Studies on the Syntheses of Possible Antifertility and ...

Studies on the Syntheses of Possible Antifertility and Anthelmintic Agents

( SUMMARY )

A THESIS SUBMITTED TO THE

ALIGARH MUSLIM UNIVERSITY, ALIGARH FOR THE

DEGREE OF DOCTOR OF PHILOSOPHY .IN CHEMISTRY

BY Mohammad Shamim Akhtar

M. Phil.

DIVISION OF MEDICINAL CHEMISTRY CENTRAL DRUG RESEARCH INSTITUTE

LUCKNOW-226001 FEBRUARY, 1987

SUMMARY SYNTHESES OF POSSIBLE ANTIFERTILITY AGENTS

In India, one of the pr ior i ty areas of research work in

medicinal chemistry relates to the prevention of pregnancy. A chemical

agent, capable of preventing pregnancy, has greater acceptabili ty u

provided it does not prolong the period of menyistr^tion and does

not cause unwanted side-effects. Search for structural lead to achieve

this objective has resulted in the synthesis and biological evaluation

of thousands of aryl-ethylene der ivat ives . These compounds, though

very effective in preventing pregnancy in experimental animals, are

not suitable for clinical use. Thereafter no useful structural lead

has been identified for the development of a non-steroidal oral contra

ceptive agent. However, various attempts made to identify possible

lead molecules have .suggested that exploratory research act ivi t ies

in the area of heterocycles may provide useful lead for achieving

this objective. The present study i s , therefore, aimed towards identi

fying simpler heterocycles capable of evoking contraceptive ac t iv i ty .

The various molecules identified for the syntheses and biological eva

luation are l isted belbw:-

n ni

IV

R^=H,Ac

R- 0^ VI

In an another approach i t was considered of interest to

design compounds which may selectively alter the permeability of

the decidual cells in uterus and may also interfere with the oxido-

reductive processes of the cell biochemistry. Representative proto

types of such compounds are:

V M2 O2 C V ^ , X X ^ C 0 2 M G

.CO2MG M(2-

vn

H

vni

•Mc2

Syntheses of compounds belonging to prototypes I-III required subst i

tuted aroyl propionic acids as the starting materials. These were

u ni

prepared by the Friedel-Craft reaction of siragfle aromatic substrate

with succinic anhydride. Esterification of the substituted aroyl pro

pionic acids so obtained followed by their reduction with sodium

borohydride gave a mixture of 4-hydroxy-4-substituted phenyl buty-

rates and 2-oxo-5-substitutedphenyl tetrahydrofurans (Prototype I ) .

These were separated by column chromatography over sil ica gel.

Ring closure of methyl-4-substitutedphenyl-4-oxo-butyrates with acetic

anhydride and hydrazine hydrate yielded compounds belonging to

prototypes II and III respect ively.

Synthetic approach towards

fourth prototype, namely 4-aryl i -

dine-3-methyl-isoxazolone-5- invol

ved the reaction of appropriate

aryl aldoximes with methylaceto-

acetate. One representative comp

ound of th is class of compounds

was reduced with sodium borohydride to obtain a stereomeric mixtures

of IVa.

Syntheses of prototypes V and VI namely 3-substitutedphenyl

2-isoxazolin-5-one and 3-substitutedphenyl-5-methyl isoxazole respec

tively required starting materials of the type A.

R'=H,Ac

N - H 9OCH3

^ - - - - -COR.

VI

o A.R^=OMe,M<2

Ar

M G O C O

COM<2

C02MG/COM<2

1

These compounds namely 4-acetyl-3-methoxycarbonyl or acetyl-4-subst i -

tutedphenyl but-3-ene-2-ones, in pr inciple , can be prepared by the

pyridinium chlorochromate (PCC) oxidation of 2,5-dimethyl-3-methoxy-

carbonyl or acetyl-4-substitutedphenylfurans. Results of the PCC oxi

dation of these furans indicated that instead of compound A 4-acetoxy-

3-methoxycarbonyl or acetyl-4-

substitutedphenyl but-3-ene-2-ones

(1 ) were obtained. This oxidation

is novel and the products being

reactive organic intermediates are

of interest for more than one reason.

For example, the reaction of the oxidation products with aqueous

acid to yield acetophenones may evoke interest for vinderstanding reac

tion mechanism. This is precisely why an appropriate intermediate

has been isolated to delineate the reaction pathway of this reaction.

Hydroxylamine reacted smoothly with the PCC oxidation products to

furnish compounds belonging to prototypes V and VI. However, the

facile loss of COCH group even at milder reaction conditions prevented ^ 1

the preparation of other compounds (R = COCH ) of the prototype

V. Besides the chemical reactivity of the PCC oxidation products ,

their behaviour under electron impact was also interesting and a care

ful study of their mass fragmentation pattern has been carr ied out.

The reqiaired starting material for the synthesis of 2,5-dime-

thyI-3-methoxycarbonyI-4-(3-cyano-2-oxo-l , 2 ,3 ,4 - ' tetrahydroqulnollno)

furan (Prototype VII) was obtained by carrying out modified Nef reac

tion of l-(p>acetamldophenyl)-2-nitro propene. The resulting compound

.C02MG

R H

M G O S C

H

vni

'C02Me

namely 2,5-diraethyl-3-methoxycarbonyl-4-(p-acetamidophenyl )fiiran

was subjected to Vilsmier-Haak

reaction and the formyl derivative

so obtained on oximation gave

2 ,5-dimethyl-3-methoxycarbonyl-

4-(3-aldoxiinino-2-chloro quino-

lino) furan. This compound on

reaction with SOCl followed by

sodium borohydride reduction

furnished the desired molecule.

Compounds representing prototype VIII were prepared by

reacting substituted aldehydes

with methyl-3-amino crotonate

in presence of appropriate solvents.

In order to change R, two synthe

tic strategies have been employed.

In the first one, appropriately

substituted aromatic or hetero

cyclic aldehydes were prepared

and in the second one, appropriate

functionalities on the phenyl ring of 2,6-dimethyl-3,5-dimethoxycarbo-

nyl-4-phenyl- l ,4-dihydropyridine were simulated for elaborating them

to heterocyclic ring systems. Aldehydes such as 4-chloro-3-nitro ben-

zaldehyde, 2-chloro-3-formyl-quinoline and 4-formyl tetrazolo [1 ,5-

a] quinoline have been prepared. The heterocyclic ring systems on

the phenyl ring of 2,6-dimethyl-3,5-dimethoxycarbonyl-4-phenyl-l ,4-

dihydropyridine have been made from appropriate 1,4-dihydropyridine

subs t ra te . For example, hydrogenation of 2,6-dimethyl-3,5-dimethoxy-

carbonyl-4-(4-amino-3-nitrophenyl)-l ,4-dihydropyridine gave the expec

ted diamine, which without further purification was reacted with

a variety of reagents such as a ry la ldehydes , sodium nitrite, benzil,

CS, and phthalic anhydride to yield 2-aryl-5(6)-[ 4-{2,6-dimethyl-

3 ,5-dimethoxycarbonyl-l ,4-dihydropyridyl ] - l (3 )-substitutedbenzyl ben-

zimidazole, 5(6)-[4-(2 ,6-dimethyl-3 ,5-dimethoxycarbonyl-l ,4-dihydropy

r idy l ) Ibenztriazole, 6-[4-(2 ,6-dimethyl-3 ,5-dimethoxycarbonyl-l ,4-

dihydropyr idyl ) 1-2 ,3-diphenyl quinoxaline, 5(6)-l4-(2 ,6-dimethyl-

3,5-dlmethoxycarbonyl-l ,4-dihydropyr idyl) ]-2-mercapto benzimidazole

. and 10-H-2-[4-(2,6-dimethyl-3,5-dimethoxycarbonyl)-l,4-dihydropyridyl]-

10-oxo- isoindolo [ 3 , 2 - a ] benzimidazole r e s p e c t i v e l y .

Biological Act iv i ty:

Various compoxinds syn thes i s ed as p o s s i b l e a n t i f e r t i l i t y agents

have been screened for t h e i r con t racep t ive eff icacy in r a t s and hams

t e r s . New leads have been generated and signif icant con t r acep t ive

a c t i v i t y in 2 , 6 - d i m e t h y l - 3 , 5 - d i m e t h o x y c a r b o n y l - 4 - s u b s t i t u t e d - l , 4 - d i h y -

d r o p y r i d i n e have been o b s e r v e d .

SYNTHESES OF POSSIBLE ANTHELMINTIC AGENTS

The ex i s t ing medical facilijties in ru r a l a r ea s and the soc io

economic s t r u c t u r e of the endemic populat ion of India cal l for the

development of a b road - spec t rum an the lmin t i c . In p r i n c i p l e , a b r o a d -

spect rum anthe lmint ic agent should possess the a b i l i t y to enter ces tode

and nematode p a r a s i t e s i r r e s p e c t i v e of t h e i r t i s sue locat ions and should

be capable of evoking se l ec t ive in te r fe rence wi th the b i o c h e m i s t r y

of the p a r a s i t e s . In o r d e r to ach ieve t h i s o b j e c t i v e , the iden t i f i ca

tion of the p h a r m a c o p h o r e , which should be a t t ached to the lead

molecule, i s e s s e n t i a l . Ear l i e r work on benz imidazo le -2 -ca rbama tes ,

c a r r i e d out in t h i s l a b o r a t o r y , suggested tha t the or ien ta t ion of the

carbonyl g r o u p ( s ) s imulated in the molecular framework of subs t i t uen t s

a t t a c h e d to pos i t ion 5(6) of benzimidazole nucleus g r ea t l y influences

antineraatodal and an t ices toda l p r o p e r t i e s . Although a large number

of compounds have been s y n t h e s i s e d to ident i fy the pharmacophore

r e s p o n s i b l e for w ide - spec t rum anthe lmint ic a c t i v i t y , t he complete

knowledge of d e s i r e d pharmacophore has not yet been a c h i e v e d . I t

w a s , t h e r e f o r e , cons ide red of i n t e r e s t to ex tend t h i s e x p l o r a t o r y

r e s e a r c h a c t i v i t y and the s y n t h e s i s of p r o t o t y p e s IX-XXII ( F i g . l )

was then unde r t aken . The design of p r o t o t y p e s XVI and XVII i s based

on t h e concept t ha t d i h y d r o p y r i m i d i n e d e r i v a t i v e s may in t e r f e r e s e l e c

t i v e l y wi th the r edox reac t ions of the p a r a s i t e energy metabol ism

and if found a c t i v e may s e rve as lead compounds for drug development

work .

The synthet ic strategy for prototype IX, namely 4 -benzoy l -

N- [5 (6 ) - (2 -me thoxyca rbony lamino)benz imidazo ly l I py r ro l i d in -2 -one r e

qu i red 4 -benzoy l -N-(4-amIno-3-n i t ropheny l ) - p y r r o l i d i n - 2 - o n e as the

s t a r t i ng mater ia l and t h i s was p r e p a r e d by reac t ion of /&-benzoyl-

V-butyro lac tone wi th 2 - n i t r o - £ - p h e n y l e n e d iamine . S t ruc tura l e luc ida t ion

IX

MG02C

N

M c 0 2 C ' ^ \ H

^

NOH

MGO2C/ ^N H

XII

MG02C'^ \ H

XIll

, C 0 2 M G

M G - ^ , ^ > - M C

MG02C

XIV

v<r.

N

N ^ N H C 0 2 M e

H

XV

Contd.

N-M(2

XVI xvn

NH

N ^ N H C 0 2 M e

Me02C

NHCO2MG

CO2MG

XVIll

H

XIX

R

^ " ^ ^ C H ^ N H ^ / NC02M(2

NHC02Me

XX

Me

^NHCq2MG

XXI

M202C-

Ar '

•N / C02Me

/ ; -N

-N

H

CO2M2

XXII

(Fig . 1)

of t h e r e a c t i o n p r o d u c t c a l l e d

for a model r e a c t i o n a n d t h e y

one i d e n t i f i e d for t h i s p u r p o s e II -Nc^v^/'N—i

w a s c o n c e r n e d w i t h t h e r e a c t i o n r y ^ y y \ / / ^ ^ \ i^ i j

of /3 - ( £ - t o l u o y l ) - Y - b u t y r o l a c t o n e s

w

, N ; Y ' ^ ~ - C 0 2 M Q

H

i t h £ ^ - a n i s i d i n e . D e t a i l e d s t u d i e s 13 15

on t h e C and N-NMR s p e c t r a

of t h e r e a c t i o n p r o d u c t in t h i s

mode l r e a c t i o n h e l p e d to i d e n t i f y

i t a s 4 - t o l u o y l - N - ( £ - m e t h o x y p h e n y l ) - p y r r o l i d i n - 2 - o n e . B a s e d on t h i s

r e s u l t , t h e r e a c t i o n p r o d u c t of 0 - b e n o y l - Y - b u t y r o l a c t o n e w i t h 2 - n i t r o -

£ - p h e n y l e n e d i a m i n e w a s i d e n t i f i e d a s 4 - b e n z o y I - N - ( 4 - a m i n o - 3 - n i t r o p h e -

n y l ) - p y r r o l i d i n - 2 - o n e .

The s y n t h e s i s of p r o t o t y p e X, n a m e l y ^ - [ ( 2 - m e t h o x y c a r b o n y l -

a m i n o ) - 5 ( 6 ) - b e n z i m i d a z o l y l ] - a c r y l i c a c i d , w a s i n i t i a t e d b y r e a c t i n g N-~-^^^^'^\^''^^^^^55i./' 2

4 - a c e t a m i d o - 3 - n i t r o b e n z a l d e h y d e

w i t h ma lon ic a c i d in p r e s e n c e

of c a t a l y t i c amount of p y r i d i n e .

The r e s u l t i n g p r o d u c t w a s h y d r o - ^

l y s e d u n d e r a l k a l i n e c o n d i t i o n s

a n d t h e £ - n i t r o a n i l i n e d e r i v a t i v e

so o b t a i n e d w a s h y d r o g e n a t e d u n d e r c o n t r o l l e d c o n d i t i o n s to

f u r n i s h t h e d e s i r e d d i a m i n e . R e a c t i o n of t h i s compound w i t h N , N - d i m e -

t h o x y c a r b o n y l - S - m e t h y l i s o t h i o u r e a y i e l d e d t h e d e s i r e d p r o t o t y p e X.

The s y n t h e s i s of p r o t o t y p e XI, n a m e l y m e t h y l 5{ 6 ) - [ 5 - e t h o x y -

c a r b o n y l - 6 - m e t h y l - 3 , 4 - d i h y d r o p y r i m i d i n - 4 - y l 1 b e n z i m i d a z o l e - 2 - c a r b a m a t e

r e q u i r e d 5 -e t h o x y e a r b o n y 1 - 6 - m e t h y l -

2 - o x o - 4 - ( 4 - a c e t a m i d o - 3 - n i t r o p h e n y l ) f-j>^ >N"

p y r i m i d i n e a s t h e s t a r t i n g m a t e r i a l . M G O O C ^ \ M

T h i s w a s p r e p a r e d b y t h e a c i d \-\

c a t a l y s e d r e a c t i o n of 4 - a c e t a m i d o -

3 - n i t r o b e n z a l d e h y d e w i t h e t h y l -

a c e t o a c e t a t e a n d u r e a . T h e p y r i m i

d i n e so o b t a i n e d a f t e r h y d r o g e n a -

t i o n w a s c y c l i s e d w i t h N , N - d i m e -

t h o x y c a r b o n y l - S - m e t h y l i s o t h i o u r e a to g i v e t h e r e q u i r e d p r o t o t y p e XI .

During the course of present investigation on the preparation of dihy-

drop yri mi dines, it was possible to isolate the intermediate formed

in the acid catalysed reaction of aromatic aldehyde with /3-ketoester

in presence of thiourea. It is noteworthy because earlier attempts

to isolate the intermediate were unsuccessful.

The preparation of the

prototype XII, namely 2-raethoxycar-

bonylamino benzimidazole-5( 6)-aceto-

xime by the direct oximation of

methyl-5{6)-acetyl benzimidazole-

2-carbamate has revealed that MGO2C despite the presence of a carbamate

moiety, the oximation of carbonyl

group was possible.

r>^

XII The synthesis of prototype

XIII (X = NH) owes i ts origin to the observation made during

the upscaling studies of the CDRI compound 83/148, a potent anthelmin

tic agent. It was observed that during the preparation of 2,5-dimethyl-

MGO2C .C02Me

8 3 / 1 4 8 :X = 0 XIII ;X = NH

XIV

N

N'^NHC02M«

3-methoxycarbonyl-4-( 4-amino-3-nitrophenyl) furan, the required inter

mediate for the preparation of 83/148, invariably a small amount of

red compound was formed in the reaction mixture and this was identi

fied as 2 ,5-dimethyl-3-methoxycarbonyl-4-( 4-amino-3-nitrophenyl)pyrrole.

The formation of this pyrrole derivative along with the corresponding

furan in the Nef reaction of l-( 4-amino-3-nltrophenyl )-2-nitro propene

in presence of methylacetoacetate and piperidine suggested that a

careful study of th is reaction may furnish a reaction condition in

which the pyrrole could be obtained as the predominant product.

10

This prompted to study the Nef reaction in presence of pr imary,

secondary and ter t iary amines and the results were monitored by

HPLC. The reactions of 2-nitro-l-substi tutedphenyl propenes with

methylacetoacetate in presence of primary amines gave exclusively

N-substituted pyrrole , while the presence of secondary amines fur

nished a mixture of pyrrole and furan derivatives and the presence

of ter t iary amine exclusively yielded furan der ivat ives . The hydroge-

nation of 2 ,5-dimethyl-3-methoxycarbonyl-4-( 4-amino-3-nitrophenyl )-

N-substituted pyrroles and 2,5-dimethyl-3-methoxycarbonyl-4-substituted

phenyl-N-( 4-amino-3-nitrophenyl )pyrroles followed by the cyclisation

of the hydrogenation products with N .N-dimethoxycarbonyl-S-methyliso-

thiourea yielded compounds of prototypes XIII and XIV respect ively .

The C-NMR spectra of tetrasubstituted furans and tetra or penta-

substituted pyrroles and their comparislon with spectra computed

by a computer on the basis of literature data have yielded useful

information.

The synthetic strategy for prototype XV, namely 2 ,2 ' -d ica rbo-

methoxyamino-5,5'-dibenzimidazolyl methanol involved the following

sequence of reactions. Nucleophilic displacement of chlorine in 4 , 4 ' -

H

XV

dichloro-3,3 ' -dini t ro benzophenone with aqueous ammonia followed

by the sodium borohydride reduction of the resulting nitroamine gave

the secondary alcohol. Hydrogenation of this compound followed by

the cyclisation of the resulting diamine with N ,N-dimethoxycarbonyl-

S-methylisothiourea furnished the required prototype XV,

Synthesis of prototypes XVI-XVII were initiated by carrying

out acid catalysed reaction of aromatic aldehydes with methylaceto

acetate in presence of thiourea. Studies on the alkylation of resulting

compound, namely 5-ethoxy-carbonyl-6-methyl-4-substitutedphenyl-3,4-

dihydropyrimidine-2-thione, gave useful information. Alkylation with

11

me thy l iod ide in presence of sodium h y d r o x i d e in methanol y i e lded

Et02C

N-MG

Et02C

XVI XVII

only S-methyl d e r i v a t i v e and the raethylation of the same subs t r a t e

in DMSO gave the d imethy la ted product namely 1 ,6 -d ime thy l -5 - e thoxy -

ca^bony l -2 -me thy lmercap to -4 -pheny l -4 - ( l - H ) - p y r i m i d i n e . Nucleophi l ic

displacement of methyl mercaptan from S-raethyl d e r i v a t i v e (mono

a l k y l a t e d p roduc t ) by N-methyl p ipe raz ine furnished XVI. In an another

novel a lkylat ion reaction, 5 - e t h o x y c a r b o n y l - 6 - m e t h y l - 4 - s u b s t i t u t e d -

p h e n y l - 3 , 4 - d i h y d r o p y r i r a i d i n - 2 - t h i o n e was r eac t ed with monochloroacetic

ac id in presence of BF - e t h e r a t e in methanol and t h i s r e su l t ed in

the formation of p ro to type XVII.

The synthes i s of prototype XVIII, namely 5( 6 )-N[ {2-methylmer-

c a p t o - 6 - m e t h y l - 4 - p h e n y l p y r i m i d i n - 5 - y l ) methyl Jamino benz imidazole-

2-carbamate invo lved the a romat i sa -

tion of 5 - e t h o x y c a r b o n y l - 2 - m e t h y l -

mercap to -6 -me thy l -4 -pheny l -4 ( IH)-

pyr imid ine wi th manganic ace ta te

as the f i r s t s t ep of the total

s y n t h e s i s . Th i s was followed by

LAH reduc t ion and the resu l t ing

compound namely 5 - h y d r o x y m e t h y l -

2 -me thy lmercap to -6 -me thy l -4 -pheny l

py r imid ine was r eac t ed wi th PBr,

to obtain the d e s i r e d bromo d e r i v a

t i v e . The l a t t e r was r eac t ed wi th

2 - n i t r o - p - p h e n y l e n e d iamine , and the reac t ion product was then h y d r o -

genated to y ie ld the d e s i r e d diamine which was then c y c l i s e d with

N,N-d ime thoxyca rbony l -S -me thy l i so th iou rea to furnish the r e q u i r e d

compound of the p r o t o t y p e XVIII.

