuOHj j, Al(^t-BuO)^, cyclohexanone.

Scheme 14 35

Biological Activity

The (")-a-bisdehydrodoisynolio acid (^8) was found to be most active in the D-seco-steroids, the racemate being half as active as the levo form. The activily of the 7-methyl ether of £8 was not much different from the hydroxy compound. The effect of substitution on the biological activiiy of bisdehydrodoisynolic acid (B) showed that acids (Z=COOH) without alkyl groups or with only one methyl

B group at C-13 or C-14 were practically inactive, ¥ith 2 or 5 Me groups or one Bt and one Me at G-15 and C-14, the maximum activity readied was 0.1-0, 2Y. Further lengthening of the alkyl chain reduced the activity. Replacement of the carbonyl group by the substituents e.g. Z=CH2C!00H, -GOCH^ or -C0GH„0C0CH^ abolished the activity, however, -CHO and -OHpOH compounds were onJ^ slightly less active and showed more prolonged action on s.c. adjninistration,

A comparative data of the estrogenic activiiy of these compounds is shown in the Table 32 0 , 36

Table 3

Estrogens Single dose, BD^QQ (rat) s,c. oral a-Bisdehydrodoisynolic aoid (-) 0.05-0,07 0.05-0.07 7-*Iethyl ether (-) 0.05-0.07 0.05-0.07 a-Bisdehydrodoisynolic acid 0,1 -0.5 0.1 -0.5 (raoemic) p-Bisdehydrodoisynolic acid ( + ) 100 100

Doisynolic acid (from estrone)(+) 0.7 -1.0 1-5 ^s trone 10 -15 20-30 Estradiol 1 -2 20-30 Equilenin 100-150 300-500

A,.B-5ec o-s ter oids

5.10-Se CO-steroids

116, 117 and 118 were reported by Batzold et al. * ^nthesis of p ,Y-ace-tylenic-3-0x0-steroids of the 5,10-seco series was achieved 25^ starting from 3p ,17p-diacetoxy ester-10- hydroxymethyl-5,6-ene (ll^) which was oxidised to the corresponding 6-keto-5(10)-ene (120) (Scheme 15) and converted to the 5,10-epoxide (121). This epoxide on treatment with £"• toluene sulfonyl hydrazide generated the 5,10-seoo-steroid (122) which was hydrolysed and converted to 5,10-seco-estr-5-ynG-3,10,17-trione (116). 37

ca^ie

116, PJ1'=0 118 117, R=Gaie, R'=H

OAc OAc

AcO' AcO^ 119 120

\1^ OAc OAc

ACQ AoO

122 121

Contd... 38

125 124

a, OrO^-Py oomplex/pyj b, 3/0 H2O2J c, ^toluenesulf0203^1 hydrazidei d, KOHj e, Jones reagen-

Scheme I5

Bi ol OiSl cal Ac tivi 1y

In the mature rat, 5,10-^eco-estr-5-yne-3,10,17-trione (116) (1-22 mg/kg/day for 7 days i.p. ), 5,10-seco-19- norpregn-5-yne~5,10,20-trione (117) (1-20 mgAg/day for 7 days i.p.) and (4R)--5 AO-seco-19-norpregn-4,5-diene-3»10> 20-trione (118) (0,8-8 mg/kg/day for 7 days i.p.) caused dose dependent decrease in the wet weight of ventral and dorsal prostate glands. This antiandrogenie behaviour of these compounds was attributed to their possible androgen biosynthesis inhibiting effect.

BtG-DiSGCo-steroids

5«6"8tl4°Diseco-steroid

2Hyie thyl-2-[ 3-( ^-hydroxy phenyl )hexyl 1 qy-cl open tan-l~one (12,7) aad the corresponding l-ol (1,25.) were prepared by Ananthanarayanan et al. through desulfurization of the 39

monothioketal (12^5j obtained from 2-methyl-2-[3--(2- methoxyphenyl)hexan-2-yl] cyclopen tan-1,3-dione (124) (Scheme 16),

0 0

Me MeC 25 124

MeO MeO 12 6 125

Hc/^^ MeO' 128 129

Contd. 40

126 •» 129 >•

Hcr"^^^ 127 a, 5io PCI-C/H2J b, Ethanedithiol, TsOHj 0, Raney-M| d, Pyrid3.ne liydrochloridej e, NaBH.i f, metliylene glycol, KOH- SCheme 16 5.6--12,13--Diseco-steroids

The sjrnthesis of 5-(£-hj''d3X)xyphenyl)-4~( 3-oxocyolo-

penVl)^®^^^® (U§.f '^l) ^^ "^^® corresponding cycaohexyl 27 derivative (1J6, n=2) was achieved via Michael addition of jg-methoxybutyrophenone to cyGlopenten-2-one (131, n=:l) and cyclohexen-2-one (13I, n=2) respectively (Scheme 17).

+ •(SHg)^-^ MeO/ ^ MeO

1^0 122. Oontd»»• 41

(CH2)„-ii->

MeO 134

d 0 •M/ ^

^ COH^)^ (CH^2') n

MeO 115

iOE^l,

136

a, K^J-Bur+,. O ; b. Ethylene gOycol, EPJlt^Oi c, BttvigBr; d, 1(Y^ Pd~C/H2J Ac OH, perchloric acidi e, BBryCH2Cl2» f, 9^^ HCOOH

Scheme 17 42

Selective tetalization of the dione IJ^ followed by Grignard reaction udth ethyl magnesium bromide furnished the alcohol 1^ which could be dehydrated to an isomeric mixture of tetraenes 1_2J, Catalytic hydrogenation of IJJ or hydrogenolysis of IJ^ afforded 2J3. which could be demethylated to 1J6, (n=:l or 2) using 3Br^,

5.6-12.13--Diseco-'Steroids

The syntliesis of 3-( 4-oxo-2-cyclooihexen-l-yl )-4-( 3- hy dr oxy cycle pen tyl) hex ane (l^) was achieved by Parbwerke , starting from 3~(4-oxo-cyclohexyl)-4-( 3~oxocyclopentyl) hexane (1.^8) (Scheme 13), Reduction of the cyclopentanone carbonyl could be achieved via selective ketalization of the diketone IJ^, Cyclohexene double bond was introduced through bromination folloired by dehydrobromination,

Bii&9^^^®i^Miild--jfeZ

In the above group of B,0-diseco-steroids only 144 vras found to possess androgenic, gestagenic, antiinflammatoiy and anabolic activiiy.

The B-seco analogues of doisynolic acid emerged as another class of potent estrogens, 4~(£-Methoxyphenyl)~5a- ethyl-6a-methylcyclohex-3-ene-a-carboxylic acid (147) and its isomer 148 were prepared hj Pouque3r et al» 29-^ , starting 43

•? t-Bu

158 t-Bu M9

a M/ OH

<- t-Bu

141 •t-Bu 140

^ OH

0 Br

a, BHT I b, H2S0,/acetone > o, Br2/0H2Cl2

Sclieme IB 44 from substituted cyclohexenone, its reduction and reaction vriLth ^-methoxyphenyl magnesium bromide (Scheme 19).

^,v CO2R

145 MeO 146

^^.oCOgR .^s^^CO^R

(+) 147 (-) 148

Scheme 19

Crenshaw et al. have ^nthesized a number of 2,3-- and 6-alkyl substituted 4-aryl~2-methylGyclohexene carboxylic acids. In this approach, l,4~addition of dimethyl copper- lithium complex on to the hexenone JJ^ led to the dimethyl- cyclohexenone (l^) (Schene 20) which xms converted to the required compounds 1^ and 1^ following the above approach. 45

G02Et a

149 150 (R=C5Hc- or H) I 151

I Q

MeO 152

a, (CH^)2Culi j b, ArMgX (AT=£-CH„OCgH^)j

0, 5f» Pd-G/H2

Scheme 2 0

BiologLcal Activj-try

BjD-Seco-compounds 1^47 and 148 were found to be more potent as estrogens than doisynolic acid 20 . Of the dimethyl cyclohexenes, trans-4--(p-^nethoxyphenyl)--2.6^6--trimethvl- A^- cyclohexenecarboxylic acid (l^l) inhibited pregnancy in mice or rat at a dose of 0,1 mg/kg/day given on days 1-5 post-coitum. 46

GyD-Diseco-steroids

The G-seco-analogues of doisynolic acid also showed promise as antifertili-ty agents-^ -^"', The derivatives of 6"hydroxy-2-naphthalene propionic acid (1^) were called allenolic adds, after S.Allen. The synthesis of a,a- dimethyl-p~ethyl-2-naphthalaie propionic acid (l^) was carried out 36 from 6-cyano nerolin 155,. Treatment of 1^5

with BrCMepOOOEt and Zn gave ethyl 3-(6~methoxynaphthyl)-3- keto~2,2-dimetliylpr;5panoate (156)' (Scheme 21). G-rignard reaction idLth ethyl magnesium bromide and suhseqaent dehydration by KHSO. led to p-ethylidene ester 3-5.8 which wae hydrogenated to the required compound 15^,

^, QBt

a .^ •> MeO- MeO^ 156 , 155 Oontd. • • 47

OEt OEt

MeO 157

a, BrCI!!4e2G02iilVZni b, B*IgBri o, KH30, MeO 159 Scheme 21 ^nthesis of deliydro allenolic aoid derivatives ]^6J. (R=02Hf- and C„H,^) xfas reported by G-opalachari and Iyer •'. The synthetic sequence involved condensation of 6-niethoxy-l~ tetralone (16^0) -with ethyl orthoformate followed by Grignard reaction with appropriate alkyl magnesium bromide to give the alkyledene derivative 1_62 (Scheme 22), Refortnatsky reaction mth 162, using a-bromoisobuiyrate gave the lactone 16_2, which on Jiydrolysis followed by reduction and dehydrogenation afforded the desired product 16^,

^GHOH

•> > MeCr •

151 0 ontd... 48

^^GER

^ MeO-^% MGO 162

MeO- MeO

165 164 a, Ethyl formate | b, RI'IgBrr o, BrCMe2C00Et/Zni d, NaOHj e, 1. NaBH^, 2. HBr g^j^eme 22 The synthesis of ll,12~seGo-iiionode]jiydrodoisynolin acid (16J) and the tvro isomers of 11,12-secodoisynolic acid 16J 31 and 1^0 was reported by Dubois et al« , The intermediate 16J; when reacted wi th MeLi gave the lactone 166_ which was hydrolysed and dehydrated to give 16J. Dehydrogenation of 167 gave the 11,12-seco-analogue of bis-dehydrodoisynolle acid 168 (Scheme 23), Catalytic hydrogenation of 16J gave the isomers 169 and 170. 49

\|^02H

^ MeO MeO' 166

\UcO2H \\^0^E H

H Me 0 ^-^ MeO"

168 169

CO2H

MeO 170 a

Scheme 2"^ 50

Biological Activi1;y

In the illen-Doisy test (Xfa-cLiiae'thyl-^-ethyl derivative cf allenolio acid 1^^_ was active at 1.5Y close. In this sex-ies the methyl ethers of a,a,p-trimethyl and a ,a-dimethyl-f-ethylidene-, a-methyl-p-ethylidene-j a- methyl-p-ethyl"derivatives showed activi-ty in the decreasing order. Isomers of 1^, having MeO group at 3»4>-7 or 8 positions were inactive while 1- and 5-isomers were more active than lis.

Compounds 16^ (R=Et and R=Pr) elicited almost equipotent estrogenic response as compared with ethynyl estradiol and mestranol. However, I65. (R=St and R=Pr) were 10 and 15 times more potent than ethynyl estradiol and mestranol as antiimplantation agents, Ihese compounds prevented implantation in rats when admirJ-Stered orally on day 1-7 post-coitum at a dose of 50 and 25 \xg/kg respectively,

11,12-Seco derivatives of doi^nolic acid 1§^> 2T§3. and IJO were less estrogenic than allenolic SLCid. 51

Discussion Seoo-steroids by virtue of their flexible nature, can take up different geometrical shapes depending upon tlie position from where the molecule has been scissored. Some of the conformations of a seco-steroid may be more helpful in binding to receptors as compared to others. The receptor binding affiniiy or its biological activity observed is the mean effect of the preferred conformations. The flexibiliV in the molecule may also lead to their binding to more than one type of receptors-^'37 . It is also likely that such a steroid receptor complex may lack the capability, partly or fully, of producing hormonal effect and thus act as antihormones. Thus C-nor~9,ll-seco-estradiol J8 (R=H) which binds to estradiol receptor with 80^ affinity has only one-fifth equivalent of uterotrophic activity of estradiol. Similarly, seco-doisynolic acid derivative 16^ shows a clear dissociation of estrogonecity and its antifertLliiy activiiy pointing to the possibility that its estrogeneciiy is certainly not the only contributor towards its antifertilily effect.

The data so far available on seco-steroids is much too insufficient to judge the requirements for high receptor binding capabiliiy and biological activi'ty which, if generated, would be helpful in the designing of much needed an ti hormones. 52

1, 2 Referenogs

1. A.Kasal, Coll., ^,, (1980), 2541-2549. 2. A.Kasal, Coll., ^^, (1978), 1773-1792. 3. A.Kasal, Ooll., 44, (1979), 1619-1G29. 4. J.S.Bindra, A. T.Ueyyarapally, R.O.G-upta, V.P.Kamboj and H.Anand, J. Med. Gliem., 18, (1975), 921-925. 5. I.V.Torgov, pare Appl. Chem., 6, (1965), 525. 6. S.N.Ananohenko, V.N.Leonov, A.V,Platoiiova and loV, Torgov, Dolk, Akad. Nauk SSR, 1^, (i960), 73. 7. A.T.Neyyarapally, R.G.&upta, S.G.Srivastava, J.S, Bindra, P.K.G-rover, B, S.Setiy aLid N.Anand, Ind, J, Chem,, 11, (1973), 325-329. 8. G.B.C.Boyce and J. S.Whitehurst, J. Chem, Soc, Part IV, (I960), 4547-4553. 9. R.G.Gupta and U.^nand, Ind. J, Chem., IJ, (1975), 759-760. 10. R. C, Grupta, S, Durani, A.IC, A^'ar^^ral, V.P,Eamboj , and H.Anand, Ind. J, Chem., 19B. (1980), QQ^-Q^d^ 11. I. S.Crossley and R.Dowell, J, Chem. Soc. C. , (1971), 2499-2502. 12. (a) P,I.Eole, S,Ray, V.p.Kamboj and I.Anand, J. Med. Chem., 18, (1975), 765-766j (b) R.COambie and T.D.R.Manning, J, Chem. Soc, C, (1968), 2603, 13. K.Lai, P.I.Kole and S,Ray, Ind. J. Chem., (1982), 0000. 53

14. S.Ial, P.L.Kole and S.Eay, Ind, J. Ohem, , IgB, (1980), 90-92. 15. L.J.Chim:., J.H.ItrgoG, E.M. Staneley, L.A.Richard ajid E.R.Ramiey, J, Med. Chem., 1J(5), (1974), 351-354. 16. K.Lai, Ph.D. Dissertation, Meorut University, Moerut (1981). 17. K.Lai and S.Ray, Steroids, ^i* (1982), 537-546. 18. G,V,Ananthanarayanan, S,N,Rastogi and N,Anand, Ind. J. Ghem., 11, (1973), 1054-1055. 19. F.S.Crossley, R.DoxTell, J, Chem. Soc, G. , IJ., (1971), 2496-2493. 20. K.Meisher, Chem. Rev,, 42, (1S48), 367. 21. (a) J.Eeer, J.R.Billeter and K.Miesher, Eelv. Ghim. Acta, 28, (1945), 1342-1354} (b) C.A. 4jO, (1946), 1850^. 22. H.S.Crossley, J, Chem. Soc, C., 12, (1971), 2491-2495. 23. Ji',H.Batzold, D.E.Govey and C.H.Robinson, Cancer Treat. Rep, 61(2), (1977), 255-257. 24. CI-I.Robinson, U,S. 4,059,630, 22nd Nov, 1977, C.A. 88, I21546h (1978).

25. F.H.Batzold and G,H.Robinson, J, Org, Chem., 41.,

(1976), 313-317,

26. C.V.Ananthana-rayanan, S.N.Rastogi and N.Anand,

Ind. J. Chem., 11, (1973), 1051-1053. 54

27« Y.DabraL, H.Ila and N.inand, Ind. J. Chem., 18B, (1979), 152-161, 28. H.Earbwerke, A-G.Sr,Pat 1,575,753. C.A. H, (1970), 25013. 29. G.iPouqaey, J.Jacques end. G-.Azadian-Boulanger, Eup. J. Med. Ghem,, 10, (1975), 543-547. 30. R.R.Orensliaw, G.M.Luke, T,A.Jenks and G.Bialy, J. Med. Chem., 17, (1974), 1262-1268.

31. J.C.Dubois, A.Horeau and H.B.Kagan, Bull, Soc. Clrlm.

PP., (1967), 1827-1833i O.A. 6J, ll6662e. 32. R.Courrier, A.Horeau and J.Jacques, Gomp. Rend., 224. (1947), 1401. 33. A.Horeau and J.Jacques, Biai. Soo. Chim. Pr., (1948), 707. 34. J.Jacques and A.Horeau, Bull. Soc, Gliim, Pr,, (1948),711. 35. R,Gopalachari and R.N.Iyer, Ind, J, Cliem,, 14B, (1976), 700-701, 36. A,Horeau and J.Jacques, U.S. 2,547,123 Apr. 3, 1951; G.A. 4^, (1951), 8045. 37. J.Delottre, J.P.Mornon, G.Lepicard, T,Ojasoo and J.P.Raynaud, J. Steroid Biochem., IJ, (1980), 45-59. CHAPOBR .II

5,6-SECO~BSTRADiaL AHD RELAIBD CQCPOUNDS 55

2.1 IHglODUGTION MD BAgIS OF \miK

Presently used contraoeptivos are a combination of an estrogen and a progestogen, xfliich pose the risk of hormonal imbalance on continuous long term use, A relatively recent approach in the field ox contraception is the development of anti-hormones. An ideal anti-hormone should bind to the hoimone receptor -vjith high affinity but without B.iay hormone action of its own. Although a number of compounds have been found to have an ti-hormone action but none has so far emerged which is totally devoid of hormone action^ these studies" -^f however, have lead to interesting SAR correlates and one can now rationalise by substructure approach, the parts of the molecule responsible for hormone or anti-hoimone action.

Regulation of hormone action

It is important for the development of better contraceptive to have a clear understanding of the biological events leading to pregnancy and the mechanism of hormone action.

Hormones are secreted by endocrine glands which influence the functioning of cells in distant and apparently unrelated tissues. Steroid hormones play their regulatory role by controlling the synthesis of specific proteins. During the metabolism of cells, protein molecules are responsible for conveying the information carried by genes. 56

The functioning of hormones is dependent upon the interactions betr^een -aie specific protein molecules within the cell, called receptors,^ The complex so formed by these interactions controls the protein synthesis by acting directly on the genetic material of the cell,.

Sex steroid hormones comprise of androgens (the male sex hormones), estrogens and progestins (the female sex hormones). During the sequence of protein synthesis the DNA molecule (car3:ying the genetic information) sends 'instructions' through messenger MA for the synthesis of proteins. At the ribosomes the RITA is translated, each group of three bases specifying one amino acids, which is added to the protein molecuJ^e, Through that mechanism the sequence of bases in a segment of IMA specifies a sequence of amino acids which determines the properties of the proteins.

It has been confirmed that the administration of steroid hormone in an animal causes an increase in the synthesis of Hl'JA in the target cells, followed by an increase in the protein synthesis. Thus, steroids are thought to induce gene expression by coupling with receptor proteins that have a positive regulatory offoct.

It has been revealed that there are receptor proteins in the target cells which bind to specific steroid hormones 57

with high affini-ty and prevent their escape. The receptor proteins for all the knoim steroid hormones have now been identified,

A series of experiments by O'Malley and Shutsung Liao has disclosed that the steroid hormone and receptor complexes after being formed in the target cells move to the nucleus and bind to the chromosomes,

Tlie chromosomes of higher animals is made up of both DNA and a variety of specialised proteins, organized in the complex substance called chromatin. It has also been sho\m that the hormone receptor complexes bind directly to chromatin isolated from the nucleus. However, receptor proteins without hormone were incapable of binding to chromatin.

It can therefore be concluded that horm,one and receptor complexes have the capacity to bind to chromatin and to increase both overall gene transcription and expression of BEA sequences coding for specific messenger MA molecules,

Thus one of the approaches for the designing of molecules would be to liave molecules that would compete for binding to the specific receptor protein with endogenous hormone but fail to initiate synthesis of the desired proteins associated with hormone action. The endogenous 58

estrogenic hoxmone is estradiol. Figure 1 shows the important features of the estradiol molecule. For high affini-iy estradiol receptor binding it is important that the stiiuctural features mentioned in Fig, 1 should not be interfered vjithin the estrogenic ligand. A systematic study of these molecules and the various derivatives for their RBA, indicate that there are factors other than estrogenecity which contribute towards their antifertilily activity. Thus, there exists the possibility of modifying the estradiol structure which would lead to complete or partial loss of estrogeneciiy and enhanced antiferiiliiy activLV«

Delettre^ has pointed out the possibiliiy of achieving this objective through designing of flexible steroid molecules, A great amount of flexibility could be introduced in steroids through the cleavage of specific, bonds which increases free rotations of the substructures in the steroid molecule, which can assume different confoimations including the ones considered essential for binding as well as for -their biological activity.

An important class of seco-steroids vrhich has shown promise as antifertLlity agent is B-seco-steroids, Our earlier work on * B-seco-estradiols has revealed that unlike estradiol molecule in which homologation at C~13 59 o 0.1 c 8 0) c a» en o

mm* D U S o E

o a. 4^ V) 0) I. s c 0) £ OJ *1*5. & 60

results in a drop in receptor binding as well as antiimplantation activities', in the corresponding 5,6-seco-estradiols a marked increase in both. RBA and estrogenecity is observed when 0-15 methyl (C-6 of bicyclo[ 4.5. Ojnonane system) is replaced by an ethyl group, This is very similar to the findings in the 19-nor-testosterone compounds (c,f, norethindrone vs. norgestrel)* Thus 13-e-thyl-5,6-seco- estradiol (I, E=R'=C2H^) (see compound 7, Table 1, Chapter I) was found to be 100^ effective in rat at 2 mg/kg, given orally whereas the corresponding 13-inethyl compound (I, RrrOgH^, R'=CH„) (see compound 5, Table 1, Chapter I) was active only at 5 mg/kg dose. In the present study we have prepared the other ti-ro isomers of (I, R=R'=C2K^), viz, cis-anti-trans compound (8) aiid cis-^n-trans product (_2),

Ij R=CHr,j GpHj-J C„Hry J R =CH™tCpHj- 61

and have studied their biological activities. To enhance the oral absorption and consequently their activi-fcy the synthesis and biological evaluation of 17a-ethynyl group (G-7 of biq5rclo[4.3»0]nonane system) incorporated seco- analogues was carried out, which is reported in this chapter.

In the B-seco-estranes, the role of the alkyl subs tit uent at 0-8 is rather important. Based on receptor binding studies with steroidal and non-steroidal estrogens , a hydrophobic pocket in the receptor has been suggested in this region. It has been discussed in the Chapter I that B-seco-compounds without any alkyl substituent at 0-2 were found inactive at 20 mg/kg, Fo biological data is available of the corresponding 0-2 methyl compounds, C-2 ethyl substituted compound (I, R=OoH(-, R'=CH,.) (see compound 3, Table 1, Chapter I) was found to be 100^^ active at 5 mg/kg as an antiimplantation agent in rat. Further, homologation to propyl (I, B.=:0^ri^, R'=OH ) (see compound 6, Table 1, Chapter I) resulted in a marked drop in RBA, antlimplantation and estrogenic activities. It, therefore, appeared that an optimum bulk requirement in this region which may be responsible for giving a proper thickness to

Q the molecule (c,f, diethylstilbestrol compounds^) is essential for their biological activity. To study the effect of tlie steric requirements for biological activity, 62

preparation of derivatives of B-h.omo~6,7-seoo-estradiol and their corresponding cis-syji-tran^ and jj.^s-ajirfci-traji^s isomers and a study of tlieir biological activity was undertaken, which is described in this chapter.

In a yet another study, sjmthesis and biological evaluation of 17a-ethynyl--5,6-seGO-19-nor"testosterone, the 5,6-seGo counterpart of norgestrol was carried out,which is also included in tiiis chapter.

2,2 ^n the sis of 2a,6p~ and 2p ,6p-diethyl •-3a/p-(£-hydraxy- phenyl) -trans-bigy clo[ 4,3.0] nonan-7p -ol and 2a, 6p - die thyl-7a -e thy nyl -3a -(^-hy dro^qy phenyl) -trans -bicyclo -• I II im •\' Mil —iim 1 • "—riiiwiir-i-i—-m jmi ii iir-im •imT^aii •• i—inM-n • IMI..JBI ^— • n n i mi .. M • ii • -• r- rr i "^ ' 1 —' [ 4. 3.0] nonan-7p -ol

The synthesis of 2a-ethyl, 6p-methyl-3p-(j2""^

The keto enol ^ was prepared by the condensation of 2-ethylcyclopentane-l,3-dione (^2) wi-th propy3. vinyl ketone (l) (Scheme l). Gupta et al. carried out this condensat ion by refluxLng the mixture of 1 and _2 in methanol, in the presence of an alkali. Much better yields of ^. could be obtained by stirring 1 and _2 in demineralized water under nitrogen atmosphere , Cyclization of the trilcetono J^, td-th £-toluenesulfonic acid in benzene, resulted in the formation of 2,6p-diethyl-bicyclo[4*3.0]non-l~en-5,7~dione (4).

Catalytic reduciion of this double bond at the dione stage was found^ to give a mixture of ois and ticgns fused bicyclononan-diones. To have trans^ fused system it was thought advisable to have the 7-keto group in 4; reduced. This reduction was carried out with sodium borohydride in a mixture of isopropanol and ethanol 13 to afford 2» Catalytic reduction of the double bond in ^ would again yield a mixture of cis_ and trans fused compounds, so the reduction vras not carried out at this stage. Condensation of ^ with p-methoxyphenyllithium in ether-TEIP at -40 gave the 2,6p-diethyl-5-hydroxy-3-(£-methoxyphenyl )bic5'-clo[4, 5«01 non~l-<3n-7p ~ol (^), which, vdthout isolation, was converted to the dienol J, by the addition of one drop of HCl, This crude compound was purified over silica gel column using hexane-benzejie gradient. Catalytic hydrogenation of 2, i^^ ethanol at RTP over lO'/s Pd-C gave a mix'ture of two isomers 64

0 ^ a + n^ (5^0^

H/ OH

<- 0' k^

N OH

^ OH -=> CH.0"'^3H^< ^ -^ 6

Oontd... 65

g g ^i. M'

HO^^'^'*^^ 10 11

t II .^^CsGH

r^^,

HO--"^^^.^ 12 66

,C=GH

•V

H H 5 17

a, Demineralized water/lT2J ^» PTSAj c, NaBH^j a, ^BrCgH,OCH„/n-BuLi| e, HGlj f, lOfo Pd-C/H^, 2 mol} g, ZOH, diethylene gil^coli h, Jones reagentj i, LiG=OH/e thylenediamine

Scheme 1 67

having ci^~a3^ti-trans_ (8) and .ois.-sxii-tra2is_ (^J stereo chemistry in a ratio of 40:60, In the nmr spectrum the isomer formed in major amount (2.) showed a M.ghly shielded CHo-CH^ resonance. This isomer was assigned the cis-syn- IR ,^£§;M. (5,) Stereochemistry, on the basis of earlier work ''^ as in this isomer, due to steric interaction between the 6p-ethyl and 2-ethyl group, 2-CHp-GH- group would be held in a plane away from the e-OHp-GH^ (S'ig, 2) and due to this s-beric ct^mpression the ring may undergo some conformat ional change, which would bring 2-0Hp--CH^ within the shielding cone of the phenyl group, This -0H„ appears at 0,136 in nmr. The forma'iion of ^ isomer in major amount could be due to relatively less steric hindrance for hydrogenation from a-face of the molecule. The tvro isomers 8 and % were obtained in pure form by crystallization from ether-hexane, Demethylation of 8 and % with potassium hydroxide in diethylene glycol gave iSie corresponding phenols, viz. 2a,6p- and 2p ,6p-diethyl-5a/p-(^-hydroxyphenyl )-tran^s-bicyclo[ 4,3.0] nonan-7p"Ol (10 and 11). Jones oxidation of 10 yielded the 7-keto analogue 12, Similar Jones oxidation of 8 and % led to the formation of 14 and 16, Ithynylation of 12. was carried out by the method of Hiraga , where lithium acetylide was used in presence of 68

FUi 2 69

ethylenediamine, this gave -fiie Ta-ethynyl product 1^ in very good yield. This compound gave in nmr a characteristic sharp singlet at 2,456 for the acetylenic proton (CsCH), Ethynylation of 14 and 16 by a similar procedure gave the respective 7a-ethynyl compounds l^, and IJ,

2«5 Sijrnthesis of 2a ,6p-diethyl-7a-ethynyl-5p-(£-«ethoxy/ hydroxyphenj'-l)-tranS'-bicyclo[ 4.3» 0]nonan-7p -ol

A controlled catalytLc hydrogenation of J in ethanol at RTB over lOfo Pd-C till 1 mol of Hp was absorbed gave the corresponding Jbrgns fused enol 18 (Scheme 2), The stereospecificity of hydrogenation could be explained by the fact that both 6-ethyl and 7-hydroxy groups are P-oriented and would thus hinder the p-face approach to the catalyst. Such effects in controlling the stereospecificiiy of reduction have been reported for liydrindane and decalone system 17' , The styryl double bond of 18 was reduced with potassium in liquid ammonia to yield 1^, The stereochemistiy of Ig has been assigned on the well established mechanism of metal-ammonia reduction and by analogy vrith the intermediates in estrone synthesis. Column chromatography of I3. over silica gel gave a colourless solid melting at 90^0j the compound reported by Gupta et al, was an oil. Jones oxidation of 12. gave the corresponding 7-keto compound _20 70

e Nl/

vx>C=CH 71

o^ C=CH

2^- 24

a, lOfo Pd-G/Hg, 1 moll b, K-NH„? c, Jones reagenti d, IiC=GH/e thy lens diaminei e, KOH, dieiiiylene glycol

Scheme 2 72

which on ethynylation by the method described for the preparation of l^., provided iiie desired 7a-ethynyl compound 21.

Demethylation of 19. yielded the diol 22 which on subsequent oxidation by Jones method and ethynylation afforded the desired 7a-ethynyl compound 2A,

2.4. ^nthesis of 2a,6p-diethyl-7a~eiiiynyl-3p-(cyclohex-2- en-4-one )-ti;ans-bicyclo[ 4, 5,0]nonan-7p -ol

1 ft Birch reduction of 19 with lithium in liquid ammonia in the presence of a pro tic solvent such as e-liianol, resulted in the reduction of aromatic ring so as to give 2^^, The oxidation of 7-OH in 2^^ cannot be carried out with chromium trioxide (Jones oxidation) as it results in the 19 20 re-aroma"bization of the ring A , Oppenauer oxidation of 2^^ m.th aluminium t-butoxide in benzene-ace tone mixture provided the corresponding 7-keto compound 26_ (Scheme 3). Ethynylation of 26_ was first carried out with lithium acetylide in presence of dimethyl acetamide-dioxan and ethylene diamine by the method of Smith et al, 12 , which was found unsuccessful. In another approach, similar to that for the preparation of norgestrel 12 , lithium acetylide- ethylenediamine complex was used in the presence of dimetliylacetamide. This too could not fetch the desired compound. In both these cases the starting material vras 73

\ OH

O^^V^

28 27

a, Li-^HyCgH^OHi b, AX"^[ t-BuO]" ) c, IiO=OH, etliylene diamine i d, HCl, Me OH

Scheme 3 74

1 C recovered back. However, application of the Hiraga's method over ^ gave the corresponding 7a-ethynyl compound 27. Addition of a drop of hydrochloric acid to 2J afforded the 5,6-seco analogue of norgestrel, viz, 2^,

2,5 SE^nthesis of 2,6p-diethyl~3-(2-methyl-4-hydroxyphenyl) bicyclo[ 4,3,0] non-1-en-7p ~ol

Condensation of ^ with 2-me thyl-4-methoxy phenyl- 13 lithium Ijy the method of Boyce and White hurst -^ gave 2,6p- dietbyl-5-hydroxy-3-( 2-fflethyl-4-«iethoxy phenyl )bicyclo[ 4,3.0] non-l-en-7p-ol (2^). Unlike ^, the 5-hydraxy intermediate 2^ was quite stable and did not dehydrate by the addition of HOI, even after heating, ^. was, therefore, refluxed in benzene with _£-toluenesulfonic anid to give the desired diene ^^O. Catalytic reduction of JO in ethanol over 10^/^ Pd-C till 1 mol of Hp was absorbed gave the trgjis fused enol ^^, Reduction of the 2,5 double bond of ^ was tried t'dth lOfi pd-C, a mixture of 5/» wet and lO/* Pd-G, Raney nickel and PtOp at pressures TBXi^lxig from R[CP to 1500 p,s.i., but without success, Steric hindrance could be a possible reason for the inertness of Ihe double bond

for the a-face approach of hydrogen during reduction (Pig. 3)» It was then thought to migrate the 2,5 double bond in Jg. to 3»4-position by refluxing ^1 with HCl in metlianol, c. f. 9,11-shift in 5a7p-dihydroxy-estra-l,5,5 (10), 8(9)-tetraene . This shift could not be, however. 75

^\CReduc-id.o n -^

12 a, 2-Metliyl-4-bromoaniso[Le/n-BuXi> b, PTSA» c, lOp Pd-C/H2> 1 mol» d, Jones reagentj e, KOH, diethylene glycol. Scheme 4 76

CH,0

Fiqs 77

brought about as the system is some\diat different in flexibility as compared to estradiol, which ha,s a more rigid framework (Scheme 4)»

A newer method of reduction utilizing hydride shift reagent triethyl silane alongwith trifluoroacetic acid " was also tried but ihe reaciion did not go beyond the intermediat, (II), which on treatment with LiAlH^, went back to the orLginal starting compound ^, The identiiy of intermediate

F_COGO 5

(II) was established on the basis of i.r., nmr and mass spectrometiy. Catalytic reduction of the 5-hydroxy compound 22 over 10^ pd-C gave the dienol ^^, As no success could be achieved at this stage of reduction of ^, the reaction sequence in the scheme was carried out without the 2,5-double bond in ^ being reduced, Jones oxidation of ^. yielded the corresponding keto compound ^2., whereas the demethylation of ^ gave the corresponding hydroxy phenyl compound ^,

'"•••-' ^ ^" 78

2.6 BKpBRIMMTiL

All the melting points were taken in sulphuric aoid bath and are uncorrected. IB spectra ( y) cm" ) were max recorded on a Perkin-Blmer 157 spectrophotometer. 'BMR spectra in CDC1„ (unless otherwise stated) were run on a Varian SM3f'0I (60 MHz) or a Perkin-Blmer R-52 (90 MHz) spectrometer using tetramethylsilane as liie internal standard. Ihe chemical shift values are given in 6 units and coupling constant (J) values in Hertz. The mass spectra were recorded on a JEOD-JMSDJOO spectrometer. The purity of the compounds was checked routinely by TDC using silica gel-G as stationary phase and chloroform or chloroform-methanol as mobile phase,

2-Ethyl-2-(3-cKohexyl)cyclopentane-l,3-dLone (j):-

To a suspension of 2-ethylcyclopentane-l, 3-dLone (2) (36.54 g» 0.29 mol) in 65 ml of demineralized water was added, at once, propyl vin^^-l ketone (58.8 g, 0.59 mol) and the mixture i-jas stirred under nitrogen atmosphere at room temperature for 5 days. It was -then extracted with benzene, dried (Na2S0,) and concentrated to yield the desired compound (j) as an oil| yield 50-0 g (76.97/0«

IR(Neat) : I710 (C=0) (broad) 79

mm : 0,85 (n, 6H, tv;o CH,)

Analysis : C,,H2QO Found : C, 69.80j H, 9.11 Reqd. : C, 69.61; H, 8.99/-

2,6p-DiethyllDic3r7clo[ 4. 3.0]non-l-ene-3,7-dione (4): -

A mixture of ^ (22,4 g, 0,1 mol), p-toluenesulfonic

acid (PTSA) (l g) and dry benzene (150 ml) was refluxed using Dean-Stark water separator; a,fter 3 h additional quantLiy of PTSA (l g) was added. The recjiired amount of water was collected in 5-6 h. The reaction mixture was cooled, washed with water, the benzene layer dried over HgSO., the solvent removed under reduced pressure and the residual oil distilled under high vacuum to give J;} yield 17.6 g (86j^), b.p.ll5°/0,01 mm.

IR(Neat) 1745 (7-0=0) acid 1665 (3-0=0) mm 0,85 (m, 6H, 2-aH2CH^ and 6-0H20H„),

Mass M"^ m/z 206.

Analysis ^13^8^2 Pound C, 75.83; H, 8,93 Reqd. 0, 75.69; H, 8.805^;. 80

2,6p-Diethyl-7p-hydroixybicyc5lo[4.3.0]non-l-ene-5-one (^);-

A solution of NaBH. (1,02 g, 0.0268 mol) in isopropanol (100 ml) and ethanol (100 ml) was added drcpwise to a stirred solution of ^4 (20,6 g, 0,1 mol) in ethanol (50 ml) kept at 0°, The reaction mixture was further stirred at 0° for 0,5 h and allowed to come to room temperature and the stirring continued for 1 h, A few drops of acetic acid were then added to decompose excess of NaBH. and the solvent removed under reduced pressure. The residual oil was taken up in ckloroform, washed with water, dried (J^FapSO,) concentrated and distilled to give ^j yield 18,5 g (89/0, b.p. 135^^/0.001 mm.

IR(Neat) : 1660 (C=0) and 3450 (OH). IMR 5 0,85 (m, 6H, 2'

jinalysis : ^2.1^2d^2 Found : G, 74.79; H, 9.61 Reqd. ; C, 74.96; H, 9.68/.

2,6p -Diethyl-3-(£-methoxyphenyl )hioyclo[ 4.3.0]nona~2,9- diene-7p~ol (7):-

A solution of ^ (10.4 g, 0,05 mol) in diy ether (100 ml) was added to a solution of j)-methoxyphenyllithium, prepared from butyllithium (14,4 g, 0,225 mol) and 2-bromoanisole (29.9 g, 0,16 mol) in etiier (400 ml) at 81

-50 to -40^ under nitrogen aimospherej sufficient tetrahydro- furan (about 100 ml) was added to dissolve ihe separated complex aru3. the reaction mizturo stirred at this temperature for 1 h. The temperature was then allowed to rise gradualism to 10*^ and kept at this temperature overnight. The complex was decomposed by adding a saturated solution of sodium ciiloride (lOO ml). The ether layer was separated, dried (NapSO,), concentrated to give 6. (12.32 g, 78,0/0. 7 was obtained hy adding idrop of concentrated HCl to 6. This crude product was chromatographed over silica gel (350 g) using hexane:benzene gradient, to afford pure 7} yield 10.4 g (34.4/0.

IR(!Teat) : 3350 (OH) MR : 0.85 (m, 6H, 2-CH2~0H and 6-GH2^H ), 3.58 (s, 3Ii, -OOH^), 3.95 (t, IE, 7a-H, J=9 Ha),

5.33 (t, IH, 9-11). Analysis ; CgQHpgO^ Found : C, 80.80| H, 8.92 Reqd. : G, 30,50| H, 8.78W

2a ,6p -Diethyl-3a•-( p-methoxyphenyl )~trans--bicyclor 4. 3.0] ~ non-2 •"en-7p -ol (8) and 2p , 6p -Die thyl -3|3 -(£-me thoxy phenyl) - jfcrans-bicyclo[ 4. 3.0]non-2-en-7p-ol (^): •°-

A solution of J (2.98 g, 0.01 mol) in EtOH (100 ml) was hydrogenated over lOf^ pd-C (0.6 g) till 2 mol of H^ was 82

absorbed. The catalyst -viras removed and the filtrate concentrated to give a mixture of 8 and ^ (2,85 g, 95/^). This miztui-^ on crystallization from ether:hexane afforded the two isomers (8 and °ij in a ratio of 40:60,

8, yield 1,05 g (36.84>.), m,p,76^.

IR(KBr) : 3350 (OH) mR i 0.65 (t, 3H, 2-CH2GH^), 0,92 (t, 3H, 6-GH2aH^), 3.18 (m, IH, 3-H), 3.67 (s, 4H, -OCH„ and 7a-H), 6.58-7.16 (m, 4H, Ar-H)

Analysis '^20^30^2 Found G, 79.52; K, 9.92 Reqd, 0, 79.42> H, 10.00/..

2, yield 1.5 g (52.63fO, m,p.ll5°.

IR(iCBr) i 3330 (OH)

NI-IR : 0,13 (t, 3H, 2-CH2CH^), 1,03 (t, 3H,

Analysis • ^20^So*^2 Found i C, 79.73; H, 10.22 Reqd, : C, 79.42; H, lO.OOfo. 83

2a ,6p -Diethyl-3g~(p-hydroxyphenyl )"t3:^ns-bic.yclo[ 4. 3.0] nonan-Tp-ol (lO)s-

A mixture of 8 (0,9 g, 0,003 mol), potassium hydroxide (10 g) and hydrazine hydrate (1 lal) in diethylene glycol (40 ml) was heated at 225° under nitrogen atmosphere for 1,5 h (the heating was slightly slowed at 150-160° to avoid excessive froathing). The cooled solution was diluted wi-fch water (400-500 ml) and neutralized with ace-bic acid and extracted with ether. The organic layer was washed with. 105^ sodium bicarbonate solution, water, dried (lTa2S0.) and concentrated. The residual oil was crystallized with benzene;hexane to afford 1_0} yield 0,65 g (75.5/0> m,p,l63 ,

IR(KBr) J 3300 (OH) MR(GDC1„ + : 0.66 (t, 3H, 2-CH2CH^), 0,97 (t, 3H, 2 drops of DMSO-dg) 6-CH2GH ), 3.2 (m, IH, 3"H), 3.6 (t, IH, 7a-H), 6.6-7.1 (m, 41, Ar-H) Mass M"*" m/z 288 Analysis ^19^28*^2 Found G, 79.00} H, 9.69 Reqd, G, 79.12| H, 9.78fs, 84

2p ,6p -Diethyl-513 -(^-hydroxyplienyl )-^jn£"'bicyclo[ 4. 3. 0] nonan -7p ~ol (11): -

It was obtained by the deme thylation of ^ vd. th EOH in diethylene glycol as described for lOj yield 69.7/^, m,p.l57^

IR(liBr) : 3300 (OH) M'IR(0DC1^-H : 2.8 (m, IK, 3-H), 3.6 (distorted t, Hi, 2 drops of DMSO-dg) 7a~H), 6.5-7.2 (m, 4H, Ai'-H) Mass M"*" m/z 288

'^ TT n Analysis "19 28^2 Pound C, 79.32; II, 9.87 Re qd« C, 79.12; H, 9.78'^

2a,6p-Diet]qyl-3a~(j3-hydroxyphe^y•l )~jtr^ans-bicyclo[ 4.3. 0 nonan-7-one (1^);~

A solution of 10 (0,288 g, 0,001 mol) in acetone (10 ml) was cooled to 15*^, Jones reagent was added dropwise under stirring till the coloar of the reagent persisted. Stirred well for 10 min. Methanol was added dropwise to decompose excess of Gr0„. The reaction mixture was diluted 3 with TTater and extracted with ether, washed successively I'd. th water, sodium bica-rbonate solution, vrater, dried (MgSO,) and concentrated in vacuj) to give 12 as an oil} yield 0.219 g (76.7^0. IR(Neat) : 1710 (0=0) 85

Analysis • C^ 0^26^2 Found J C, 79.34} H, 9.01, Reqd. : 0, 79.68| H, 9.15V-"

2a f 6p "DiG thyl-7a -e thynyl-3a-(^-hydroxy phenyl)-trans- bicyclo[4. 3.0]nonan-7p-ol (r^):-

Acetjaene was passed through a solution of 0,5 g lithium in ethylenediamine (100 ml) for 1 h and a solution of 12. (0.286 g, 0.001 mol) in tetrahydrofu.ran (10 ral) was added to the aceiylide solution during 30 min, with stirring at room temperature. Acetylene was passed for further 2 h and the mixture was neutralized with ammonium chloride (5.0 g), diluted with water (500 i-nl) and extracted mth ether. The organic layer was washed with 2^o cold HpSO,, saturated sodium bicarbonate solution and water successively, dried over NapSO, and concentrated to afford l^S yield 0.214 g (68.59>'0.

IR(Neat) : 3300 (OH) MR ; 2.45 (s, IH, CsCH) Mass ! M^ m/z 312 Analysis 1 ^21-^28^2. , .- Found G, ai.02; E, 9.21 Re qd, !' G, 80,73? H, 9.03/^ 86

2a,6p -Diethyl~3a"(£-methoxyphonyl )-tran^,"'taicyclo[,4, 3.0] nonan-7~one (14-.): -

It was prepared from 8 in 89?^ yield by a similar procedure as described for iiie preparation of 12,

IR(Neat) s 1750 (0=0) Analysis • n T^T n . ^20-''28 2 Pound ; G, 79.61} H, 9.21 Reqd. s G, 79.96j H, 9.39/».

2a, 6p -Die thyl ~7a -e thynyl-3a -(£-me thoxy phenyl) -trans- bicyclo[4. 3.0]nonan-7p-ol (l^)s-

Preparation of 1^ was carried out from 14 by a similar method as described for the preparation of 1^| yield 72.5/°.

IR(Neat) 5400 (OH) MR 2,42 (s, IH, C=CH), 5.68 (s, 3H, OCH^)

Mass M"*" m/z 526 Analysis a^oH^,.22^^50 o2 . ,. Pound C, 81.14; H, 9.57 Reqd. C, 80.94} H, 9.26^.

2p ,6p -Diethj'-l-5p -(^-methoxyphenyl )--;fc.-mns--bicyclo[ 4. 5.0' nonan-7~one (16);-

^nthesis of 16 was performed by Jones oxidation of ^ by the method described for the preparation of 12^} yield 92/.. 87

IR(lTeat) 1730 (G=0) Analysis ^20-^28^2 Pound G, 8D.23J H, 9.51

Re qd. C, 79.96; H, 9.39?t.

2p ,6p -Die•tih.yl-7a-e 1ib.ynyl-3a~(£~me thoxyphenyl)-trans-- bicyclo[ 4. 3.0]nonan-7p -ol (IJ): -

It was prepared from 16, in 82^^ yield, by following a similar method as desoDibed for the preparation of IJ^.

IR(iT8at) 5400 (OH) MR 2.53 (s, IH, C=GH), 3.67 (s, 3H, OCH^)

Mass M"*" m/z 326 Analysis ^22^30^2 Found C, 81.28J H, 9.40 Reqd. 0, 80.94; H, 9.26?i.

2, 6p -Die thyl-3-(£~nie-'aIioxyphenyl )-trans,-bicyclo[ 4. 3. 0] non- 2-ene-7p-ol (18);-

A solution of J (2.98 g, 0.01 mol) in EtOH (100 ml) was hydrogenated over lOji Pd-C (0.5 g) till 1 mol of Hg had been absorbed. The catalyst was removed and the fi.ltrate concentrated to give 18 as an oilj yield 2,7 g (90,3/0.

IR(Neat) .' 3350 (OH) 88

MR ; 0.66 (t, 5H, 2-QE^GE^), 0,93 (t, 5H, 6-CH2CH ), 5.65 (s, 4H, -OGH and 7a-H), 6.6-7.2 (m, 4H, Ar-H) Mass • iC m/z 3OO iUaa3.ysis ; GgQH^gOg Pound : 0, 80.21; H, 9,52 Reqd. : C, 79.96} H, 9.39/=.