NH

N^NHC02M<2 XVIII

12

NHC02Me

Syntheses of molecules belonging to prototype XIX, namely

methyl-5{6)-l4-(2,6-dimethyl-3,5-

dimethoxycarbonyl-1,4-dihydro-

pyridino) ]-N-substituted benzitnida-

zole-2-carbamates, were achieved

by following sequence of 3-Nitro-

4-substitutedamino-chlorobenzal-

dehyde was reacted with methyl-

3-amino crotonate to obtain the

corresponding 1,4-dihydropyridine

der ivat ive . NMcleophilic displace

ment of chlorine by amines followed

by hydrogenation of the resulting

o^-nitro aniline derivatives gave

the desired o^-phenylenediamine derivatives which were then cyclised

with N,N-dimethoxycarbonyl-S-raethylisothiourea to yield XIX.

XIX

R

The compounds representing prototype XX namely N,N-dime-

thoxycarbonyl-N-substitutedbenzyl

auanidines were prepared by react

ing appropriate benzylamines with

N, N-dimethox year bony 1-S-me thy 1-

isothlourea.

CH2NH NCO2MG

NHCOaMe

VNHCO2MG

XX The synthetic strategy for prototype XXI namely methyl-4(5)-

methyl-5( 4)-(3 ,4-methylenedioxy-

phenyl )-4 ,5-dihyd^oimidazole-2-

carbamate, involved the reaction

of 2-nitro-l-(3,4-methylenedioxy-

phenyl )propene with hydroxylamine

hydrochloride as the first s tep.

The compound so obtained was

hydrogenated and the resulting diamine was cyclised with N,N-dirae-

thoxycarbonyl-S-methylisothiourea to yield compounds of the prototype

XXI.

The synthetic strategy for the prototype XXII, namely 1,6-

biscarbomethoxy-2-methoxycarbonylamino-7-methyl-5-(p-methoxyphenyl)-

4,5 ,6 ,7- te t rahydro- l ,3-diazepln.e required 3-amlno-2-^ (p-methoxyphenyU

nltroethyl Jcrotonate as the starting material. This was prepared by

XXI

13

react ing 2 - n i t r o - l - ( p - r n e t h o x y p h e n y l )propene with methyl -3-amino c r o t o -

na te . This compound was reac ted wi th hydroxy lamine to obta in 3 -ca rbo -

methoxy 5 -n i t ro -2 -ox i -M(2

mino- (4 -p -me thoxypheny l ) » P O - i M s

pentane. Evidence for KyloO P / "N

the ass igned s t ruc tu re

MeO ^ >^xjj

was obta ined from s p e c

t roscop ic data and

from the r e s u l t s of

chemical t ransformat ions .

Similar to the reac t ion

of 1-( p- rae thoxyphenyl ) -

propene with methyl-3-araino c ro tona te , reac t ions of o the r 2 - n i t r o -

1-subs t i tu tedphenyl propenes with the same reactant gave 3-amino-

2-[ ( / a - s u b s t i t u t e d p h e n y l )n i t roe thy l I c ro tona tes , which in turn r eac t ed

wi th hydroxylamine to yie ld 5 -n i t ro -2 -ox imino -4 - subs t i t u t edpheny l

pentanes . Oxidation of the l a t t e r c lass of compounds with e i t h e r mono-

p e r p h t h a l i c ac id or m-chloro perbenzoic acid y ie lded 3-ca rbomethoxy-

3 - h y d r o x y - 5 - n i t r o - 4 - s u b s t i t u t e d p h e n y l pen tan-2-ones . Oximation of

t he se compounds gave 3 - c a r b o m e t h o x y - 3 - h y d r o x y - 5 - n i t r o - 2 - o x i m i n o -

4 - subs t i t u t edpheny l pen tanes . On the bas i s of Dreiding model the

s t e r e o c h e m i s t r y of these d ias te reomers was ass igned as R R ( t h r e e ) .

In o r d e r to obta in the p ro to type molecule, 3 -ca rbomethoxy-4- ( p -methoxy-

phenyl ) -5 -n i t ro -2-ox imino pentane was hydrogenated and the produc t

without fu r ther pur i f ica t ion was cyc l i s ed wi th N ,N-d ime thoxyca rbony l -

S-methyl i so th iourea to y ie ld XXII,

Biological Act iv i ty

Compounds r ep resen t ing var ious p r o t o t y p e s were eva lua ted

for t h e i r an t i f i l a r i a l a c t i v i t y against L. cciAinii, cestocidal a c t i v i t y

against H. nana and nematocidal a c t i v i t y against A. cZLj^anicum and N.b^a-

'4-ttten-4-i'4 infec t ions . Compounds exh ib i t i ng signif icant an the lmin t ic

a c t i v i t y were screened for t h e i r a c t i v i t y against developing s tages

of p a r a s i t e s . Sens i t iv i ty of pharmacophores in "benzimidazole carbama

tes" for e l ic i t ing anthelmintic ac t iv i ty has been discussed and a comp

ound e x h i b i t i n g spec i f ic a c t i v i t y against hook-work has a l so been

iden t i f i ed . Compounds r ep resen t ing p r o t o t y p e s X, XII, XVI, XVII,

XVIII, XX and XXII d id not e x h i b i t any anthe lmint ic a c t i v i t y .

Studies on the Syntheses of Possible Antifertility and Anthelmintic Agents

A THESIS SUBMITTED TO THE

ALIGARH MUSLIM UNIVERSITY, ALIGARH FOR THE

DEGREE OF DOCTOR OF PHILOSOPHY IN CHEMISTRY

BY lAohammad Shomim Akhtar

M. Phil.

DIVISION OF MEDICINAL CHEMISTRY CENTRAL DRUG RESEARCH INSTITUTE

LUCKNOW-226001 FEBRUARY, 1987

Jfl'S - 'if ••0 l> *

T3594

TO

MY PARENTS

AND

TEACHERS

T«lex : 0 5 3 5 - 2 8 6 T«l«gr«m : CENDRUG Phone : 32411-18 PABX

No.

srR jrftrw. Tte inw #<> 173

rtisH* 226001 (TTOT)

CENTRAL DRUG RESEARCH INSTITUTE Chat tar Manz i l . Post Box No. 173

LUCKNOW—226001 (INDIA)

Date 23Kd FzblaoAij 1987

CERTIFICATE

Thi6 i6 to czfitily that thz wofik zmbodizd in tki6 thz6i6

ha6 bzzn ccxAtizd oat by Mt. M. Shamim AkhtoA, hA.Phii., undz^

ouA 'tupzxvi'iion. He ha6 {,a<ilMzd thz izquin-Zmznt^ ^01 thz dzg^zz

oi Doctor oi Phiio'iiophLj o^ thz AttgoAh Mu6tLm UnivZAMty KzgaxdinQ

thz natuAZ and pAZ6CA.ibzd pziiod ol in\JZ6tigationai woik. Thz

woAk inciadzd in thi'i thzM6 uniz^6 othZAwi^z 6tatzd. i6 ait oii-

ginai.

<N\ . S -M-^E^ (M.S. AHMAV) (A.P. BHAVURl)

ACKNOWLEDGEMENT

I •ê.zi plza^zd to izcofid my i.ndzbtnz^^ and pA.o{,ound ê-n^z

0^ gâtitudz to Vi. A. P. BhaduA^., Sc<.tnt<6t E-U, Mzdicmal Chzm4.6tA.Lj

Viv-tMon, CzntA.al VKUQ Rz^zaxck iMtitutz, Lucknow ÔA. hi6 indi6pzn-

6abiz Qindancz, con6tn.uctiK/z cn.itici6m, a^zctionatz behav-couA and

Z^zxia6ting zncouAagzmznt which 6u6tainzd my zô^.t'S at all thz ^tagz^

o{j thz oAduoa^ andzitaking^.

I wi6h to convzy my 6inczn.z thanks to Pxo{,. M.S. Ahmad,

Chaiiman, VzpoAtmznt o^ Chzmi6tn.y, AttgoAh Mwtilim LlnivzA.Mty, AligoAh,

•lôt hi6 kzzn intZA.Z6t and mvaluablz hzlp izndzizd during thz tznuAZ

0^ thi'i woik.

Thank'^ oAZ daz to Vi. M.M, VhoA, f.N.A., DixzctoA., Czntn.al

Vn.ug Rz^za/ich In'ititutz, Lucknow and VA.. V.S. Bhakuni, FNA, Vzputy

ViKZctoi Ion. providing lahoKatoiy iacilitiZ6.

1 am al6o gâtzûl to all thz mzmbzn6 o^ PoAaMtology and

Endocrinology Division o{, thi^ In^titatz ô-t providing valuablz âizzn-

mg data.

I am thankful to Vi. K.P. Madha6udanan, Sciznti'it, ôx provi

ding mz thz opportunity o{, Ma^'f^ Spzctromztric ^tudiZ'i.

1 would tikz to acknowlzdgz Pro^. E. Rti'dzr, Pharmaczutical 13 In^dttitutz, Bonn, ôr pro\îding UtzratuAZ ba^zd computzr data o^ C-

NSAR.

I am indzbtzd to thz 6ta{,i o^ Rzgional Sophi'iticatzd In^trumzn-

tation Czntrz, C.V.R.I., ôr providing ^pzctro^copic and analytical

data.

My warmz-it thanks arz daz to Vr.iMi'i,^) M. Szth ôr many

hzlpiai di^ca^îon^, zncouragzmznt and a^^zctionatz bzhaviouA daring

thz zntirz cour6z o^ thi4> ^tudy.

A gatorz oi thanks to Vr. U.L. ShoAma •{,or 6pznding hi6 vaia-

ablz timz in zxprz^îng hi^ {înz 6zn6Z o^ art.

1 atio acknowlzdgz thz 'Stimulating companion6hip and nzzd-

lui cooperation oi A^hrai, A^hwani, Rahul, Sangzzta, Winzzt, Ranjan,

Anita, Vibyzndu, Raju, Sham-ihad and Pzrwaiz.

http://Chzm4.6tA.Lj

I would be {,cUtinQ in my duty i{, I do not mucoid my /iumfa£e

appfizciation ioi thz conîttant dncouAagzimznt, in my all zndzavouA'^,

I izczivzd ôm my paA.znt6 and otkzn. {,amiiy mzmbzX'i.

finally thz ^nancial a^î^tancz in thz ô^m oi {,zliow6hip

oi Council o{j Scizntiiic and Indu^tîal Rz6zaA.ch (CSIR), Nzw Vzlhi,

i6 gâtz^fully acknowlzdgzd.

^<^^

(M. Shamim AkhtoAJ

LIST OF ABBREVIATIONS

Unless o the rwise s ta ted a l l a r a b l e numbers in BOLD l e t t e r s

re fe r to compounds d i scussed in t h i s d i s s e r t a t i o n .

AlCl^

bs

^^C-NMR

d

DMSO

DMF

Ether

Hz

IR

Jra

Jo

LAH

m

m . p ,

PMR

q

s

t

TEA

UV

Aluminium Chlor ide

Broad s inglet

Carbon nuclear magnetic resonance

Doublet

Dimethyl su lphox ide

Dimethyl formamide

Diethyl e the r

Hertz

Inf rared

Jmeta

Jor tho

Lithium aluminium h y d r i d e

Molecular ion

mult iplet

Melting point

Proton magnetic resonance

quar t e t

s inglet

t r i p l e t

Tr i f luoroacet ic ac id

Ul t ravio le t

CONTENTS

Page

Preface . . . i

CHAPTER 1

1.1 : Non-s tero ida l contragesta t ional he t e rocyc l e s . . . 1

1.2 : Basis of work . . . 9

CHAPTER 2

2 .1 -2 .5 : Syntheses of compounds belonging to p ro to types

I-VIII . . . 12

CHAPTER 3

3.1 : Exper imenta l . . . 37

3.2 : Tables . . . 47

3.3 : Biological a c t i v i t y . . . 69

CHAPTER 4

4 .1 -4 .5 : S tudies on chemotherapy of helminth infec t ions :

Ret rospec ts and p ro spec t s . . . 72

4.6 ; Basis of work . . . 76

CHAPTER 5

5. L-5 .9 a : Syntheses of compounds belonging to p r o t o t y p e s

IX to XXII . . . 100

CHAPTER 6

6.1 : Exper imenta l . . . 138

6.2 : Tab les . . . 154

6.3 : Biological a c t i v i t y . . . 189

REFERENCES . . . 199

PREFACE

In India, the pr ior i ty areas of research work in medicinal

chemistry are two; the first one is concerned v i th parasi t ic infec

tions while the second one relates to the prevention of pregnancy.

The existing medical facility in villages and the socio-economic

structure of the population call for an easy access to a broad-spec

trum antiparasitic agent and if a cheap chemoprophylactic is also

made avai lable , the diseased and those who are succeptible to

the disease would be benefitted and would help in restr ict ing the

endemic areas to a managable situation. Prevention of pregnancy

by a chemical agent would be more acceptable to those who are

motivated for family welfare if the agent causes minimum physiologi

cal insult and if the period of menv/stmtion is not enhanced.

In the light of these concepts the present study has been

init iated. Approach for developing an agent to prevent pregnancy,

therefore, is concerned with the non-steroidal molecules. The failure

of tr iarylethylenes to provide a non-steroidal remedy has prompted

a world wide search for new structural leads. The f irst part of

this dissertation is directed towards this objective. The second

part of this dissertation is aimed towards identifying pharmaco

phores which are responsible for evoking anticestodal and antinema-

todal act ivi t ies i rrespective of their tissue locations. Care has

also been taken to evaluate active compounds for chemoprophylactic

activi ty since this may help to identify the pharmacophore respon

sible for chemoprophylaxis.

Synthesis of desired prototype molecules is often associated

with interesting chemical reactions and a few of the molecules synthe

sized might also offer an opportunity to enrich the present state

of knowledge provided their spectral properties are carefully studied,

The present dissertation has taken a note of this situation.

CHAPTER 1

1.1 : NON-STEROIDAL CONTRAGESTATIONAL HETEROCYCLES

1.1.1 : Monocyclic heterocycles

1.1.2 : Blcyclic heterocycles

1.1.3 : Polycyclic heterocycles

1.2 : BASIS OF WORK

CHAPTER - 1

1 . 1 NON-STEROIDAL CONTRAGESTATIONAL HETEROCYCLES

C h e m i c a l a g e n t s w h i c h p r o v o k e p r e g n a n c y f a i l u r e

by c r e a t i n g a s i tuat ion in which the p r o p e r endocr ine requ i rements

have been d i s tu rbed to cause d i r e c t l y or i n d i r e c t l y an t i zygo t i c ,

b l a s t o l y t i c , ba l s to tox ic or abor t i fac ien t a c t i v i t i e s , a r e te rmed as

contragesta t ional agents . Control of popula t ion , a pr ime need of

the present age , may be ach i eved by these agents but the small

and defini te incidence of se r ious s ide-e f fec t s resu l t ing from these

agents should be careful ly s tud ied since i nh ib i t i on of pregnancy

must lead to minimum phys io log ica l in su l t . The most common type

of contragesta t ional agent effect ive by oral route of admin i s t r a t ion

(oral con t r acep t ives ) r e p r e s e n t a combination of an es t rogen and

a p roges t in since a p roges t in or an es t rogen alone i s c l i n i c a l l y l ess

a c c e p t a b l e . However, the need for a second generat ion of ora l con t ra

c e p t i v e s has been r e a l i s e d because agents which do not e x h i b i t any

hormonal or ant ihormonal effect but a re capab le of evoking s e l ec t i ve

physiologica l imbalance in the u terus to cause pregnancy fa i lu re

would be more a c c e p t a b l e . A d e s i r a b l e choice for designing a such

contrages ta t ional agents a r e he te rocyc les and t h i s has p rompted to

review the e a r l i e r r e p o r t s of the cont rages ta t ional eff icacy of h e t e r o

cyc les and a br ief r e p o r t of the same i s p re sen ted h e r e .

1.1 .1 Monocyclic heterocycles

The most conspicuous non-hormonal con t r acep t i ve agen t s ,

developed by Omodei-sale et_ aj^ , a r e r e p r e s e n t e d by 3 , 5 - d i s u b s t i -

t u t e d - l H - 1 , 2 , 4 - t r i a z o l e s (!.)• These i n h i b i t pregnancy in hams te r s

at a dose of 5 mg/kg P .O . A more effect ive compound of t h i s s e r i e s .

N NH f? -J R = Cj_^-alkyl; R = H. F, CI,

MeO, Etc , Cj_^-alkyl;

2 R = H, F, CI, Cj_^-alkyl or

alkoxy; R R = OCH O

2-

namely 3 - ( 2 - e t h y l p h e n y l - 5 - ( 3 - m e t h o x y p h e n y l ) - l H - l ,2 , 4 - t r i a z o l e {2),

deve loped by Galliani et_ al_ , e x h i b i t s a n t i f e r t i l i t y effect by acting

on the u te roplacenta l unit (UPU), resu l t ing in r e so rp t ion or expuls ion

of the concep tuses . This compound i s ac t ive whe ther given o r a l l y .

OEt OMe

M(2

subcutaneous ly , in t r amuscu la r ly or in t r avag ina l ly but the requ i rement

of a ve ry high ora l dose l imi t s i t s usefulness . Another d e r i v a t i v e

of 2. (R = CH ; R , R , R = H) also e x h i b i t s s ignif icant pregnancy 3

terminat ing a c t i v i t y . The ED values a r e 0.3 mg/kg /day s . c . in

hams te r s and 2 mg /kg /day s . c . in r a t s . Instead of t h r e e ni trogen

atoms in the he t e rocyc l i c r i n g , py razo l e d e r i v a t i v e (3 ) wi th two 4

ni t rogen atoms a lso e x h i b i t s pregnancy terminating a c t i v i t y and

the r e p o r t e d ED values a r e 3 mg/kg /day in hams te r s and 25 mg/kg/

day in r a t s .

Oral admin i s t r a t ion of a r e p r e s e n t a t i v e of s ix -membered

he t e rocyc l e s namely 2 ' , 3 ' , 5 ' - t r i - 0 - a c e t y l - 6 - a z u r i n e (\), at a dose

of 300 mg/kg on days 8, 9, 10, 11 and 12 of pregnancy in r a b b i t s ,

have been r e p o r t e d to i n h i b i t pregnancy in a l l cases whi le d e r i v a

t i v e s of 2 - [ « < - ( o r t h o s u b s t i t u t e d benzimidoyl) benzy l ] p y r i d i n e (5 )

have been found to i n h i b i t ovula t ion .

AcOHoC R^N

1.1.2 Bicyclic Heterocycles

Compared to monocyclic heterocycles, more number of bicyc

lic heterocycles have been evaluated for their ant ifer t i l i ty ac t iv i ty . 7

Of these 3-(2-benzofuranyl )-3-alkyl-2,2-dimethyl propionic acids a

(6^), substituted 2-anilino benzoxazoles (7 ) and 2-benzoyl-3-phenyl 9

benzothiophene (or benzothiophene oxide) derivat ives (8) have

shown pregnancy inhibiting act ivi ty in r a t s .

R R2 ^^C02R^

7

R = H, Me; R = H, OMe;

R^ = Me, Et; R^ = R* = H

R = H, a l k y l , a l k o x y ,

a l k y l o x y , CO H

R,R = OH, H, a l k o x y ;

2

R = H, CI , Br , OH, a l k o x y ;

R = H, Py r ro lod inoe thoxy ;

n = 0, 1

8

Derivatives of 2-aroyl-3-phenyl benzothiophenes {9) at 1 mg/day

S.C. and 1,2-diphenyl-l ,2,3,4-tetrahydroquinoline (2£) at 50-100

mg in ra ts have been reported to prevent pregnancy.

R = H, 4-OMe, 4-OH, 4-Cyclo-p h e n t y l o x y , 3-OMe, 3-OH, 2-OMe, 2-OH, 3-Cl , 4-Cl;

R^ = H, OMe, OH

Bicycl ic compounds such as indazole d e r i v a t i v e s (JLi.) ^^^ 1 ,2 -d iphe 13 . .

ny l indoles (^2^) a lso e x h i b i t con t r acep t ive a c t i v i t y .

NHR R

11

R = Me, Ph, CH :CHCH , Et; 1 I i

R = Me, H, HC; CCH^, H^CtCHCH^Ph

R = Cyc lohexy l , Me2CHCH2Ph

R = H, CN

10I>™

R MeO; R = Et NCH^CH^

A chroman der ivat ive namely 3 ,4 - t rans -2 ,2 -d lmethy l -3 -pheny l -4 - [2 . -14 (3 -pyrro l id inoethoxy)phenyl ] -7 -methoxychroman (13) prevents con

ception at a single oral dose of 1,25' mg/kg in rats and mice and

2.5 mg/kg in dogs and rhesus monkeys immediately postcoitum. It

also possesses estrogen, antiestrogen and antiprogestational propert i e s .

M G O

13

The isoquinol ine d e r i v a t i v e , l - N - m e t h y l p i p e r a z i n e - 2 - c h l o r o - 4 - n i t r o i s o -15 quinoline (î.)» e x h i b i t s an t i - implan ta t ion a c t i v i t y in r a t s at '^ 50

mg/kg wi th maximum a c t i v i t y on days 4,5 and 6 of p regnancy . Another

c lass of b i cyc l i c n i t rogen he t e rocyc le s such as 3 - l2 . - (W-subs t i tu t ed

aminoalkoxy)benzoyl l indo les (15) and o i - m e t h y l a r y l a m i d o - P - n a p h t h y l ( 1-

raethylamino-2-methylbenziraidazolyl) e t h e r s

lantat ion a c t i v i t y in a lb ino female r a t s .

17 (16) pos ses s an t i - imp-

0-eCH2] R^ .NHCOPh

15 16

R = H; R = NEt^, NMe^, N(CHMe ) , p i p e r i d i n o , morpholnio , p y r r o l i d i n e ;

R^ = R^ = CHÔ^S n = 2-3

: R,R = CH CHÔCH^CH^

: R = Me; R = Ph

A b i cyc l i c su lphur containing h e t e r o c y c l e , 2 - P h e n y l - 3 - a r o y l benzoth io-

phenes ( r z ) . a t 1 mg /kg /day S.C. in r a t s for 15 days complete ly 18 i n h i b i t s fetus development

17

1.1.3 Polycyclic Heterocycles;

A wider variety of tr icyclic and/or polycyclic heterocycles

exhibit ant lfert i l i ty ac t iv i ty . Tricyclic nitrogen containing compounds 19 such as 18-24 are reported to be useful contraceptives.

18

^ H

19

20 21

22 23 24

Of the polycyclic oxygen containing heterocycles, benzofurobenzopyra-

nones (25 ) cause ^83 .3% inhibition of fetal implantation at 10 mg/kg

i . p . in ra ts and several derivatives of the nitrogen containing

heterocycle 2^ are luteolytic in rabbi t s at a dose of 1 mg/day for 1^ A 21 14 days

25 26

R = H, aminoalkyl

Several subs t i t u t ed indeno, naphtho and cyc lohep ta py razo le s (27) 22

e x h i b i t a n t i f e r t i l i t y a c t i v i t y in r a t s at a dose of 1-100 tug/day

and t r i azo lo i so indo le d e r i v a t i v e s (2^) are pos tco i ta l con t racep t ives

in r a t s . The ED va lues range between 1-10 mg/kg s . c .

R'

R' m^ hi 11 N-^R

27 28

R = 4-pyr idy l

R = R = H;

n = 2

R = Subs t i tu tedpheny l

Nitrogen and sulfur containing heterocycles such as thieno l 2 , 3 - g ] 24 indazoles (29) at 50 mg/day , 2H[ 1 ]-benzothiepino [4 ,5-C] pyrazoles

(30) at 2 mg/day and methanobenzo [b] thiophenes (21) at 5 mg/

kg exhib i t ant i fert i l i ty a c t i v i t y .

Eight membered tr icyclic nitrogen containing heterocycle such as 27 dibenzodiazocine {^) and the angular benzo [4,5] pyrano [2 ,3-

C] pyrroles (3^ ) with two different heteroatoms are reported to

be useful contraceptives.

32 33

X = 2,8-Cl2; R = 2-thienyl R,R = H, a lky l , a lkoxy, OH, halo, acyloxy;

2 R = H, a lky l , a r a lky l ,

aminoalkyl

Another class of compounds namely pyrazolo [5,1-a] isoquinoline

(34; 2 mg/kg/day in hamsters) and aminoindolobenzodiazepines

(35) have been found to possess contraceptive efficacy.

34

The con t racep t ive eff icacy of the members of the n i t rogen containing

t r i c y c l i c he te rocyc les (36-38) a r e as follows ' .

36

H

H S»5 H m-(OCH, )C,H.

3 6 4

CH,OH C,H_

CH3 C^H^

H S"5

CH2-CH3

CH = CH

CH = CH

CH^-CH^

CH^-CH^

CH,

ED

Hamster

3 .5

1

> 5

- 1 0

>3

10

50 (mg / k g / d a y)

Rat

20

5

25

- 2 0

20

>5

1.2

N-^Ph

BASIS OF WORK

3.5 1.8

16

The tex t of the p receed ing sect ion suggests t ha t h e t e r o c y c l e s

a re capable of terminat ing pregnancy but the a v a i l a b l e r e p o r t s do

not pro jec t a c l ea r p i c t u r e about t h e i r con t r acep t ive eff icacy by

ora l route of admin i s t r a t ion and the r e s u l t s of the follow up act ion

in la rge number of cases have p o s s i b l y remained u n r e p o r t e d . However,

the d e s i r e d de t a i l s concerning the prof i le of a n t i f e r t i l i t y a c t i v i t y

a r e a v a i l a b l e for t r i a z o l e d e r i v a t i v e s and t h i s p rompted an e a r l i e r

effort in t h i s l a b o r a t o r y to a s c e r t a i n whe the r the i somer ic t r i a zo l e s

(A&B) and the rep lacement of one of the ni trogen atom In t h e s e com

pounds by o the r he teroatoms could influence the con t r acep t ive e f f i cacy .

10

It has been found tha t i someric t r i azo le s (A&B) and oxadiazole d e r i

v a t i v e s (C) e x h i b i t con t racep t ive a c t i v i t y but t h e i r eff icacy by 31 32 oral route of admin i s t r a t ion is ex t r emely poor '

The present s tudy r e p r e s e n t s an extension of the work in i t i a t ed

e a r l i e r in t h i s l a b o r a t o r y and is d i r ec t ed to exp lo re the con t racep

t ive efficacy of o the r s impler he te rocyc les possess ing only one phenyl

r i ng . The s t r e s s on having a single phenyl r ing in the molecular

framework of s imple r he t e rocyc l e s is based on the assumption tha t

tw.o phenyl r ings in s imple r he te rocyc les at h igher doses may lead

to o e s t r o g e n i c i t y . In view of these c o n s i d e r a t i o n s , the syn theses

and the evalua t ion of con t r cep t ive efficacy of p r o t o t y p e s I-VI were

under taken .

u ni

«^a?r

R'=H,Ac VI

11

Another approach in the present s tudy for obtaining second

generat ion con t racep t ive agents r e l a t e s to the design of compounds

which simulate two d i s t i nc t type of pharmacophores in t h e i r molecular

s t r u c t u r e s ; the f i r s t one i s concerned with i t s a b i l i t y to a l t e r mem

brane pe rmeab i l i t y whi le the second one i s concerned with i t s capa

b i l i t y to in te r fe re ox ido- reduc t ion p r o c e s s e s . The choice of these

pharmacophores i s based on the assumption tha t compounds capable

of e l i c i t ing these a c t i v i t i e s in uterus would lead to inh ib i t ion of

pregnancy. A s i tuat ion such as t h i s would generate f resh l eads and

th i s approach a p p e a r s to have not been employed by e a r l i e r w o r k e r s .

In the present s tudy two r e p r e s e n t a t i v e c l a s s of compounds namely

1 . 2 , 3 , 4 - t e t r a h y d r o q u i n o l i n e d e r i v a t i v e s (VII) and 1 ,4 -d ihyd ro p y r i d i n e

d e r i v a t i v e s (VIII) have been ident i f ied for s y n t h e s i s and biological

evaluat ion as a n t i f e r t i l i t y agen t s .

V Me O2 C V . ^ / > V ^ C 0 2 M G

.CO2MG H

•Ms

vn vni

CHAPTER 2

2.1 ; Syntheses of 2-oxo-5-substitutedphenyl tetrahydrofurans,

5-Substituted phenyl-2,3-dihydrofuran-2-ones and 6-oxo-

3-substitutedphenyl-l ,4,5 ,6- tetrahydro-lH-pyridazines

2.2 : Syntheses of 4-Arylidine-3-methyl-4,5-dihydroisoxazolin-

2-ones and 4-(3 ,4-Dimethoxybenzyl )-3-niethyl-2 ,3 ,4 ,5 -

tetrahydroisoxazolin-5-one

2.3 : Syntheses of 5-Substitutedphenyl-2-isoxazolin-5-ones and

3-substitute dp henyl-5-methyli so xazoles

2.4 : Synthesis of 2,5-Dimethyl-3-niethoxycarboxyl-4-( 3-cyano-

2-oxo-l ,2,3 ,4-tetrahydroquinolino)furan

2.5 : Syntheses of 2,6-Dimethyl-3,5-ditnethoxycarbonyl-4-substi-

tuted-1,4-dihydropyridines

12

CHAPTER - 2

2.1 Substituted aroyl propionic acids (jL.- ) were identified as

the starting materials for the syntheses of compounds belonging to

prototypes I-III . Benzoyl propionic acid (Jl ) was easily prepared

by the Friedel-Craft reaction of benzene with succinic anhydride

I II m while 4-( 3-nitrophenyl)-4-oxo butyric acid (2) was obtained by the

nitration of \_ with fuming nitr ic acid. Esterification of these acids

with dry methanol in presence of BF -etherate furnished the methyl

esters (3^-^) which on reduction with sodium borohydride gave a

mixture of 4-hydroxy-4-substituted phenyl butyrates (^-^) and 2-

oxo-5-substituted phenyl tetrahydrofurans (Z"^' Scheme 1) . These

were separated by column chromatography over silica gel using chlo

roform, hexane (1:1) as the eluent. The dehydro derivat ives of

7 -8 , the second prototype of the present study, were obtained by

the ring closure of 2"*. with acetic anhydride (9-10; Scheme 1).

Similar ring closure of 2.~JL with hydrazine hydrate furnished 11-

12, the representatives of prototype III (Scheme 1).

2.2 Compounds representing the molecular framework of prototype

IV were prepared by reacting appropriate aromatic aldoximes (19-

24) with methyl acetoacetate in methanol

(Scheme 2 ) . The resulting compounds

(25-30) were obtained as non-separable

mixture of Z and E isomers. The stereo-

meric mixture was evident from the

PMR spectrum which indicated the p re

sence of two singlets for ethylenic proton at around 8.50 - 9.00

ppm.

One representative compound of prototype IVa was prepared

by the sodium borohydride reduction of the benzylidene derivative

CH3

13

CO2H

1:R = H 2:R=N02

C02Me

1:R=H j4;R=N02

CO^MG

5:R=H 6;R=N02

9:R=H 10:R= NO2

ri:R=H 12;R=N02

7:R=H 8.:R=N02

a,Dry MeOH/BF3cth2ratG; b,NaBH4/M<20H; c,Ac20; d,NH2NH2H20/MeOH

ScherriG 1

14

13R=H

14:Rr3,4-diOMe

1 5 . R = 4 - O H - 3 - O M G

16.R=4-NH2-3-N02

17:R=4-NHAc

18.R=4-NH2

a •> R

19-24

Substitucnt R

as in 13-18

Mc

Substituent R

as m 13-18

2 6 . c

OMe

6 OMe

M2 HH^'

HH^ O

H

31

a, NH2OH / McO H; b, AcCH2C02Me / MeOH;

c ,NaBH4/M20H

Scheme 2

15

26 in methanol (Scheme 2 ) . The compound 31^ so obtained was a mix

ture of isomers, which could not be separated by preparat ive TLC

or column chromatography.

2.3 A retro-synthetic analysis for obtaining compounds repre

senting prototypes V and VI suggests the preparation of starting

materials of the type A (Fig. A), which in turn can be obtained

V:R = H,Ac VI

from tetrasubstituted furans (B). Since substantial quantity of tetra-

substituted furans were available in the laboratory, efforts were

directed to obtain compounds of the type A from these furans (B).

Oxidation of substituted furans (C) have been reported earl ier to 33 34 yield oC,|3-unsaturated ketones (D; Fig.A) ' . In the present study

it was observed that the oxidation of 3-methoxycarbonyl-2,5-dimethyl-

4-substituted phenyl furans (32-36) with pyridinium chlorochromate

gave compounds which exhibited sixteen mass units more than the

expected mass of 0(,/3-unsaturated ketones (G). The ir spectra of

these compounds showed carbonyl bands at around 1660, 1720 and

1760 cms . The nmr spectra indicated the presence of two acetyl

groups and one methoxy group besides the required aromatic protons.

On the basis of these data, three tentative structures E, F or H

may be proposed for these oxidation products. Structures E and

F were ruled out on the basis of C-nmr spectra of the oxidation

products. The two quaternary carbon atoms of three membered ring

in E and the quaternary sp carbon atom in F are expected to appear

between 65-70 ppm but the C-nmr signals for the two quaternary

carbon atoms at around 160 ppm and the absence of any peak between

65-70 ppm supported the structure H (37-41; Scheme 3) . Since these

compounds were the enol acetates of the desired starting material

(A), they were reacted with aqueous acid . Instead of the desired

ketones (A; R = OMe) substituted acetophenones 42-46 were obtained.

The possible mechanism by which these compounds (37-41) broke

down to substituted acetophenones is described in Scheme 4. The

16

R- s COCH3

COR'

O

.1 A R = O M G . M ( 2

R COR'

B R^rOMG. Me

'f v . . o

SR' FCC

•^ ^^^o o^^^^

D

R- s O.

H3COC COR^

E R ' ^ = O M G , M 2

COCH3 R

COCH3 COCH3 CORI

O

.1 FR'=OMe,Me

COCH3

C02M(2

COCH3

G

Fig A

17

Rv ^C02Me PCC

}2_

li 36

Rv /COM(2

CH2CI2 M G O C O Q'Z^OMe - >

37-41

Subst i tuent R as in 32-36 R = 3 ,4 -d ime thoxypheny l

R = 3 ,4 -methy lened ioxypheny l

R = 4-methoxyphenyl

R = 4 -e thoxy-3 -me thoxy-pheny l

R = 4 -benzy loxy-3 -me thoxy-pheny l

(Scheme 3)

R

O

Me 42-46

Substi tuent R as in 32-36 47

CO2MG

R ,C0M(2

H / Me

Rv R h—Me

O^ O

-CO2

C02Me O C02Me

(Scheme 4)

18

c o n v e r s i o n of 2 * °^ a c t i v a t e d s u r f a c e to jn[^ p r o v i d e d a d d i t i o n a l e v i

d e n c e fo r t h e p r o p o s e d m e c h a n i s m . T h e s t e r e o c h e m i s t r y of 37 -41

h a s b e e n w o r k e d ou t b y c o m p a r i n g PMR s p e c t r a of t h e s e c o m p o u n d s

w i t h t h e s p e c t r a of STj-bl^ ( T a b l e 8 , p a g e 54 ) . The d o w n f i e l d s h i f t

of t h e -OCOCH s i g n a l s in 37-41 i s o n l y p o s s i b l e if t h e a c e t y l g r o u p 35

a t C-4 a n d m e t h o x y c a r b o n y l a t C-3 a r e o r i e n t e d in a c i s g e o m e t r y

T h e r e a c t i o n of o x i d a t i o n p r o d u c t s ( 3 7 - 4 1 ) w i t h h y d r o x y l -

amine h y d r o c h l o r i d e in me thano l f u r n i s h e d one of t h e p l a n n e d c o m p

o u n d s (R = H) of p r o t o t y p e V (Scheme 5 ) . T h e f a c i l e l o s s of t h e

COCH g r o u p e v e n a t m i l d e r r e a c t i o n c o n d i t i o n s p r e v e n t e d t h e p r e p a -^ 1

r a t i o n of o t h e r c o m p o u n d s (R = COCH ) of t h e p r o t o t y p e V. A d d i

t i o n a l e v i d e n c e for t h e a s s i g n e d s t r u c t u r e s ( 4 8 - 5 1 ) w a s o b t a i n e d

b y an u n a m b i g u o u s s y n t h e s i s of one of t h e r e p r e s e n t a t i v e compound

(49) b y r e a c t i n g 47 w i t h h y d r o x y l a m i n e h y d r o c h l o r i d e .

X - a

32 li 11 40

:

:

:

.

R

R

R

R

3 , 4 - d i m e t h o x y p h e n y l

3 , 4 - m e t h y l e n e d i o x y p h e n y l

4 - m e t h o x y p h e n y l

4 - e t h o x y - 3 - m e t h o x y p h e n y l

48-51.

S u b s t i t u e n t R a s in 37 -40

a , NH^OH/MeOH

(Scheme 5 )

T h e c o m p o u n d s b e l o n g i n g to p r o t o t y p e VI n a m e l y 3 - s u b s t i t u t e d -

p h e n y l - 5 - m e t h y l i s o x a z o l e s ( 6 2 - 6 5 ) w e r e p r e p a r e d b y r e a c t i n g 4 -

a c e t o x y - 3 - a c e t y l - 4 - s u b s t i t u t e d p h e n y l b u t - 3 - e n e - 2 - o n e s ( 5 7 - 6 1 ) , o b t a i n

e d b y t h e p y r i d i n i u m c h l o r o c h r o m a t e (PCC) o x i d a t i o n of 3 - a c e t y l -35

2 , 5 - d i m e t h y l - 4 - s u b s t i t u t e d p h e n y l f u r a n s ( 5 2 - 5 6 ) , w i t h h y d r o x y 36

l a m i n e h y d r o c h l o r i d e in m e t h a n o l ( S c h e m e 6 ) .

19

R ^ .COCH3 a

R COCH3

M H3COCO COCH3

52 : R = 3 ,4 -d i ine thoxyphenyl 57 : R = 3 ,4 -d ime thoxypheny l

53 : R = 3 ,4 -me thy lened ioxy phenyl

58 : R = 3 ,4 -methy lened io-xypheny l

54 : R = 4-methoxyphenyl 59 4-methoxyphenyl

55 4-e thoxy-3-methoxy-phenyl

60 : R 4 -e thoxy-3-metho-xypheny l

56 : R = 4 - b e n z y l o x y - 3 -methoxyphenyl

61 4 - b e n z y l o x y - 3 -methoxy phenyl

62 : R = 3 ,4 -dimethoxy

63 : R = 3 ,4-methylenedioxy

64 : R = 4-methoxy

65 : R = 4-ethoxy-3-methoxy

a, PCC/CH^Cl^; b , NH^OH/MeOH

(Scheme 6)

20

This faci le loss of two carbon atoms from 4-ace toxy-3 -me thoxy-

c a r b o n y l - 4 - s u b s t i t u t e d phenyl bu t -3-ene-2-ones under ac id ic and bas ic

condit ions p r o v i d e d temptat ion to s tudy the behaviour of t h i s c l ass

of compounds under e lec t ron impact . Since t h i s c lass of compounds

is nove l , i t s chemical r e a c t i v i t y and behaviour under e lec t ron impact

a re expec ted to con t r ibu te towards the ex i s t i ng knowledge in t h i s

f i e ld . Only the r e s u l t s of the mass spec t romet r i c s tudy a re d e s c r i b e d

h e r e . The EI and CI behaviour of t e t r a s u b s t i t u t e d e thy lenes (37-

40, 57-60) under pos i t i ve and negative ionizat ion condi t ions employing

techniques such as co l l i s ional ac t iva t ion (CA), metas table scan , exact 37 mass measurements and DÔ CI have been s tud ied . The pos i t i ve

ion EI, p o s i t i v e ion CI (CH ) and negat ive ion EI spec t r a of ^

and ^ a re shown in F i g . l and 2 . The metas table t r ans i t ions o b s e r v e d

under p o s i t i v e ion EI condit ions by high voltage scan a re a lso i n d i

ca ted in the EI s p e c t r a of ^ and 3^ (F ig . la and 2 a ) . The molecular

ions are not suff ic ient ly s t ab le to give intense M peaks and they

fragment by loss of CH CO and CH CO. A / - h y d r o g e n rearrangement

followed by the t rans fe r of the acy l group from C to C cou ld

lead to a s t ab l e (M-CH CO) ion (Scheme 7 ) . In the compounds 57-

60, the (M-CH CO) ion decomposed by the loss of r a d i c a l s such

as CH^*, CH^CH^*, HO". CHÔ* , CH^CHÔ" and CH^CO' whi le in the

compounds 36-39 loss of CH CO from M ' induce fur ther loss of neu

t r a l molecules (CH OH and CO) as shown in Scheme 8. A c h a r a c t e r i s

t ic difference i s o b s e r v e d in the p recu r so r ion s p e c t r a of the a roy l

ion in two groups of compounds (57-60 and 37-40) . The major p r e c u r

sor for a roy l ion in 57-60 is the (M-CH COCH ) ion, whi le in 2 1 "

40 arCO"^ i s formed mainly from (M-CH CO-CH OH) ion (Scheme 8 ) .

Metastable t r ans i t i ons were a lso o b s e r v e d for the d i r ec t decomposi

t ion of the (M-CH CO) ion to ArCO in a l l t he compounds thus su

ppor t ing the generat ion of a carbonyl function on C by the t ransfer

of the acyl group during the loss of CH CO from M .

The CA fragmentation pa t t e rn of the s t ab l e M " r e sembles

the 70 eV EI induced fragmentation of the molecule. However, the

r e l a t i v e abundances of the fragment ions a r e different in the two

s p e c t r a . This difference i s a t t r i b u t a b l e to the difference in the

r e a c t i v i t i e s of the decomposing and non-decomposing M ' ions due

to t h e i r having di f ferent s t ruc tu re s ' . The (M-CH CO)* ion gives

r i s e to the base peak in the CA s p e c t r a , wh i l e the most Intense

21

z u

»^SN31NI T3b

22

\

• X X .8

J U L * =

S—.

o o

—r-o

I c o

X

s

8

r-o o

—r— o

o a

i

AJjSN3iNI • 13b

23

CH30

C H ^ C

CHjO'

I T

•^O-COCHj

71 2 233

CH3O

1/1 203 m/i 1»1

(Scheme 7)

m / i 1CT 0C«3

n/x t77

24

CH30 CH30

0 nu II ^^

OCH3 - C H j C O * ^ " ' ' ^ " Y ^ \ ° « = " 3 ~'^'^3°" , O'"'

COCH3

5 , M —

m / i 292

-CH3C0

( M - C H j C O ) '

m/z 2S0

•/-CH3 •^

OCH3

(Scheme 8)

25

ion in the normal" EI s p e c t r a is ArCO . This i s unde r s t andab le as

the a roy l ion i s produced in the source from seve ra l p r e c u r s o r s

such as (M-CH^CO)"^, (M-CH CO-CH )"*• and (M-CH^CO-CH CO)"^.

As obse rved from CI(CH ) spec t r a of 57-60 and 37-40, a p a r t

from MH and (M-H) ions , the major fragment ions a r i s e by the

loss of neutra l molecules such as CH CO. CH CO H and CH O (Scheme

9 ) . The methyl e s t e r s 37-40 a r e c h a r a c t e r i s e d by the presence of

peaks corresponding to (MH-CH CO-CH OH)"*" and (MH-CH CO H-CH OH)"^.