2a ,6p-Die-tihyl-3p -(^-methoxyphenyl )"jfcr||is--bicfy-clo[ 4. 3.0] nonan~7p-ol (l^J:~

Potassium metal (4.5 g, 0,115 g atom) was added in small pieces to a stirred mix'ture of 18 (3.0 g, 0,01 mol), dry dioxane (125 ml), dry ether (15O ml) and liquid ammonia (500 DLL) kept at -50°, The solution was further stirred for 2 h, the excess of potassium amide was d.ecomposed by the addition of FH/Gl and ammonia was allowed to evaporate off. Water was added to the reaction mix-bure, saturated m.-'!ii UaOl and extracted with ether. She organic layer was washed wiiii saturated FaGl solution, dried (ITapSO.) and concentrated. The residue was chromatographed over silica gel (75 g) using benzene as eluant to furnish a ciystalline prc^dunt 12} yield 2.4 g (79.6f«), m,p.90°,

IR(Iffir) ; 3350 (OH) 89

mm. : 0.68 (t, 3lh 2~aH2aH^), 0.96 (t, 5H, 6-GH2CH^), 3.2 (m, 1H,3-H), 3.67 (s, 3H, DCH ), 6.6-7.2 (EI, 4H, Ar-H)

Mass M"^ m/z 302

Analysi s ^20^30^2 Found C, 79.67} H, 10.18 Reqd. G, 79.42} H, 10.00^..

2a ,6p "Die thyl-3p -(^-methoxyphenyl )-_tr£ms-liicyclo[ 4. 3. 0] nonan -7-one (2j0) s -

A treatment of 1^ with Jones reagent, as described for the preparation of 12, gave 2£ as an oil in 92,7/= yield.

IR(Neat) : 1740 (G=0) : 3.2 (m, IH, 3~H), 3.65 (s, 3H, -OGH^), 6.6-7.15 (m, 4H, Ar-H)

Mass. M"^ m/z 300

Analysis °20^28''^2 Found G, 79.70} H, 9.22 Pue qd. C, 79.96} H, 9.39^^.

2a, 6p -Die thyl-7a ~e thynyl -3p -(£-me thox yphonyl) -trans- bicyolo[ 4. 3.0] nonan-7p -ol (^): -

It was prepared as an oil. from 20 follomng the method described for the preparation of 1^, in 80^^ yield.

IR(Neat) : 3450 (OH) 90

MR 0.69 (t, 3E, ^-CHgCH^), 0.94 (t, 3H, 6-

CH2GH^), 2.42 (s. Hi, CECH), 3.25 (m, IH, >-K), 3.68 (s, 3H, -OGH^), 6.6-7.2 (m, 4H, Ar-H) Mae M"^ m/s 326

Analysis ^22^*30^2 Pound G, 81.27; H, 9.31 Reqd, C, 80.94; H, 9.26fb.

2a f 6p"Die thy 1- 3p- (p-liy dr oxy phenyl Ktrans-'blay clo[ 4.3.0] nonan-7p-ol C^):-

It -was obtained by the demethylation of l^ wi'feh. KOH in diethylene glycol as described for 10, in 57.7f= yield. m.p.l49°.

IR(KBr) 3350 (OH) MR 3.2 (m, IH, 3-H), 6.5-7.1 (m, 4H, Ar-H).

Mass M"^ m/z 288

Analysis *^19^2S^2 Found G, 79.45; H, 9.93 Reqd, G, 79.12; H, 9.78^^.

2Q;>6p"I)iethyl-3p-(j?-hy droxy phenyl )--_trans-bioyclo[ 4.3.0] nonan~7-one i2^)i-

Treatment of ^ with Jones reagent following the method used for iiie preparation of ^} afforded ^2 as an oil in 77^/^ yield. 91

IR(Neat) : 1750 (C=0) : 0.69 (t, 3H, 2-CH2CH^), 0.97 (t, 5H, 6-CH2GE ), 3.21 (m, Hi, >-H), 3.65 (t, IH, 7a-H)

Mas s s M"^ m/z 286

Analysis • ^19^26*^2 _ . Found ! C, 79.74i E, 9.24 Reqd. : 0, 79.68; H, 9.15f^.

2a ,6p~Diethyl-7o:-ethynyl-3p-(j>-liyd.roxyplienyl )-trans-

r^ Pw 11 ••-*;>- iDicyclo[ 4,3.0]nonan-7p~ol (^4):-

Application of a procedure similar to the one described for the preparation of _21 produced ,24 as a colourless solid in 74/o yield, m,p,99^.

IR(ia3r) 3300 (OH) roiR ! 2.4 (s, IH, G=CH)

Mass ! M"*" m/z 312

Analysis ' °21^28^2 3?o und ! C, 80.94', H, 9.19 Re qd. ! C, 80,73i H, 9.03

2a ,6p-Diethyl-3p-( 4-me thoxycyGlohexa-l,4-diene )-jfcra^'' bicyclo[4. 3.0]nonan-7p-ol {2^)t-

A solution of ^ (2,0 g, 0,0066 mol) in dry ether

(200 ml) was added to liquid ammonia, folloi^d "by small 92

pieces of lithium (2.5 g). After half an hour, absolute alcohol (35 ml) was added dropwise during 15 min. When the blue colour had disappeared, the ammonia was evaporated off. After the removal of most of ammonia, cold Ti^ater vras added carefully the product was extracted with ether, saturated NaCl solution, dried (Na2S0.) and concentrated to yield ^ as an oil; yield 1.41 g (70.2^0.

IR(!Ieat) J 5350 (OH) miR : 3.45 (s, 3H» OCK^), 4.55 (distorted t, IH, C(OCH,)=CH"), 5.4 [distorted t, IH, C(0CH^)"CH2-CH=a] Mass M"^ m/z 304

Malysis ^20%2^2 Found C, 79.26) H, 10.73 Reqd, C, 78.90; H, 10.59/».

2a,6p-Diethyl-3p-( 4-methoxyGyclohexa-l,4~diene )-tran,&:: bicyclo[ 4.3.0]nonan-7-one (26):-

A mixture of ^ (1,0 g, 0. OO33 mol) in dry acetone (10 ml) and dry benzene (I5 ml) was heated to boiling in an oil bath, maintained at 85 . A solution of aluminium tert-butoxide (1,5 g, 0.00042 mol) in dry benzene (10 ml) was added in one portion to the boiling solution and re fluxed for 4 h. After cooling the reaction mixture was poured in water, extracted with ether, washed with water, 93

dried (Na^SO.) and concentrated in vacuo to afford 26 as an oil; yield 0.74 g (74.6'/.).

IR(Neat) : 1720 (0=0) ! 3.45 (s, 5H, OCH^), 4.5 [distorted t, IH, G(OCH^)=CH-], 5.37 [distorted t, IH, C(O0H„)-GH2-QH^

Mass } M^ m/z 302

Analysis ' '^20^30*^2. Pound : C,, 79.71; H, 10.28 Reqd, J C, 79.42; E, 10.00>^.

2a,6p"Diethyl-7a-ethynyl-3p~( 4-metlioxycyclohexa~l, 4--diene)-

_ !•- •• r •- - "I ^ - - -| - 1 -- 1 'ir -n " - r. - - • i ir - -i n - - i i i - - n •• i • --i ^ i - - - -^ -r t 'i H -ii - - trans-bicyclor 4.3.0]nonan-7p~ol (_27)s-

It was prepared as an oil from _26 by a similar procedure as described for 1iie preparation of IJ in 70^b yield

IR(Neat) s 3400 (OH) ; 2.45 (s, IH, C-GH), 3.53 (s, 3H, -OCH^), 4.58 [distorted t, IH, C( OCH^)=CH-], 5.44 [distorted t, IH, G( 0CH^)-CH2-C^Ii=G]

Mass : M^ m/z 328 Analysis • ^22^32^2 Pound { C, 80.69*, H, 9.99 Re qd. ; C, 80.44; H, 9.82w 94

2a:,6p-Dieth7l-7a~etliynyl-3P"(cyclohex-2--en-4-one )"tr£tnS" «-.*—Ifcrifcl • l%l 1..^..1^l •ll^ni III *•!•••• W, ..li 1^1111.1 M.II.I ^—— — .1*.*. nil ^1 I.I •^lll^..i^»M«.l i>»M^lMiH 11.111 11 •••11 > •l.iail «IWI» II » II •! II -f • biqyclo[4, 3.0]nonan-7p^ol (18):-

To a solution of 21 (IO5 ng, 0,00033 mol) in msthanol (5 ml), 2 drops of cone, HOI were added and the solution was heated at 60 for 15 min, The reaction wavS cooled and poured into cold water, extracted with ether, washed -vrith bicarbonate solution and water. The organic layer was dried (NapSO^^) aiid concentrated under vacuo to provide ^28: yield 0.071 g (70,6^i).

IR(Neat) 1720 (0=0), 3350 (OH)

Mass M"*" m/z 314

Analysis ^21^'30^2 Found 0, 80.52; H, 9.75 E.eqd, 0, 80.25i H, 9.55>^.

2, 6p-Die thyl- 3-hy dr oxy- 3- (2~me thyl- 4-me thoxy phenyl )bicyc lo- • 1*1 n •ii-o ^— — »i«î —la—iîiiM i'. i^tiiiti wii wiiia, ^ I— i»iii«pii.wi.».fi.«.i I, ,,,. wmwmn\ • iîai i ii».ii.i ^.i if »i m *»> » •! 11 ••^•••fci i—.•••i — •» .WWIP.^ [4.3.0]non"l-en-7p-ol {2^);"

To a solution of 2-methyl--4-methoxyphenyllithium, prepared from bui:yllithium (19.2 g, 0,30 mol) and 2-methyl-- 4-bromoanisole (42.21 g, 0.21 mol), was added on solution of 3 (10.4 g, 0.05 mol) in dry ether (100 ml) at -40^ under nitrogen atmosphere. Further, reaction w^s carried out similarly as described for the preparation of J7. The crude product thus obtained vras ciystari-ized from benzenejhexanej yield 13.6 g (82.4^i), m.p. 123-124°. 95

IR(KBr) : 3400 (OH) i 0.9 (t, 6H, 2-CH2CH^ and S-GE^CE^), 2.4-3 (s, 3H, ilrCHj), 3.65 (s, 4H, OCH^ and Ta-^), 6.3-6.9 (m, 3H, Ar^)

Mass : M m/z 330 Analysis • ^21^30^3 Found J 0, 76.42| H, 9.28

Reqd. 5 C, 76.33; H, 9.15f».

2,6p--])iethyl- •3- ( 2-me thy 1-4-me th-oxy phenyl )'bicyclo[ 4. 3.0] nona-2,9-dier le- 7p-ol (JO):-

A solution of _23 (6.6 g, 0.002 mol), and p-toluene sulx.:3iiiG cucld • (1 g) in benzene (100 ml) was refluzed for 2 h. The mixture was washed with water, solvent layer dried (NapSO.) and ooncentrated to provide ^0 as an oil; yield 5.25 g (84.13/0.

IR(lTeat) ! 3350 (OH) NiyiR i 0.88 (m, 6H, 2-CH2CH, aid 6-CH2OH ), 3.68 (s, 3H, OGH^), 4.08 (t, IH, 7a-H, J=9Hz), 5.43 (distorted t, IH, =CH), 6.45-6.9 (m, 3H, Ar-H) + Mass : M m/z 312

Analysis ; C2^H2Q02 Found : C, Sl.Olj H,9.19

Reqd. : C, 80,73; H,9.03fi. 96

2, 6p -Die thyl" 3- (2-me thyl- 4-me th oxy plieiiyl) b icy c 1 o [ 4.3.0 ] non- l-en-76"-ol Cli)'"'

A solution of J^ (3.12 g, 0,01 mol) in ethanol (100 ml) was liydrogenated over 10^^ Pd-G (0.3 g) till 1 mol of Hp '^SiS absorbed. Ooncentration of the filtrate obtained after removal of the catalyst gave Jl as an oil; yield 2.9 g (92.35^^).

IR(lTeat) : 3400 (OH) i 0.73 (t, 3H, 2-CHpCH^), 1,03 (t, 3H, e-Cll^OE-^)y 2.1 (s, 3H, ArCK^), 3.63 (s, 4H, OCH^ and Ta-H), 6.47-6.9 (m, 3H, ir-H) Mass : M m/z 314

Analysis ! C2]_H^Q02 Pound : G, 80,56} H, 9.86 Reqd. J G, 80.21} H, 9.625^.

2, 6p-Die thyl-3-(2-me thy 1-4-me thoxy pheny 1)b icy clo[ 4.3.0] non- l-en-7-one (^):--

• i-^im^git^aw^^jMi*-*^'-^!!^ Jones oxidation of ^ following a similar procedure as described for the preparation of j.2 gave ^2 in 74/^ yield,

IR(Neat) ; 1740 (0=0) mR : 3.67 (s, 3H, CCK^), 6.5-6.9 (m, 3H, Ar-H)

Mass : M^+ m/z 312 97

Analysis • *^21^26^2 Pound : C, 31.11} H, 9.21 Keqd. : G, 80.73; H, 9.05;^.

2,6p~Diethyl-5-( 2-methyl~4« hydroxy phenyl )bioyclo[ 4.3» Ojnon- rijiii•>>••. •.^>..-% i-*T I^.nnM^îî-i ifci ji%LWî«%..jiwi%, I—iiMi.i ÎJW l-en-7p-ol (^)!-

A mixture of J,l (0.628 g, 0.002 mol), potassium hydroxide (7 g) and hydrazine hydrate (1 ml) in diethylene glycol (50 ml) was given a similar treatment as mentioned for the preparation of _10, to ohtain ^5_2 8-^ a colourless solid, yield 0.550 g (56fO, m. p.l50°.

IR(KBr) 5400 (OH) 2.5 (s, 511, ilrcCH^), 6.4-6.9 (m, 5H, kc-E) Mass M"^ m/z 500

n TT rs Analysis ^20^28 2 Found C, 79.76J H, 9.21 Reqd. C, 79.96j H, 9.59/». 98

2• 7 BIOLOqiOM) AGi'IVITT

iJJ. the 5» S-secoestradiol ds.rivatives pi-epared were assayed for tlieir receptor binding affinity for es'brogen receptors and antiimplantation activity in the Ifi.vision of Endocrinology of Central Drug Research Institite, lucknow, according to the follotTing methods*

22 -* ^S"!^yQg,QJi_J^eo6ptor_Bin^d^^ Tlie relative binding affinity of the compound for estrogen receptor was determined by competition assay, employing radiolabelled estradiol ([•^H]-E2)> as the reference compound. The test ligands and [^HJ-Bp "^^^-'"^ incubated ''4^) with cytosol estrogen receptors obtained from immature 20-21 days old rat uteri. ALiquots of the uterine cytosol (200 \ili cone* 1 uterus per ml} prepared in ISA buffer (10 mM Tris, 1.5 mfi EDTA, 0.02/a sodium azide, pH 7*4) were incubated in trip].icate with a fixed concentratLon of radiolabelled estradiol x-dth or without various concentrations of the competLtor -substances dissolved in 60 |il of the EBA buffer containing IMP as cosolvent (final concentration of MF in the incubation medium never exceeded 5^), for 18 h at 4°. At the end of this period, dexti^an coated charcoal (DCG) (5^9 Noi-lt A, 0.5^» dextran) suspension in 100 |j.l of 3EA buffer was added into each tube, which were briefly vortexed and allowed to stand for 15 min..DCG was precipitated by 99

cen-ferifugation (800 g x 10 min.) and the supernatants counted for radio-activity in 10 ml of a dioxane-based scintillation fluid. RBA of the test compound was computed from a graph plotted between percent bound radio-activity versus log concentration of the test substance. At 5Qf» inhibiHon, the log of the competitor concentration relative to ths,t of estradiol, gives the affinliy of the test compound relative to estradiol. Tliis when multiplied with 100 gives the percentage value designated as RBA.

RBA = "'"^^ concentration of reference compound _ log concentration of test compound

2. Antiimplantati on AotLvity

An tiimplantati on activity of the B-secosteroids was studied in sperm positive female albino rats mated to coeval males of proven fertility. The compounds were administered orally as a suspension in gum acacia to colony bred adult mated female rats (150-170 g) on days 1-5 or 1-7 post-coitum using five to seven animals in each group. The animals were examined by leprotomy on day 10 of pregnancy for the number of implants. ThQ results were scored as positive only if implantations were totally absent in both uterine horns. 100

Results of i±Le testing are ^ven in !Ikble 1.

_!fa,ble_ 1

G ompd* Dose Route of No. of day percent No. mg/kg/day adrninist- of pre g- efficacy (rat) rati on nancy post- coitum i

8 10 oral 1-7 100 9 20 oral 1-7 28.5 IP. 10 oral 1-7 100 11 20 orsr-al 1-7 80 M 10 oral 1-7 100 5 oral 1-7 66 il 10 oral 1-7 80 21 2 oral 1-7 80 2i 1 oral 1-5 100 i5 2 s. c. 1-5 16.6 26 2 oral 1-5 16.6 29 10 oral 1-7 100 2 oral 1-7 50 IQ 2 oral 1-7 100 S 2 oral 1-7 100

^ 10 oral 1-7 100 12 10 oral 1-7 100 2 oral 1-7 40 101

JUscu^ssijoai

The results show i±iat £4 lias about 25«48fo RBA as compared to that of d-estradiol-lTp ("^2^' ^-^ decrease is quite significant, even if we presume that the drop may be because 24 is a raoemate and its RBA may be an underestimate to at least 50fei it is known tSiat l-estradiol-17p has quite 23 low receptor affiniiy '^.

Ihis could be die to the increased degree of rotational and vibrational freedom in the region of ring C as compared to that of ^2* ^'•'•^-^ points out iiie need of molecular rigLdity in this region for a Mgh RBA.

The earlier work' on the comparative affinities of 6-CH, (I, S=02He, R'=CH^) and G-GgH^ (22) had revealed that the latter has almost three-fold greater (25.85/» of ^2^ affinity for the receptor. This suggested the presence of an additional binding interaction due to additional methyl (6-CpHt-) as compared to the former, perhaps, vdLth a hydrophobic pocket in the vicinity where 14-GH^ of E2 binds. This additional receiotor interaction, thus, partly offsets the loss of affinity owing to cleavage of ring-B of estradiol, as is clear from the norethj,sterone-norgestrel relaUonship .

Hie present studies have indicated that iiiere is, but slight, increase in the RBA of ^ by the introdictlon of a .7a--ethynyl group (c. f. 24; )• 102

The pregnancy inhibiting results reveal that 2A is the best candidate molecule in the 5» 6-seco-estradiol series, being lOOfo active at a dose pf 1 mg/kg when administered orally to female albino rats on 1-5 day post-coitum. Testing results at &rther lower dose are in progress.

The 3-methoxy analogue of 2J. (c.f« 21) was found to be only QCf/o active at an oral dose of 2 mg/kg when administered 1-7 day post-coitum. This emphasizes the need of -bvio hydroxy groups viz* phenolic and 7p-0H for better pregnancy inhibiting results.

imongst the 5»6-seco-norgestrel series both 25. and _26 were ineffective in inhibitirig pregnancj?- when adudnistered subcutaneously and orally respectively at a dose of 2 mg/kg on 1-5 day post-coitum. The result of progestrational assay for the 5> 6-seco-analogue of norgestrel (c. f. _28) is yet awai te d.

Other important compounds vrere the representatives of B-homo-6,7-seco-estradiol series. ^ and ^1 were found to be 10055 active at an oral dose of 2 mg/kg. Testing at further lower dose is also being carried out at present. The activity of these compounds could be explained on the basis of their resemblance with the -ya-methyl analogue 7 of estradiol which ha^ better activity as compared to EJ. 103

However, it would not be jusiified to draw a conclusive remark on ihe structure activity relationsMp of this series as the data available at present is not sufficient. 104

2,7 References i

1, S.Durani, A.K.Agarwal, R.Saxena, B. S.Setty, E.G.Gupta, P.L.Kole, S.Ray and N.Anand, J. Steroid Biochem. , 11,

(1979), 67-77. 2, S.Diirani and N.Anand, Int. J. Quantum Cliem,, XZ, (1981), 71-83. 3. W.Herimann, R,¥yss, A.Riondel, D.Philbert, G, Teutsch, E, Salciz and E.Baulieu, G.R. Acad. Sc. Paris, 2^±, Series ill, (1982), 933-933.

4. ¥.0'Malley and W. T. Schrader, Sci. Am., _2j4( 2),

(1976), 32-43. 5. J.Delettre, J.P.Mornon, G.Lepicard, T.Ojasoo and J.P.Raynaud, J. Steroid Biooliem., 12, (1980), 45-59. 6, R.C.Gupta, S.Durani, A.K.Agarvjal, V.P.Eamboj and N.Anand, Ind. J. Chem., l^^B, (1980), 866-890. 7. J.P.Raynaud, T,Ojasoo, M.M.Bouton and D.Philbert in Dru^ Design, Vol,VIII, Academic Press (1979), pp 169-214. 8, Einer-Jensen N,; Acta pharmac, tox., 2^, Suppl. 1, (1968), 1-97, 9. J.Grundy, Chem. Rev., 5J., (1957), 281-416. 10, A,T.lleyyarapally, R.O.Gupta, S.C.Srivastava, J.S. Bindra, P.K.Grover, B,S.Setty and N.Anand, Ind. J. Chem,, 11, (1973), 325-329. 105

11, S.W, Ananchenko and I.V, Torgov, Dokl. Akad. Nauk.SSSPL,

12.7, (1959), 553. 12, H. Snith, G,A.Huges, G.H.Douglas, G.R.Wendt, G.G.Buzby, Jr., R.A.Bdgren, J.Fisher, T.Foell, B.Gadsby, D.Hartley, D.Herbst, A.B.A.Jansen, K.Ledig, B,J. MacLoughlin, J.McMenamin, T.W.Pattison, P.C.Phillips, R.Rees, J.Siddall, J.Siuda, I.L.Smith, J.Tokolics and D.H.P.Watson, J. Chern. Soc., (1964), 4472-4492.

13, C.B.C.Boyce and J, S.White hurst, J. Chem. Soc,

(I960), 4547-4553. 14, Z.G.Hajos and D.R.Parisch, J. Org. Chem., 2Sl) (1974), 1615. 15, J,S.Bindra, A. T,Ney7arapally, R.C.Gupta, V.P.Kamboj and N.Anand, J. Med. Chem,, 18, (1975), 921-925. 16, K.Hiraga, Chem Pharm. Bull., IJ, (1965), 1289-1294,

17, H. anith, G.A.Hughes and B.J.McLoughlin, Experentia,

la, (1963), 177-178. 18, A.L.Wilds and N.A.Nelson, J. Am. Chem. Soc, J3., (1953), 5360-5366. 19, H,J,Ringold, G.Rosenkranz and F, Sondheimer, J. Am. Chem, Soc, 18, (1956), 2477. 20, R.V.Oppenauer, Org. Synth., 21, (1941), 18-20, 21, D.N.Kursanov, Z.N.Parnes, G.I.Bassova, IT.M.Loim and V,I,Zdanovich, Tetrahedron, 2^, (1967), 2235-2242. 106

22. J. A.Katzenellenbogen, H.J. Johnson, Jr. and H.V. Mysers, Biochem., 12 (1975) 4065. 23. (x.A.C]aernayaev, T.I.Barkova, V.V.Egorova, I.B.Sorokina, S.N.Ananchenko, G-.D.Ha*tetradje, H. A. Sokolova and Y.B.Rozen, J. Steroid Bioohem., 6 (1975), 1483' CHAPTER III

CEVBLOPMBNO} CF SPERMICIDAL ACSFTS 107

3.1 SPERjyilOlDAL AGENTS: AE OVBRVIM

The oral contraceptive 'pills' have no doubt revolutionized the practice of birth control, but at the same in view of their continuous administration needed in their use have raised many problems regarding their safely and acceptability due to high motivation required. Search for a safer, simple, effective need based method is therefore continuing. One such approach is the use of spermicides as topical contraceptives. As the name suggests these are agents that inactivate or kill sperms and prevent fertilisation and thus provide pregnancy protection.

The spermatozoon is a high3^ specialized cell. As shown in the Fig. 1, it consists essentially of a head containing the paternal hereditary ma-terial, mid-piece and a tail which provides a means of locomotion. The latter two constitute the 'flagellum'.

The process of fertilization involves an effective union of sperm and ovum. The biochemistry involved in the process is still not clearly understood. The two main considerations that may lead to fer-fcilization are motiliiy and fertilizing capacity of the sperm. 108

Outer acrosomal ton9 Galeo Acrotome capitis Inner acrosomal tone } HEAD - ^luclear membrane Nucleus Po5t-nuclea^ cap NECK - Centrjole Axial filament complex Mitochondrial sheath MIDDLE ^ PIECE Plasma membrane

TAIL

Tall sheath

End piece

FiG. 1 DIAGRAMMATfC REPRESENTATION OP A SPERMATOZOON 109

Although the freshly shed spermatozoa, after ejaculation, lodged in the vagina, are morphologically and metabolically immature with low motility, they reach the u-terus ascending -tiirough the cervical canal, within minutes after ejaculation by some as yet rather poorly understood mechanism.

Estrogen, progesterone, oxytocin and -fiie sympathetic and parasympathetic nervous system impulses and hormones have aH been implicated as regulators of spermatozoon transport in the female tract . Estrogen enhances the rate of ciliaiy beat to some extent, whereas progesterone alters the uterine motility markedly. Oxytocin speeds up the spermatozoa to the fertilization site. However, it has been observed in most of the mammals studied that the spermatozoa are not ready to fertilize ova until they have been in the female tract for about six hours after mating.

Physiological changes occur in -tiie spermatozoa while passing through the female tract and give rise to 'capaci tated' spermatozoa. Certain detrimental forces extensively reduce the number of spermatozoa passing up cervix. Spermatozoa finally penetrate zona pellucida of the ovum, for the formation of zygote. Only one spermatozoa penetrates the inner most vitelline membrane in normal case, though several come into contact with each ovum. 110

This is an important factor to block polyspermy; the precise mechanism for this is not yet clear. Thus, in the process of fertilization an effective blockade of sperm function either by immobilizing it or by interfering with its maturation can result in contraception. In the female, spermicides are used in the form of vaginal creams, jellies or foam tablets.

Vaginal contraceptive preparations or spermicides are amongst iiie oldest and simplest methods used to prevent conception. Egyptians dating back to 1850 B.C. were the first to use these vaginal contraceptives. They mixed honey and natron (native sodium carbonate) with crocodile or elephant dung to form a paste for using intravaginally. Later on oils, honey, cedar gum, various fruits, herbs, juices, alums and rock salts, based primarily on their strongly acidic or astringent properties were used as . . , 2 spermicides , Among the first commercial spermicides were quinine pessaries developed by Walter Rendell, an English pharmacist in 1885 , These preparations consisted of soluble cocoa butter plus quinine sulfate. Since then a number of compounds such as boric acid, tannic acid, lactic acid, ricinoelic acid, sodium sulfate, formalin and urea have also been used '^, Later on in 1939,, hydroquinone was shown to be better than quinine. Ill

These spermicidal preparations can be classified as follows;

(1) iia&^al-^rodUSis;

The natural products represent one of the oldest class. Quinine salts (1) were the first to be used for this purpose-^''* , later, emetine h;i'drochloride (2) was also found to be active, but the former (^1) showed better 7 activity .

C=CH

HOv.

.2HC1

OGH. OCH.

.2H01 H^C-^/^^X. 112

More recently the saponin soheffleroside i"^) derived from Schefflera J3aj2itata and the saponin from ,%jeindu,s mukorosii have heen shovm 9—11 to possess marked sperm 12 killing activity. A triterpene glycoside, Samaninr-D (A) , isolated from the flov^ers of ^±ihOQolobiAm saman Benth. was also found to have a high order of activity.

iPuc ose-galactose-gluconi c~ Cr\;/t\^

Me-x^Me

R,R,,R2=glucosyl, arahinosyl, 4 xylosyl, rhamnosyl.

This was followed by the demonstration of spermicidal activity in saponins of Madhuca Jbutzracea, M.latifolia, Mimusops hexandra> M.elen^i seeds, Albizzia procera roots and seeds, Acacia concinna hark and Anagallis arvensis whole plant -^ -^, The saponins mostly belonged to 113 spiroketal steroid and oleanane series of triterpenoids.

An orally effective male contraceptive, gossypol (^), isolated from Thespesia ppigulnea and cotton seed was found to be an effective spermicidal agent. The polyvinyl pyrrolidone derivative of ^ I'^as more active than the parent compound 17

OHO OH OH 0111

18 lutenurin , another plant product showed high degree of sperm kiJJLlng activity besides its antimicrobial activiiy. It did not show an;^'' vaginal irritation and had an instantaneous spermicidal effect in a dilution of 1:1000. OleanoglycotoKin-A and lemmatoxin derived from l^hvtolaQQa dqdecandra, were reported to be 5 times more potent as spermicides than the saponin (_4), and the latter was more effective against human sperms. Foaming capacity of these saponins can be retained for a longer duration by mixing it with polyvinyl alcohols. In the initial experiments saponins were found free of irritating property. 114

(2) Synthetj^c Spermiqidesj

These can be broadly classified into two groups?

(i) NitrOi^enous compounds Recently it was observed 20 that chloroquina {j6) was even more active than quinine (2-) as a spermicide, being active at a concentration of 3.6x10 ^M, as coippared -A to 5x10 M' for quinine. RHCHMQ(CH2)J^TSt2

Spermicidal activity for cetylpyridinium chloride (J) 21 or bromide was reported in late thirties, which were

Cl^^GH2)i5CH^

1 known for their bactericidal activity.

Potent spermicidal activity has been reported for 2-mercaptobenzthiazole (_8) and 2-mercaptobenzimidazole 22 23 (_2) , comparable to that of gramicidin , a kno-^cn spermicidal agent. 115

High, spermicidal actLviiiy lias been reported for N-n-buVlbenzisothlazolone (jX)), being effective at concentrations as low as 0,0001562/9} 1,2-benzisolthiazole derivatives could be included in various spermicidal preparations at a concentration of O.l-ljS due to their high potency 24^ .

if—Bu

Recently, F-alkylated l,>-dihydroxy"-2-alkyl-2- aminopropane (11) -', F ,N -disubstituted piperazines (12) 27 ' and a-ace-tylenic amines (_12) were reported to show spermicidal actLvi-ty.

H-pOH R-|_ = H or alkyl Rn •\ f Rp = alkyl !!• •0 —R. R„ = methyl or ethyl R. iH2O H 11 OHCO2H Cl^ \y_OHRlTHCH2CH2 6HCO2H 12 Ha=C.GHR.ITHR' 12 116

(ii) Non-nitrogenous compounds Sperm inmiobilizing activiiy was also observed po PQ in polysaccharides , S-J-chloi-opropane-lja-diols (j^) and lipid peroxides-^ , The polysaccharides are known to OH HO-CH2-6H-GH2-CI

(M)

exert their action by inhibiting liyaluronidase and blocking fructolysis while lA inhibits glycolysis in sperms. The potassium salt of copolymer of jo-hydroxyplienylmethacrylate showed activity^3 1 , especially when ascorbic acid was used as stabilizer. A well known analgesic antipyretic drug, aspirin, was capable of exerting spermicidal activiiy by "52 maintaining pH 4.5 of vagina . Some highly acidic agents such as lactic acid, boric acid, tartaric acid and citric acid in combination with other spermicidal ingredients also find use as vaginal contraceptives'^-^33,

(3) Oompounds of Heavy Metals: In late thirties spermicidal activity was observed in some knoiwi bactericides e.g, mercuric chloride, 34 phenylmercuric nitrate (PMN) (15.) or phenylmercuric 117

acetate (PMA) (J.6)^ ^ , A number of spermicides were later prepared with PMA as base materia]., which had greater 0 II HglTO, ^,^;^ /HgO-G-CPI^

spermicidal effectiveness than quinine sulfate, Eie use of PMA is generally not advised due to knovm toxicily of mercuric compounds. The world contraceptive panel 36 classified'^ mercury containing compounds in category tT;'To, that is 'effective' but not generally recognized as safe.

A number of heavy metals such as copper, iron, zinc and cadmium, lithium and magnesium were capable of immobilizing sperms-^"^*"^ . X& X4.'fe9 studies revealed that iron in lo;f concentrations (0,153M in FeCl„) could immobilize human spermatozoa.

(4) Surface .^.ctiye, ,Agents; A major important advancement in the development of spermicides was the demonstration of activity in non- ionic surfactants or surface active agents. These compounds immobilize the sperms by disrupting the integrity of the sperm membrane and inhibiting the oxygen uptake and fructolysis. The main surfactants which are in clinical 118 use are nonylplienoxypolyethoxyetlianol (nonyl-9 or nonoxynol-9, XI) ^^^ menfegol or j-methanylphenyl- 40 polyoxyethylene ether (TS-88) (IS) Incorporation of 3-9 0-(aH20H20)^H Me

'y-{0QE2^E^)^0E

17. CHMe^ parts of water soluble alkylphenoxyethanol (jL8) having 6-7 ethoxy groups into an emulsion base yielded combinations which were spermicidaJ. in pH range 6,8-8,6. _17 and _18 are now the principal active ingredient in most of the spermicidal products commercial3-y available. TS-88 is one of the simplest, safest and most effective and economical contraceptive.

Another surface active agent which possesses potent activity is Laurenth-9 [a poly(oxy )"ethylene lauryl ether] . It ira-S lethal to human spermatozoa within 20 seconds in a concentration of 1:2000, A contraceptive cream containing Laurenth-9 and 7-chloro-4-indanol (clorindanol) {1^) was subjected to toxicologioal and

01 19 spermicidal properties and reported to immobilize 119

spermatozoa within 20 seconds even after 100 fold dilution.

The screening results of a large number of non-ionic, anionic and cationic surface active agents have revealed that appropriate oil solubility and good surface tension lowering properties were desirable for spermicidal activity in noil"ionic and anionic surface active agents, Cationic compounds exhibit activity, although they have low fat solubility. Benzalknnium chloride (^2^0) and trimeiiiyl ammonium bromide, cationic surface active agents are active

+ OH cr f CH^ 11 R R=CgH^^ to C^gE^^ CH. 20 with almost the same order of activity as the most active anionic and non-ionic surface active agents. Surface active agents and bactericides together often have a synergistic effect, making the combined product a more effective spermicide than any one of the ingredients alone.

(5) Aero sin Inhibitors; Certain synthetic inhibitors of acrosin when mixed with spermicide were reported to be more effective preparation than the spermicide alone. These compounds readily penetrated through the acrosomal membrane of 120

spermatozoa and brought about certain 'biochemical changes whinh inhibited fertilization. _p-Nitrophenyl-j'- guanidinobenzoate (NPGB) was the most potent inhibitor of human acrosin, KPGB v/hen mixed with K-Y-jelly in a concentration of 0.09 mg/ml, showed marked contraceptive activily in primates.

In a more recent study 43 the use of acrosin inhibitors for inhibiting human spermatozoa was disapproved. A majoriiy of the tested acrosin inhibitors, except proteinase inhibitor Trasylol and synthetic inhibitor 4^nitrophenyl-4- guanidinobenzoate, could not penetrate the acrosomal membrane of testicular, ejaculated and uterine human spermatozoa. These two inhibitors caused moderate to complete inhibition of the gelatinolytic actLvily of the spermatozoa if applied in concentrations of 1-10 mmol/liter.

The spermicidal products used today are available in the forms of creams, jellies, foams in pressurized containers (aerosole), foaming tablets and suppositories.

Spermicides appear to possess a number of advantages and if used properly may be more tlian 95/o effective?

''Being locally used they are not likely to interfere with the various biochemical events of the human system. 121

'•'They are easy to appljr and require low motivation. Moreover, they may not interfere with the milk production during lactation period of women.

''They may provide certain degree of protection against various veneral diseases caused by Neisseria gonorrhoeae and .Tro_£ei^ma ^gaJLLidim and some other sexually related infections.

Spermicides can also be used in males. Development of such agents as may take care of the drawbacks mentioned below appears an important area in the battle against population increase.

Two major drawbacks associated with the use of spermicides are that: (1) they must be applied shortly before coitus which some women may dislike, and also affect privacy and (2) they have relatively greater failure rates as compared to sterilization, oral pills, lUDs or condoms. 122

3.2 INORODUCOIOIT MP BASIS OF WORE

Until some ten years back there appeared little hope for promoting tlie use of vaginal spermicides in the family planning programmes, as an answer to the ever mounting population. However, the limitations encountered with the more commonly used contraceptive methods such as the oral progestational pills and the lUD have brought back into focus this simple and old fashioned method 44 , Spermicides could be used liien other methods are not available e.g. if the pill has been missed for several days, after an lUD has been expelled or for the first few months after vasectomy. In fact, spermicides are generally used not so much due to their advantages but out of the disadvantages vjhioh users find or fear in other methods.

Some of -Hie most common spermicides available today suffer from the drawback of their effect being pH dependant. This underscores the need for developing newer spermicides which may take into account the drawbacks of the present day spermicides.

Since very few compounds are knovm as spermicides and still less is known about the biochemistry of 123

spermatozoa, on the basis of wliich. new corapounds could be designed, one of the ways for generating new leads was the random screening of available compounds. In such a screening programme carried out earlier in tliis Institute, ll-substi"buted 3"aminpacrylophenones synthesized as anti inflammatory agents ' were found to possess marked spermicidal activity. In particular various IT-substituted a-aminome thylac ly lophen one s (I) and a,a-diaminome thylac e to- phenones (II) and their quaternary salts 47 possess

spermicidal actlviiy, which was about 10 times that of nonylphenoxypolyethoxyethanol (nonyl-9) (JL7)» ^^^^^ novel lead seemed worthy of exploration and prompted the synthesis of U-substituted a-(2-aminoethyl)acrylophenones (III) and >-F-substituted 2-methyleneindan-l-ones (IV) as potential spermicides, which is discussed in this chapter. 0. / 0

III 124

3.3 ^nthesis of N-substituted a-( 2-aminoethyl)ao2•ylo- phenones (^-^)

The starting materials ydilo'ô'bu-'qyroph.enone (_21-^) re quired for the qynthesis of N-sabstituted o;-(2-aminoethyl) acrylophenones (38-^0) were obtained either commerciaD.lj'' or prepared by the Friedel-Crafts reaction of substituted benzenes and y-chlorobutyryl chloride. The intermediate compounds y-aminobutyrophenones (22-_27) were obtained by re fluxing y-chlorobuiyrophenone with an appropriate amine in acetone in the presence of potassium carbonate. The condensation of y^îô^jutyrophenone with paraformaldehyde was first tried by re fluxing the mixture of the t\fo in acetic acid but no success could be achieved. This is quite surprising since the analogous compounds viz. 47 IT~substituted a-aminomethylaciTlophenones (I) were obtained by reflujcing 1 mol' of paraformaldehyde with p-aminopropiophenone in acetic acid or directly by re fluxing a mixture of acetophenone and two mol of paraformaldeh^'-de vrith 1 mol of an appropriate amine in acetic acid (Mannich reaction). However, the reaction was carried out successfully by using higher boiling pentanoic or hexanoic acid instead of acetic acid, to furnish the required compounds Q&-J5.0). The condensation of yânîiioûtyrophenone with paraformalde hyde first gives an intermediate (V). The hydroxy 1 group 125

from Y is then dehydrated under acidic conditions to give the desired compounds QS-^jO) (III), The structure of these compounds was assigned on the basis of their nmr and ir data.

0—ti

H V

21, n=^ 25-27 22, R=Br 25, R=CH_S 5 b 24, R=H N/

H^ (III)

a, M.T'/ I h, HCOpH, Hexanoio acid

Scheme 1 126

Characteristic signals at 5.2 6 and 5.5 6 for G = C appeared as narrow doublets (J=l, 5 Hz) due to their allylic coupling with G-2 protons in the nmr spectrum of the compounds ,58-^_0. The carboiiyl group in ir appeared around 1650 cm" .

3,4 Etynthesis of 3-amino-2-substi-!:uted me-bhylene indan-1- ones (6^72)

¥ith a view to study the effect of the relative disposition of the side chain and ihe aryl ring on the biological activity of the acrylophenones, synthesis of 2-substituted methyleneindan-1-ones (,M-_^'72) (IV) was undei^ taken, Indan-1-one (^51) was prepared by the qyclization of p-phenylpropionic acid ^jith AlCl-^ respectively by the literature method^4. 8 , Bromination of_^1 with N-bromosuccinimide in presence of catalytic amount of benzoyl peroxide afforded 4-9 the corresponding 5--bromo derivative ^J, Similarly cyclization of p--( >-methoxyphenyl)propionic acid to ^2 and subsequent bromination yielded ^4. The presence of bromine at G-3 position was confirmed by nmr spectroscopy. The signal for 3-JH appeared as a quartet due to .cis- and _trans_. coupling, wj.th C-2-proton around 5.45 6 (q, IH,

-Gjpr, JQis=3.5 Hz and J^^.^ns'^''' ^^^ ^^^ '^^^'^ ^°^ ^'^ ^^ ^'^^ ^ (m, 2H, Ojip). Reaction of ^ with different amines gave 127

.<^^ a -^ E-^^^:::^ R' •Br 31, R=H 52, R=oaH_ 5i, R=OCH^

0 r^>^ <=^^p.

/ R,y^^ ^N 64-72 (IV) 55-62

Q OH, r<;^ 1 f R^"^/^ NT-"

6J, H=H

a, OS, CCl./Benzoyl peroxidei b, M , Aoetone/K2C0„i c, 4fo KOH (alcoholic)

Scheme 2 128

the respective 5-*aminoindanr-l-ones (.55-^0), Similar reaction of ^4 with piperidine and N-phenylpiperazine yielded 61 and J2, ildol condensation of the amino derivatives {^^ and_^§_6) with different aldehydes in the presence of 4^ alcoholic solution of potassium hydroxide furnished the desired benzylidene derivatives C§,5-J0). Similar condensation reaction of £1 and _62 with j)-methoxyben2aldehyde yielded _71 and ^22 respectively. In the nmr of all these compounds (^"29 andJ2) the C=CH proton was submerged in the multiplet for aromatic protons. However, in _21 it appeared separately- as a doublet, due to a]-lylic coupling with 3-iJ at 7.58 6 (d, IH, G=CH, J=1.5 Hz). The '^-E appeared as a doublet around 5.0 6 (d, IH, C~3jx, J=1.5 Hz). In the ir spectrum of these compounds (^6_5-_J2) the oarbonyl peak appeared around 1690 cm"-''.

Surprisingly, the condensation of amino derivatives (_^-j60) with various substituted benzaldehydes could not be carried out aiccessfully. In all these cases the starting material was recovered back. However, if much stronger alkali solution vras used, only polymeric, tarry substance was recovered. The reason for -tiais failure could not be assigned. 129

Condensation of 3-piperidinoindan-l-one (,^) with paraformaldehyde in acetic acid, after refluxing for 5 h resulted in a mixtare of tifo isomeric compounds 6J and 6A (R=H) in a ratio of TOs50 as established hy the peak intensities in nmr spectrum. In the nmr spectrum of this product there appeared a singlet at 1,18 6 for •fe.e methyl protons of J^ and a doublet at 4,46 6 for 3-J and two ssts of doublets at 5.76 6 and 6,05 6 (J=1.5 Hz) for the O^CE^ of j64. Attempts to separate these two products in pure form were unsuccessful. 130

3-5 EXpERIMSNTikL.

Melting points were taken in a sulfuric acid bath and are uncorrected. IR spectra ( smsx in cm" ) were recorded on Perkin Elmer 157 model and IMR on a Varian A-60D, Varian Bl\I56QL or Perkin^Blmer R-52 model in CDCl™ unless othervd-se stated, using 3MS as internal standard. The chemical shift values are given in 6 units and coupling constant (J ) values in Hertz. The purily of the compounds was checked routinely by TLC using basic alumina as stationary phase and chloroform or chloroform-methanol as mobile phase.

Y-Piperidino-_p-fluorobu1yrophenone {_^5)

A mixture of Y-chloro-^fluorobuiyrophenohe (21) (2.0 g, 0.01 mol), piperidine (0.79 g, 0.01 mol) and potassium carbonate (2.76 g, 0.02 mol) in aihydrous acetone (50 ml) vras refluxed for 48 h. The reaction mixture was cooled and filtered to remove the suspended solid. The usual work up of the filtrate furni^ed the required compound (2_5)» which was converted to its hydrochloride, m. p.l79^ (lit.^^ 180-181°)! yield (free base), 1.2 g (48/0-

IR(neat) : 1680 (C=0) 131

NMR : 1.4 [m, 6H, NCH^CGH^) -], 1.66-2.58 [m, 8H, QE^OE^lS{CE^)^]j 2.68-3.21 (m, 2Ii, C0CH2)» 6.66-7.45 [m, 2H, ilr-H (o to F)], 7.55-8.11 [m, 2H, Ar-H (m to P)] Analysis ; 0^ H^QKfO.HCl Pound s G, 63.251 H, 7.14j N, 4.79 Reqd. j G, 65.04| H, 7.00| N, 4.9(^»

£11 the compounds (£5-^) described in Table 1 were prepared hy the above method. The spectroscopic data of these compounds are given in Table 5» a-N-Piperidinoeiiiyl-^fluoroacrylophenone (_28)

A mixture ot 25 .HCl (2.85 gi 0.01 mol) and paraformaldehyde (0,9 g, 0.01 mol) in hexanoic acid (25 ml) was heated to reflux. After 2 h paraformaldebyde (0.45 g» 0.005 mol) was again added to the solution and refluxed for an additional period of 1 h. The acid was removed under recluced pressure and the residue crystallised from alcohol- ether mixture to give ^ as hydrochloride; yield 2. 0 g (67.5/0, m.p.(HGl) 206°.

IR(ITGat)(free t 1660 (C=0) base) NIffi(free base)s 1.3 [m, 6H, HOE^iOE^) ], 2.1-2.75 (m, 8H,

GH2GH2l^r(0112)2), ^'^^ f^* -^^^ -'=^H(1^^§ "^ CO) J=1.5Hz], 5.64 [d, IH, C=C-H(cis to CO) J=1.5Hz], 6.8-7.1 [m, 211, Ar-H(o to F)], 7.6-7.9 [m, 2H, Ar-H(m to F)] 132

Analysis ; C^gH2QB"H0.HCl Pound : C, 64.74l H, 6.86| N, 4.61 Reqd. : C, 64.531 H, 6.72| IT, ^,l^o,

ill the compounds (^-^0) described in Table 2 were prepared by a similar procedure. The spectroscopic data of these compounds are given in Table 5.

3-Br omo indan.-1- one (55)

A mixture of indan-1-one fjl) (13.2 g, 0,1 mol), N-bromosuccinimide and benzoyl peroxide (1 g) in anhydrous COl. (350 ml) was refluzed for 4 h and then allowed to stand overnight in a refrigerator. Succinimide was removed by filtration and the filtrate was concentrated under reduced pressure to give _52. The resultant product was ciystallized from CCl^i yield 15.6 g (73-9^0, in.p.55° (lit/^ 54.5-55°).

IR(Neat) : 1700 (0=0)

NMR(CC1.) : 3.09 (m, 2H, -CH2), 5.45 (q, IH, 3~H,

'^cis^^-^^^' '^fe^ns"'^^^^* 6.83-8.05 (m, 4H, Ar-H).

3-Bromo-5-methosyindati-l-one (_54)

It vras prepared by the bromination of 5-methoxyind~.an~ l~one by the procedure mentioned for the synthesis of J_J in % yield. 133

IR(Neat) : 1700 (0=0) raR(GCl ) , ; 3.02 (m, 2H, -CH2), 3.85 (s, 3H, OCH^), 5.4 (q, IH, 3-H, Jcis=5-5Hz, J^^^=7Hz), 6.73- 7.05 (m, 2H, 4-H and 6-H), 7.38-7.63 (m, IH, V-H).

3-N-.piperidinoindan-l-one (J5,)

A solution of 3-l)romoindan-l-one (53) (2.1 g, 0.01 mol) in anhydrous benzene (50 ml) was cooled to 5-10° and a solution 01 piperidine (1.7 g» 0.02 mol) in anhydrous henzene (5 ml) was added dropwise with stirring, Ihe reaction was allowed to continue for an additional period of 30 minutes The separated piperidine hydrobromide was filtered off and the filtrate was concentrated to give the desired compound (55)1 yield 1,5 g (70.45/0, m.p.(HCl) 187°.