The CI ( D O ) spec t r a of t he se compounds show (MD-CH CO)"^ and

(MD-CH CO H) ions indicat ing t h e r e b y that proton a d d e d is not

lost in these p r o c e s s e s . Moreover , metastable peaks cor responding

to these losses a re a lso o b s e r v e d in the CI(D O) s p e c t r a (Table

1 ) . The re tent ion of D in the el iminat ion ion ind ica tes t ha t a mecha

nism such as the one shown in scheme 9 opera tes for loss of ace t ic

ac id resul t ing in a cyc l i s ed p roduc t . In the methyl e s t e r s 37-40

the fur ther fragmentation of t h i s ion r e s u l t s in the loss of CH CO,

Yet another noteworthy feature in the CI spec t ra of these compounds

i s the presence of (M+43) i ons , p resumably due to se l f - acy la t ion 40

reac t ion . The acy l ion is produced from the sample i t se l f as a

r e su l t of which the (M+43) ion abundance increases when the sample

p r e s s u r e inc reases or when acetone i s used as the reagent for CI .

With ace tone-d , both (M+CH,CO)"^ and (M+CD,CO)"^ a re o b t a i n e d . The

(M+CH CO) ion is found to decompose by the loss of CH CO giving

r i s e to MH ions (Scheme 10b ) . A metastable peak i s seen at m/z

240.5 in CI (ace tone , CH or D O ) spectrum of ^ cor responding

to t h i s decomposi t ion.

The CA spectrum of the MH"*" and CI(CH spect rum of 59^

show cons iderab le difference in the r e l a t i v e abundance of the fragment

ions (Table 2 , F i g . l ) . Loss of H from MH (Scheme 10c) l eads

to the most abundant ion in the CA spec t rum, whi le (MH-CH CO)

g ives r i s e to the base peak in i t s CI(CH ) spec t rum. The (MH-Ar-

COCH ) ion i s ve ry prominent in the CA spectrum showing t h e r e b y

tha t the acyl migration i s an important p rocess even under CA condi

t i o n s .

As in the case of p o s i t i v e ion EI s p e c t r a , t he molecular

ions in the negat ive ion EI s p e c t r a a re a lso not v e r y abundant excep t

in iM and 40. However, in the e lec t ron cap ture spectrum of 59 (rvin

26

CH30

C H 3 0 - > f « ^ " V ^ O-COCr.3

3 , M H ' m/2 307

-CH2CC cm

(Mh-CM2C0)* rr./j 265

OCOCH3

C H 3 0 - ^ ^ ^ \ J ^

3 .MH+ m/z 307

CH3O

( M H - C H , C 0 2 H ) m/ z 2 i T

7,(MH-CH2CO) m / i 261

0 I

^ ^ - C H 3 -CH3OH O s C v - ^ O H

7 — * CHaOYTy^O

CH3O'

rr.'i 2iS

_Ch, -CH3OH CH30

CH3O

7 . ( M H - C H 3 C C : -

rr .'7 263

(Scheme 9)

27

T a b l e 1; M e t a s t a b l e P e a k s O b s e r v e d in t h e D O CI S p e c t r a of ^ ,

^ , 2 7 a n d 3 9 .

Compound T r a n s i t i o n m

o b s d C a l c d

52 MD' (MD-CH CO )'^ 199 .8 1 9 9 . 7

m/z 248+60)

58 (M+CH,CO) ' MH"^ 2 4 0 . 5 2 4 0 . 5

MD"*"

( rn /^ 308

(M+CH CO)"^

(m/z_

MD"

(m/z_

MD"

(m/z_

MD"*"

( m / z

319

278

324

324

m/z 277+42)

59 MD' (MD-CH CO)"^ 2 0 0 . 5 2 0 0 . 3

m/z 236+42)

1 2 MD' (MD-CH CO)"^ 2 4 5 . 5 2 4 5 . 4

m/z_ 282+42)

(MD-CH CO^H)"*" 2 1 5 . 0 2 1 5 . 0

ni/_z 264+60)

(MD-CH CO H)"^ (MD-CH CO H- 2 0 2 . 3 2 0 2 . 1

CH^OH)"^

(m/z 264 m/z 231+33)

39 MD"^ (MD-CH CO)"^ 216.0 216.0

(m/z 294 m/z 252-42)

28

CMj.

M j ^ O C « , ^ CH, 0

'4- ~ 0CH3

1

OCOCH3

m/i 07

0 ^ ». 0

1 i: 1:

CH3COO COCH3

CH3O CH3

m/2 2 77

-H2

CH3O

(MH -M j ) mjz 275

(C)

iScheme 10)

29

T a b l e 2 : Ion A b u n d a n c e s ( I ) in t h e CA S p e c t r a of 59

m/z_ P E I , M"*", P C E ( C H J . M H " * " NCI ( C H ^ ) , M '

ra/z 276 m/z 277 m/z 276

35

28

43

44

59

77

83

85

92

107

108

109

121

127

135

141

149

158

159

161

175

177

191

201

203

215

216

217

219

233

234

235

261

275

30

—

—

14

—

—

12

—

15

—

—

—

50

—

16

—

15

—

—

13

20

—

24

—

—

18

26

45

100

—

—

._

20

25

15

18

15

44

10

13

18

35

50 14

26

26.3

15

20

12

23

13

38

54

16

18

30 100

30 31

45 25

25

100

30

CO2M2

under CI(CH,) condit ions) the base peak i s due to M * . Fragmentation 4

occurs mainly by the loss of CH^CO* , CH^CO^H, (CH^CO^H+'H) or

(CH CO H+CH ) as shown in scheme 1 1 . Similar fragmentation pa t t e rn

of the M ' of 59^ i s also o b s e r v e d under col l i s ional ac t iva t ion (Table

2 ) . The methyl e s t e r s 37-40 fragment less r a p i d l y than ST-bO^. The

presence of ace toxy group in these molecules r e s u l t s in the formation

of abundant ace toxy ion, a t m/z 59 which g ives r i s e to the base

peak in the spec t r a of 57-60 (M-ArCO) is a c h a r a c t e r i s t i c fragment

obse rved in t h e i r s p e c t r a . It i s in te res t ing to note tha t the comple

mentary fragment ion, ArCO , g ives r i s e to in tense peaks in the

pos i t i ve ion s p e c t r a . An acyl group migrat ion from C to C as under

pos i t i ve ion condit ions would exp la in the formation of (M-ArCO)

ion (Scheme 11) . This rearrangement ion is a l so o b s e r v e d under

col l i s ional ac t iva t ion of the M * (Table 2 ) .

2 .4 The s y n t h e s i s of p ro to type VII involves Nef react ion of 2-

n i t r o - l - ( p - a c e t a m i d o p h e n y l ) Propene

(66) as the f i r s t s t e p . The r e s u l

ting compound namely 3-methoxy-

carbonyl -2 , 5 -d ime thy l -4 - ( p - a c e t a - iN^ '

midophenyl) furan (6^) was sub jec - M G — ^ V'~~-M<2

ted to v i l smier -Haak reac t ion and

the formyl d e r i v a t i v e {66) so VII

obta ined on oximation gave 69^ which was then reac ted with SOCl

to y i e ld 7^. Sodium b o r o h y d r i d e reduc t ion of ^ in methanol furnished

d e s i r e d compound ^ 1 (Scheme 12) .

2 .5 Compounds represen t ing p ro to type VIII (79-86) were p r e p a r e d

by reac t ing a p p r o p r i a t e l y subs t i t u t ed a l d e h y d e s (72-78 and bS^) wi th

methyl-3-aminocrotonate in presence of a p p r o p r i a t e so lvents (Scheme

13) . In o r d e r to change R in comp

ounds 79-86, two s t r a t eg i e s have

been employed . In the f i r s t one

a p p r o p r i a t e a l d e h y d e s were gene

r a t e d and in the second one , a p p

r o p r i a t e funct ional i t ies on the

phenyl r ing of 2 ,6 -d ime thy l 3 , 5 -

dimethox year b o n y l - 4 - p h e n y l - 1 , 4 - d i h y

d r o p y r i d i n e were simulated for e l abora t ing them to h e t e r o c y c l i c r ing

s y s t e m s .

31

0

OCOCHj

0CM3

C"3 0 ^ « « ^ 3

-COCH3 -CH3CO2H

^**3^^°~ C H 3 \ / 0 C H j v ^ O

6 Jk * 0 ^ " ° COCM3

(Scheme 11)

32

"""10 M G - ' ^ N 0 2

66

a

CO2MG

M G

\^

a , CH COCH2C02CH^, Et^N/MeOH; b , DMF/POCl^; C , NH^OH/MeOH;

d , SOCK/Benzene; e , NaBH./MeOH 2 4

(Scheme 12)

33

R-CHO

72-78 k 68

V M G ' N

H

79-86

^Me

72 : R = 4 'Ch lo ro -3 -n l t ropheny l

73 : R = 4-acetainidophenyl

74 : R = 4-amino-3-n i t rophenyl

75 : R =

CHO

4-ch lo ro -3 -n i t ropheny l

4-acetamidophenyl

29 : R

80 : R

81 : R = 4-amino-3-n i t rophenyl

82 : R =

R^ R' R' = M ( 2 , R 2 = C 0 2 M G

76 : R =

77 : R

N - ^ C I

H H OH OH

OH OH H H

''••''- 0 C X CI

H H OH OH

-CH2OH 3, ^ , ^ -CH2OH

OH OH H H

78 : R = 85

86 : R =

a , Me thy l -3 -aminoc ro tona te / app rop r i a t e solvent

(Scheme 13)

M G - ^ ^ O

34

Appropriately substituted aldehydes were prepared as follows:

Nitration of 4-chloroben2aldehyde with cone. UNO gave

72 while 2-chloro-3-formyl quinoline {Tb), obtained by vilsmier-

Haak reaction of acetanil ide, was reacted with sodium azide to obtain

78 (Scheme 14).

o 76

a, DMF/POCl ; b , NaN 78

(Scheme 14) In order to build heterocyclic ring systems on the phenyl

ring of 2 ,6-dimethyl-3 ,5-dimethoxycarbonyl-4-phenyl-l ,4-dihydro

pyridine, the compound ^ was hydrogenated in presence of Raney-

nickel to obtain the corresponding diamine (90; Scheme 15) and

a

H , Raney-Ni/MeOH

90

(Scheme 15)

the methanolic solution of compound 72 was reacted with aqueous

methylamine and aqueous ammonia under pressure to yield ^ and

89 respect ively. Reaction of 72^ with benzylamine at atmospheric

pressure yielded 88 (Scheme 16).

35

M G 0 2 C

79

NO2

C02Me a

• ^

NO2

MG 02C\^'^^\X02M(2

87

88

89

a. Appropr i a t e amine

(Scheme 16)

MG-H

' M G

R = NHCH

R = NHCH^Ph

R = OH

Ring closure in 90 was evoked ei ther by reac t ing i t with

substituted benzaldehydes in presence of acetic acid (91-95; Scheme

17) or by reacting it with sodium nitrite in p resence of aqueous

acetic ac id . The latter y ie lded 9b^ (Scheme 17) . Synthes i s of a

9 0

HN-N \ \

MGO2C CO2MG

H 92

93

94

95

Substituted benzaldehydes/AcOH;

(Scheme 17)

R = Phenyl

R = p-methoxyphenyl

R = 3 ,4-dimethoxyphenyl

R = 4-methoxy-4-hydroxy-phenyl

R = 3 ,4-methylenedioxy-phenyl

b , NaNO^/aq. AcOH

36

compoxind ^ structurally related to prototype VIII, v/as achieved

by reacting ^ with benzil in presence of methanol (Scheme 18)

whereas the reactioft of 9£ with carbon disulfide yielded ^ (Scheme

18). The tetracyclic heterocycle ^ was obtained by refluxing metha-

nolic solution of 90^ with phthalic anhydride (Scheme 18).

M ( 2 0 2 C \ ^ > \ / C 0 2 M G

CO2MG

99 9 8

a, Benzil/MeOH; b , CS ; c .Phthall ic anhydride/MeOH

(Scheme 18)

CHAPTER 3

3.1 : EXPERIMENTAL

3.2 : Tables

3.3 : BIOLOGICAL ACTIVITY

37

CHAPTER - 3

EXPERIMENTAL

3.1 The melting points were determined on a sulphuric acid

or an electrically heated block and are uncorrected. All the reactions

were checked by thin layer chromatography over silica gel G or

alumina (basic or neutral) plates using iodine vapours. KMnO spray

or Dragendorf's spray as the developing reagent. The structures

of all the compounds were routinely checked by IR and PMR Spectro-

scopy-IR spectra were recorded on Perkin-Elmer 157 infracord and

Beckman Acculab-1 grating instruments and values are expressed in

cm . PMR spectra were recorded on varion EM 360L or Perkin-Elmer

R-32 using TMS as internal reference (Chemical shift in ppm).

C-NMR spectra were recorded on CFT-20 spectrometer operating

at 20 MHz, 8192 data points were collected and a sweep width of

5000 Hz was used. Pulse delay of 0.5 sec. was essential for record

ing the signals of carbonyl carbons. N-NMR spectrum was recorded

on a Bruker WM-400 FT NMR spectrometer using formamide as the

reference. Mass spectra of these compounds were recorded on Jeol-

D 3000 instrument. The second place after the decimal point in the

required value of C,H,N analyses have been approximated.

jg-Benzoyl propionic acid (1)

This compound was essentially prepared by the method re-

ported earlier

p-O-Nitrobenzoyl) propionic acid (2 , Table 3)

To fuming HNO (d; 1.54, 20 ml) held at 0°C was added

2. (5 gm) in portions to prevent the reaction temperature to rise

above 8-10°. After complete addition of the compound it was stirred

at the same temperature (8-10°) for an additional 1 hr. The reaction

mixture was then poured on to crushed ice under stirring. The sepa

rated solid was filtered and washed with ice-cold water ('^5°).

The compound (2 ) was dried and recrystallized from minimum ethanol.

Methyl-4-substituted^phenyl-4-oxo-butyrate {y-±. Table 3)

A solution of 1_ or 2 (5 gm) in methanol (20 ml) and BE -

etherate (0.1 ml) was refluxed for 3 hrs. After completion of the

reaction, water was added to the reaction mixture and extracted

38

with e t h y l a c e t a t e . The organic l aye r was washed with water t h r e e

t i m e s , d r i e d on anhydrous sodium su lpha te and removed under reduced

p r e s s u r e . The r e s idue on t r i t u r a t i o n with hexane gave the d e s i r e d

compound ^ o^ i_ which was r e c r y s t a l l i s e d from e t h a n o l .

Methyl -4 -hydroxy-subs t i tu ted^pheny l butyrate (5 -6 . Table 4) and

2-oxo-5-subst i tuted phenyl tetrahydrofuran (X"8.» Table 5)

To a solut ion of 3_ or 4 (0.01 mole) in methanol (8 ml) was

added sodium b o r o h y d r i d e (0,02 mole) in por t ions under cooling

(0-5°) and s t i r r i n g . The reac t ion was al lowed to continue at the

same tempera ture for 20 mts . The react ion mixture was then d i lu ted

with water (20 ml) and e x t r a c t e d with chloroform (50 m l ) . Usual

work up of the organic l aye r y ie lded a mixture (^-6^ and T_-B) which

was s e p a r a t e d by column chromatography over s i l i c a gel using a

mixture of ch loroform, e t h y l a c e t a t e (80:20) as e luen t . The comp

ounds ^-6^, ob ta ined as c r y s t a l l i n e , s o l i d s , which were r e c r y s t a l l i s e d

from chloroform-hexane mixture whi le compounds 7^-|[ were o i l s .

5-Substituted phenyl-2,3-dihydTofuran-2-one (9-10)

A mixture of ^ or 2 (0.01 mole) and ace t ic a n h y d r i d e (0.04

mole) was re f luxed for 3.5 h r . After removal of the solvent under

reduced p r e s s u r e , water (20 ml) was added to i t and e x t r a c t e d with

e t h y l a c e t a t e . Usual work up y ie lded dark r e d so l id which was

pur i f i ed by column chromatography over s i l i c a gel using chloroform-

e t h y l a c e t a t e mixture as the e luen t . The compounds 9-10 so ob ta ined

were r e c r y s t a l l i s e d from chloroform-hexane m i x t u r e .

Yield 70%; m . p , 82-83" ; M" at m / i 160

IR (KBr)

PMR (CDCl^)

So»8°2

11 IR (KBr)

PMR (CDCl^)

CioHyNO^

1800 (CO)

3 .36-3 .38 (d ,2H,CH2) , 5 .68-5 .74 ( t , IH, CH),

7 .25-7 .63 (m, 5H, Ar-H) .

Requ i r e s : C, 75.00; H, 5.00

Found: C, 74.62; H, 4.81

Yield 65%; m . p . 105°; M" at m/z 205

1790 (CO)

3 .44-3 .46 ( d , 2H, CH^). 5 .91-5 .97 ( t , IH, CH),

7 .42-8 .33 (m, 4H, Ar-H)

Requ i r e s : C, 58 .53; H, 3 . 4 1 ; N, 6.82

Found: C, 58.70; H, 3 .88 , N, 6.60

39

6-Oxo-3-subst i tuted'^phenyl- l ,4 ,5 ,6- tetrahydro- lH-pyridazine (11-12)

To a solut ion of 2 °^ A (0-01 mole) in methanol (5 ml) was

added hyd raz ine h y d r a t e (0.02 mole) and s t i r r e d at room tempera ture

(25°) for 1.5 h r . Addi t ion of water (10 ml) to the reac t ion mixture

y ie lded 11 or JJ^ as so l id which was r e c r y s t a l l i s e d from hot metha

nol .

Yield 65%; m . p . 120-21°; M" at m/z 174

1680 (CO) 11 '• IR (KBr) :

PMR (CDCl ) :

S0«10N2°

12 :

IR (KBr) :

PMR(CDCl,+DMSO-d, ) : i 0

SoV3°3

2.40-2 .98 (m, 4H, ZxCH^), 7 .20-7.70

5H, Ar -H) , 9 .12-9.40 ( b s , IH, NH)

Requi res : C, 68.96; H, 5.74; N, 16.09

Found: C, 69 .21 ; H, 5 .98; N, 15 .84.

Yield 60%; m . p . 160°; M" at m/z 219

1680 (CO)

(m.

2 .40-2 .60 ( t , 2H, CH^) . 2 .98-3 .19

CH ) , 7 .55-8.21 (m, 5H, Ar-H & NH)

Requi res : C, 54.79; H, 4 ,10; N, 19.17

Found: C, 54.90; H, 4 .45 ; N, 1 8 . 9 1 .

( t . 2H,

Table 6) Aryl id ine-3-methyl -4 ,5-dihydro- i soxazol in-5-one (25-30,

To a solut ion of the r e q u i r e d oxime (19-24; 0.01 mole) in

methanol (10 ml) was added methyl ace toaceta te (0.02 mole) and

ref luxed for 3 h r . Solvent was removed under reduced p r e s s u r e and

the r e s idue was t r i t u r a t e d with water (20 ml) to y ie ld the r e q u i r e d

compound (25-30) as so l id which was r e c r y s t a l l i s e d from aqueous

methanol .

4-(3 ,4-Dimethoxybenzyl ) -3- inethyl -2 ,3 ,4 ,5- te trahydro- i soxazol in-5-one

(31.)

To a suspens ion of £6 (2:47 gm, 0.01 mole) in methanol

(10 ml) was added NaBH (0.57 gm, 0.015 mole) in por t ions vmder

s t i r r i n g and cooling ( 5 - 1 0 ° ) . It was s t i r r e d at the same tempera tu re

for 1.5 h r . Water was a d d e d to the reac t ion mixture and e x t r a c t e d

wi th e t h y l a c e t a t e . Usual work up of the organic l a y e r y i e lded a

r e s idue which on t r i t u r a t i o n wi th petroleum benzene furnished 31

as a c r y s t a l l i n e s o l i d . This was r e c r y s t a l l i s e d from c h l o r o f o r m - p e t

roleum benzene m i x t u r e .

21 IR (KBr)

PMR (CDCl^)

40

Yield 55%; m . p . 130°; M" at m/z 251

1738 (CO)

1.93-1.98 (m, 3H, 3H^), 3 .00-3.90 (m, lOH,

CH , 2XCH, 2X0CH^). 6 .50-6 .98 (m, 4H, Ar

il , NH)

C , , H , , N O , : Requi res : C, 62 .15; H, 6 .77; N, 5.57 13 17 4

Found: C, 62 .48; H. 6 . 3 1 ; N, 5.81 2 , 5 - D i m e t h y l - 3 - m e t h o x y c a r b o n y l - 4 - s u b s t i t u t e d ; ^ p h e n y l furans (32-36) .

Table 7)

To a suspension of a p p r o p r i a t e n i t r o s ty r ene (0 .01 mole)

in methanol (10 ml) was added t r i e thy lamine (0.02 mole) and methyl

ace toace ta te (0.02 mole) under s t i r r i n g and cooling ( 5 - 1 0 ° ) . After

15 minutes the react ion mixture was a l lowed to come to room temp

e r a t u r e (25°) and the s t i r r i n g was continued for 24 h r . Removal of

solvent under reduced p r e s s u r e at 25°C was followed b y the addi t ion

of wate r (20 ml) and the react ion mixture was fur ther s t i r r e d at

room tempera tu re for 1 h r . The s e p a r a t e d so l id was f i l t e r e d , washed

with w a t e r , d r i e d and r e c r y s t a l l i s e d from chloroform-hexane mix tu re .

4 -Ace toxy-3 -me thoxyca rbony l -4 - subs t i t u t ed '^^^ pheny l bu t -3 -ene-2-ones

(37-41,, Table 8)

To a well s t i r r e d solution of t e t r a s u b s t i t u t e d furan (32-

36, 0.01 mole) in d r y dichloromethane (50 ml) was added pyr id in ium

chlorochromate (0.04 mole) and the reac t ion mixture was s t i r r e d

at room tempera tu re (30°) for 10 h r . It was then d i lu ted with e the r

and the organic l aye r was pas sed through a column of s i l i c a ge l .

The column was f ina l ly e lu ted with chloroform-hexane (1:2) and

the usual work up of t h e organic l ayer y i e lded the d e s i r e d compounds

(37-41) as s o l i d s excep t 2*. * 21. which were o i l s . The so l id comp

ounds (2Z» .*?. * Ai) were r e c r y s t a l l i s e d from chloroform-pet ro leum

benzene.

Subs t i tu ted acetophenones (42-46)

To a solut ion of subs t i t u t ed bu t -3 -ene-2-ones (37-41; 0.5

gm) in methanol (1 .5 ml) was a d d e d aqueous h y d r o c h l o r i c ac id (10%,

0.5 ml) and the mixture was ref luxed for 2 h r . The solvent was

removed and the r e s idue was e x t r a c t e d wi th ch loroform. The usual

work up of organic l aye r gave t h e r e q u i r e d acetophenones ( 42 -46) .

These were iden t ica l in a l l r e s p e c t s wi th the authent ic ace tophenones .

41-

Isolation of methyl piperonoylacetate (47)

A solution of 2^ (1 gm) in chloroform (3 ml) was added

to a short band of silica gel and left to stand for 48 h r . There

after it was eluted with a mixture of hexane-chloroform (1 :1) . Re

moval of the solvent furnished oily residue which on tri turation

with hexane gave 4[7 as a solid. This was recrystal l ised from chlo-

roform-hexane mixture, and was found to be indentical in all res

pect with the authentic sample prepared by the method reported ,. 42 earlier

3-Substituted phenyl-2-isoxazoliii-5-ones (48-51, Table 9)

To a solution of subsituted but-3-ene-2-ones (37-41; 0.001

mole) in methanol (10 ml) was added hydroxylamine hydrochloride

(0.003 mole) and sodium acetate (0.003 mole). This mixture was

s t i r red at room temperature (20°) for 6 h r . Water (10 ml) was then

added to the reaction mixture and the separated solid was f i l tered,

washed with water, dried and recrystal l ised from methanol.

2,5-Dimethyl-3-acetyl-4-substituted'^ phenyl furans (52-56, Table 7)

These were essentially prepared by the method described

for compounds 32-36.

4-Acetoxy-3-acetyl-4-substituted^~*'phenyl but-3-ene-2-ones (57-61,

Table 8)

These compounds were prepared by the method described

for compounds 37-41.

3-Substituted phenyl-5-methyl-isoxazole (62-65, Table 10)

The method described for compounds 48-51, was followed

to obtain these compounds.

2-Nitro-l-(£-acetaniidophenyl) propene (66)

This was essentially prepared by the method reported ,. 43 earl ier

2,5-Dimethyl-3-methoxycarbonyl-4-(£-acetainidophenyl) furan (67)

The method of preparation of these compounds was essen

t ia l ly the one reported for compounds 32-36.

42

2,5-Dimethyl-3-methoxycarbonyl-4-(2-chloro-3-formyl qiiinolino) furan

(68, Table 11)

To a precooled flask in an ice-bath for 15 minutes was

added POCl (15.4 ml, C.Ol mole) and it was allowed to stand at

this temperature for another 15 minutes. Dry DMF (6.3 ml, 0.01

mole) was then added under stirring and after 5 minutes the comp

ound 62 (2.87 gm, 0.01 mole) was added. The resulting mixture

was heated at 70-74° under stirring for 16 h r . Finally it was poured

onto crushed ice , and then separated solid was f i l tered, washed

with water and dr ied. It was recrystal l ised from chloroform-hexane

mixture.

2,5-Dimethyl-3-methoxycarbonyl-4-(3-aldoximino-2-chloroquinolino)furan

(69, Table 11)

A solution of bS^ (3.4 gm, 0.01 mole) in methanol (15 ml),

hydroxylamine hydrochloride (1.38 gm, 0.02 mole) and sodium acetate

(1.64 gm, 0.02 mole) was s t i r red at room temperature (25°) for

3 h r . Water (20 ml) was then added to the reaction mixture and

the precipitated solid was f i l tered, washed with water, dried and

recrystal l ised from aqueous DMF.

2,5-Dliiiethyl-3-methoxycarbonyl-4-(2-chloro-3-cyano quinolino) furan

(70. Table 11)

To a suspension of b9^ (1.79 gm, 0.005 mole) in dry benzene

(8 ml) was added SOCl (5.9 ml, 0.025 mole) and the resulting mix

ture was heated at 90° for 3 h r . Finally it was poured on to crushed

ice and the resulting mixture was allowed to come to 90°. After

holding it at this temperature for 45 minutes, it was cooled to room

temperature (25°) and allowed to stay at this temperature for 6

h r . The separated solid was then f i l tered, dried and recrysta l l i sed

from aqueous DMF.

2,5-Dimethyl-3-methoxycarbonyl-4-(3-cyano-2-oxo-l,2,3,4-tetrahydro-

quinolino) furan (71)

To a suspension of 25. (0.35 gm, 0.001 mole) in methanol

(5 ml) was added NaBH (0.15 gm, 0.004 mole) under cooling (5-

10°) and s t i r r ing . It was then s t i r red at th is temperature for 1.5

h r . water was added to it and the mixture was extracted with ethyl

acetate. Usual work up of the organic layer yielded an oil which

43

on t r i t u r a t i o n wi th petroleum benzene furn ished a s o l i d . This was

r e c r y s t a l l i s e d from aqueous DMF.

71 : Yield 92%; m . p . 145-46°

IR (KBr) : 1675, 1700 (CO). 2200 (CN)

PMR (TFA) : 2.00 ( s . 3H. 5-CH^), 2.40 ( s , 3H, 2-CH^),

3 .27-3 .37 ( d , 2H, CH^) , 3.69 ( s , 3H, CO^CH^) ,

3 .89-4.07 ( t , IH. CH), 6 .74-7 .18 (m, 3H, Ar

il)

C , „ H , , N , 0 . : Requi res : C, 66.66; H, 4 .93 ; N, 8.64 18 i o i. 4

Found: C, 66 .51 ; H, 4 .58 ; N, 8 .35 .

4-Chloro-3-nitro benzaldefayde (72)

To fuming HNO (d ; 1.54, 10 ml) he ld at 0° was added

£ - c h l o r o benza ldehyde (1 .4 gm) in por t ions so t h a t t empera tu re r e

mains 0 -5° . After complete add i t ion of the compound i t was s t i r r e d

at the same tempera tu re (0-5°) for 2 h r . The reac t ion mixture was

poured on c r u s h e d ice and the s e p a r a t e d so l id was f i l t e r e d , washed

with water and d r i e d . It was r e c r y s t a l l i s e d from chloroform-hexane

mix tu re .

4-Amliio-3-nitro benzaldehyde (74)

This was p r e p a r e d by following the method r e p o r t e d in 44

the l i t e r a t u r e

2-Chloro-3-formyl quinoline (76)

This compound was p r e p a r e d by the method r e p o r t e d e a r -, . 45

l i e r

4-Formyl tetrazolo [ 1 , 5 - a ] quinoline (78)

This was p r e p a r e d by the method r e p o r t e d in the l i t e r a -45a tu re

2 ,6 -Di inethyl -3 ,5-dimethoxycarbonyl-4-subst i tuted- l , 4 - d i h y d r o p y r i -

dines (79-86, Table 12)

A mixture of the a p p r o p r i a t e a l d e h y d e (72-78 & Ml, 0.01

mole) and methyl -3-amino crotonate in an a p p r o p r i a t e solvent ( e t h y

lene glycol for 79-83, 85 and methanol for 84, ^ ) was hea ted on

water ba th for 24 h r . Thereaf ter water was added to the reac t ion

mixture and s t i r r e d at room tempera ture (25-30°) for 2 h r . , t he

44

separated solid was f i l tered, washed with water and dr ied. In the

case of ^ the work up was different. In this case dry ether was

added to the reaction mixture and the solid obtained after trituration

was filtered and dr ied. The compounds 79-83 and 8b^ were recrys ta-

llised from aqueous methanol while ^ was recrystal l ised from aqueous

DMF.

2,6-Dimethyl-3 ,5-dlmethoxycarbonyl-4-substituted ^^ phenyl-1,4-dihydro

pyridines (87-89, Table 12)

A mixture of the compound J9^ (0.01 mole) and aqueous

methyl amine or aqueous ammonia (0.04 mole) in methanol (20 ml)

was heated in steel bom at 150° for 24 hr . The volume of the reac

tion mixture was reduced to 10 ml and to t h i s , water (20 ml) was

added, and the mixture was allowed to stand at room temperature

for 3 h r . The separated solid was f i l tered, washed with water and

dr ied. These compounds were recrystal l ised from aqueous methanol.

2,6-Dimethyl-3»5-dimethoxycarbonyl-4-{4-benzylaniino-3-iiitro phenyl)-

1.4-dihydropyridine (88, Table 12)

A mixture of the compound 79 (3.8 gm, 0.01 mole) and

benzyl amine (6.3 ml, 0.06 mole) was heated at 110° under stirring

for 8 h r . Water was then added to the reaction mixture and extrac

ted with ethyl acetate. Usual work up of the organic layer gave

an oil which on trituration with petroleum benzene, yielded a solid.

This was recrystal l ised from chloroform-petroleum benzene mixture.

2,6-Dimethyl-3,5-dimethozycarbonyl-4-(3,4-diaiiiinophenyl)-l,4-dihydro-

pyridine (90, Table 12)

To a suspension of 81 (1 gm) in methanol (100 ml) was

added Raney-Ni (~0.2 gm) and hydrogenated at 2.5 kg/cm^ for 2

h r . The catalyst was filtered off, solvent removed under reduced

pressure and the residue tr i turated with water. The solid so obtained

was f i l tered, dried and recrysta l l ised from aqueous methanol.

2-Aryl-5(6)-(4-{2,6-diinethyl-3,5-dimethoxycarbonyl-l,4-dihydropyri-dyl)l-l(3)-substituted2benxyl benzimidazole (91-95, Table 12a).

A mixture of 90 (0.001 mole) and substituted benzaldehyde

(0.002 mole) in glacial acetic acid (2 ml) was heated on a water

bath for 4 h r . Water was then added to the reaction mixture and

45

e x t r a c t e d wi th e t h y l a c e t a t e . Usual work up of the organic l aye r

y ie lded a r e s idua l mass which on t r i t u r a t i o n with petroleum benzene

furnished 90-95 as s o l i d . These were r e c r y s t a l l i s e d from aqueous

methanol ,

5 ( 6 ) - l 4 - ( 2 , 6 - D i m e t h y l - 3 , 5 - d i m e t h o x y c a r b o n y l - l , 4 - d i h y d r o p y r i d y l ) l benz-

triazole (96)

To a solution of 90 (0.662 gm, 0.002 mole) in ace t ic ac id

(2 ml) was added water (2 ml) and to t h i s mixture was added a

solution of sodium n i t r i t e (0.414 gm, 0.006 mole) in water (1 ml)

under cooling (5-10°) and s t i r r i n g . It was s t i r r e d under cooling

for 15 minutes and then i t was hea ted at 80-90° for 1.5 h r . The

reac t ion mixture was poured into c rushed i c e . The s e p a r a t e d so l id

was f i l t e r e d , washed wi th water and d r i e d . It was r e c r y s t a l l i s e d

from aqueous methanol .

5-I4 - (2 ,6 -Di ine thy l -3 ,5 -d l ine thoxycarbony l - l , 4 -d ihydropyr idy l ) ] 2 , 3 -

diphenyl quinoxaline (97)

A mixture of 9£ (0.331 gm, 0.001 mole) and benzi l (0 .21

gm, 0.001 mole) in methanol (5 ml) was ref luxed under s t i r r i n g

for 5 h r . Water was then added to the reac t ion mixture and the

s e p a r a t e d so l id was f i l t e r e d , washed wi th w a t e r , d r i e d and r e c r y s

t a l l i s e d from aqueous methanol .

97 : Yield 65%; m . p . 172-3°; M* at m/z 505

IR (KBr) : 1680 (CO)

PMR(CDCl,+DMSO-d,) :2 .29 ( s , 6H, 2xCH, ) , 3.51 ( s , 6H, 2xCO,CH,) , i o —3 2 3

5.10 ( s , IH, CH), 7.15-7.65 (m, 13H, Ar-H) ,

8.38 (s_, IH, NH)

^ 3 l " 2 7 ^ 3 ° 4 • Requi res : C, 73 .66; H, 5.34; N, 8.31 Found: C, 73.66; H, 5 .37; N, 8 .66.

5 ( 6 ) - I 4 - ( 2 , 6 - D i m e t h y l - 3 , 5 - d i m e t h o x y c a r b o n y l - l , 4 - d l h y d r o p y r i d y l ) ] - 2 -

mercapto benzimidazole (98)

To a solution of 90 (0.331 gm, 0.001 mole) in ethanol (2

ml) and wa te r (2 ml) was added carbon d isu l f ide (2 m l ) . The r e a c

t ion mixture was heated under s t i r r i n g at 60° for 6 h r . Water was

then added to i t and the s e p a r a t e d so l id was f i l t e r e d , washed wi th

w a t e r , d r i e d and r e c r y s t a l l i s e d from aqueous DMF.

46

98^ : Y i e l d 70%; m . p . 2 6 8 - 9 ° ; M"^ a t m/z 373

IR (KBr) : 1660 (CO)

PMR(CDCl,+DMSO-d, ) : 2 . 2 0 ( s , 6H. 2xCH-) , 3 .50 ( s , 6H, 2xCO,CH ) , 3 6 J £ . 5

4 .85 ( s , I H , C H ) , 6 . 6 9 - 7 . 0 5 ( m , 3H, A r - H ) .

C H N O S : R e q u i r e s : C , 5 7 . 9 0 ; H, 5 . 0 9 ; N , 11 .26

F o u n d : C , 5 7 . 6 8 ; H, 4 . 8 1 ; N, 1 1 . 5 8 .

10-H-2 - [4 - {2 ,6 -Dimethy l -3 ,5 -d in i e thoxycarbony l ) - l , 4 -d ihydropyr idy l l -

10-oxo-isoindolo [ 3 , 2 - £ ] benzimidazole (99)

A m i x t u r e of ^ ( 0 . 3 3 1 gm, 0 . 0 0 1 mole ) a n d p h t h a l i c a n h y

d r i d e ( 0 . 1 4 8 gm, 0 .001 mole) in me thano l (5 ml) w a s r e f l u x e d for

10 h r . Wate r w a s a d d e d to t h e r e a c t i o n m i x t u r e , t h e s e p a r a t e d s o l i d

w a s f i l t e r e d , w a s h e d w i t h w a t e r , d r i e d a n d r e c r y s t a l l i s e d from

a q u e o u s m e t h a n o l .

9 i : Y i e l d 55%; m . p . 2 2 5 - 6 ° ; M"^ a t m/z 443

IR (KBr) : 1680, 1650 {CO)

PMR(CDCl,+DMSO-d, ) : 2 . 22 ( s , 6H, 2xCH 1 , 3 .50 ( s , 6H, 2 x C 0 , C H , ) , 3 o —5 Z 3

4 .90 ( s , I H , C H ) , 7 . 0 1 - 7 . 8 0 ( m , 8H, A r - H .

NH) .

C H N O : R e q u i r e s : C , 6 7 . 7 2 ; H, 4 . 7 4 ; N, 9 . 4 8

F o u n d : C , 6 7 . 5 8 ; H, 4 . 9 2 ; N, 9 . 0 5 .

3.2 TABLES

Physical and Analytical data of Compounds, which have not

been included in Experimental section, are described

under vertical columns and their spectral

characteristics have been recorded in

horizontal sequence.

47

(0

c <

•o d o fa

T3 (1) 1-1

"3 cr 0)

a;

3: ^

u

o

i n

U

l -H

3 •

So' o

s

T3

a u • o

e

o

a:

a:

o

O 3 U O

i n

in O 2 -~

O <M

o

i n

(M i n

O Z

c l

o t--

00 I o

in

X I u

3:

rg <fl

CO I

m

= 1 U

O

I

o XI

I O

s Q +

I—I

U Q U

CO

i n

o o

03

o

O U

« X pn

» n

+

,, m w ifl S

i n

<N1

X I u

fM

X

I

(M

i n

o

vO

: ; E

fVJ 1—1

o O- 00

X U

•N I - O

u m

Kl XI h-t U * |

r- ,^

( ' 1

XI

u M

X

* -t-"

a-rPi

I i n fM

< fM

X

^

00 1

i n

00

(M

xi U

X fM

^ • ( ->

"'"' f—t

CO

fM 1

i n —. vO

a - fM • - *

^ •• ^ - s

^ fM

i -H

"E

•J ft)

+ s ro

•—t

u Q U

OS S Oi

'* O u • ^ ^

i n fM t~

,, m Ui ta

2 .. X

1 u < »

X i n

^ e

^ • j "

• i n i n

i n O 2

(—1

X l -H

i -H

u

o r

vO 00

..,

fM .

fM t< XI u

X (\J

^ 4-»

"tr 00

fM 1

O t~-

fM

.. ^

<

N O

09

i r

X fM

E — o

CO 1

r^ O f - H

u Q U — o: S cu "* O u ^^ o ro r-

f—t

• 00

* .— X

1 u < i n

X l-H

* E

o

. - I 00

u f^ ffl * "^

Oil * l ^ S u e

48

•o c 3 O

in

>. la c <

CO in

U

T3 (U l-i

"3 o-a:

u

^ ^

o • '. o - S o S

,: u

O

00

T3 C 3 O • a o 6 2 o u

o -

^l

u

X

^ XI o I <N 0

• i n

. ^ - v

r X

u *

X (M

• *^

fM

OO

<M I

•T-l +

U ^

X

IN I

00

U a u a S 04

XI I

<

X

00 I o

; : XI

o . •~' X

o o

- » 0 u

i n fNJ t^ • — 1

•• ** 0) 2 %, OSI -HI

^ 1 0 r -

^ ^

^^ PO

XI u rvl

0 U

* X f o

49

3CI

o

X) c 3 O

in

o r--

I

O o

I o m

in >. nj

C o i -H

N

u

K

•a a 3 o . a o E 2 o u

CO .-H +

Q U

o — 2 I-o~ o

X 'J o ^^

u

in

XI u ^

X (\J

. E '— o ^ IN]

1

. t--o <M

N

^ t~ +J

^ a 2 Qu

U] M flj

s in

XI I l-i

<

H

o u

o 00

w ~ XI OQ

^ . 00

OSI •

<M O 2

u Q U

OS S a,

^ O u

i n

1—1

•• *- < (Q « •^^f

cc I-H

n)

s JC X

1

u <

X V

^ E ^ o <VJ

GO t

Psi

m 1 • f -

50

4)

in >•

c <

XI

o

<a i-H

3 • E - o • '. o ^ S o

u

T3 c 3 O . a o E 2 o o

u

XI 0) u • 3

cr

a: u

00

o

00

00

00 in

o -. 2 t-O- 00

a: '- u

fM

in m I

11 I

<

00 1

I

u <

in

I

XI u

X

U Q U « S a.

O U

in

00

in

(M

vO

-.O

O 2 — po (»-

a: psi pn ^-' i-H

u

00

V E

•*>) Q 1

• *

* ro

> X O x: •!->

03

ai '-

XI u •s

>o «M

XI u o 0 0

•—t

o >£>

1 1—I

oo •

>o

» . - pn

XI U O X

(M

* X ' O

* w ^ o o

sO

, 0 0

• -^ N

X <M

II

E I - )

N

X a-

ti

c 1-)

T ^

1 1-1

X I u •T-"

PO

m "-' o f M

I -H

XI XI

o •X)

t~

1

^ XI F - ->—1

.. u CQ Ui ^ ^ r

cc -^

1 t i

< <N

« X I -H

Ul

^"^

• t~-

^ ^ X o»

II

o ^

r-V f M

N

"E

+ s V}

s •• '"" X u

oo

o (M

O O

o 00

o 2 --

I-H pn I-H m

X <M eg '-'

U

0 0

r-1

t~-f M

>. X 0 I H

T3 >• X

1 «9*

f-l «>»l

>. X 0 £ + j

V E 1

51

1 u o

^ -

w

X I-H

^ tf)

<NJ

- O i n

oo

o9

i n

CO

'

XI r t " U O

•• a: f>-i

a,

Ul

o oo

CO

•

E >->

* X a^

II

o

X I

XI -

1

11 Q 1 •

m • o ^ ^

N

X

II

0

x\

CM

M

^^ E

CO

+

M <9 s

X

u

o ' J *

t>-i -H

I -H

o

I -H

i n

o o

• i -H

•^ < J

r o

^

i n

1 "

O CO - - ^

2 p -

X <^ I-H ^—' I-H

u

I-H 0 0

0 0 1

o 0 0 f M

E::: < c

?l «Nj|

•» X I -H

^ w —' 0 0 t -

r -

N

X

II

0 >->

' x*| 1 u 

^

p -l -fNJ

N

"E 4 - t

+ s « w m S

X u

* X I -H

• in

( M

o

o8

O

Kl X U

X m

* in

-

o <NJ

r g

r g

II

E 1-5

N

X

CO o

I -H I -H

o CO

• " J "

fM I -H

^ 0

P -•«!»<

• I-H I-H

I -H

(T-

•V

r o

pn

O ( M

2 — (VJ 'S ' I-H • ^

X ( M m —

II I-H

O o

vt l T) XI

, , i n

O IH • - •

C/3 <

s •*

IX

• • I

.. ^ o u

o m

•• )H

0

I-H

X I-H

•c •a

r r i r

00 t

P>J

00

1 r g

>o <M

1 <d

4-1

u o < -o

1 " ^

^ E

X I 1

< <\

o l Ml

•• X m

^ E

o r g

CO 1

O CO

• p -

— m

X I u o X 2

. X

^ «

i n

m

XI

u . X pn

m

o m

r g

••

, 1 "

r g

~E

4H 10

+

s ,. (0

^

6 w S D 06

O H

o

Q PO p -f - H

•a

o r g

•-H

•• M

m :>::

a HH

5

X 1

I H

<

X

E

i n 0 0

" 0 0 1

o i n

CO

X I

u <

«0

1 ^ u

f - H

i n

r o ^H

I -H

I T

f -H

I-H

i n N O

>o 0 0

• p n I-H

i n o

•^

Tt"

i n

r g

o r g

2 — o rg

1-4 — '

I -H

u

o p -

i n 1

p p |

i n r j

O C

E <

1 4.