IR(ICBr)(HCl) : 1700 (G=0) MR(free base); 1.1-1.83 (m, 6H, NaH2(0H2)3)> 1.96-2.45 [m, 4H, IHOE^)^], 2.46-2.7 (m, 2H, -CH2), 4-38 (q, IH, 5-H, J^=4.5Hz, Jtrans=^-5Hz), 7.1-7.75 (m, 4H, Ar-H) Analysis : C^.E^^O.HCl Found J G, 67.071 H, 6.91| N, 5.35 Reqd. ; C, 66.80| H, 6.76? I, 5.56fo.

The compounds {5^-§2) (Table 3) were prepared by a similar procedure starting from _5,3 or ^54. 134

2-Ben25''lide ne- 3~ piperidinoindan-1-one (65 )

p^.d*«iWw%««9*im">i*r«'i«B*'vuita«iawNn.Mi(..7iaiM To a mixture of 5-W-pip'eridinoindan-l-one (^JJ (2,15 g> 0.01 mol) and bmzaldehyde (1.06 g, 0.01 mol) was added ^'jo alcoholin potassium hydroxide solution (10 ml) and stirred at room temperature for 2-3 minutes. The separated solid was filtered and vrashed with cold dilute ethanol to furnish 6_5f yield 2.4 g (79.2/0, m,p.l41° (lit^^ 141-145^).

IR(EBr) ; 1690 fC=n). fflffi % 1.4 [m, 6H, NCH2(CH2)J, 2.16-3.0 (m, 4K, N(CH2)2]» 4.96 (d, IH, 3-H, J=1.5Hz), 6.76.- 8.21 (m, lOH, G=C-H and Ar-H)

Analysis ; ^21^21^^^ Found : C, 77.76f H, 9.03i N, 6.18 Reqd. : G, 77.88J H, 9.15j N, 6.05f».

ill the compounds (66-7_2) described in Table 4 were synthesized by a similar procedure from the corresponding 3-amino derivative of ^ or _54- The spectroscopic data of these compounds ere given in Table 5» 135

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in- coj en' inj. inf i^n1 o ins inS VJ3! 149

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M o H CMi 151

5*6 BIOLOGICAL AOTIYIGJT

Th.e general pharmacological screening of the compounds was carried out in the livision of Pharmacology and Hie spermicidal activity in the lUvision of Endocrinology of the Central Drug Research Institute, lucknow.

PharmacaLcgiCc.l Activity

Methods Gross behavioural effects and toxicity (ALDco) of the coapounds were studied in albino mice by intraperitoneal administration of the compounds suspended in gum accacia using five animals per dose* The antiinflammatory activity was tested against carrageenin induced oedema in mice at 1/5th of liDcQ dose p, o* by standard method-^ . The . diuretic activity of the compounds was tested in rats according to the literature method accept that chloro- thia2d.de (125 mg/kg) was used as standard instead of urea. Activity of the compounds was expressed as ^ activity of the standard*

Effect on blood pressure and interaction with histamine, adrenaline, isoprenaline and acetyl choline was studied in anaesthetised (pentobarbitone 35 mg/kg, i.v.) oats, using 5 mg/kg i.v« of each compound. 152

Spermicidal Activity Method

The animal spermatozoa were collected from the cauda epididymis and vasa deferentia of admit male albino rats of the Institute's animal colony, and suspended in physiologj.oa' saline. Ihe human sperms were obtained from masturbated semen sample from healthy human volunteers (2 or 3 parity) with normal consistency and good quality were allox-red to liquify for 50 minutes at 37°C. Specimens sho^Ting good sperm numbers and motility were used during -this stuc3y.

The compounds were dissolved in physiological saline at different concentrations. Two drops of human semen suspension were placed on a slide and to this were added tvro drops of a solution of the compoundj control slides were prepared by adding two drops of physiological saline. The contents vrere mixed with a glass rod for five seconds and examined under a phase contrast microscope (hereafter referred to as spot test). The results were scored positive if lOOfb of the spermatozoa became immotile instantaneouslyj even if one or t^-ro spermatozoa showed sluggish motLlity> the test was scored negative. The minimum effective concentration of positive compounds was repeated twice on different semen samples or rat sperm suspension to confirm the activity. 153

Results

None of the oompoands screened for pharmacological activity showed any notevroriiiy activity.

All the compounds wore screened for their spex^micida: activity and iho results of only such compounds which show noteworthy spermicidal activity are included in Tahle 6.

J^iS. J

Compound Spennicidal Compound Spermicidal No. activlV^' 0 ITo. activity* S 0.5 14; >0.5 19 0.1 15 > 0.5 12 >0.5 1§ 0.05 li > 0.5 11 >0.1 0.025 0.5 •t-iBO-^ .i9 12 > 0.1 50 0.1

^Spermicidal activity in human semen. Ihe values are given as ft minimum effective concentraiion*

Ihe screening results showed that the substituted indanones (IV) had no spermicidal actj.vity» thus showing that incorporation of the side chain into a cyclic structurt abolished Vae activity. In the N-substituted a-(2-amino- 154 ethyl)aoryloph.enones [4-tertiary amino-a-methylene-butyro- phenones] (III) the piperidiiio oompounds had a higher order of activity than the others and amongst them a-K-piperidino- ethyl-j-hromoacrylophenone (42.) was the most active. But now the activity of this compound was almost l/lO that of the corresponding propiophenones (l), the prototype, based on whose structure these compounds were designed, thus showing ttiat lengthening of the side chain led to a lowering of activity. 155

3 - 7 RSJWlJNCB_S

1. T.I'Iann, in ' The Biochemistry of Semen and of the Male Reproductive Tracts Gh. II, Methuen and Co.Ltd., London, (1964), 17. 2. IT.E.Himes, in 'Medical History of Contraception', Williams and Willikins Company, Baltimore (1963), 521. 5. J.R.Baker, in ' I!he Chemical Control of Contraception' Chapman and Hall, London (1935), 151* 4. M.D.Aktiebolag, Ger. Pat. 9556^0, (Jan. 3, 1957); C.A. 5^, 10674e. 5. C.Umezaki, D.S.Fordney-Settlage, Contraception, JLO, (1974), 135.

6. J.R.Baker, J. Hyg., 3,? (193?)» 171 7. P.O.Brown-Woodman, I. G.White, Theriogenology, 8> (1977), 199. 8. A. A.Kassem, A. Ahdelbary and S. A. Hour, Bull Fac Pharm. , 14, (1977), 199. 9. G.K.Jain, J.P.S.Sarin and N.M.Khanna, Ind. J, Chem., Sec.B (1977), 1139. 10. B.S.Setty, V.P.Kamhoj , H. S.Garg and F.M.Khanna, Contraception, 14, (1976), 521. 156

11. B.S.Setty, V.P.Kamboj and N M.IOaanna, Ind. J. Exp. Biol., 15.» (1977), 251.

12. I.P.Varshney and N.Khanna, Ind. J. Pharm^ Sci , _40,

(1978), 60. 15. R.Banerji, G.Misra, S.E.Nigam, S.Singh and R.C. Sazena, J. Steroid Biochem., 9, (1978), 864. 14. R.Banerji, A.K. Srivastava, G-.Misra, S.K.Nigam, S.Singh, S.G.Nigam and R.C.Saxena, Indian Drugs 1J(1), (1973), 6-8. 15. R.Banerji and S.K.Nigam, J. Ind. Chem. Soc, ^,(10), (1980), 1045-4. 16. S.M.Gameron, D.P.Waller and L.J .D.Zaneveld, Fertil. Steril., 17(2), (1982), 275-274. 17. D.P.Waller, L.J.D. Zaneveld and H.H. S.Fong, Contraception, ^2(2), (1980), 185-187- 18. S.A.Viohkanova, M. A.Rubiuohik and V.Fitontsidy, Non. Khoz. Sb., (1964), 254 (Russ)j C.A. 6J, 12159e. 19. S.J. Stolzenberg and R MParkhurst, Contraception, 10, (1974), 155. 20. N.P. 5trifnnac and G.S.Bernstein, Contraception, ,25(1), (1982), 69-87. 21. V.ITreka, Gasopis Lekaru Ceslcych., 89, (1950), 651. 22. M.L.Tarakhovskii, V.V.Rybakova, E.P.Mesynov and p.S. Pel'kis, Fiziol. Ak'tiv- Veshohestva, (1969), 115| C.A. 72, 156559m. 157

23. I.I.Sokolovskaya, L.P.Drozdova, M.G.Golysheva, A.I. Korotkov, Y.I.Maksimov and V. A.Lebedev, Izvest. Timiryazev Sel'skokhoz Akad (1956), 195$ 0 A. 51, 11442. 24. A.Butti, G.Gazzani, Ger. Pat. Ofien 2,629,018 (27 Jan., 1977)l CA. 8,6, 145938X. 25. B.L.Zenitz, U.S.Pat. 3,315,682 (11 April, 1967)j C.A. 67, 11187a. 26. U.A.Sonurlikar, B.Shanker, P.A.Kirke and IT.B.Bhinde, Bull.-Hoffkine Inst. ^, (1977), 946| C.A. 8j, 141224a.

27. M.B.Walter, J. Michel, Belg. Pat. 868,594 (16 Oct.

1978)I C.A. JO, 103380m.

28. P.Buckner, W.Herbrand and ¥.Rockt, Deut. Med. Woohsclir

78, (1953), 907.

2 9. J. N. Fredrick, Ger. Pat. Of fen. 2,743,858 (6 April, 1978)? C.A. 89, 24722a

30. J.Roy, M. Thaddeus and S.Richard, Fertil. Steril.,

S' (1979), 531. 31. S.I.Kotenko, V.Y Pochirok, M L.Tarakovskii, AG Fadeioheva, Farm Zh. 2^5, (1970), 53 f^Russ, )| C.A. r^_, 69786k. 32. R.M.Haller, Ger. Pat. Offen 2,044,401 (11 March, 1971); C.A. 75, 25386s. 158

33. Population Reports Series H, No.3» 'Vaginal Contraceptives, A Time for Reappraisal?' (January, 1975). 34. T.Fikola and M. Slavica, Acta Pharm Jugoslav., 1^, (1966), 17^ 35. V.Kozlik, K.Hejedly, A.Vesela and E.Bayerova, Pharraazie, 2^0, (1965), 106. 36. United States Pood and Drug Administration (USFDA) Advisory Review Panel on OTO Con-braceptives and other Vaginal Drug Products, Rockville, Maryland, USFDA, (Nov. 1978), 6 p. 37. K.Loe^dt, Contraception, ^j (197L), 219. 38. R.I .May hew, I.G-.Nunn and R.L. Sundberg (to G-enex-al Aniline i Film Corp.), U.S.Pat. 2,774,709 (18 Dec, 1956 )j C.A. ^, 4639e. 39. W.G.Mende and V.R.Berliner (to Ortho Pharmaceutical Corp.), U.S.Pat. 2889250 (2 June, 1959)| C.A. ,5J, I8399g. 40. I.Atsumi and I.Toshio, Contraception, j6, (1972), 401. 41. (a) D.A.Berberian, W.G,Gorman, H.p.Drobeck, F.Coulston and R.G, Slighter Jr , Toxicol. Appl. Pharmacol,, 7, (1965), 215. (b) D.A.Berberian, ¥.G.Gorman, H.p.Drobeck, F.Goulston and R.G,Slighter Jr. Toxicol. Appl. Pharmacol., 7, (1965), 206. 159

42. L.J.D. Zanevald, S.Beyler, D.S.Zimand A.K.Bhattacharya, Biol. Reprod., 20, (1979), IO45. 45. W.B.Scliill, M.Peifel, H-Fritz and^ J.Hammerstein,

Int. J. Androl., 4(1), (1981), 25-38. 44- Population Reports, Series H, No. 5, 'Spermicides - Simplicitj/- and Safety are Major Assets' (September 1979). 45. R.C.Gupta, Ram pratap, S.K. Chatter jee, R.C.Srimal and N.Anand, Ind. J. Chem. 1^5B, (1977), 641. 46. Ram Pratap, R.C, Gupta, R.0.Srimal and N. Anand, Ind, J. Cliem., 19B,(1980), 695. 47. P.Nautiyal, Ph.D. Dissertation, Luoknow University, ludcnow (1981 )• 48. D.B.Bruce, A.J.S.Sorie and R.H.Thomson, J. Chem. Soc.

(1953), 2403. 49. C.S.Marvel and G.W.Hinman, J. Am. Chem. Soc, J65

(1954), 5435. 50. Science-Union et Cie-Sooiete Prancaise de Recherche Medicale ("by Laszio Beregi and Pierre Hugon^ Pr.1,301, 863, Aug 24, 1962, Appl. June 29, 196lf C.A. 5,9, 8759. 51. Istituto Luso Parmaco d'Italia S r.l. Brit 1,075,156 July 12, 19671 Ital. App.Aug. 27,1963s C.A. 68,69041k. 52. Aktiebolag Perrosan, Brit 1,000,781, Aug. 11,1965? C.A. 6_2, 13279 53. G.Maury, E.Ming ¥u and N,H.Cromwell, J. Org, Chem , ^2, (1968), 1901. 160

54- R.O.Srimal and B.N.Dhawan, Ind, J, pharmacology, 14:» (1972), 172. 55- D. S.Bhakuni, M.L.Dliar, M.M.Dhar, B.I.Dhawan and B.N. Mehrotra, Ind. J. Expt. Biol. J, (1969), 250. GHAP!EBR ly

JiiTTljOBPRESSMT A(MITS 161

4.1 A SHCRT REVrSW OT JCTTIDEPRBSSMTS

PsycotherapoutLo agents that normalize the mental state of a depressed individual are termed as antidepressants or ' thymoleptics'r Mental depression is a complex disease and is more like a syndrtsne TCL"tihL a multifactorial causation.

Ihe drugs presently available for depressed states are far from satisfactory. The ' tricyclics' are the most commonly clinically used group of aniidepressants, but even to these only about l/5rd of the patients respond satisfactorily. Further they have some major anticholinergic side effects such as dryness of mouth and constipation and serious toxic effects such as hepatotoixicity. Ihese factors underscore the need for new and novel antidepressants.

Ihe oliance discovery of the mood elevating effect of the antitubercular drug iproniazid (1, Fig. 1) in early 1950's, led Kline and co-workers to use it ra.th some success 2 in depressed patients. Simultaneously Zeller _et al. discovered that iproniazid (1) was an inhibitor of the enzyme 162

CH~NH~NH-C-f N

S^^is-^sO

FIQl 163

monoamine oxidase (MAO). Following -fee discovery of iproHiazLd, a series of monoamine ojcidase inhibitors (MAOI) and non- inhibitors were derveloped as antidepressant drugs, which is discussed below.

(l) Monoanine CKidase Inhibitor (MAOIJ as intidepressants

Monoamine oxidase, an enzyme >Jidely distributed in the body* oxidatively deaminates physiologically active primary amines. For example, tyramine is oxidised to p-hydroxyphenyl- acetic acid, epinephrine to 5,4-dihydromandelic acid, dopamine to 3»4-dihydroxyphenylacetic- acid, tryptamine to indoleacetic acid and 5-hydroxytryptamine (5-HT or serotonin) to 5-hydrcK:yindoleacetLC acid.

The possible role of monoamine such as norepinephrine (NE), dopamine and serotonin in the central and peripheral nervous system was suggested by Brodie'^*\ Tlie monoamine depleting action of reserpine"^5 ' 6 , a potent tranquiliser and hypotensive agent and the reversal of its biochemical and v ft pharmacological effects by the MAOI, iproniazid (j.) indicated an important role of these monoamines in controlling the GNS functions. Ihe MAOI couj-d be broadly classified into hydrazine derivatives and non-hydrazine deriva-tives. Some comprehensive reviews ' are available -vjhich deal •^d.th these two class of compounds. 164

(i) I^drazine Derivatives

Some hydrazines and iiydrazide derivatives were found to be potent MAOI. ThRS H-benzyl-N'-(SHnethyl-^- isoxazolyljcarbcxylhydrazide (isocarbosazide) (g) which exhibited 7 to 33 times the potency of iproniazidjis used clinically 11 *1 2 . In contrast 3,5-dimethyl-4-'isaxazoylcarbo- xylic acid isopropylhydrazide (j) was inactive.

OdlMHCH^Ph V\ /«Q^™(CH3)2 '2

H^O- VQ /

Nialmide (_4) is 3*-12 times more potent than iproniazid and produces less liver toxicity 13•^ . A hydrazone like compound {5) was very potent inhibitor of MAO .

0=C-CH20H2NHNHC-^ ^N 0 ^^

Hydfazides exhibited different brain versus liver MAO inhibitory activity* Bic isonicotinoyl derivative of JB-516 (6) had a high affinity for liver MAO and much less of an enzymo inhibitory capacity towards brain MAO1 5 • 165

CC2TMHCH(0H )0H2Ph

(ii) Non-hydrazine derivatives Many non-hydrazine compounds also show algnifLcant MAO inhibitory ac-fcivity. Some of the important representative members of this class are the harmala alkaloid -harmine (J), and some related p-carboline derivatLves- tryptoline (8), the indole alky lamines 17'- a-methyltryptamine (9) and a~ethyltryptamine (etryplam) (jLO)» the propargylamine - TO pargyline (^1)1 oyclopropylamine deriva"fcives 12 and

.^^

.^^^^ OH, R H 9, R=CH^ i 10, R=CH2CH 11 1^ and some of the aminopyrazines 14 and 15 "* •

^6%°\ /NH2

12 13 166 .N OH, n ^^CH.

•^^ n N"^N ^N(GH^)2

14 15

Several reviews on MAOI are availalDle 24-27

(2 ) !&.-iqy.clic Antidepressants

The first major antidepressant, imipramine (lj5, Fig. 2) was obtained by the isosteric replacement of sulfur in phenothiazines \i±th an ethylene bridge. Eiis modLacaiion

which changed the structure from a flat two-dimensional phenothiazine ring system to a three-dimensional dibenzazepin ring system, was found to be of paramount importance.

The clinical effectiveness of imipramine (1^) led to the investigation of "a number of tricyclic compounds. Structural variation of imipramine can be classified as follows:

(i) Variations in the non-aromatic portion 29 Horn £t _al. observed that desipramine (17) which is a metabolite of imipramine (16) was tv^enty times 167

^^N-CH2CH2CH2(^ CH 16

C,9"24^2

flQZ 168

as potent as latter (l6) in the hypothalamus, but in the corpus striatum 17 was only one-sixth as potent as 1_6. However, removal of "both the methyl groups slightly reduces the antidepressant activity. Higher alkyl amino compounds

12 —5 were found to be devoid of benzoquinolizine-antagonist effect^ . The N-oscide was only one-third as potent as the "51 parent compound in tetrabenazine-antagonist test . Optimum activity of the compounds was observed when the basic nitrogen was separated from the nucleus '\iY a propyl unit* Branching of the propyl chain (IB) has little effect on antidepressant activity^ ^^. Active compounds

GH^ CH2CH(GH,)0H2l^ IB CH_ also resulted viien the dimethyl amino propyl chain was substituted with a quinuclidine {"13) or a tetracyclic compound (20)-'^ . 169

(CH2)2N CH. 20

Metapramine (21) has antidepressant and psychostimulant activity with a rapid onset of action^ . EEGE.

Introdactlon of 10,11-double "bond does not change the activity of the parent compound. Ttm.3 2^ has antianxieiy

{GH2)^IHGH and sedative properties in addition to moderate antidepress!on action-^.

Va,rious analogues of imipramine with hetero atom substituted at 10 or 11 positions have been made. The 170

diazepins (_23) is as active as imipramine as an antagonist of reserpine induced effects-^ . 0 ox-re spending thiazepines £4 also possessed antidepressant activity • The oxazepines 2^_ had sLgnificant antihistaminio action^ .

(CH2)^N(CH^)2 _22, Z=NH

25,, X=0

Replacement' of the ring nitrogen with a carbon bond yields dibenzocycloheptadiene compounds, amitriptyline (_26), a therapentically useful antidepressant. Ihe major pharmacological and clinical actions of 26 are similar to

CH(CH2)2N(CH )R 26, P=OH^ 27, R=H

28, R=GHGIL,—7—N-CH„

29, R=F0(CH2)2^(011^)2 171

to those of imipramine'^ . Nortriptyline (27), a metabolic product of ami try pty line 2^ retains ihe aatibenzoquinolizine action of "liie parent compound and is five times more potent as a clinical antidepressant^^. IJhe t\fo most important compounds with structural variations in the side chain are 28^,44^ and 29 (Hoxiptillne)^^,

The dibenzocycloheptene analog ^ of ami tripts^line 46 (26) has oLinical and sedative properties . Introduction of a 10,11-double bond into nortripiyHine (2J) causes an increase in the antidepressant activity in a tetrabenazLne- 47 antagonism test in mioe^ . 03ius ^ was found to be about 1.4 times potent than nortriptyline {2jj, vrhereas the primary amine ^ had almost similar activity.

29, R=CH(CH2)2H(CH^)2 ^, E.=CH(CH2)2NHCH

_22, R=CH(CH2)NH2 172

Reduction of the olefinLc side chain in dihenzocyclo- heptene (dibenzocycloheptatriene) derivs.tives, in general, enhances the antidepressant activity. Ihus protriptyline JJJ, the side-chain reduced counterpart of ^ had 9-10 times more potency thaji ami trip inline {26) as a tetrabenazine

y H OH2GH2OH2H OH, 22. agonist in mice^.47' and it exhibited enhanced clinical ,48 activity without causing much sedation 53 is presently in clinical use.

A spiro-4-aminocyclohexenyl derivative ^ inhibits upta,ke of norepinephrine and serotonin and lacks antl- cholinergic activity^ . Octriptyldne ^ which was obtained by the incorporation of the 9,10-ettiylene bridge of nortriptyline in'bo a cyclopropane, has amitriptyline (_2J) like properties in animals 50.

GGH(CH2)2NHGH ,

M 173

DoKepin ^f a nixtare of geometrical isomers, has an-td.depressant activity comparable to that of amitripiyline (£6) but has a more sedating effect. It has found olinical

CH(GH2)2l'f(GH„)2

51-53 use particularly in cases of agitated depressed patients The thioanalogue, prothiadene ^ also showed antidepressant activity* In animals J7 caused \jeak depression of the QNS, inhibited the action of reserpine and marked antihistaminic properties 54-.

Ihe antidepressant property of Ihe piperazLnyl - substituted dibenzoxepin ^ was found to be somewhat lesser than ami triply line (26), upon oral administration to rats in an antireserpine test 55 . The piperazinyldibenzothiepin ^ was also found to possess central depressant and 46 antagonist of reserpine induced effects 174

r if "^r"^ "^^^^^-^V^^:^ s/^^ CH„ 5 ^' X=0 39, z=s Hie antidepressant behaviour of propazepine AO was observed to be only one-half that of imipramine (]-6), but it was better tolerated in man. Some other dibenzodiazepines such as jP. and j2 were inliibiters of pseudo-oholinestrase^5 6 . ])ibenzo[b,f][l,4] thiazepine, 4^ showed neurotropic properties. The amincmethyl derivatives of 11-diazepinones e.g. _44 were potent antihistaminic agents •vriLth antianaphylactic properties-^5 7 . Dibenzazepin 45 is used clinically as a selective antidepressant tti-th mood elevating actions 58 ' 59 . It is of special interest to note in thi.s series that amino group is separated from the tricyc.njLc ring through a ti-ro carbon unit rather than the normal propylene linkage .

/^^N^ \ CH^CHoN{CH„) ^2"^^2' y2 i5» ^^=^ 41, X=NH 46, R-^=GIL _42, Z=1T0H 43, X=S 175

OonTersion of the side chain to an additional ring gives a novel class of antidepressant dibenzazepines. A 61 prototype of this class is mianserin (47) ""•

The antidepressant action of tricyclic compounds vjith a central six-memhered ring could be imparted only if the system does not attain planaritjr. Introduction of some bulky groups, such as methyl, was found to be helpful in this direction. Thus melitracene J8 and the corresponding 62 dimethylacridan (dimethacrin) 4S are clinically used

H„C 0H„ ^X'- 3 \/- 3

CH(CH5)^N(GH„) 12/2 y2 43 antidepressants._£9 was similar to imipramine as a reserpine 63 or benzoquinoHzine antagonist in mice and rats . 9-Meth;7ldihydroanthracene, fluotraoen 50 is a clinically acceptable antidepressant -s-dth minor side effects ^*^^, Another 9-substituted dihydroanthracene, 176

danitracen ^ is a very potent clinically effective antidepressant *'. OH ^'••..^Z- OHo

H (CH2)^(GH^)2

It has been observed, in general, that tricyclics with 5- or 8- memberod central ring are less potent as antidepressants in comparison to those with 6- or 7- membered central rings. Maximum antidepressant activity is encount ered \Then aromatic rings of the tricyclic nucleus are at appreciable angle to one another. It is for this reason that 7- membered central ring is favoured over 6 or 8 membered compounds. However, iiiere are certain compounds which have noteworthy psychotropic activity.