1 el w |

* X I -H

^ U]

- O

~--

f-H un

P -1

i n ' T

p -

^ ,.- X I

1

3

• X r g

•. E

PO

> o

i n i n

N O

. •—

PO

X

• in

—

I -H

•

r g

N

"E

(d

+ 2

as

XI 1

O CO 2 Q

0 6

Qu

•* ' • ^

o u i n o » > 0 I-H

• • I H

CQ

• H

XI 1

I H

< • 8 r g

X r g

E"

I -H

0 0 1

I-H r g

0 0

X I u

52

in >• (0 c <

T3 C 3 O

z "

X n;

u2

T3 (U U

3 CT (U

rvj oo

U [ -

Id

«3

3

E • o 2

o S

• o

B

•X3

i n

OC

CC

•a a . B o O 2

u

(Nl

in O

o —.

X o

U

vO

i n

U CM

O

0) B I I

m w

i - H

^0 •

ro

* .""»

< * 1

I I u rg

X pn

w

00

N

^ B

•*->

nt

+ s .. U] (0

I

u <

r^ XI u i n

n

T ^

m

* E -'

CM ro

• f~ 1

O

NO

o •

<VJ

^

^ rsj

XI u

'^rn ^ x\ u

X fo

* + J

*—' >o NO

• i -H

1 f - H

i n

1—(

„ ^ - s

fV)

• O" ""^ I - H

ro a

^ 1

o I-H

• ^

M

.. m

XI u o

ro * 1 — 1

u Q U Ofi S IX

••

o u

i n

,. ^ M ^

V4 CQ m " > • - '

« 1—*

X ro

» (0

[ » !

00

pn

^^ m XI

U O u

• X pn

. • ^ «] O

— ro

rg pn

>0

o

<\1

in O

(M —. ro vO

X vO pg po CM —

u

i n

00

O

o u I >» c - . «

CQ X E I o I

TT rt pP|

o

o oo

XI u u

X C O

in

i n •

«3

j ^

X 1

<

• * H

I - H

NO

i n

o

u I

tM i n «r r-I

in

o i n

a

IM

^ '~-

PO XI u 1 in

p-r» - J M

P O

at

(n

XI 1 u < •

X rrt

. e

o O^

•O 1

t -i n

rg XI

- . 8 X fM

u Q U

S 0^

^ o O ^' U

CD pn

I - H

•. u

OQ m -.mtf

B:| H H |

XI u o

X* p>^

• M

—'

. <o >o m

00

O

O o --~ (M 00

X CO

1—k ^ ^

O

00

o

X u o u >. X o s: + j

u 1

V

ifli ^r^\

>. X O

Si *-> 0) E 1

CO

53

o

u 1 ( \1

« X

^ w

i n f-H

.<£

„ « w (0

, '—«. X i-i

<

** ^ i n

-* 1

CO

• i -H

«• "

' T

1

O^

m

^ -—

po

XI

8 f O

^ - 4

u Q U ^ • r '

cc s cu

• « k

"^^ O U -'

o

f - H

., »-l

CQ u: ^ • • ^

OS HH

* X f o

* in

^ <M 0 0

• PO

^ ..-

f<-l

XI

8 u

« t

X rr,

. • oo

M 0 0 > - <N

o I—I

i n

i n

r o

XI u

1 i n

X PO

• w

PO <T>

1—4

« -—*

PO

X I U

e

oo

•o 1

sO

• >o

f M

XI u

•k

X

1

0 0 ( M

N O

<N ' i '

• i n

o PU ^ - .

X in pg po (SJ —

U

o in

i n

X u o u

I >. o c -M 0) >, «

CQ X E I o I

<n

o 0 0

rr\

o i n pn

N

^

pn

Xl + u o u

» X

. (/I

— 0 ^ I *

• Pv3

* "" PO

U (M

M

X rr,

2

,. U) in

« 2

.« -—» X

1

< fc

X i n

E

o i n

• 1

i n

rsj

^ pn

Xi u

i i n

X pn

M

O

XI 1

<

X PO

* e

rvi

0 0 i n

^ •«

(v-1 <N

U Q U i h V

OS

^ a.

.« «—^ o u ^ v '

i n 0 0 vO f - H

• • u

CQ « • . M '

OS w

X I

8 X ( M

M

m ^ . f f

o 1—1

i n

^ -^

pp|

XI

8 • k

X fT,

54

(0

c •<

2 < ^

X : i

2 c

00

V

(d E H

T3

U

3

o OE:

n) 3 •

o

a

S

a:

u

•a a . S o o 2 u

i n

o -00 (M

u

o

oo

u o u > X o

0) B

t-

II X

r- CO i-H i n

i n

« XI

I l-i <

•8

o

i n

U l I

00

&• • i n

o

• i n

^ "^ en

>T< |

u ^ O '^ u

•T^ " ^ ^

>*

0 0

I fM

^- in

X Ul f-

rg o O •

V S

U l I u <

(M

m

(M in

O^ i n

i n

(M sO

^ P O

XI U o u

* X

X ? II

o • - 1

•

X u l o u o

O" --'

i n i -H

• 0 0

O ^

flj

+ <£

in «) S

pn XI I )-i

w < • ^ i n

o 00

i n

o

U l I

i n

C7»

X

u 8i o

f—I

U Q U

OS S

: - -^. *

CO

I

Ul Ul p\l

. f ~ o ^ CO rt CO 1

• 00

o ^ XI

pn

o 8

X

^ y.

CO

;:; '^

u Q U i-i

e c I

u

N X PO

II

>o X -o (J o O - U l

" i n

• t>-

po •—I X

-> E pn ' ^

XI u O X

ul o X U l

I

u ^ ^ « « • • * ^ ^

0 0

O rg

i n

i n

0 0

oo in

O — --H O

X <^ in - ^

U

i n

X U

o u

>» Si - t J

V S

1 ' J '

» pn

eol fO

>. X o •^ TJ V c (U

« X m

. w *—'

oo i n

pn

-^

o l-J

XI 1 u <

o

oS

fSJ pn

XI U o u o *

X m

^ fi

(M T

* rg

* • — '

X (M

» T3 •o ~-' rg m

• r~

1 o fM

• r

. *«» a

- T 3: XI u o u *

X m

a,

m

o 00

r-H

(y-

II

0 i - j

^ XI

1 i i

< in

" X f—t

^

u u ' - OS

a,

___ o u *-* i n >0

1—1

•s

o fVl t -1—1

« o o -o 1 - ^

•• • " • ^

• M

<d o 2 06 H H

pn

o

(M -£>

-o

__-

O

" o <M

00 O r~{

. • o 00 o I -H

• "" Ul

1

* M

•"T

^ •

^ O

m « X -^ Ul ul

Pvl 1

o u

1 P O

* cr

• — •

•a* T~\

fM i n

> >—*

r^

• in

'—'

pn rg

• fM vO r-H

* ^ Ul

1 <r\ u

X Ul o u o

• cr

>, '

(M i n

o fM

« « u

1 (-H

cr

o o 00 I -H

_,. n

u Q

xT ii U o

a

X <\j

(0

^ pn o

' i n

* ^ - v

u E c 1

u pn I -H

• a

- N

X pn

II

E '^

pn . XI U

N X

IN) O^ O U II

< •K

in

oo T T

pg i n »-H

o m

• 00 1 "

o pn

I-H pn P H

^ o

r X

' 8 •4-1

[ ^

o •

(V )

o I -H

«, ^-^ u IH

<

X)

I -H

pn

pg I -H

pg

55

1

^ m i n

o o

fSJ N O

m

VI

o N O

• ro

* ro

XI U o u o .

X f n

(\J

T3 T3

0 0 P -

r-1

vO

p -

N

X (M

II

o

o

U)

« (0 S •* .

p

X u o u o •0

u p^

•X)

o — »0 (M 1—1 ^

X fM i n ^— 1—<

r, U

O i n

m

0» 00 1—1

• pn

X u <M

o. ul • (0

—

X u o u

>. ^ o X! + j

u S

i n

XI u o u

X

X

XI I i-i <

Pvl O

" - 5

u 2

o u

o

o INJ

in

•« !S I ™

• •»

. X -— pg

pp>

XI II

8 E

X

^ 00

X

o

Xj I u <

Id

Z Xi o U

OE:] O HH U CM

o PvJ

O O

i n

o

o

U

00 in

in

I

X u

Pv o u

X O o ^

ji **

a B ^ ppi

X ppl

. Ul

'-' r o i n

X 1

t-i

< i n

x" 1—1

m XI u o u

» X

Cf-oo o

1 (T-t~-

O

X 00 U 00

X Pvl

XI u X PO

f p l

^ X PsJ

PJ

N

~i

t ~

o — O vO PM rp|

X f^ P- ^—

1 PM I "

•

1—)

o PsI

I oo

< ^ lO

XI -^

8 ^ X

U Q U

a.

O u

o 00

o 00

p<l

XI I

•^ u

XI u o ;i3 u

X CO

I

PPI 0 0

,i? xi S o i-i O

N

X

i n

N O

p j i n

v £

o -pj oo p j <y-

X <*i p j ~ ^ pv]

u

i n i n

X u

Pv o u

o ">. N

c V QQ

Ti<

X o x: • f j

(0 E 1

CO

X m

w — a-i n

^ - N

X I

< t«

X i n

m X U o u o ^

X r o

w ._ 1—»

«»" • ( M

« ^ - v

X I O o u .

X p-1

PO

^ t~-

1 p j 1—1

• t—

* .- X I 1

)-• < M

X pn

• E

i n

vO 1

0 0 i n

Pvl . XI

o

o

'J" a; PJ

S 8

i n 0 0

1—1

•1

o vO

<o 1—1

•• +J <4 4) 2

K| H H |

. Ul

^ 0 0

• PO

^ PP)

X U

p j

o u

0 0

a-pp|

N • " " ^

E 4- *

R>

+ s

• a

to V) (4 S

56

o

o i n

oo 00

o

—I o

1—I

U

CO

pg

O 1-1

I u

<

•a

lf>

o

ir>

X I u o u

1 •a

o

rg

X I u o u

O

X I I u

JX

i n

C <

— T3 U — U

CO

I

U l I

X u o u

I

o pn

. ^—« ro X Ul o Q

o

* rf—^

Ul 1 u <

• (0

i n o in

S ; * O

U l r I

I—I

a I m

in - • m O^ ^~ -H U l

I u

— <:

X

u 8i o ul

I PM

00

O

-* O X a-

in pg I—I •>—

U

X m

V)

— sO (NJ

(M

« - ro

XI u o u 1 ro

« X ro

(/) '»«' 0 0

c-i -H

^ , -

r^ X I u o u ^

X ro

«k

(0

o t ^

•-) ^

XI 1 u < vO

•8

<M

X

^ T3 •a

o

r-( t—

fM

f -

« ~. ISJ

. X

II

0

<->

x"| tH

* r r

• o ro

• '— pp

X Ul o u o "

cr

•»j-i V

- T3

u u

2

U l I u <

CO

fM in

o

f M

N

CO oo

o o i n

i n

X o i n t^ •-H (M

U —

o in rt +

X fO

00 IM

X I U O u

I

ro

00

X I u o u

X ro

O' o

u U

a. S

X u o u

0)

B Q >^

^ o • JS

ro + j

O

u i n i n

o

O m >0

Q

J? ^ XI E U

o

X

in 00

X ro

E

X

X I o

u o

oo

in

o

in ro

i n

X

I I

po

n

PO X PO

.5 2-

«_ ul

X

8 u

« S I (1)

* « PO 1-1

o u

:: o

o CO

5 8 xTH u

ro «0

E c I

X U

i n

N X PO

CO

i n

•"• •" - ^ ^

PO

X I

SI a Psl

U l I u <

PO

X U o ul o

ul I

fM

- CO PO

X ^. ^.

X u o u

o • J

O

u o in

in 00

o in

(1) 2 ^1

57

I '— '— I

X

O • - 1

I Ul o o

I

00 " pn . ty o

I O 0 0 LTl > 0 > 0 TJ< ^

T3 T3

o

N X r o

II

E

*— p

a: u o u o

ul I u < IM

N

a-

f M

i n

o Pvl

ul I

in w

. n]

pn

Q U

0 0 C I

o U I p^

ppi • - <

N . X

^ ^ fM pn

XI II 8 X PO N

X

— II CO O

XI I

x T ^ 8 -O <M

p^ in

( M

<N U •V O

o -^ ( M

oo ^ pn I—I •

8'V'

a-i n

• m i n

f«^ i n

• r->o 1—1

pn

f»1

i n

1 l-l



^ ,—. Ul

<

» T3

i n i n

o

pn

X U

o ul o •a

Ul 1

PVJ

U]

i n r-

pn

vO

O o

X pn

u

o

X u o u

>< X X O 0 J : •^ <o b) E

1 I ^ pn

X pn

Ul

-'

o

XI 1 u <

m

X pn

X|pn

U V

XI

u ^ X

I M

XI o-u ""

^ I—1

X ' . m ^

I

4-> O^

rn in in

I XI

'no .. X

—« pn

8i

u i i x i o

t ^ 1—1

•8

o o^ >o r-i

• o i n >o 1—1

•• '-' Id 0)

z

u o u o

. X pn

• (Q

" i n pn

• I N

o <Na pn

N

" B 4-> Id

--t+ l ' ^

XI

8 CO

« I « i-H pn 2

a-

O

i n

O — rg rg l\J 00

X f^ ro '-'

U

f M in

X

u o u

>.

o I -H

>< N C 0)

03 1

>>

o x: <-• (U E 1

X pn

in

—' •o< pn

r g

^ ^

pn

XI u o u

1 m

• X m

• w

~--fM O

r g

. ,-

pn

XI u o u

^ X pn

'—. XI u <

A

X i n

e —' 0 0 m

• 1

t—t r—4

r*"

j — ^

XI 1

i i

<

X m

• E

o o^

>£) 1

r-i n

• »—1

,,

U o

^ IE »—1

u u ' • — '

oi S ou

• a t

-- o u ^m^

o

1—1

o9

i n 0 0

I -H

« o i n

• — 1

PM

• in

o I -H

• i n

» '" pn

XI

8 «

X pn .

f M . 0 0

M " ^

N

^ 2 E 0 0 ^

'^' 15

: : - H S Id ID Z

X y w o «

* o s

58

•o c 3 O fa

c -X 0 SI • J

El I

1-1 < in •8

X

I U <

in in

I o

321 U O X

X ^ ^ IM

IN

""" E

00 - ,

ml »i-i I

U

I <M

« W

^ o t^

• ro

,, ^ - s

m « «) S , —. N

K p j

II

E •-1

• t

N X

^ <> l -H

u Q U

tf S

.. ^ o u o

t^ 1—1

"" u to Ui

Ofi

II

o •n

X u < o

• X 1—I

•0 "O " • ^

o ^ t ^

1 00

. f ^

•

o

00 in

oo

i n

oo i n

o in

I

o

X\ u o

X

O

o

I I u

X (M

00

O ^ 2 in X fM

O ^- ' I—I

U

oo

i n

t ^

• ^

00

INJ

O z

00

1

>o f - H

1

J3 •!->

s 1

«J« •

pn

>. X o

T3 (U c (U

l -H

NO T3

1

o M

s Q +

pn ^ H

u Q U * t a^

o: S 04

• * ^•^ O o - -i n 00

1—t

^

Ui

>-t

• i n o <SJ

N

— E

+ j

n

•• V) U}

2

••

1 ^

X '^

u

pp>

I

X

o

<-•

2

59

o

X] I u < in 08 m

oo

in

0=1

X

T3 o-

o CO

>0 N

• E " *

CO

X\ U

<M

ro XI u 0

* X m

• t

w • * - *

>o r

• m

• ^ og

XI U

«. X

« 1/1 <0 S

.» - N X ro

II

E •-J

» N

X (T-

II

0 >-)

X

-o

I t-l <

«a

X (M

— -a

u •"" Q o

I

O '

X

II

E

CQ N « X

l-H II

in

i n

PM

O 2 ^

- T ' f^ X (M

(M ^ l-H

u

in

vO •"S*

1 i n ^ l -H

>s X 0 si •M

u 1

1 "

>. X 0

X!

<-• v E 1

t--in ro (M

N

xT ^ u *. X* +_

•«-> """^ 00

^ • .—1

1

m

1—t

^

in

S

• •> „—K

XI 1

< ^

X vO m

T3 1

0 W S Q +

• E •^ 0

(\) ^ ' " r - " U Q U ^ cn 2 OU

, ^ 0 u

0 0

1 I "

>o

• " • ^

p^ XI u

x" Psl

^ c r

l-l 03 I

lO

60

V

S

o -^ z=(

ir

Xi

in

C <

o •

o S

0)

u

o . a o E Z o u

T3 C 3 O

T3 a> u 3 a (1)

u

^ " ' ul

00 00

o

i n

U i n

i n

m o z f^

f—1

X CsJ

.—» er t—1 (M

"-' u

in oo

o r-

1

Q 1

* * r<

>. X O Si •4->

0) e

^

X I 1 u <

•k

X ro

m >m^

<M O

• O^ >o

I

1^

Ul I

in

po

• o XI ^ U

o rv4

X

Ul I

* ' • ^

m X I u o X

X

IT-o~

m X

8'

Ul in (Q S

u 1

<

f<1 00

• PO

• '^ m XI u ^

X

M

M

*^ >o Csj

pva

i n o • vO

i n

• i r« l

X Ul ••i

m l

cy -e^

-i n

^ f - >

•^

1—1

* (NJ I *

O^ ^

^ 1-^

r-o 1—1

. O o • o*

1—1

1-1

(NI

O

i n

a-CO

O

i n

O — Z ri

(T- O X <M

u

00

I

I

I ° E "O I «

* «

« s Ou

u E d

^ o 00

M «

XI I

<

X

O

I

XI u

XI

X

13-

in

p<^

XI u

X m

m rN4

• «M

•• ' • " *« ro ^ u

ro O eg

N

^

u *. K

cul S

^ r-

O

(NJ OO

00

(NJ

O Z —

-H 00 X ^

u

m I

(N)

o

X o

•M

61

o

XI I l-l

< • 8 pn

fM

i n

i n

i - i

3=1 U

oo

I *-•

0 0

• +

XI

o

XI

8

X

o

in in

..

II

E •^

o o vO

m TT

>o

i n

I

XI u o

oo 00

• r^

. • po

XI o

X f«^ .

• p^ pn (M

N

^ 4-1 (d

4-

rg

X PO

o 00

XI I u < •a PsI

X PM

^ -a pn "O

XI ^

u rg

•^ I 0 0

. m

PO (g

• X

u Q O

II

E

X

po

o 2 -^

in pn r - l pn

X <N l-H

u

00

•>»• 00

1

oo

>. X 0

ji • J

w 1

>. ii o •C •!-•

4) E 1

o PO

(M

_ pn

XI u X

(0

s ..

X 1

<

X* ppi

J E

m po

(M

I 00

in ye

1^ ==• ^ u u Q U « s CLi

* X f - l

w

62

T3 C

o

A

&

CO

<:

T3

OS

U

U

o •

o 2

i n

fM

o

o

00 00

fM

U

oc

o 2

^ • r-H («-i

K •»• C O ro t- i "-*

U

o i n

o I

oo

O X u

o . a o E Z o u

X I o

CM o u

X

i n

X I u

1 (NJ

X

m "

CM ^

' ^ N

X I c U

I i n

X m

* <n ^ 0 0 • — 1

• (NJ

J^ CO

f ^

U

2

•• V) (0

(d S

.*, o ' ^

o X u X

Q U

Oi

^^ «o O U

o o

Sl I u <

eo

o 0 0

i -H

•• u a Ui I w *

QCI -•

« B

vO

o 0 0

o >o

• l~-

i n

o

00

o C O

X I U

(M O u X

i n

X I U

I (NJ

X <»1

00

i n ( M

• o o

•^ O

(NJ

z u i n

1—t

1 X 0 0 f—1

U

N O

CO

..—* i n

• CO i n f o

*^

• -*

XI u 1 i n

• X m

m

o 1—1

(NJ

•• ^^ < Cb

• CO i n m

M

19

+ s

I-H (NJ

X o 2 II

X u

B: 2 Qu

•«

O u ^

(0

u (« 2

X 1

u <

9-

O O

- e u

03 w (NJ

o: • I-H 00

i n

in (J'

(NJ

(NJ (NJ

o U

X pn

X I u

I (NJ

m 1 "

f O vO

(*1

O (NJ

2 ^ U

X 0 0 1—4

u

CO

o

oo r-

1 >o t>-1—1

2 U

"^ i n

• o m -'

U 1

i n

X PO

• in

~-' o I - H

(NJ

-^ < Cb

EH • W

Q: S cu

• a .