Olie aminoalkylated fluorene _52_ produced distinct clinical antidepressant effects 68 .

^N ,r^^

0H(CH2)2N(GH^)2 52 177

A series of dibenzo[a,b][l, 4]-cyclooctaaienes (53-55) was compared ^•dth amitrip-fcyline (££) in several tests, but was found to be less potent * .

H^C/ 53 54

I 53 (CH2)3N(0H )2

(ii) Variatd.ons in the aromatic portion

Nuclear sabstitulion has a variable effect on the antidepressant aotivity. Substitution at 5-position has little effect. Thus chlorimipramine (56) is somewhat

r^^'^-V~~~\

(CH2)^(CH^)2 56 less potent than imipramine in reversing sedation caused a tetrabenazine-like compound"^ ' ' . Substitution at 178

other places generally decreases activity.

A (aibenzo['b,f][l, 4] thiazepin ^^ *^ has neurotropic properties. In mice, 5J antagonized reserpine induced hypothermia and ptosis in a oomparahle manner. Tixe related .46 tertiary amine 58 has little aotivi-ty on the CJtJS

Presence of a carbonj'-l group in position 1- of the side ffiiain of chl or promazine analogues gives clinically acceptable antidepressants. 'SOT example, both chloraoizLne (52) and its trifluorcmethyl analogue fluoracizine (60) -^* ' ^ are therapeutically effective antidepressants.

C0(CH2)2l^(C2Hc)2

59, X=G1 60, X=aP^ 179

1-Ohloraprcmazine (61) was found to impart imipramine-like actions in mice and rats. In this case the substitution in posiHon 1- of the phenothiazLne nucleus may interact sterically with the side chain to hinder attaimient of planarity by the tricyc3.ic nucleus.

<^N

CII2)^T(GH,),

Some amitriptyline analogues 62 and ^t benzothienocyclo- heptanes, bearing an unsaturated piperidylidene side chain,

62, R=H 6J, R=CH^ possess pharmacological properties, similar to known anti depressants. Thus pizotifen _62 exhibited imipramine-like effects , however, it is a potent central antagonist of serotonin and fails to inhibit norepinephrine reuptake 180

is vivo '* ' • Anoliher related compound _62, the lO-methyl derivative, has aLmilar pharmacological properties with the serotojoin antagonist cyproheptadine and the antidepressant mianserin (£7) .

A clinically effective antidepressant (64) » has a structure quite different from the other tricyclics, "but possesses antidepressant properties similar to those of

O -I imipramine (l^) .

-OH. i

( 5) MOP.o~ and Bicycli c Gompounds

Yet another class -which showed weak/strong thymolep-trf.c activity belongs to dimethylidene series _65» Pluvoxamine (_66) exhibited exclusive 5-HT uptake inhibitory properties op in rat brain , both in vivo and iji vijtro, and was found to

rr-^^ r^^^

CHCHgR 0(GH2)2NH2 66 ^ . ^0H„ CH- 65 181

be somewhat more effective than imipramine (16) •^.

Siiiiilarly, zimelidine (_67)» 4-hromo derivative of j55 a A exhibited potential usefulness in clinical trials . A closely related compound _63 was more potent than chlorimipramine {%) in reducing blood serotonin and initial clinical studies suggest good antidepressant activity.

f^^ rr^^ r^^^ N-^

l"l CHGH2N(CH-,3^)2 HCH2N(GH~)2 67 68

A somewhat related 3-aminopropylidenyl benzocyclooctane o c (^9) showed antireserpine actions in mammals •

0H(CH2)2^UGK } y2 69

A newer class of antidepressants include phenyl subsiituted tetrahydroisoquinolines that incorporate a diphenyImethyl moiety in their structure. Clinical studies have revealed the effectiveness of nomifensine (JO) Another compound 4-ph.enyltetrahydroisoquinoline (Jl) was e qui potent with imipramine (16) in preventing reserpine- induced ptosis in mice 89^ . TIio aminotetralin 12 was capable 182

of inhibiting the uptake of norepinephrine, serotonin and dopamine in rat brain tissue 90 .

?6^5

OH.

70 W^%aim C^6^H5

f^P UHOH.^

,91-93 Tofenacine (7j)» a benzhydrol ether, was found"' to have clinical properties of an antidepressant. A cyclic benzhydrol ether, fenazasine (74), inhibited neuronal uptake of norepinephrine and tyramine 94

^6^ 0{OE2)^^llOE, n

A diphenylmethane analogue in which one of the phenyl rings has been hydrogenated, viz. gamfexine (75) produced 183

clinioal antidepressant actions 95

^^^^ 5 ir^0H2)^l(C!-:„) 75a 75 96 Bupropion (7,5a) represents a new kind of anti depressant drug and reverts tetrabenazine-dnduced sedation in mice like indpramine (l^), but it does not affect various conditioned avoidance responses in rats. Bupropion (25a)-was found to be effective in th.e treatment of depression in man '»-^,

Benzobic^rclononadiene 2§. is five times more potent than cliloriiaipramine (^) as an inhibitor of serotonin uptake whereas it has no apparent effect on norepinephrine

^ ,jJill2-9''^3^3 E

76 11 uptake'^•^. Cyclazocine (rz) t the analgesic benazocine, improved depressed patients, however secondary effects are common. 184

AntLdepressant-act is claimed for some T-aminobenzo- oycloheptene derivatives e.g. 78 and homologous compound ^gl03,104^

All these drugs discussed above act by increasing the availability of catecholamines and indole amines at the appropriate sites of the brain. IHiis is achieved either by inhibition of reuptake of biogenic amines by the parasjoiaptic terminals, as is the case with trioyclic antidepressants ov by preventing the inactivation of the biogenic amines, as is the case -vrith MAOI. 185

4*2 BAg^IS G?^¥_ORK

The inadequacy of the drugs currently available for the treatment of depressed state underscores the need for developing new drugs- The search for new drugs could he based on new and novel lead structures.

Daring the course of a Tri.de spectrum primary screening 105 of the compounds in this Institute -^, it was observed that Jbrans-2-[H-(2-hydroxy-l,2, 3,4~tetrahydro-l-naphthyl)]iminothiazolidine (8_0) emerged as a now class of representa'iive molecule possessing significant antidepressant activity.

^^^^^^ OH N H

¥ith a view to study the effect of structural variations the synthesis and bioevaluation of various analogues of 80 was undertaken. As most, of the known tricyclic an'bidepre- ssants have a central seven memborod ring, it seemed of interest to homologate ring B of 8_0 and sftudy their antidepressant activity. This prompted the synthesis of _94 186

(Scheme 2). To see the importance of hydroxyl group and a heteroatom in 8_0, the synthesis of iminothiazolidine derivatives "without OH grdup (86af 8 6b and 86d) and x-rith oxygen as a hetero atom (8Jc) was carried out. These studies are described in this chapter.

4. 5 _Sj_n_tlie^s_îs^ ôf Iminjothjâ^zo3J.Jijie^ djeriyativeg The iminothiazolidine derivatives 8Ja-d and _94 were synthesized starting from the corresponding benzocycloalkanones 81a-d (Scheme l). 5-Methoxy- and 6*.methoKytetra- lones (81a and 61b) were obtained commercially, while chrananone (31c) and benzsuberone (81d) were synthesized by the qyclization of phenoxypropionic acid and 5--phenylvaleric acid respectively, in presence of polyphosphoric acid, according to literature methods.

5-Methaxytetr alone (81a) on re fluxing with hydroxylamine hydrochloride in aqueous ethanol, under basic conditions yielded the corresponding oscime SJa. Catalytic reduction of 8^a over 10^ Pd~C furnished the corresponding amino derivative 8^5^. Treatment of amine 8_3.a idth benzoylisothiocyanate in dry acetone afforded N-(5-methcix:y-.l,2,5, 4-tetrahydro-l-naphthyl)-N'-benzoylthiourea (Sjt-a)- The hydrolysis of 8Ja with 10^ aqueous sodium hydroxide solution gave the corresponding tliioarea derivative 8^a, which on condensation VTith bromoethylamine hydrobroraide in ethanol gave the 187

R -> R- ^^^2)n 0

3 la-a 82a-d

^ (CH^)^

84a-d II ii S 0

X R •^ R-i C0H2)„ ^

N 'N 8^a-d \ y

1| NligOH.HCl J 2. H2[Pd/G] j 3. CgH^OaNrCS

4. 10f» NaOE I 5. Br(0H2)2^^H2.HBr

a, R=5-0CH^, Z=GH2, ^^"^ * ^* R=6-0CH„, Z=CH2, n=l 1 c, R=H, X=0, n=l I d, R=H, Z=CH2, ^^^^

Scheme 1 188

desired oompound, 2-[]iir-( 5~inethoxy-l, 2, 3,4-•feetrahydro-l- naphtllyl)]iininothiazolic^irLe (8j5a). A similar sequence of reactions starting from other benzooycloalkanones, viz. 31b, 8^1c and 81d afforded the respective irainothiazolidine derivatives (86b-d).

The synthe sis of trans~2-.r I-( 6-.hydroxy-6,7,8,9- tetralTydro~5H-benzpoycloheptene )]iminothiazolidine (J^) I'J'as carried out from amj.no alcohol 90f follo\d.ng Scheme 2. _90 was obtained by the treatment of bromohydrin Q9 T-dth 106 aqueous ammonia solu-feion f £9 in turn was obtained by the reduction of benzsuberone (81d) i-riLth sodium borohydride to alcohol 87 109 and subsequent dehydration wixh potassium "LfR bisulfate to provide 6,7-'(3ih;^'-dro-5H-benzocycloheptene (88). Reaction c£ 88 with K-bromo succinimide in the presence of MSO-water under nitrogen atmosphere, furnished the bromohydrin 8^ . Two ooapling values for the doublet for 5-H at 3-9 6 [J5.6(trj^?)='""''"' '^5,6(^r^H^l ^^ pmr spectrum showed ^ to be a mixture of tvro isomers. The structure of 90 was further confirmed by the corresponding aoetoxy derivativs _95« The signal for 5-H in the pmr spectrum of 25 appealed as a triplet, which could be due to the participation of NH-proton in coupling. However, the triplet collapsod to a doublet #ion Iho Nlt- proton was exchanged with D^O, oonfiaming the partLcipatLon of m in Coupling with 5-H. lEho t^'TO isomers of 90 could 189

a ^

.^^^

NHo ^^ NH-C-ITH-C-Ph 90 0 91 h +

0 b~G-CH„ NH-0-ITH-O-Ph NH-G-CH^ ^ I! 5 S 0 0 92 95

91 •^

OH NH~0~NH2 S

a, NaBH-i b, KHSO,, » c, l^BS, IMSO-xfater» d, NH^OHj

e, CgH^CONCSi f, 1(^0 FaOHj g, BrCH2CH2im2«HBrj h, AQ2'^/'BJ

Scheme 2 190

not be separated at this stage. Reaction of this mixture of 90 with benzoylisothiocyanate in dry acetone again gave a mixture of two isomerj.c benzoylthi.ourea derivatives _91 and ^ in a ratio of 65:35» Biese trfo could be separated through fractional crystallization. Ihe signal for 5-H in nmr of 91 appeared at 5*3 6 as a doublet (J=10Hz) and 6-H appeared at 5.7 6 as a multiplet, while in J[2, the 5-H appeared at 5»6 6 as a doublet (J=6Hz) and 6-.H at 3.95 6 as a broad mU-ltiplet. The upfield position (5. 3 6) of 5-H in _91 as compared to that for 5-H in 92 was in accordance •tidth the characterization of trans- (^) and ,cd.^s- (92) isome?-3 'oj Khanna e^ aJ.. . Hie stereochemistiy of the t\7o isomers _91 and 92 was lUr'uher confirmed by NOE experiments. Irradiation of 5-H resulted in 345^ increase in the intensity of 4-H and 15/9 increase of 6-H in the ci^s-isomer _92 vrhereas in the _trans-isomer _91 no significant change was noticed. In the mass spectrum of both these isomers (_91 and _92) the parent ion peak corresponded to the dehydrated product (M'^-18 m/z = 322).

Hydrolysis of _91 -t-Jith IQfo aqueous sodium hydroxide solution afforded the thiourea derivative _92, which was confirmed by the disappearance of the ketone peak in the ir spectrum of _93« Eic ^cA^s-isomer _92, hoi-jever, behaved in an entirely different manner. Ihe hydrolysis of _92 id-th 2^ 191

alcoholic sodium hydroxide solution yielded a mixture of -irf'To compounds _% and _97» which, were separated by fractional crystallization using chloroform. The less soluble fraction

2io 92 NaOHale .

NIL 8 q5 • • 0 • 96 97 was formed in 8C^ yield. The nmr spectrum of this compound shCTfed it to be the cis-amino alcohol J^. A clean doublet at 3.92 6 was observed for 5-H of _96. Further confirmation for Ihe identity of _% was provided by mass spectrum (M m/z = 177) and elemental analysis. The product ^2. obtained in minor quantity showed a clearcut peak of ketone at I70O cm ". The elemental analysis of liiis product 97 showed the absence of sulfur. In the nmr spectrum of _97 5-H appeared at 5.026 as a doublet (J^ r- 4Hz) and 6-H as a multiplet centred around 3.87 6. Further confirmation by mass spectrum

(M'^-IS m/z = 202), proved this product to be a urea derivative _97«

The reaction of 22 -v^th bromoethylamine hydrobromide in ethanol at 85 for 24 h afforded the desired compound Jbrans-2--[N-( 6-hydroxy-6,7>8,9-tetrahydro-5H-benzocyclo- heptane)]iminothiazolidine (_94). 192

4.4 Syntliosis of 3"(4-oliromanyl)-2-inGthylmorcapto-4(5H)-

pyrimidinone and 3-( S^TjSyQ-tetrahydro-SH-benzocyclo-

heptene )-.2-methylmercapto-4( 3H)-p7riinidinone

A large number of pyrimidinone derivatives synthesized recentls'- in this Insti-fcute were found to exhibit either stimulant or depressant action in their gross behaviour, besides some other pharmacological activities. Ihis prompted us to synthesize some pyrimidinone derivatives (]^ and 105)» utilizing some of the thiourea derivatives (8_5c and 3^5d) as intermediates.

The thiourea derivative 85.C on refluxing with methyl iodide in absolute methanol afforded the N-substituted isothiouranium salt _^ (Scheme 5) which witlaout isolation was converted to the N-substituted S-alkylisothiourea 100, 112 by treatment with sodium bicarbonate solution . Ihe desired pyrimidinone derivative 1.02 was obtained by the reacHon of 100 with ethyl propiolate, according to loiomi method -^« In the i_r spectrum of 102 the peak for carbonyl group of pyrimidinone ring system was observed at 1650 cm" . Similarly S-methylatl on of 8^d, followed by cyclizatLon with ethyl propiolate furnished the corresponding pyrimidinone derivative ro^. 193

\^ R - - C I S S-CH_ 8,5c, R=H, Z=0, n=l 58 =0, n=l 8^d, R=H, Z=0H2, n^2 99i , x= =CH,> , n=2

NH-G=KH I

100, X=0, n=l

102, X=0, n^l 101, X=0H2, ^=^ 103, Z=CH2» n=2

a, OH„l/mot]ianol| b, Sodium •bicarbonatoj c, aH=CG020^1^

Schomc 3 194

4.5 Synthesis of 5-[ (H-su^^stituted3-amino-2-hydroKyp^opyl)-

oKimino] -6,7,819-te trah5'-dro-5H-benzocy oloheptene

A recent publication '^ has disclosed the importance of aromatic oxime ethers as p-cholinergic blocking agents. This prompted to utilize the oximino deriva-fcives 8^ of the benzocycloalkanones (8J.) for the E^mthesis of some title compounds (105"-107)« 114 Utilizing the procedure of Leclerc ejt al. 5-^oximino-6,7,8,9--tetrahydro-5H-benzocycloheptene (8__2d) was treated with sodium methoxide/methanol. After removal of the solvent the re si clue was taken up in anhydrous H<1P and was allowed to react with epichlorohydrin. Ihe vjork up of the reaction yieHed the epoxy derivative 104.. In the nmr spectrum of I04 a multiplet at 3«2 6 was observed for 0-CH- proton and at 4*15 6 a multiplet for NO-CHp protons. Opening up of this epoxide _104 with various amines gave the final compound, vi z. 5-[ (N''-substituted3-amino-2-hydroxypropyl)- oximino]-6,7»8,9-'tetrahydro-5H-benzocycloheptene. (12.5-1£7). 195

// NOH 82d, (E=H)

104-

N-OCH2OH-CH2-N OH

105, V - W

106, /= -JT N-Ph

107, N^= -ITH-C(CH^)^ a, 1. Na 0"Mo/mGthanol, 2. opichlorohydrin/K'IFj 7 b, W \

Sch^neJ. 196

4.6 ^SKpERBfflFTiL

Hie pmr spectra in CDC1-, unless otheririi se stated, were recorded on Varian A-60D and Perkin Elmer R-52 PICl spectrometer using ©IS as internal reference, the chemical shift values arr-e expressed in 6 units and -Sie J values in Hz. IT. spectra were recorded on Perkin-Slmer Infracord Model 157 and 577 and the i) „ valuies are expressed in cm" . Melting points were determined in sulfuric acid bath and are uncorrected, ill the compounds were routinely checked for their purity by thin layer chromatography using alumina or silica gel G as stationary phase and chloroform-*methanol (98:2) as mobile phase.

5-Methaxy cKimino-1-tetralone (82a)

A mixture of 5H3iethoxy-l-tetralone (81a) (9«fi3 g, 0.055 mole), hydroxylamine liydrochloride (5»97 g» 0,(B6 mol) and sodium hydroxide (11.0 g, 0.275 mol) in ethanol (100 ml) and water (40 ml) was refluxed for 30 minutes. Eie reacHon mixture was cooled and poured into a mixture of hydrochloric acid and water (200 ml). The precipitated axime £2a was filtered and washed with water and driedj yield 10.0 g (95.2^)j m.p.l55°. (lit.^^^ 155^).

NMT. : 1.8 (m, 2H, Ar-CH2CH2'~^' ^'^ ^^^' ^'^'' ^"^^

CHgOHgCHg)* 3»7.(s, 3H, OCH„), 6.6-7.5 (m,

3H, Ar-H). 197

115 6-Methaxy cKimino-l~tetralone (82b)

Tliis was prepared from 811) "by adopting a similar procedure as described for 8£a in 87.82/9 yield, m.p.l^O •

NME : 1.76 (m, 2H, Ar-CH2CH2CH2), 2.7 (m, 4H,

Ar-GH2CH2CH2), 3.7 (s, 5H» OCH^), 6.45-5.8

(n, 2H, 5- and 7-Ar-H), 7-73 (

4-CKiinino chromanone (82c)

It was prepared from 81o in 88.2fj, yield according to the method described for 82a, m.p. 142° (lit. ^ 144-145°).

MR i 2.96 (m, 2H, OCH2CH2), 3.68 (m, 2H, OGH2), 6.5-7«9 (m, 4H, Ar-H).

5r-CKimino~6,7j8,9-tetrahydro-5H-benzocycloheptene (82d)

It v;as prepared from 81d in 81.4/» yield according to the method described for £a, m.p.lC6°..(lit« 107-1C6^). mR(CCl.) ; 1.68 [m, 4H, CH2(0H2)2CH2] , 2.65 [m, 4H,

CH2(GH2)2CH2], 6.6-7.5 (m, 4H, Ar-H).

l-Jknino-5-methoxy tetralin (8Ja)

82a (9.55 g, 0.05 mol) was dissolved in ethanol (100 ml) and hydrogenated over Rane3r-.nickel at 40 lb. pressure -till the absorption of hydrogen had ceased. The reaction mixture was filtered to remove the catalyst and the 198

filtrate was concentrated to give an oil wliicli iras parified by acid-base treatment to give S^a, 3/-ield 9« 0 g (95/o)» This oil was converted to the corresponding hydrochloride, m. p» 247°; (iLt.^^'^ 250"^).

IR(Neat) : 330O (NH2) ]^Tm(free base) • 1.46-2,18 [m, 6H, ((^^2)^]* 2.64 (m, 2H, inig), 3.75 (s, 3H, OCH^), 3.9 (m, IH, OMH2), 6,56-7.51 (m, 3H, Ar-H). ai7 l-jAniino-6-methoxy-tetralin (83b

This was prepared from 8^b by a, similar method as described for 8_3a, yield 92.7f»» m.p. (hydrochloride) 2:3T»

IR(Neat) : 330O (NHg) mill i 1,58-1.96 (m, 2H, -^CE^OE^^iE^)t 2.5-2.91 (m, 4H, Ar-CH2 and NHg-CH-GHg), 3*1 (s, 3H, OCH ), 6.45-6.8 (m, 2H, 5- and 7-Ar-H), 7.73 (d, IK, 8-Ar~H, J=9Hz).

4-Aminochranan (gjc)—^

Reduction of 82c by a method similar to that described for SJa furnished S^o as an oil in 92f» yield.

IR(Neat) ; 3200 (UHg).

HMR : 1.25-1.7 (m, 2H, 00H2-CH2)» 2.02 (m, 2H,

IJH2, DgO exchangealDle), 3.7 (m, IH, NH^CH-),

3.8-4.4 (m, 2H, OCHg), 6.5-7.3 (ra, 4Ii, Ar-H). 199

5-imino-6,7>8,9-tetrahydro-5H-'bonzocyGloh.eptene (83d)

It was prepared from 82d by a similar procedure as described for 8Ja, yield 76.08'/^, m.p*10B^ (lit.^-'-^ 106°).

IR(ICBr) : 3200 (NH2) miR : 1.0-2.1 [ra, 4H, CH2(CH2)2CH2] , 2.4-3»0 [m, 4H, 0H2(CHo)2CH2], 4.01 (t, IH, CHLIH2),

6.5-7.5 (m, 4H, Ar-H).

F-(5-^Iethax:y-l,2, 5,4-tetrahydTO-l-naphiiiyl)-R' -benzoyl- thiou.rea (84a)

A mixture of benzoyl isothiocyanate (8.15 g» 0.05 raol) and acetone (15 ml) was added drop'vri.se to a stirred solution of 8Ja (8.85 g, 0.05 mol) in 40 ml of acetone, at room temperature. After the addition was complete, the reaction mixture was'refluxed for 30 min, cooled, poured onto ioe and extracted with ether. Ihe organic phase was washed vdth water and dried (Na^SO.) and concentrated to give'a

(m, 2H, ir-CH2-), 3t78 (s, IH, OCH^), • 5.5 (m, IH, 0H~NH)r-6.6-7-96 (m, 8H, Ar-H),

8.33 (s, IH, NHCO, D2O exchangeable), 10.15

(d, IH, GHNHCS, D^O exchangeable).' 200

Mass ; M"*" m/z = 340

Analysis • ^19^2(^2^2^ Pound i 0, 67.54* H, 6.15J W, 8.07 Reqd. : G, 67.051 li, 5.831 N, 8.237^

N-( 6-Methozy-l, 2,3,4-tetraliydro-l-naphthyl)-n' -'benzoyl-

thiourea (84b)

It vras prepared from 8^b by a similar procedure as that for the preparation of 84;a in 82^ yield.

IRdJeat) : 167-e-(C=0}, 32 X (HH) FMR : 1.51-2.51 [m, 4H, GEl2(CH2)2CH], 2.45-2.98 (m, 2H, Ar-CH2-)» 5.75 (s, 5H, OCH-), 5.67 (m, HI, CMH), 6.45-7.95 (m, 8H, Ar-H), 8.55 (s, IH, moo, H^O exchangeable), 9.92 (d, IH, CHMHCS, D2O exchangeable).

Mass M\/Z = 540

Analysis ^19^20^2^2^ Pound G, 67.4O1 H, 6.2I1 N, 8.11 Reqd. C, 67.05* H, 5.88* N, Q.2jfo,

H-( 4-.Ghromanyl)-N' -benzoylthiourea (84c)

Hiis was prepared from 8Jc by the method described for the preparation of 84a, yield 97*S/** m.p.95°.

IR(KBr) J 1660 (0=0)^ 5200 (FH) 201

N14R : 2.25 (m, 2H, 0CH2GH2)* 4.15 (m, 211, OGH^), 5.56 (m, IH, OMH), 6.62-7-9 (m, 9H, Ar-H), 9.05 (s, IH, NHOO, D2O exchangeable), 10.0 (d, IH, OmH-OS, D2O exchangeable).

Mass M"*" m/z = 312

Analysis ^lft6^^2Q2^ Pound C, 65.72, H, 5.25» N, 9.13 Re qd. 0, 65.38» H, 5.12| N, Q»91fo.

N-( 6,7»8,9-Itetrahydro-5H-benzocyclcheptyl)-.l\r' -benzoyl-

thiourea (8J-d)

It was prepared by the condensation of 8^3d vjith benzoyl isothiocyanate by a method similar to the one described for 84a, yield 84. 7^^, m. p.l09°.

IRdOBr) 1680 (0=0), 3230 (IIH) NMR 1.4-3.3 [m, 6H, Ar-CH2(CH2)2GH2] , 2.6-3.1 (m, 2H, Ar-CH2), 5.65 (t, IH, CPMH), 6.6-7.9 (m, 9H, Ar-H), 8.05 (s, IH, NHCO, D2O exchangeable), 10.5 (d, IH, GffiTHGS, D2 0 ex change ab le ).

Mass M"^ m/z = 336

Analysis ^19^2(^20^ Pound C, 67.53* H, 5.8I1 N, 8.63

Reqd. C, 67.85, H, 5.95, F, 8.33f». 202

II-( 5-Me-thaxy-l, 2, 3, 4-tetrahy dr o-l-naphthja) tliiourea (8^5a)

A mixture of 84a (1.5.6 g, 0. 04 mol) and lOfo ilaOH (15 ml) was re fluxed with stirring for 1 h, the reaction mixture was cooled and the separated solid was filtered, washed with water to furnish the desired pi-oduct 85a, yield 8.9 g (94.6/0 » m.p.lS3°.

IR(lCBr) : 5300 (NE and NH2) NMEdMSO-dg) : 1.87 [m, 4H, 0H2(GH2)2-CH] ,2. 32-2, 46{m, 2H, Jlr-CHg), 3.75 (s, 3E, OGH^), 5.^> (m, IH, CH-FH), 6. 67-7.58[m, 5H, Ar~H and IH2 {NH2 D2O exchangeable )j ,7,38 (d, IH, OMH, DgO exchangeable). Mass : M"^ m/z = 326 Analysis : C^2H^gF20S Pound : C, 61.33* H, 6,92| N, 11-69 Reqd. : C, 61. Olj E, 6.77* N, 11.8 6fi.

N-( 6-MethcDcy-l,2, 3, 4-tetrahydro-l-naphthyl) thiourea (8^b)

It was prepared by the debenzoylation of 8_4b, by a method similar to that followed for the preparation of 84a, yield 9(^^, in.p.l63°.

IR(KBr) : 3IOO (NH and NH2) 203

NMR(CDC1 + 2 1.5-2.06 [m, 4H, CH2(CH2)2CH] ,2-5 5-2.7 drops of (m, 2H, Ar-GR^), 5.6 (s, 5H, OGH^), 5^2 6 MSO-dg) (m, IH, CMH), 6.2 (s, 2H, NH2» B2O, exchangeable), 6.4-6.65 (m, 2H, 5- and 7- Ar-H), 7.1 (d, III, S-Af-H), 7-65 (bs, IH, NH.j, DgO exchangeable).

Mass M"^ m/z = 526

Analysis ^12%^^202 Found 0, 61.57J H, 6.98| N, 11.71 Reqd. C, 61.01» H, 6.77| N, 11.8 ^b.

i\[- ( 4-Ghr oman.vl) tlii ourea (85 o)

It was obtained from 84c by a procedure similar to that for the synthesis of 85a, yield 70fo, m.p.l68°.

IR(KBr) : 3100, 3200 (NH and I^E^) NMR(CDC1_ + : 2.1 (m, 2H, OCHgCHg), 4.1 (m, 2H, OCH2), 5 2 drops of 5.52 (m, 3JR, CHNH), 6-23 (s, 2H, NH2, mso-dg) D2') exchangeable), 6.6-7*6 (m, 4H, Ar-H) 8.1 (m, III, GMH, D2O exchangeable^.

Mass M"** m/z = 2!X

Analysis ^lA2^^2^^S 1^ omid G, 57,47| H, 5.59» IT, I3.63 Re qd. G, 57-69| li, 5.76| li, l3.46/fl. 204

S-{6f 7?3,9-5tetraiiydro-5H-T3en20cycloh.eptyl)thiou.rea (3^5,d)

It was prepared from 84(3. "by a similar method as described for S^^?^* yield 82f., m*p.l42°.

IR(ICBr) : 3300 (HH and NH2) mm : 1.0-2.3 [m, 6H, Ar-QHg(GH2)3] » 2.4-3.0 (m, 2H, ir-CHg), 4.7 (m, IH, CHNH), 6.0 (bs, 2H, NH2» D2O exchangeable), 6.5-7-5 (m, 4H, ir-H), 8.11 (bs, IH, CMH

Mass : M"*" m/z = 22 0

Analysis : C]_2%6^2^ Found : C, 65.71» H, 7.52f N, 12.55 Reqd. : C, 65.45> H, 7.27» N, 12.72fa.

2-[N-(5-Methoxy-l,2,3,4-tetrahydro-l-naphtlTyl)]iminothiazolidine (86a)

A mixture of 8_5a (4*72 g, 0.02 mol), bromoetliylamine hj'-- drobroinide(6.15 g, 0. 03 mol) and ethanol (50 ml) x^as re fluxed for 24 h T/dth stirring. The reaction mixture x-ras concentrated, diluted lATith water (3OU.40 ml), and made basic by the addition of aqueous ammonia solution. Eie separated solid was filtered, washed with water and dried to furnish 8_6a, yield 4.1 g (78.25^01 m.p.156%

IR(KBr) : 3300 (ITH) 205

Km : 1.55-2.13 [m, 4H, 0H2(0H2)2CH] , 2.43-2.83 (m, 2H, kv-OE^), 3-ll-3»46 (m, 2H, ITOHg), 3.78 (s, 3H, OCH^), 4.06 (m, 2H, ' 3CH2), 4.7-5.11 {m, IH, GH), 6.58-7.55 (m, 3H, ir-H), 7.66-7.B8 (m, IH, im, D2O exchangeable).

Mass : M"*" m/z = 262 Analysis : C^4%6H20S Found . G, 64.40J H, 6.24i F, 10.53 Reqd, : C, 64.121 H, 6.10$ If, 10.68-/' •

2-[H~( 6-Methoxy-l,2,3, 4-tetraiiydTO-l-naphthyl)] iminothiazolidine (86b)

Application of a similar method on 8_5b afforded the desired compound 86b in 42.1^ yield.

IR(Neat) : 3300 (NH) mm i 1.4-2.2 [m, 4H, GH2(0H2)2GH], 2,3-3*1

(m, 2H, Ar-OHg), 3*35 (m, 2H, N.GHg), 3.65 (s, 3H, OCH ), 3.95 (m, IH, oCHg), 4.6- 5.0 (m, la, CH), 6.3-7.2 (m, 3H, Ar-H), 7.3-7.9 (m, IH, NH, DgO exchangeable). Mass : M"** m/z = 262 Malysis j G^^%5N20S

Pound 0, 64.37J H, 6.29» M, 10.74 Reqd. 0, 64.121 H, 6.IO1 N, 10. 206

U^(4-.0hromany 1)im3.nothiazolidine (8_6c)

Eiis compound was obtained from 85c in a similar manner as B6a, yield 73^» m.p. l?!*^.

IR(liBr) : 3100 (I«i) iTMR(CDCl + i 2. 0-2,4*(m, 2H, OCH2OH2), 3«13->66 (m, 2H, 2 drops of NCH5), 3,83-4.46 (m, 4H, OGHp and SCHp), n-lSO-dr) -^ ' • 5.05 (m, m, OH), 6.66-7.58 (m, 5H, Ar-H

and ICi).

Mass ; M"^ m/z =234 Analysis : G^2%4^^2^^ Pound : C, 61.27» H, 5.75} N, 11.81

Reqd. : C, 61-53? H, 5.98| N, 11.9^».

2-[N-( 6, 7j8, 9-!retrahydro-5H~'benzocyclolieptyl)]imino-

thioa2olidine (86_d)

It ¥as prepared from 85.d "by a similar procedure as the caie described for Ja, yield 72^^, m.p. 156°.

IR(KBr) ; 3250 (im)

NMR : 1.0-2.0 [m, 6H, Ar-GH2(qH2)^] , 2,7 (m, 2H,

Ar-GK2), 3.15 (m, 2H, HC)H2), 3.8 (m, 2H,

SCHg), 4.75 (m, IH, CH), 6.75-7.4 (m, 4H,

Ar-H). Mass : M"^ m/z = 234 Analysis : Ci^H^s^gS Found t 0, 66.871 H, 7.85» N,11.72 Reqd. : 0, 66.66j H, 7-69j N,11.96f». 207

S-Hydroxy-e, 7.8,9-tetrahydro-5H-TDenzoq7 clolieptene (8 7)

It was prepared according to literature method, m.p. 100°(lit.^°5 lDO-101^).

6,7-DiIi5^dro-5H-TDen2oQyclolieptene (88)

It >ras prepared by the knovm method, b.p.96-99/7 ram (lit.-^^^ 120-122^18 mm). rom : 1.94 (m, 2H, ArCH2CH2), 2.2-2.9 (m, 4H, jir-0H2CH2GH2), 5.8 (m, IH, Ar-GH=GH), 6.4 (m, IH, ir~GH=OH), 6.7-7.1 (m, 4H,

iir-H).

6-Br cm o-5-hy dr oxy-6,7,8,9-te-brahy dr o-5H-TDen zo cycloheptene (8^) To an ice-cooled mixture of 88 (1»4 g, 0.01 mol), dimethyl sulfoxide (lOO ml) and water (4 ml) was added N-bromosuccinimide ^d-th vigorous stirring under nitrogen atmosphere. After being stirred for flirther 1 h, the reaction mixture was poured onto crushed ice in small portion; with vigorous stirring. A colourless precipitate was formed which was filtered and thorouglily washed with ice-cold water to furnish 89, yield 2.0 g (82.9^0» m.p.87*^.

IR(KBr) : 3300 (OH) 208

MR(ODCl^) : 1.2-2.3 [m, 6H, (CH^)^], 4.2 (a, IH, CHBr), 4.78 {d, IH, CHOH, J=9Hz), 6.82-.7-54 (m, 4H, ir-H). Analysis : ^n-iHi'^^Br Pound : C, 55. OJj H, 5.51 Reqd. : C, 54.77t H, 5.39f».

5-Anino-6-hy dr oxy-6,7 >8,9-te trahy dr o-5H-ben zo cyclo- lieptene (90)

A mixture of 8^ (2.41 g, 0.01 mol) and aqueous ammonia (15 ml) was stii-red overnight. Tne resulting precipitate was filtered and washed with hexane to furnish JO as a colourless crystalline compound, yield 1»44 g (81.4/0J m.p.l58°.

IR(ISr) : 5100 (NHg), 5400 (OH) urn. : 0.9~2.3 [m, 4H, (CH2)2l» 2.7 (m, 2H, Ar-CH^), 3.17 (m, IH, CHOH), 3*9 (two set of d, IH, CMH2, J5,6(,te:^is)= 10, J^ eidsr^'^^^' ^•6Q-'7«28 (m, 4H, Ar-H). Mass : M"^ m/z = 177, M^ -- 13 m/z = 159

Analysis ; ^r^S^'"^ Pound : C, 74-73J H, 8. 61> N, 7-76 Reqd. : 0, 74.57} H, 8.47» N, 7.90fo. 209

trans- and ci^-N-[5-(6-H}'-drox3'-~6,7»8, 9-tetrahydro-5H-

a^M»* ••*•-'»-* << •» i»i \mi »WM benzocycloheptene)]-N'-'b8nzoylthiourea (_91 and 92)

Ihe mixture of _91 and J£ was prepared from £0 by a similar method as described for tbe synthesis of QAt yiold (mixture) 80f^. The two isomers viz. trstns- _91 and j3i_s-92 were separated by fractional cnrystallization from acetone, in a ratio of 65:35«

91, m.p«17S°

IRdCBr) : 1700 (0=0), 3400 (NH and l^E^) MR : 1.32-2,24 [m, 4H, Ar-aH2(GH2)2]» 2,8-3«2 (m, 2H, ir-CH2), 3» 7 (m, IH, CHOH), 5.3 (cl, IPI, CMH, J=10Hz), 6.55-7.5 (m, 4H, i3.--H), 8.3 (s, IH, KHGO, D2O exchangeablo), 10.2 (d, IH, CIMHCS, J=10Hz, DgO exchangeable)^

Mass M"*" -18 m/z = 322

inalysi s ^19^^20^^2''*2^'^ Found 0, 67.57i H, 6.03J IT, 8,15 Reqd. C, 67-58J H, 5.82| N, 8.23/«. o

IR(KBr) : 1680 (0=0), 3200 (KH and FHg) NMR : 1.3-2.2 [m, 4H, Ar-0H2(0H2)2]» 2.56-3.2 (m, 2H, Ar-0H2), 3» 95 (m. 210

IH, CHOI-I), 5.6 (d, IH, Omil, J = 6Hz), 6.8-7.9 (ra, 4H, Ar-^), 9.0 (s, IZ, HHCO, DgO excliaiigeable), 10.5 (d, IH, CETHCS, J=^.Hz, 3D2O exchangeable), Mass : M"^ -33 m/z = 322

Analysi s ' ^19^^2./2'^2^ Pound : C, 67.79; H, 6.15j N, 7-93 Reqd. J C, 67-58 J H, 5.82| II, 8.23fp.

^trans-N-[ 5- ( 6-Hy dr oxy-6,7»8,9-te trahydr o-5H-benzocycl o- heptene)]thiourea (93)

Debenzoylation of 91 by the procedure described for the preparation of 85.a afforded 9J in 79^^ yield, m.p.lll°.

IR(lCBr) 3000 (NH) NMR(0DC1 3^ 1.11-2.4 [m, 4H, Ar-CH2(0H2)2]» 2.63- niSO-dg) 3.05 (m, 2li, ^r-GH2), 3.6 (m, IH,

CHOH), 4.94 {a, IH, CHMH, J=10Hz),

6.5 (s, 2H, ^E^), 6.8-7.3 (m, 4H, Ar-H), 10.3 (d, IH, CHNHCS, J=10Hz).

Mass M^ -18 m/z = 218 Analysis C^2Hi6fl20S Pound C, 61.33> H, 6.881 N, 11.58 Reqd. C, 61.01} H, 6.78} N, 11.8^.. 211

jtrai]^s--2-[F-( 6-Hyd.roxy-6,7»8,9-te trahydr o-5H-"b•^nzo-

Gycloheptene)]iminotl^iazoli^±ne (_94)

Preparation of 94 was carried out fran 93 by 'the same method as the one descrihed for 8 6a, yie3.d 5 6fof m.p.l79

IE.(ZBr) : 3200 imi) NMR(CDC1^ + % 1.05-2.1 [m, 4H, Ar-CH2(CH2)2l» 2.6- 2 drops of 2.9 (m, 2H, Ar-CH2), 3.14 (m, 211, MSO-dg) iJOHg), 3.76 (t, 2H, SGH2), 3«85 (m, IH, OHOH), 4.3 (d, IH, CMH, J=8Hz), 6.85-7.57 (m, 4H, Ar-H). Mass M"*" m/z = 250 iUialysis Found C, 62.73> H, 7.45» N, 10-93 Re qd. C, 62.4o» H, 7.20» N, 11.2C^».

'J-Ace tamido-6-ace toxy-6,7,8,9-te trahydr o-5H- benzocycloheptene (95)

A mixture of 9_0 (0.177 g, 0.001 mol), acetic a?::'.h3rdride (2 ml) and pyridine (0.3I6 g, 0. 004 mol) vras stirred for 10 h. The mixture was then poured over crushed ice-vrater mix "cure, extracted with chlorofoim, washed succesively with dilute HCl, water, dried (RagSO.) and concentrated to produce 95 as a colourless solid} srLeld 212

0.2 g (87.53^^)» mp.l69°.

IR(lCBr) ; I64O and 1750 (0=0)

MR : 1.5-2,15 [m, lOH, Ar-GH2(CH2)2» NHOOGH and OGOOH ] , 2.55 (m, 2H, Ar-CHj)? 4r8 (m, IH, OGK), 5*25 (t, IE, EHOH, J=10Hz), 6.5 (d, IH, JTH, J=10Hz, D2O exchangeable), 6.8-7.25 (m, 4H, Ar-H). Analysis ; C^^H^gNO, ?ound : C, 68.72| H, 7.15j N, 5.44 Reqd. : 0, 68.96j H, 7-27» N, 5.5^/^- cis- 5-itoino-6-hydroxy-6,7,8, 9-tetrahydro-5H-'benzocyolo-

•u^r jitt. •! ^MiU'i'i'fc.-'ifci 11' .«L.^» III heptene (_9_6) and ci^"N-[5-( S-H^^di-oxy-S,7»8,9-tetrahydro-

5H-TDenzocyclohepi;ene)]urea (_97)

A mixture of ^2 (0.540 g, 0.001 mol) and 2fo etlianolic ITaOH soluiion (1 ml) was re fluxed for 50 minutes. The separated solid was filtered, washed iiioroughly tdth water to give 0.18 0 g of a mixture "of _9§ ^^^ 97. The two were separated by fractional crystallizaiion from chloroform in a ratio of 8 0:20.

96, yield 0.125 g (70.62fO-

IR(KBr) : 5120 (FHg), 3500 (OH). 213

mm. : 1.1-2.5 [m, 4H, (CH^)^], 2.75 (t, 2H, AT-CH2, J=5.0Hz), 3*25 (m, IH, CHOH), 4.02 (d, IH, OPMH2, J=4Hz), 6.75-7.8. (m, 4H, Ar-H).

Mass M"*" m/z = 177, M"^ -13 m/z = 159 Analysis ^lA5^^^ Pound 0, 74.681 H, 8.551 IT, 7.76 Reqd. C, 74.57> H, 3.47J N, 7,30fo,

97, yield 0. 030 g (13.65/0.

IR(lC3r) : 1700 (0=0) : 1.05-3.2 [m, 6H, (CH2)^], 3-85 (m, IH, CHOH), 5.02 (d, IH, CHTH, J=4Hz), 6.75-7.85 (rn, 4H, Ar-H). Mass M"*" -18 m/z = 202

Aaalysis ^12^16^2 "^2 Found G, 65.72| H, 7.45» N, 12.51 Re qd. 0, 65.45i H, 7.27| N, 12.72/a.

1T„ (4-Olir cmany 1) -S-me thy 11 so thi oure a ( ]A)0 )

A mixture of (85c) (2.06 g, 0.01 mol), methyl iodide (2.84 gj 0.02 mol) in absolute methanol (50 ml) was refluxed for 4 h.. Concentrated invaouo ejd crystallized vdth ethanol to give the corresponding N-substituted isothiouranium salt 98, which. ira,s treated with sodium bicarbonate 214

solution to give the desired oorxipound 100, yield 1.7 g

(78/0, m.p.135-156^

NI'iR : 2.':>-2,2 (m, 2H, OOH^GH^), 2,5 (s, 3H, SCH ), 4*15 (m, 2H, OCH^), 4.8 (m, 2H, WE, Dp 0 ex changeable), 6. 5- 7« 2 5 (m, 4H, Ar-K). Analysis : Cj^^H-j^.1^2^^ Found i 0, 59.75» H, 6.48| N, 12.37 Reqd. :• C, 39.45» H, 6.30» N, 12.61/.,

•N'-[ 5-( 6,7,8,9~ Te trahy dr o-5H-hen zocyclohe ptyl)] ~S-me thyl-

isothioui-ea (1^3-) It Tfas prepared from 84<3- "by a similar procedure as the one for 100, yield 72/^, m.p.l^O^. mm : 1» 14-2.15 [m, 6H, Ar-CH2(CH2),] , 2.5 (s, 3H, SCH ), 2.6-3.0 (m, 2H, Ar-CHg)* 4.95 (m, IH, GMH), 5.1 (bh, 2H, mi, D2O exchangeable), 6.3-7.3 (m, 4H, Ar-H). Analysis : 02^11221^2 OS

Found '. 0, 71.33> H, 6.69j N, 8.01 Reqd. : G, 71.00» H, 6.50^ N, 8.28-^. 215

3~(4-Ohromanyl)-2-.metla.ylmercaptO'4( ^lO-pjTiinidinone (102)

A mixturo of 100 ( 0.444 g, 0.002 mol), etliyl p^c-opiolate (0.196 g, 0.002 mol) and absolute methanol (20 ml) was stirred at room temperature for 24 h. The reaction mixtire was concentrated and triturated with hexane-ether. The separated solid i-jas filtered to oh tain the required compound lA)2, yield 0.54 g (62fO» m.p. 99 .

IR(KBr) ; 1650 (0=0) NMR : 1.9-2.2 (m, 2H, 0qH2CH2)» 3.24 (a, 3H» SCH-), 4*13 (m, 2H, 00H2)» 4.85 (m, IH, CM), 6.34 (d, IH, COCH, J=7Hz), 6.58-7.6 (m, 5H| kc-E and NCH=OH).

Analysis $ ^14%4-'^2^2^ Found : G, 61.47| H, 5.231 N, 10.05 Reqd. : C, 61. 31| H, 5. lOj IM, 10.215:^.

3-( 6, 7y8 ,9-Tetrahydro-5H-ben2;ocycloheptene )-.2-

m ^IBB^II HI i«ii '•• >i f methylmercapto-4( 3H)-pyrimj.dinone (103)

It was prepared from 101 by a similar method as mentioned for 102, yield 58'>, oil.

IE,(Neat) : 1660 (C=0)

NMR. : 1.1-2.2 [m, 6H, Ar-CH2-(CH2),], 2.^2 (s, 3H, SGH ), 2.56-3.2 (m, 2H, Ar-CH^ 6.2 (d, IH, G0C§, J=8H2), 6.76-7-28 (m, 5H, ilr-H and NCIfcGH). 216

Analysis : CigH^gNgOS FoLuid ; C, 66.S8i II, 6.06» F, 9.98 Reqd. : C, 67.15J H, 6.29i II, 9.79;'^.

5-[ (BpaK:y-2-propyl)oximino] - 6,7 j8,9-tetraliydro-5H-

•benzocyoloheptene (104)

To a solution of sodium methoxide (prepared from 0.138 g, Na and 15 ml of absolute methanol), oxime 82_d (1.0 g, 0.0058 mol) was added during a period of 5 minutes. Metlianol was removed in vacuo and the dry residue talKsn up in anliyd.rous DMF (5 ml) and a solution of epichlorohydrin

(0.555 St 0.006 mol) in anh^j^drous J2"IP (10 ml) was added dropi-Jise and the reactL'.ui mixture stj.rred for 1 h, du:d.ng which a precipitate of HaOl was formed. Hie mixture was extracted ^-rith chloroform, washed thoroughly with xirater to remove most of the HIE, dried (MgSO.) and concentra-bed in vacuo to afford 104 as an oil, yield 0.71 g (54v»)« mm : 1.34-2,04 [m, 4H, ^r-CH2(GH2)2]»

2,4S~2.9 (m, 611, Ar-aH2(CH2)2C'S2 ^^

GH — GHg)* 3.2 (m, IH, !TuOH), 3.88-

4.45 (m, 2H, ITOCH2), 6.88-7'45 (m,

4H, Ar-H). 217

5-[ (N-'Morpholino-3-liy(3roxy-2-proHy-l)-CKimino] -6,7>8,9- wii>Miiiiii»^'wi mil - •- 1 -i - - - ^,. .1. in 1 -|i m.i.i ii»ii *i I mil T" ii T lil ' i i ir imn - —— - - > r • —-- --- ' - ' *"•' tetrahydro-5H-benz;ocycloh.eptene {l_05)

I mil 4—%^ A mixture of Ihe epoKy compound 104 (2«31 g, 0-01 mol) and morpholine (0.87 gV 0. ">! mol) in absolute methanol (40 ml) was re fluxed for 48 h- After the reaction period was over, the solvent was removed under "\racuam to fUrnish the crude product as an oil, •vfoich was purified 'oy column chromatography to provide the pure compound 105, yield 1. 6 g (55/0*

NMR : 1.29-2*25 [m, 4H, Ar-CH2(CH2)2] » 2,32-3.0 [m, lOH, Ar-CH2(0112)2022 and N-(GH2)^], 3.52-3.76 [m, 2H, 0(012)2] » 3.85-4.3 (m, 3H, OOH2OHOH), 6.7-7.82 (m, 4H, Ar-H).