. ^ Z u

o 0 0 (NJ fNJ

^ ^ o u o

• o

N

" * e

• 4 - *

«3

+ 2

•• in (0

• *

•«—s

X 1

IH

<

X ^

* E "^ I -H

(NJ

00 1

--^ f -I H

03 ixj

» | «l

• ^^* XI u

63

(M

a o

in

C . c 0) A a o

c 1 ro

o

U

c o

XI u o u

DC

o o 00

K l U X

( \ j

. X ^

o ( M

( M

• •

t ^

-!-> (d

+ S

•« (0 Ul

2

• * -

X I 1

u <

J ^ X u Q U K S cu .

, , , • 1 ^

O O *"

o 0 0 >o t - ^

,,

1-1

CQ txi

«

tf\

, E •~' OO >o t ~

1 0 0

f -

• ^ • ^

X z

• X • - I

• M

1 -o

i n

o

OO

00

pn >0

in O

fM

z — r j OO (M m

X <^ U

^0

m o

1

<o <NJ

1 (d •J 0) u <

^ >. c V J3

a o •o

O X I u .' X •::

(VJ < N O

. « a " vO

. X (0 (^ ^ -

« i n "O r- -O

* ^^ pn

. p^

^^ -^

= 1 T U rH X i n

<VJ

r-•>

X vO ^^

ti

- X w r\j

— II

f — . E

(M "-D

• a.

' ^ N T X

X I o-u

0 0 vn r o

N

" E

- * - » Id

O II .

X o ^ Z -5

X <*i X I

1 u (0 <

^^ i n •a •-H PO

I M • «.

(M X (VJ

-"» «o _r •a -a

1 "O O — w 2 o D < .

« 2 &

t ^

0 0 i -H

• r*

,..'-.. o -^ X I

u o <M

" ^ 1—1

*

o ^

<o rt fVJ

o * ^ i n U

OQ U! ' ^

in in Id 2

^^^ X I z X fSl

• X r g

^ in

Si ( M

-^ 1—1

<?•

O^

• 0

0 0

o OO

,_ M

X

II

E "-3

<k

N

X o» I I

3 f* osl XI o >H 1 u •-J

i n

i n

po

fM

o in

i n

sO

po

X "Z

U

00

PM PO

1 1—1 pn <M

0 c •«H

E <

1

^ - t

>. c «

X !

a o u *-> c 1 r^

64

I O

CO I

o

"»• a.

.-^ f n

3=1 u O o K r g

X •£>

in

o i n

m « - ( * i

X I u X <\1

« X >o

r o

N

B

•M (d

^ S

••

S

^ - H

fN3 X I z

* X (M

. u JD ^^ ^H TT

0 0 1

0 0 f\J

XI 1

O CO :^ D +

OT

J3 ^ r<^

r-f -

m ,

U o-wr

K ::§ a,

""

o u " ^ O

^H

*• u CQ M

r-

• —~ X I

1

i-i

< ^

X f« l

^ E

^~^ r o O^

vO

1—1

I—I

o rg

0 0

O pg

2 — <M • * C I f M

X >^ 0 0 ^ ^ r j

U

o

I

o

X I

u

X

o

X I

u rg

O U X

X

X I u X ^

. X P J 1—t

^ n

U t a ^

• m <M i n

ta

~i

<

OS

i n

0 0

pg

o

Psl

O pg

2 ^ "-" in U

i-H 0 0

X <*i o ^^ pg

U

OS

a,

O u

U Id

S

O ^ H

X 1 I-I

<

pg

1

o I N

\ _

i n

X I

o in

X I

u Ps

o u X

pg

X

0 0 in

(*1

X I u X PO

i t

X >o

* w

< > « • • > '

o» PM

• >o 0 0 f O

N • — .

e pg +J

^^ on QU

w VI IQ

o u

o i-H

i-i

in

CO

o pg

< O 2 —

in in pg t^

I-H

u

in in

Pvl p\i

I

XI 2 «

< xo-. I -

X -

5 CM

X o

X

- X

- X

-ox -ox

I

X I

o o u X

PJ

X

= 1 U X

( M

« X N O

* (0

i n

r<^

N

'-^ E

_ < M « Q

—' cc S O i

n Id

S

••

. . X I — u X

£ X o in

O U

i n

(Q

0 0

m in

I O pg

X I

u

;^\ > - " I OS X I 1-H U

65

i n

fvi a-

00 I

o in

= 1 U

i n

i n

= 1 U I

i n

o

U o u X

X

i n

in

m

i n

i n

o

o

o

<M

3=1 U

00

i n

o

o

o

i n

O 2 ^

rt in (M r

oo ~-' »—»

U

o 00

o in

I C

i 7 a

co

U

x m

^ T) — ' J '

INI 1

0 0 0 0

• f \ l

. '—

m 11 U X

pg

^ as •x>

,, in 10 ta S

• ik

I 1

i-<

< •

X r^

> E

o^ po

« r~

1 oo i-H

2

m w ^ H

u Q U ^ ^ f

cc S OU

• « „*->,

O U N.v'

o o

1—1

.. ^* i-i

03 ^ >»«• OS 1—1

XI

oo o

• 1

0 0 o^

• m

^^ X O

^ a i - i

, (0

>. o 0 0

1 • -*

i n

i n 00

o 2 —

i n I-H (\j i n

^ — (M

U

m O

I

o IM

I C

© C -

I E ii

eo eo

* <\I

x\ u

^ X

* T3

ir> <j-

t ^

^ -

321 U

„ I rH

U}

cr CO

(NJ I—1

(*1 o sO

1 in in

• ^a

^ - v

K U

o

66

i n

m XI u o u X

l -| ^

in

in -M in nJ

CO X I U X

(VJ

^ X >o

(Ol (0 n)! S

JZ. X 2

- Xi I

(NJ <

<M X

U £ Q U ^ —' m K| • 0 1 i^

O . O ^ ^ XI o ^ - 5

l l < — CQ Ui o _ 00

Oil •

i n

i n

O (M

2 — 00 rg

X <

CJ

ON

I oo

>> X O

73 O ^

^^ m O,

00

XI u o u fM

X

i n

" ^

XI o fM

. X

VI

>o p l

• fM

N

^

I O w S Q +

XI 2

XI 1

<

X

u a u

a.

O u

o ON

JZ m

CQ I « ON —• fM

a:| •

o

00

sO

o pn

2 1—t

fM X

t ^

^-^ 1—1

PO ro • " — '

u

m fM

O C

E

(f-(T-

"a*

«t

, ^ ro

XI u

fM o u (NJ

X >o

• tfl

*—' o vO

m

* <*• XI

u X fM

X* >0

* </}

"-' o (M

(M

•• .^™*

-l

03 Ui

«

CO m

N

E -M R>

V) V)

<-. fM

XI 2

* X (NJ

^ in X>

1—1

t»-

• t~ 1

o vO

r-

^ ^^ XI

1

u < ^

X r^

. E

fM i n

t«-

1 1—»

f ^

r

» *»-% XI 2

^ M>4

PsJ

f»

vi

1 -o

=1 u

0)

67

0 0 c8

m CO

i n 00

0)

u

OS

u

o . a o E 2 o u

u

m r-

00 fM

m t ^

O 2 —

rv] o

1—1 ^ - '

CO

O

( M

00 in

I in

a> 04

X >o

i k

M

^^

0 0 1—1

•

<NJ

,, ^^

• tn —'

(VJ "J"

0 0

* X I

1 u

O < T3

1

o W

s Q

• X. m 1—1

»

+ e f—(

U Q U

S Ou

•» O O • " '

o CO

>o 1—1

.. ^-^ u CQ Ui! ^ M *

CC H H

^^ (M i n

1 o 0 0

sO

^ • N

<> X I U

« X «M

» V)

>o fo

1 • 1 "

00 in

u INJ

o u

M

X

^ (/]

~-'

m f o

• f ^

«h

<-. CO

XI

t^ o i n

+ S ^ U]

w nl

s ^

.-^ X 2

fVJ 1—1

o

o ^

r-

CO

• i n

•»1<

00

<a3

O 2 —

m f-m vO

X ^

u

sO m 1

in ro <—1

>> »< O

4)

s 1 ' T

t—1

c V

.c a

xj u

(N )

o u

» X vO

^ to

-

• ^

i n

• m

°a

X I 2

oa

X 1

<

X <M f—(

e

XI u

X XI u

«

o IM

• (M

(M

M

in '•^ i n (M

U Q U ••mi'

o: 2 a.

• A

<^~* o u >^

o CT»

-o

X u

* X 1—t

u >. r-o

• «

*""* - xT ;: 8 U X,

CO <M

«i X HI >0

o

t - -1

I N i n

• -o

oo o^

• vO vO

00

o m

Z —• m (M

in —' po

U

( M

<N

O pg ( M

1

4) E

Q

V

* m

f—1

>. C 0)

>. »< o x: •«-•

68

o

r g t ~

m

0 0 > o

a

CO

i n

r o

, —«

U <M

O u X (M

X sO

M

" • ^

o i n

^.^ Xl 2

I i - i

* M

r -

^ Xl ^ <

* X o ^

» E

i n <M

1 o i n

•X) ' T

r -

i—t

m

i n

O 0 0

i n

i—i

o

r -

o i n

i n

=:| u

X

XI u

X o

(0 O

XI U

« X U - ' Q 5i «

O o i n

o o

XI u o »<

X

iH r-t

CQ

OS! • *

00

o m

2 — m 0»

X I " f n ~^ m

U

CO i n

o

S o

X S ««" f o

a

a

vO

f n

„ ,—*

m

X I u f M

o u

• t

X >o

en

.

o i n

r o

«a

C3 T

• m

* ' • ^

X I u x: (VJ

« X vO

X I 2

• s

X I 1

<

* X o 1—1

,»

s ^ > o

• t - -

i n r o

O

» --^

fM

XI u

* X

i -H

1 "

• r~

CT-i n

^

f—I

• vO o

i n O

f -0 0

• ^

i n i n

• sO

XI u

fSJ

• ( M

•• —*

i n ( M

• i n

* sO ' ^

T3 1

o w S Q +

X u

• X •—1

en " t—(

U Q U

s cu ^ o u

i n 0 0

<-i

•• *"* 1-4

CQ Ui

- OS w

n ^^

TT

> ^

X

8 (VJ

« X ^ 0

. (0

<VJ

o 0 0

• m

00

o 2 —

o> in (M O^

X " ro "—' m

U

i n

o 00

I

fO rt Q,

10

o o

m

X I u (NJ

0 u X r g

. X • ^

. (fl

fM

^-' 0 i n

r o

s8

m ' C

f»i

, «—

m

X I u X fVJ

X

rn

-— X I 2

<«

X I 1

u <

» X 0 f—)

, J

E " • ^

1—t i n

f -

1 i n T

• >o

^..^ 0

f M

XI u 0 X

r g

» M

••

n (M

^^

J ^ <^ 0 Q U '*m^

CU s cu •• 0 u

0

o» >o i -H

.. ^^ u

CQ :>E3

^M."

a: «

oo

i n

a«l

0 00

• i n

^.^ ( M

X I u

* X <M

. M

0 0 f -M

• u >

69

3.2 ANTIFERTILITY ACTIVITY OF COMPOUND SYNTHESISED

Almost a l l the compounds s y n t h e s i s e d as p o s s i b l e an t i f e r -

t i l i t y agents have been screened for t h e i r e a r l y abor t i f ac ien t a c t i v i t y

in the Division of Endocrinology of Central Drug Research I n s t i t u t e ,

Luc know.

Evaluation of Early Abortifacient Act iv i ty Material and Method:

Adult male and female hams te r s and r a t s of p roven f e r t i l i t y ,

maintained under uniform husbandry condi t ions having a regula ted

l igh t and da rk pe r iod were caged for overn igh t and the vaginal smear

was examined following morning. Finding of spermatozoa in smear v/as

cons ide red as day 1 of p regnancy . From days 4-8 of pregnancy in

hamster and days 6-10 in r a t s the compounds were a d m i n i s t e r e d s u b -

cutaneously .

On day 12 of pregnancy in h a m s t e r , animals wre lapro tomized

and examined for imp lan ta t i on / r e so rp t ion s i t e s . Required a rea of the

r e so rp t i on s i t e s was f ixed in Bov in ' s fluid for h i s to log ica l examina

t ions . Results have r e v e a l e d tha t if compounds were admin i s t e r ed

before day 4 of pregnancy in hamster or day 6 in r a t t hey were

not equa l ly effect ive as in the pos t - implan ta t ion s t age . The r e s u l t s

of biological a c t i v i t i e s of var ious compounds s y n t h e s i s e d a r e d e s c r i b e d

in Table 13.

RESULTS AND CONCLUSIONS:

Simpler he t e rocyc l e s possess ing only one phenyl r ing d id

not show any r emarkab l e con t r acep t ive effecacy. The p r e s e n t s tudy

has a lso r e v e a l e d the pregnancy i n h i b i t o r y effect of compounds b e

longing to a new c lass of he t e rocyc l i c sys tem namely 2 , 6 - d i m e t h y l -

3 , 5 - d i r a e t h o x y c a r b o n y l - 4 - s u b s t i t u t e d - l , 4 - d i h y d r o p y r i d i n e . Th i s new

lead i s worth exp lo i t ing and ca l l s for evaluat ion of o the r compounds

of t h i s s e r i e s .

a o

70

c 0) u o. >> y c a c tX) <D U

Ou

I eW>

in ( M

dp

o >o

o» o ( M

d«» sO >o

cW> v£> sO

dP

m 0 0

p ^

•^ 2

olP O o

<*= o o

dP vO

>o <* Q> O

i

c C 00 ID U a I c o 2

. ^ - ^ f-H * *

o 2

T3 (0 in

(d —

^ o c

c c E OJ 4-> U «s a » e

ID eo 00

in eo

71

<*p i n 00

dp o o

<*p i n 00

00

t

00

1

00

1 oo

1

Psl

O

O

O

O o o

eo

CHAPTER 4

STUDIES ON THE CHEMOTHERAPY OF HELMINTH INFECTIONS:

RETROSPECTS AND PROSPECTS

4.1 : The hea l th haza rd of helminth infections in India

4.2 : Development of exper imenta l models

4.3 : Biochemical s t ud i e s

4.4 : Mode of Action

4.5 ; Future development

4.6 : BASIS OF WORK

72

CHAPTER - t\

•STUDIES ON THE CHEMOTHERAPY OF HELMINTH INFECTIONS:

RETROSPECTS AND PROSPECTS"

4 . 1 Helminthiasis, a group of several diseases caused by the

invasion of human body by the roundworms (nematodes), flatworms

(trematodes) and tapeworms (cestodes), may be broadly classified

as gastrointestinal or enteric helminthiasis, schistosomiasis and fila-

riasis. Of these, filariasis alone affects the health and general well-

being of nearly 400 million people in tropical and subtropical regions

of the world . Of the 900 million people around the world living

in the areas endemic to filariasis, 236 million population belong

to India, The picture of endemicity in this country is becoming alarm

ing day by day since 22 million microfilaeramic carriers and about

16 million cases with clinicopathological symptoms have been reported.

The continuous spread of the disease and protracted suffering and

disability caused by the parasites to the infected population have

attracted greater attention and priority has been laid for combating

helminth infection not only in this country but also by the members

of the World Health Organisation.

The enteric or gastrointestinal helminthiasis is also highly

prevalent in different parts of the tropical world. In India it is

endemic in rural masses and the urban population with low standard

of living and poor sanitation. The incidence of ancylostomiasis ranges

from 40% in plains to 90% or even more in the area of tea estate

46,47

and coffee plantation ' . The ever growing dimensions of this dis

ease in this country is evident from the recently published statistical

figures. In Uttar Pradesh alone 20.1% of rural population is affected

by hookworm infection

Helminthiasis, in principle, can be controlled by improving

personal and public hygiene and in the event of less statisfactor y

sanitation conditions, chemoprevention is possibly the most attracting

approach. Any chemical which may evoke selective toxicity to para

sites or is capable of stimulating immunosurveillance mechanism of

the host would be of interest for the treatment of the infected popu

lation. Present state of art in immunology possibly permits immuno-

-diagnosis but specific immunotherapy of various diseases caused

73

by helminth infections is not yet possible and this has prompted

to analyse the retrospects and prospects of the chemotherapy of

helminth infections.

4.2 The retrospective analysis of the chemotherapy of helminth

infections reveals the attempts made by earlier workers to develop

suitable experimental models for the evaluation of possible anthel

mintic agents.

DEVELOPMENT OF EXPERIMENTAL MODELS

A. Filariasis: The search for experimental host-parasite models

has led to investigations on parasites such as L'ttomoAutde s caiin« 49 (Vector, Liponij^^u bacoU), Vipe.taiomma \jitzaz (Vector, Qfini-

thodO'i.ou-t moubata) and Bn-ugia malayi ' (Vector, Aede S azqijpti

The suitable hosts are cotton rats {Stgrnodon hi'^p^tdu^t,) Ma-bto-

my^ nataizn6A.6, albino rats and gerbila { h\e.n.<lonz^ unQuicutatu6 ) .

The difficulty of breeding, handling and maintenance of

cotton rats, the natural infection fori-. CCUiinil, the most commonly used

filarial infection have led to the identification of mostomys as 52

alternative host , Subsequently, mastonys has been found to be

more suitable as a host for V. viteae. and 8. maiayi infections.

The latter can also be infected to cat , rhesus monkey and langoor. B. Intestinal Helminthiasis:

Hookworms and tapeworms are the main targets in intestinal 54

helminthiasis. Albino rats and golden hamsters as laboratory animals

for W. bA.a6ihznM^ , (a trichostrongylid) and Ancijio^toma ce.bjian<curr]

(hookworm) respectively, have been found to be more suitable but

certain advantages and susceptibility of M. nataizn6i6 to N. b^aMtiZn-

-J-t-i infection have furnished yet another model for hookworm infection,

for detailed study of infectivity longevity and fecundity of A.cZytani-58 cum , laboratory rodents have been used. The laboratory models

for tapeworm infections such as Hymznolzpi'f^ nana ' in mice and

H. diminuta, a natural infection in rodents, have been found to be

satisfactory.

The development of suitable experimental models for the

evaluation of possible anthelmintic agents has provided impetus for

the screening of large number of compounds. However, the need for

the replacement of this emperlcal approach has been realised much

74

e a r l i e r and in o r d e r to p rov ide a sc ient i f ic ra t iona le for the drug

des ign , two d i s t inc t app roaches have been made. The f i r s t one is

concerned with the s t u d i e s on the b iochemis t ry of the helminth p a r a

s i t e s and the second one r e l a t e s to the understanding of the mode

of act ion of ac t ive compounds d i scovered during random sc reen ing .