5-.[N_(N^„PhenylpiperazLno-3-liydroxy-2-propyl )-cximino] -

6>7j8,9-tetrahydro-5H-benzocycloheptene (106)

It was prepared from 1.04 and N-phenylpiperazLne by a similar method as that for lj)5, in 51^ yield as an oil. mm : 1.35-2.2 [m, 4H, Ar-CH2(CH2)2] » 2.35-2.95 [m, lOH, Ar-GH2(GH2)2% andR-(CH2)j, 3.04-3-85 [m, 4H, (0H2)2-NAr], 3.9-4.4 (m, 3H, OGH2CHOH), 6.74-7.8 (m, 9H, Ar-H). 218

5-[ (lT-t-Butylamino-3-hydroxy--2^propyl)-oi5Ciinino]-6, 7>8,9-'

te-fcraIiyclro-5H-T3enzocycloheptene (JLOJ)

It >7as prepared in 58^ yield from 104 and J-bu-fcylatnine as an oil by a method similar to that for the synthesis of 105* mm : 1.06 [s, 9H (CH-),], 1.28-2.15 [m, 6H, Ar-CH2(GH2)^], 2.3-5.02 [m, 7H, Ar-CHgC 012)2^112 and GH2NH] , -^02^4.5 (m, 3H, OOHgCHOH), 6.7-7.84 (m, 4H, Ar-H), 219

^»7 Biological Activity

Most of the compounds were tasted for thei?:- gross behavioural affects in the Division of Pharmacology, Central Drug Research Institute, Luoknow, employing simils-r procedure as described in Section 3«6> Chapter III.

Results

None of the compounds tested showed any noteworthy antidepressant activity. 220

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5, (1967), 347-352.' SUMMARY Ihe work embodied in the thesis is concerned \r±tli the synthesis of some potential post-coital antifertLliiy agents, speirmicides and antidepressants. Eiis work is described in four chapters. In the first chapter the present status of seco-steroids is reviewed as this topic has not so far been properly reviewed. The second chapter describes the synthesis of some 5>6~seco-estradiols and their antiimplantation activities. In the third chapter the synthesis and bioevaluation of N-substituted a-(2-aminoethyl)acrylophenones and 3-amino~2-substitated methyleneindan-1-ones as possible spermicides has been described. Hie fourth chapter consists of the synthesis and pharmacological evaluation of some iminothiazolidine derivatives as potential antidepressants.

haptor I

The sex steroid hormones, estrogens, progestogens and androgens play an important role in the regulaiion of mammalian reproductive system. Compounds which could interfere with the action of ihese hormones could in principal block conception. One approach to ihe design of such agents is to build analogues of these hormones, which lack one of the bonds in different rings of the tetracyclic structure of the steroid (seco-steroid)j such compounds having greater flexibili-ty would thus be abJ.e to assume a larger number of discreet conformations, only one of which would mimic the prototype and would iiius have altered affinity for the receptors involved and hopefully a changed biological activity profile. In this approach a number of 5»6-seco-steroids have been synthesized and iiieir oontracop-cive activity studies are described in Chapter II.

_G^;2ter^_II.

Synthesis of 5* 6-seco-estradiols In eai-lier work -^ carried out in this laboratory 2a» 63-die thyl--33~(P"hyclroxyphenyl)-'tranS"biqyclo[ 4» 3» O]nonan- 7P-01 (1) was found to inhibit implantation in rats at 2 mg/ kg when administered orally. !Ehe RBA for liiis molecule (1) OH

was 25.85^ as compared to estradiol* In this chapter >ie have described the synthesis of other two isomers of 1 vL a. 2a, 6p-diethyl-3a~(p-hydroryphenyl)-trans-bicyclo[ 4- 3* 0] nonati-7p-ol (ll cis-anti-trans) and 26, 6B-diethy 1-36(p- hydroxyphenyl )-tran3-biqyclof 4« 3* 0] nonan-7p-ol (I2, cis sy n-trans) (Scheme 1). With Ihe hope of increasing the oral absorp.tion and consequently the activity ''che synthesis of 7a-ethynyl derivatives of IJL, and 1 lias also been cai-.tied out. Synthesis of the corresponding _^methoxyphenyl derivatives of 11 and 12 (i.e. 1_6 and IS, respectively) has also "been carried out.

Synthesis of 17a-ethynyl-13|3~ethyl-5» 6-seco-estradiols

The ketoenol _6 was prepared by the condensation of 2-ethylcyGlopentane~l,3-iione(3) with propyl vinyl ketone (2), As a refinement over the method of Gupta _et al. this condensation was carr!.ed out by simply stirring the two reactants in demineralized water for 5 days under nitrogen atmosphere, unlike the earlier method where the tvro were re fluxed in methanol in the presence of an alkali. Cyclization of the triketone (J.) thus obtained with p-toluenesulfonic acid gave 2,6p-diethylbicyGlo[ 4. !5.0]non-l- en-5, 7-dione (_5). Reduction of 7-keto group of _5 with sodium borohydride in a mixture of isopropanol and ethanol afforded _6. treatment of _6 X'/ith j-methoxyphenyllithium in ether-IHF at -40° furnished 2, 6p--diethyl-3-hydroxy-;5-(^ me thoxy phenyl )bi cyclo[ 4» 3* 0] non-l-en-7p-ol (J) t wb.ich VTithout isola-bion was converted to the dienol 8 by the addition of one d.rop of HCl. Oata3,ytic hydrogenatLon of 8 in ethanol over lOfo Pd-C at RIP, till 2 mol of hydrogen was absorbed furnished a mixture of two isomers (_9 and jX;). These two isomers were obtained in pure form in a ratio of 0 0

a 0^ 0^0^

OH 0

cfx^

Contd.. • -I-

11 12

^,> CaOH

Contd. 6

,,xC=CH

0 17 3B a, Demineralized watenVN2* ^' PTSAi c, NaBH.j d, ^BrOgH^OOH /n-BuLii e, HG1» f, IC^ Pd-.C/H2, 2 molj g, KOH, diethylene gLyoolj h, Jones reagentj i, LiC=GH/ethylene diamine

Sc±Leme 1 40:60 by fractional crystallization from a mix tire of ether and hexane. In the mnr spectrum the major isomer showed a highly shielded CH^ resonance (O.I3 &)• This isomer j^ was assigned cis-syp.-trans stereochemistry on the basis of earlier >Jork . The reason for this was -fciiat due to sterio interaction between 6-CHoCH„ and 2-.CH^CH„ group the latter would be held in a plane away from the former. As a result of this steric compression the ring may undergo some conformational change which would bring 2-ethyl chain x-dthin the shielding cone of the phenyl group. Formation of 1/^ in major amount could be due to less steric hindrance for the a-face attach of hydrogen during reduction. Demethylation of 2 Q-^cL _10 separately vdth potassium hydroxide in dieth^ylene glycol/hydrazine hydrate under nitrogen aimosphere gave the corresponding phenols viz- 2a, 6p- and 2(3, 6p-diethyl-3a/p- (£~hydroxyphenyl)-_ta;_ans-bicyclo[ 4» 3« 0]nonan-7p-ol (11- and 12) «

To synthesize the correspond-ing 7a-ethynyl derivative (C-I7 of estradiol molecule), 9 was oxidised to the 7-keto analogue (ij) by Jones reagent and subsequently ethynylated with lithium ace tylide; ethylene diamine under an atmosphere of acetylene by the method of Mraga . Similar oxidation and ethynylation reaction of 9 and 1^0 afforded the corresponding 7a-ethynyl-3P-(^methoxyphen3''l) derivatives 16 and 18. 8

With a view to increase the onral absorptivity of 1, the synthesis of the corresponding Ta-ethynj^l compound 2A was carried out a.coording to Scheme 2. Controlled catalytLn hydrogenation of 8 in ethanol over ICffo Pd-0 at RTP till 1 mol of hydrogen \^as absorbed gave 13, which on Birch reduction gave 2a, 6p-diethy3.-3p-(jg-methoxyphenyl)-_tr^s-bicyclo[4»3«^-'l nonan-7p-ol (2^)» Jones oxidation of _2_0 and subsequent ethyn;^''lation furnished the corresponding 7a-ethynyl compound 2^2 (Scheme 2).

Demethylation of 2^0, Jones oxidation and subsequent etliynylation of the product thus obtained (2j) prodaoed the desired compound 2J-.

Synthesis of 2a,6p-dietliyl-7a-ethynyl-3j3-{cyclohex~2-en-4-

one )-.ferans-bicyclo[ 4» 3« 0] nonan-7p-ol

Reduction of the aromatic ring iii 2^ was carried out with lithium in liquid ammonia in the presence of a protLc solvent such as ethanol to give 2^ (Scheme 5). Oxidation of 23 could not be carried out >d.th Jones reagent as this 5 results in the rearomatization of ring A . Opponauer oxidation of 2_5 gave the 7-keto analogue £6. EthynylatLon of _26 and treatment of the product _27 thus obtained gave 28, the desired 5>6-seco analogue of norgestrel. OH v^ C=CH

a, lOfo Pd-G/H2, 1 moll b, K-NH^j o, Jones reagentj d, Li^C=CH/ethylene diamine, e, KOH, dj.ethylene glycol.

Scheme 2 10

20 -> H

H H

OH .^•C=CE ,v^C=CIi

28 27

a, Li-WH /G2H50H» b, il"^[t-BuO],+r ^ /benzenoj c, LiCsGH/ethylGne diamine J d, HCl/mctlianol

^Sch^ne^^J 11

Synthesis of 2, 6p-diethyl~5-(2-4ae-tihyl-4—hydroxyphenyl)

•bicyclo[ ^^ 3. 0] non-l-en-7p~ol

Condensation of _6 with 2-.methyl-4-meth(K:yphenyllithiuim gave 2, 6p-die thyl-5-hydroX3r-.3-( 2-methyl-4-me -ftioxyphenyl) bicyclo[4-. 5«0]non-l-en-7p-ol (29). Unlike 7 this hydroxy intermediate (_29) was quite stable and ihe dehydration could be carried out only after ref),uxing 2^9 in benzene with _P-toluene sulfonic acid. Catalytic reduction JO in ethanol over 1(^0 pd-C at RTP till 1 mol of "B^ was absorbed resulted in the formation of ^. Further reduction of 2, 3-double bond was tried with lOjJ Pd-C, a mixture of % wet and 10^ Pd-C, Raney-nickol and PtOp at pressures ranging frnm RIP to 1500 p.s.i., but without success. Steric hindrance could be a reason for the inertness of th.e double bond towards reduction. The migration of this double bond to 3,4—position was then attempted but this also could not be carried out successfully. Iriethylsilane in GP_GOOH, a hydiride donor could also not reduce ihis p?>"oduGtf the reaction with this reagent went only to the intermediate stage J^. Forcing conditions for the forward reaction gave back _2i» "fc^^ stariing material. 12

;29 19

^ li ;c Reduction

Jl a, 2^ethyl-4-'bromoa]QisolG/n-BuIiij l3, PTSAi c, 105^^ Pd-G/H2» 1 molj d, Jones reagentg e, KOH, diethylene glycol.

Scheme 4 13

F^COOO 5

Jones oxidation of Jl gave the 7-keto analogue JJ wherGas demethylation of ^1 led to the oorr<3spending hydroxy-phenyl compound _33 (Scheme 4)»

One of the oldest and simplest method for fertility regulation is the use of spermicides for vagina. !I!b.ough the faiT-Ure rates encountered mth these agents are as high as five percent, they find their use mainly out of the disadvantages which users find or fear with other methods. In the begining of tliis chapter a hrief review about iihe known spermicides has been presented. Mongst tlie currently'' used spercn-cides nonylphenoxypolyethoxyethanol (nonyl-9 or nonoxynQl-9, ^) and p-meiiioxyphenylpolyoKyethylene ether (raenfogol or TS-88, J^6) are the most important representative

Y"0-(aH2CH2Q)^M (OCH2CH2)gOH

^9%9 cme. 36 35 14

Some of the most common spermicides available today suffer from the drawback of rather low order of activity and their action is pH dependent* This emphasizes the need for developing more active spermicides which would also be effec'tive in a broad pH range.

In earlier work from this laboratory it was reported that N-substituted a-aminomethylaorylophenones C^Ta) ajcc- diaminomethylacrylophenones (37b) and their quaternary salts exhibit strong spermicidal activity, which are a novel class of spermicides. In a study of structure-function and

R- R-

X^ structure-activity relationship of this novel class of spermicidesjf the synthesis of N-substituted a-(2-aminoethyl- acrylophenones (^) and 2-substi'buted methyloneindan-1-ones

^^^ R'

^^^^^ ^"^^V-^ •N

59 (_2l) was undertaken and their activity is described in this part. 15

Synthesis of N-substitiited a-(2-aminoethyl)acryloplienones

To carry out the synthesis of ^ various substituted Y-chlorobutyrophenones JJO were used as starting material. Ihs corresponding y-B.minohu.1;yroph.enones (ja) used as intei-nnediates were obtained by refluxLng 4£ with an appro priate amine in the presence of potassium carbonate (Scheme 5) The condensation of 4^ with paraformaldehyde was carried out by refluKing the Uio in hexanoic acid, instead of acetic acid vfnich was used for the synthesis of J7a. In the presen-c case a higher boiling acid had to be used since acetic acid could not fetch successful results. Thus the reaction of J_l with paraformaldehyde in hexanoic acid gave the desired compounds _^_.

R /^^

R=P,Br,CH S or H / IT =Pi peri dine, Morpholine, \ Dj.ethylamine or N- P heny Ipi pe ra zi ne R ^''^^ ^ / a, 11 i b, HG02H/hexanoic acid

Scheme 5 16

Synthesis of 3-aminc>-2-substL tuted methyleneindan-l~ones

The synthesis of target molecules J^ was achj.eved starting from various indaii-l~ones i^)» feenzylic bronrLnatLon of J£ vrith N-bromosuccininD.de in 001. in presence of benzoyl peroxide yielded aryl substituted 3-bromoindan-l-ones(£5) (Scheme 6), Reaction of _43 witli different amines resulted in the formation of "the corresponding 3-aniino deriva-feives J.4* Aldol condensation of J4 and various substituted aromailc aldehydes in # alcoholic KOH afforded the desired compounds ^%

Condensation of paraformaldehyde with j44 was carried out by re fluxing the two in acetic acid for 4 b.. Eiis gave a mixture of two isomers ^_ and J^ which could not be, however, separated* The nmr spectrum showed one to be having exocyclic double bond (^) and the other with endocyclic double bond ^5*

Chapter J^Y

At the start of "this chapter a brief account of the known tricyclic antidepressant agents has been given. Most of these compounds ha,ve an associated anticholinergic activity which is a serious limitation \fith. them. These drugs suffer from some other drawbacks such as hepatotoxicit^ and a very late onset of action. Hence, there is a need for 17

42 15

•N-

Q r:;:^NXV^^

N' 45

R H or CH^O 5 ./ M Pipei-idine, morpholine, diethylamine, t-butylamne, \ H-methylpiperazine or lT-ph.enylpiperazine.

R' = OgH^CHO, o-FCgH.GHO, _p-.(CH 0)CgH^CHO, p-(N02)GgH.CH0 or H a, IBS, GCl./Benzoylperoxidej b, M , Ace tone/K^^^^J c, 4^9 KOH alcoholic.

Scheme 6 18

non-tricyclic antidepressants-

Daring a course of. broad primary screening of compounds in this Inst1.tute, Jferan£^-2-[N'-(2-.hydroxy-l,2,3,4- tetraliydro-l-naphtli3'-l)]iminothiazolidine was found to possess marked antidepressant activity , comparable to that of imipramine, an important member of the tricyclic anti depressants. This novel lead proved worthy of further exploitation and prompted the synthesis of some iminothiazolidine derivatives (4_6 and _59).

Synthesis of imino'ciiiazolidine derivati.ves

To achieve the sjmthesis of various iminothiazolidine derivatives (_46 and 39) different benzocycloalkanones (47) were used as starting material. Treatment of _47 ^rf.th hydroxylamine hydrochloride in ethanol in the presence of an alkali under refluxing conditions 5^elded the corresponding oxime J8, which on catalytic hj'-drogonation over lOfo Pd-C furnished the araino derivatj-ve _^9 (Scheme 7). Condensation of J;9 1^rith benzoylisothiacyanate in anhydrous acetone afforded the benzoyl thiourea derivative 30, Debenzoylati on of _5P -vdth IGfo NaOH solution gave the tliiourea derivative _51. Oyclization of the thiourea ohain in Jl with bromoethylamine '•"'^robromide was carried out by allowing the mixture of these tr-ro in ethanol to reflux for 24 h. This resulted in the desired compound _4.6. 19

a -> (CH^) 2^n

R R-

NH-q-WH-C-Pli I! 50 s 0

N, R—

NH-O-NH.

a, KHgOH.HCl j b, H2[Pd/0] j c, CgH^CONCS » d, IC^o NaOH I e, Br(CH2)2NH2.HBr

R = 5-00H_, 6-0CH„ or H, J 5 X = GH2 or 0, n = 1 or 2

Scheme 7 20

The synthesis of trans~2»-rN~( 6~hydTOxy..6,7»8,9-- tetrahydro-5H-benzocyolohepterie)]iininotMazolidine (^9) was carried out according to Scheme 8. The starting mate::'ial •ben2;ocyclohepten-5-one (iDenzsuherone, 47, X=CHp, R=H and n=2) was treated with sodium borohydi-ide so as to provide the alcohol {5^)» Dehydration a£ _52 with potassium bisulfate afforded 5^» The bromohydrin _54 was obtained by the reaction of 52 ^'Jith E-bromosuccinimide in dimethyl sulfoxide-water under nitrogen atmosphere. ^4, when allowed to stir vdth ammonia solution gave the amino alcohol _55 via an epoxide as intermediate. Trans opening of this epoxide gave the 5- amino-6-hydrox3i^-6,7,8,9-te traliydro-5H-benzocycloheptene (J5 ) • The nmr spectrum of this product _55, showed it to be a mixture of two isomers which could not be separated at this stage. Condensation of _55 T-Jith benzoylisothiacyanate gave a mixture ^'^ J^XSIS 9^*^ .Pi.s benzoylthiourea deriva'bives {5jS and 31 respectively). These tifo isomers were separated by fract ional crystallization from acetone-water mixture. The stereochemistry of j56 and 57 was confirmed by NOB expe^r-iments Hydrolysis of 56_ and subsequent cyclization of the thiourea derivative 30^ with bromoethylamine h;ii^drobromide afforded the desired compound 5J*

Tlie ci^-isomer _52> however, behaved in an entirely different fashion. The hydrolysis of 57 with 2

RH-C-NH-C-Ph II II 56 + S 0

-^ OH NH-0-im-O-Ph S 0 57

g 16 -7^ ->

a, NaBH^j b, KHSO^, A | c, NBS, IlviSO/Waterj d, NH^OHi f, 1055 FaOHi g, Br(CH2)2l^H2.HBr

Scheme 8 22

NaOH solu-tion resulted in the formation of two products ^ and 61' 60 was characte-ized as tLie ^ca-s-amlno alcohol and _61 as a urea dei-ivative. Elemental analysis of 61 showed the absence of sulfur. 'Ihe identity of these two products was confirmed on the "basis of nmr, ir and mass spectra and elemental analysis..

Hydrolysis 57

60 8Ci5^«

Synthesis of pyrimidinone deriva-bives

A large number of pyrimidinone derivatives synthesized in tliis Institute were found to possess stimulant or depressant activity. This prompted to utj.lize the thiourea deriva-tives 31 for the synthesis of some pyrimidinone derivatives (^)»

S-Me-thylation of some thiourea derivatives (57, Z=:0, R=H, n^l and ZriCHp* E.=H, n=2) was carried out by reflijzino it in absolute methanol with methyl iodide (Scheme 9), so as to produce _62» The N-substituted isothiouranium salt _62 was formed as an intermediate which was converted to 6^ by bicarbonate treatment. Oyclization of 6^ vrith e^hyl 23

propiolate in meiaaanol afforded 6J. [ 3-(4-oromaac;l)^2- methylJneroapto-4( 3H)-pyrimidinone aiad >-(6,7,8,9-tetrahydro- 5H-l3Gn2;oc3'-cloheptcnG )-2^methylmercapto-4( 3Il)-P3nriinD. dinone].

:i (CH^), >

NH-C=NHI o2 I s 0 51 62

b V

,^^^ ^ ^ OO^'^'n (OH,2'') n NH-C=:IJH OH S,^ N Ti S-CH, 63

a, Mel/metlianol| b, NaHGO-i c, CH=C-COOC^H^

Scheme 9 24

Synthesis of 5-[ (N-substLtuted 3-amino-2-.lTydroxyproi)yl) oxindno] -6,7»8, 9-te trahydro-SH-benzoq^.'"cloheptenes

1 reoent publication has shown the importanco of aromatic oxime ethers as effective p-adronergic blocJsers. This prompted to synthesize the title oompoi:uids {SS),

Reduciion of the ciximino derivative (_48) of benzsuberono with sodium methoccide/methanol and subsequent reaction i/dth epichlorohydrin in anhydrous IMS' gave the epoxide 63* Opening up of this epoxide 65 TJith various amines gave the title compounds 66.

a ->

N-O-CH2CHCH2N 66 OH ^ + - a, 1. NaO Me/methanol, 2, epichlorohydrin/nviFj b, M^ (morpholine, t-butylamine and N-phenylpiperazLne

Scheme 10 25

Bi olq^oal Activi tv;

Most of the oompounds have been evaluated for their "biolc^oal ao"'civity. In the antifertiliiy testing £4 was found to be iiie most promising molecule of ihese stu(S.esj it showed 100^' antiimplantation activity in female rats at a dose of 1 mg/kg when adain:.stered orally. Testing at further lox^er doses is in progress.JJ- had 25.49^ receptor binding affinity as compared to estradiol. 11 and 12 have also sham lOOJo antiimplantation activity at 10 mg/kg when administered orally to rats. Two oiiier interestingly active molecules were ^ and ^1» Both these compounds showed 100/3 antiimplantation activity in rats at an oral dose of 2 mg/kg. Screening of 11, 12^, XQ ^^ -S ^^ lower doses is under process.

None of the oompounds showed any noteworthy activity in spermicidal and antidepressant screening. 26

REFEREHCBS

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11, (1979), 67-77. 3. R.O.Gupta, S.Durani, A.K. Agarwal, V.P.Kamboj and N.Mand, Ind. J. Cliem., 19B, (1980), 866-890, 4. K.IIiraga, Chem. Pharm. Bull., 1^, (1965), 1289-1294. 5. H.J.Ringold, G.Rosenkranz and F^Sondheimer, J. Am* Ohem. Soc., 78, (1956), 2477. 6. P.Nautiyal, Ph.D. Dissertation, lucknow University, Lucknow, (1981). 7. U.K.Sliukla, Pii.D. Dissertation, Kanpur University, Kanpur, (1979). 8. E.A.G-upta, Ph.D. DLsserta-iriLon, Meerut University, Meerut, (1981).

9. G-.Leclerc, A.Mann, 0. G.¥ermuth, N.Bieth and J.Schwartzr

J. Med. Chem., 20, (1977), 1657.