4.3 BIOCHEMICAL STUDIES

The anaerobic h a b i t a t of most of the in tes t ina l he lmin ths

has provoked s t u d i e s on g l y c o l y s i s and the i n h i b i t o r for the key

enzymes of the g lyco ly t i c pa thway in helminth p a r a s i t e s such as

Chandiz/iziia kaujkingi, L. ccuiimi and a^coAciia gaiU has been s e a r

ched . Cyanine 863 , an ac t i ve inh ib i to r of these enzymes , has

not deve loped into a drug because of i t s t o x i c i t y . The b iochemis t ry

of Se.taAia CZX\/i, the p a r a s i t e of the Indian water buffalo, has been

s t u d i e d . A phosphoenol p y r u v a t e (PEP) succinate pa thway requ i r ing

a th io l function for the enzyme a c t i v i t y has also been mapped . Adult

and microf i la r iae of 5 . CZn.'^i a r e ve ry different in biochemical

composi t ions . Unlike mic ro f i l a r i ae , adu l t s have h igher contents of

glycogen and glucose but have lower contents of f ruc tose , l i p i d s ,

nucleic ac ids and phosphorous . Recent obse rva t ions emphas ize t r a n s -

cu l t icu la r abso rp t ion as an ac t i ve t r anspor t p rocess and inh ib i t i on

of t h i s t r a n s p o r t can s t a r v e the p a r a s i t e and p r o b a b l y k i l l i t . P r e

sence of CAMP dependent and independent prote in k inases in B^tuQia ma-

latji and N. b->iaMUe.nM6 ' p l ay a regula tory ro le and the a c t i v i t y

of high molecular weight p h o s v i t i n kinase have been i n h i b i t e d by

suramin and h e p a r i n , indica t ing a r e l a t i onsh ip between drug action

and inh ib i t ion of t h i s enzyme. The p a r a s i t i c in tes t ina l he lmin ths

such as A. iumbxicoidz^ and A. gaiU p ro tec t themse lves from p r o t e o

ly t i c and o the r deg rada t ive enzymes secre ted in the lumen, wi th

the he lp of t r y p s i n and chymot ryps in i n h i b i t o r s . Different organs

of A. gatii contain s e v e r a l amino ac id metabolising enzymes such as

Histamine ammonia- lyase , th reonine d e h y d r a t a s e , glutamate d e h y d r o g e

nase and alanine and a s p a r t a t e amino t r ans fe ra ses of which alanine

and a s p a r t a t e t ransaminases have been found most a c t i v e . Transami

nases for alanine and a s p a r t a t e have been r e p o r t e d i n S . CZn.vi , i.coAi-

nii and v. vitzaz whi le arg in ine and isoleucine t ransaminases a re

a l so p resen t i n S . cz^vi. The p resence of var ious biogenic amines

in helminth p a r a s i t e s has been ver i f i ed as adul t worms of L. caxinii

75

contains a l l the four amines t e s t ed which a r e h i s t amine , 5 -hyd roxy

t r yp t amine , dopamine and no r - ep ineph r ine but dopamine is found

to be absent in the microf i l a r i a ! stage which possess h igher amine

content than adu l t s . S. ce^v-t, N. biaîUtnî^, A. iamblicoidz^

v a r , homini't) and A. galti contains al l the four neuro amines but

with varying level of 5-HT and NE in different p a r a s i t e s . The p r e

sence of both type A and type B monoamine ox idase (MAO) in A.gatii

and a l i p h a t i c polyamines such as spe rmid ine , spermine as well as

put resc ine in S. ce.ivi, A. qalii, C. digonopoâ have been shown

4.4 MODE OF ACTION

Biochemical composit ion ot p a r a s i t e s , the enzymes involved

in metabolic reac t ions as well as s u s c e p t i b i l i t y of the p a r a s i t e s

to an the lmin t ics have been inves t iga ted to unders tand the mode of

action of these drugs and to ident i fy target for chemothe rapy . I nh ib i

tion of c h o l i n e s t e r a s e s , in ter ference with fumerate r e d u c t a s e , i n h i b i

tion of mi tochondria l NADH oxid iz ing a c t i v i t y , in te r fe rence with p h o s -

phofructokinase and folate metabolism and d i s rup t ion of ce l lu la r in te

g r i t y a r e the major ta rge t s i t e s for the anthe lmint ic d rugs . Anti-

f i la r ia l drugs have been found to effect the var ious metabolic a c t i v i

t i e s of the p a r a s i t e s which could e i t he r be d i r e c t t a rge t s of the

drug or the secondary effect of t h e i r ac t ion . The capac i ty to syn the

s ize glycogen and p ro te in from glucose and val ine by adul t L. caiinii

i s a l t e r e d by DEC. It a l so affects the ox ida t i ve and metabolizing 70

enzymes of the host . Consequently admin is t ra t ion of DEC to the

host has been fovmd to a l t e r i t s immunological r esponse and to cause

a d v e r s e reac t ion in microf i laraemic h o s t . This drug has shown to

cause the loss of shea th from L. CCiA.inii and the e lec t ron microscopic

s tud ie s have r e v e a l e d tha t the cut ic le of microf i la r iae of 0 . volvulus

a l so undergo changes . Levamisole a l t e r s the ae rob ic glucose metabo

lism of BKugia pahanii and i t has been suggested tha t the effect

of t h i s drug on the metabolism of microf i lar iae a re secondary to 72

the obse rved p a r a l y s i s . Suramin i n h i b i t s the l ac ta t e and malate

dehydrogenase of 0 . \Jol\jaiuA . It a lso i n h i b i t s p ro te in kinase

of the same organism and c i rcumstant ia l ev idence e x i s t s to presume 74 tha t Suramin might have more than one s i t e of act ion . Nic losamide,

Praziquantel and Mebendazole Inh ib i t glucose up take by the p a r a s i t e

and an inc rease in lac ta te formation ind ica tes a change-over to homo-75

lac ta te fermentat ion . Proposed mode of act ions of drugs containing

76

benz imidazole -2-carbamate nucleus (Mebendazole, Albendazole , Fenben-

dazo le , Flubendazole , Oxibendazo le , Pa rbendazo le , e t c . ) th iabendazole

and cambendazole r evea l t h e i r act ions mainly through inh ib i t i ons

of microtubule po lymer i sa t ion and fumerate r e d u c t a s e .

Current ev idence is inadequate to determine whe t he r or

not DEC is capable of invoking a p r o p h y l a c t i c action in the lympha t ic 76

f i l a r i a s i s in man . More than thousand compounds have been eva lua ted

as poss ib l e new macrof i l a r i c ide by the spec ia l programme of WHO

alone and a few compounds which have been ident i f ied for c l in ica l 77

s tud ies a re d e s c r i b e d in f i g . a .

The chemotherapeu t ic agent employed for the t reatment

of nematode and ces tode infections and t h e i r s t r u c t u r e to a c t i v i t y 78 r e l a t i o n s h i p have been r ev iewed . Of the var ious chemothe rapeu t i c ,

agen t s , methyl benz imidazole -2-carbamates occupy a unique pos i t i on .

This is p r i m a r i l y because of t h e i r wider spectrum of anthelmint ic

a c t i v i t y but no a t tempt a p p e a r s to have been made to rev iew the

present s ta tus of benzimidazole anthelmint ics for the t rea tment of

var ious helminth infections for assess ing the spect rum of anthelmint ic

a c t i v i t y . The p resen t s ta tus of benzimidazole an the lmint ics h a s ,

t he r e fo r e , been p resen ted in t ab l e la to p r o v i d e a b i r d ' s eye -v iew

of t h e i r e f f icacy .

4 .5 The t r end of r e s e a r c h a c t i v i t i e s in the area of chemopreven-

t ion of helminth infections suggests t ha t in the coming decade^ p o s s i b l y

more emphas is would be l a id on the development of chemoprophy lac -

t i c s and immuno-regula tors . The l a t t e r v/ould be more concerned with

the development of agents which would enable an immuno-incompetent

host to regain i t s immunocompetence. The c l a s s i ca l chemotherapeu t ic

approach may furnish exc i t ing and in te res t ing data on n i t ro group

bear ing h e t e r o c y c l e s . Development of new lead molecules as GABA

agonist i s a s t rong p o s s i b i l i t y and the de ta i l ed unders tanding of

the mode of act ion of ive rmec t ins would be p o s s i b l y a c h i e v e d .

4 . 6 BASIS OF WORK

The socio-economic problems and inadequate publ ic hea l th

fac i l i t i e s of t h i s country cal l for the development of a b r o a d - s p e c t r u m

77

OCH3

NHCSCH2CH2CO2H

s

CGP 24914

OCH3

NHCSCH2CH2CO2H

S

CGP 20376

O2 N - ^ ^ ) - N - ^ ^ N - C - N ^ - C H 3

CGP 6140

T Furapyriaidone: N-(5-nitro

-2-furfurylidone) amino

tetrahydro-2(lJl) Pyrimidone

^ ^ XH2CHOHCH9OH

^ N 0 2

Desmethylmisindazole:

1,2-Propanediol, 3(2-

nitro-lH-imidazol-l-yl)

OCH3 H

H5C2\J/CH3

73>s

Contd.

78

,CH(CH2)3 CH3

Ive rmic t ine (A mixture of Components I and I I

Component I : 5-0-demethyl-22,23-dihydro avermectin Ala

Component I I : 5 -0 -demethy l -25-de ( l -me thy lp ropy l ) -22 ,23-

d ihyd ro -25 - ( l -me thy l e thy l ) avermectin Ala) ,

'XKl H

NHC02Me

ALBENDAZOLE :

FENBENDAZOLE :

OXIBENDAZOLE :

PARBENDAZOLE :

R = SCH-

R = SC,H, 0 J

R = OC^H^

R = ^"9

THIABENDAZOLE

CAMBENDAZOLE

: R = H

0

: R = NH-C-OCH(CH3)2

LEVAMISOLE N

PRAZIQUANTEL Contd.

79

Na03S

^3^~\ >-CONH SOjNa

NH I CO

NHCONH

S03Na

S03Na

Suramin (Hexa-sodium 3,3'-Ureylene-bis-8-(3-benzimido-£-toluido)-

1,3,5-naphthalene sulphonate)

0 Et / ~ \ II /

Me-N N-C-N. \ , Et

CN

H 0 2 C - C - N H - H ( > ^

CI ^NH-C-C02H

DEC (l-Diethylcarbamyl-

A-methylpiperazine)

Lodoxamide N,N'-(2-Chloro-5-

Cyano-m-phenylene)

0=C'

u

' IV^NHC02Mc

— N

Mebendazole (Methyl 5-benzoyl

-2-bensiniidazole carbamate)

o=c

H Nv^NHCO^Me

Flubendazole (£-fluorobenzoyl)-

2-benzimidazole carbamate

Fig. af

80

Table Iti: Spectrum of anthelmintic activity of caimon antinematode and anticestode drugs

Drug/Dose

1

Albendazole

400 mg ( s i n g l e )

200 mg ( s i n g l e )

400 mg ( s i n g l e )

30 mg/kg ( s i n g l e )

800 mg (Multiple)

- do -

10 mg/kg (Multiple)

- do -

- do -

100 mg/kg (Multiple)


400 mg ( s i n g l e )

10 mg/kg (Multiple)

15 mg/kg (Multiple)


Parasite

2

KicoA-U iuirhJiicoidoji,

TAichuA-ii tAickuAa.

SViongyloide^ -^WiocoAaJtU

EnteAobiiu veAjnic.aloA'ii

Co-piZZaAia. hzpcutica.

Hym.nolzpiyi nana.

TajtYua. 'iag-Lnata

0-6tZAtagia o^UeAtagl

Coop^Aia oncophoAa

CoopeAicL zuAnabada.

0p-c6-tfeoAdtc4 vivzxxirU.

VicAocoztium taxiaoMXim

Ancylo^toma daodznalz

TA'ichuALi g£oba£o-6a

¥(Uoia he.p<vtica

TAlchinoJLla. pA<Uido^piAaJiLt>

Host

3

Man

Man

Man

Mice

Man

Man

Calves

Hamst e r s

Ewes

Man

Goat

Sheep

Mice

1 —

Anthelmintic efficacy

(%)

4

93 .3 89.7 89.7

100

100

99

69

92

99.99

99.9

99.9

95 .3

94.46

79

41-50

95

36

Bibliography Number

5

79

79

90

81

82

83

84

85

86

87

88

89

81

1

20 mg/lcg (Mul t ip le )

5-10 mg/kg (Mul t ip l e )

7.5 mg/kg (Mul t ip l e )

2 .5 or 3.8 mg/ k g . ( M u l t i p l e )

2.5 mg/kg (Mul t ip le )

4 . 8 mg/kg

2

VicAocoatium dznciAiticum

k^caAidloi Qolti, RcUttiztincL t£.tA.(igona, R. zckLnobothiicUa.

CoopoAia. panctata, NejiiatocLUcui ipauthlQZA, HazmonckvUt ptojioJ., TAicho^tAongyliUi axlz, T. coZuhAi^oAmii),

W. hzZvztyLanuUi, Oe^ophago-dtomm xadiaXim

HazmonchiU coYitoKtuA, TAicko^^tAongyluA cobxbKi-

GalgzAia ovina, UoA^hallaLgia. nwukaZU.

VictyocauZa^ iiJioAla.

FoUicioia gigantioxi

3

Sheep

Fowls

Calves

Sheep

Sheep

Sheep

4

98.61

100

99-100

'V 99.8

99

63.2

5

90

91

92

93

93

93 (Multiple)

FENBENDAZOLE

50 mg/kg iKldnAjnzlla. ^pVujJLtUt Mice 95.1 94 (Multiple)

7.5 mg/kg StAongyloidU UX^tZAi, > 98.5 95 (Multiple) PaAaAicafii^ zquoAum

50 mg/kg TOKOCXlACi CCUtU, (Multiple) Ancyto-itomcL caninum, Dogs 98-100 96

TAlchuAi^ vuZpi^

10 mg/kg KiCOA-ii -iutun Pigs 100 97 (Multiple)

10 mg/kg CyothO'ittomM, (s ingle) CytLco.Ue.phaiuU,

CyticocycMU, Donkey 100 98 CyticodontophoAiu, CkatiAO'itomuin,

82

1

10 mg/kg (Single)

- do -

- do -

7 .5 mg/kg (Mul t ip le )

- do -



15 mg/kg (Multiple)


7.5 mg/kg (Multiple)

100 mg/kg (single)

- do -

20 mg/kg (Multiple)

35 mg/kg (Multiple)

- do -

- do -

2

TAiodontophoAu^, StAongyixjU) Zqainu6,

S. vuJigaA.u>

HabAonejna. majuut

H. mtucae

0>s>teJita.Qia o^tuJitagl, CoopeAla ^pp.

TA-Lcho^tAongyZwi axoJ.

VicJiocoe-tum ddndAiticum

VoAomphi^tomm -6pp.

faudOA-ola. hzpatlca.

Monizzia -6pp.

StAongylu^ zdzntatoA, TA.icJionejna. 4pp.

Taenia. >6p.

V<jpytidium canimm

ToxocoJia my^ttax Tox<ii,cjaAU> IzoniiM., UncinaAia. ^znoczphaZa

MzX(utAongyZuA 4pp.

GZoboczphaZu^ uAO^ubuiatiu

TJuchuJiti, 4,vuy6

3

Donkey

Donkey

Donkey

Hei fer

Heifer

Sheep

C a t t l e

Sheep

Sheep

Ponies

Dogs

Dogs

Dogs & Cats

Pigs

Pigs

Pigs

4

55.5-100

81.8-100

47.6-100

100

90

99.3-100

87.2

50

100

100 99.7

>90

<50

100

93-100

70-100

0-100

5

98

98

vS

99

100

101

101

101

101

102

103

103

104

105

105

105

83

1

MEBENDAZOLE

5 mg/kg (double)

70-100 mg/kg ( s i n g l e )

10 mg/kg (Mul t ip le )



- do -

1.5 g/10 kg

- do -



25 rag/kg (Mul t ip le )

100 mg (Mul t ip le )


100 mg ( s i n g l e or M u l t i p l e )

25 rag/kg (Mul t ip le )

0 .5 g/kg ( d a i l y )

2

N. da6m4

P, awb^guuA

Ancy£o4toma cxmimm

TA.'idiinziZa. ndl^oni

VicAocoe.tUm dzntAlticxm

VaudcJ-olx hzpatica.

VictyocjCiuiuu> illoAla.

UorUzzia

RaltUztincL Q-cSUnoboth-Kidd, R. cz^tLcAjUu^

Toxocjvia. auU.

TazrUcL taZruazioA.mii,

SubaZwia. bAumpti

Ent&AobiiLi \}QAmlcuZaAiA>

VlcXyocaaia& oJinilzldu.

A^c/VL-u lujihKlcoldu Tt-cciiuA-ci txidnJbjJia. StAongytoido^ MZKCoAotid

Cy^ttixLtKCMAi tznuicoltu

3

Mice

Rabbits

Dog

Rats

Sheep

Sheep

Sheep

Sheep

Hens

Cats

Chicken

Man

Donkey

Man

Lambs

EdunococcoA mitfctc£ocu£aA.o»iice

4

100

100

100

98

90.6

79.4

99

84

100

100 99 50

100

100

75-100

100 100 75

100

90.5

5

106

107

108

109

110

110

111

111

112

113

114

115

116

117

118

119

84

1 2 3 4 5 _

8.8 mg/kg StAOngylvui vuZgoAti, Ponies 99-100 120 (Mul t ip le )

10-100 mg/kg AncyZo^t^toma ce.yl(ini.ctM Hamsters 99.7-100 121 (Mul t ip le )

100 mg NzcatOA OiJieA COJUUS Man 94.9 122 (Mul t ip le )

20 mg/kg OîtVitagicL oêjitagi, (Multiple) Haejnonchui contoAtiu,

TA.ickO'UAOngylvUi ^pp., Sheep 100 12 3

ChahOAtia ovlna.

- dO - 02A>ophago^tomum uena£o4tun Sheep 90 i23

- do - tionizzie. ZXparUZ, Sheep 80 123

TA-LchuALi ovina.

100 mg ( s i n g l e ) TZAnidzni, demimXiU, Taznioi ÔLg-Lnata., Man '^100 124

MonitLÔAmLf) monltiôAmiyi,

- do - Myme.nolzp'U nancL Man '^40 124

FLUBENDAZOLE 600 mg/kg K^COAti, iutvbAlcoidz^ Man 97

(double) 125

300 mg/kg TAichuA-U tAicJliuACL Man 65 .1 125 (double)

5 mg/kg A. COH/Conen^^ Mice 65 .1 125 (Mul t ip le )

50 or 100 mg/kg V. Vlinactl Mice No e f f e c t 126 (Mult ip le) Ht/meno£epc4 Yuxna.

50 mg/kg T. ^plAotu Mice 100 126 (Mul t ip l e )

1.5 mg/kg Oeôphago-&tomum dojfvtatum, (Mul t ip le ) KiCOAlA 4uum, JAldnWlU> Pigs 100 127

4ac4, Ue.t(UtAongyluuii apAl

85

1


- do -

50 mg/Kg (Multiple)


- do -


OXIBENDAZOLE


2

StAongyioidi^ Jiayuomi

StzphanuAvui dzntatu^

Eckino-ittoma. capAorU

G. uAoûbaiattx^

CcLplttaAloL ^pp.

LZtomoôidz^ caA.ou.-c

L. caAiyuA.

3

Pigs

Pigs

Mice

Mice

Mice

Man

Man

4

88

85

100

67-100

25-86

99

85

5

127

127

128

129

129

130

130

15 rag/kg (single)MccW^4 4uujn Pigs 100 131

40 ppm, in food OdôphcLQO'itomxm dzntatwn Pigs 66.4 131 (20-29 days)

10 mg/kg StAOngyloidZ^ UX^tSAl Foals > 99 132 (Multiple)

40 mg/kg K^CjlKdia gatU Chicks 98.4 133 (Multiple)

15 mg/kg TAidlOAZma Ponies 97.7 13 4

(Multiple)

- do - StAOngylvUi vlllgoA-U, Ponies 100 134

OxyuAJU e.qai 15 mg/kg Hazmonchu plaxizi, Cattle > 99 135

(single) TJUcho-tttAongliu axil, CoopzJiia 'lipp. T. cotubAlioAnuu, BunO'&toimm pltUbotomm, OzôphagO'&tomum Koudlatim

15 mg/kg 0-titllAtagia. O^WLtougl C a t t l e 96 135 ( s i n g l e )

http://caA.ou.-c

86

1


- do -


15 mg/kg (Mul t ip l e )

- do -

- do -

- do -



THIABENDAZOLE


- do -

-do -

- do -

- do -

- do -

2

TA<chuAt6 4p .

UorU^zia -ip.

CoopoJiia oncopkoAa (Larvae)

NmatodiALU 4pp.

StAongytoiddyi pa.p>itto-i>Lui

CapiZZofiia. faou-ci

8ano4-tomiun pklzbotomum

TAlchlmlla ^pltoLU

PAob4>tmcLyA<Ji vlvipoAa, PauiaicaJi'Ui zquoAum

TAlchuAi^ txidviuAa

Kiccui-U bxTthKicioiikJi

EnteAobiu^ voAmicalaAi^

TA.ichO'i>tKongyivii, 4pp,

Necctto-t ojnet^anaA

HymznoZzpLit nana

3

C a t t l e

C a t t l e

C a t t l e

C a t t l e

C a t t l e

C a t t l e

C a t t l e

Mice

Ponies

Man

Man

Man

Man

Man

Man

4

> 80

> 71

98

83

94

92

94

100

'V 100

12

78

83

100

50

Not e f f e c -

5

135

135

136

136

136

136

136

137

138

139

139

139

139

139

139

53 mg/kg


50 mg/kg (Multiple)

OsteJitag-ia (UAcumcincta

O&WUajQAjoi o^UeJitcLQA., CoopfUcL 4pp.

StA.ongyluu> vaZgoAtii,

t i v e

Sheep 91

Heifer 100

Horses 100

140

100

141

1

87

50 mg/kg (Multiple)

CytLcocyciu^ naMatwi, Cyatho^tomm coAonatum, C. ccLtLntxXum, Cyticoîte.-phwm^ loYiQibuJucubi^, C. go Mi., Cyticocyciwi imignz, CyaUho-itomum labiatum

Horse No 141 effect

50 mg/kg ( M u l t i p l e )

50 mg/kg ( s i n g l e or d o u b l e )

300 mg/kg

CAMBENDAZOLE






- do -


20 mg/kg

20 mg/kg

15 mg/kg

VAOCUIU^ me(unen4-c4

StA.ongyla6 ^teAcoAotU Ancylo-Ucma, daoduuxJiz

VicAocoztium dzndAitiam

SiAongyloldz^ ux^WU.

P-cctocoeXoun Zanctotatum

AncyZoôma caninum

StAongyloidu ^^ZAcoAatu

VijcAocodUxm de.ndAiticvum

T)iixihoÛfLon.QyluA axzi ChaJboJiticx. ovina

TxidUneMd ^piJuxULU

PaAdicaA'U zquoAum, OxyuALt zqal

Hojzmonchu (lontoAttu

CoopzA-ia. pijcJUnatcL, C. Punctata, Otôphago^^tatum AadiatiM

Man

Man

Sheep

F o a l s

Sheep

Dogs

Man

Ewes

Ewes

Mice

Horses

Zebu

Zebu

100

84 55

< 100

> 99

9 5 . 1

70 .5

95

9 6 . 7

99

100

100

100

100

142

143

144

132

145

146

147

148

148

149

150

151

151

88

15 mg/kg

20 mg/kg

Buno-itomum phtzbotomm Zebu

0-iXeAtagioL (UACumc-mcta, sheep CaplltaAia tongipe^, Banoômum tAigoncQ.ph(ilum CoopeAla. cwiticeJ.

Inffec-t i v e

100

151

152

- do -

- do -

30 mg/kg

OXFENDAZOLE

7 . 5 mg/kg

12.5 mg/kg ( s i n g l e )

7.5 mg/kg (Mul t ip l e )

- do -

- do -

2 mg/kg (Multiple)

- do -

- do -

- do -


- do -

- do -

- do ~

- do ~

TAichoÛfiongyiwi vWiinwi

NimatodiAiu ^paMUgoA

Vixityocatxliu vivipaAoai

HazttzA<jUL4t capiilivi-u

CapiJULoAija. hzpatica

k^cjoJiiAi 4uu»n, Oeôphago-yf^mum dzntcutum

StAongyloidzAi AoruorU

TA.ichuA4Jt >itu^

Chabzfitia ovina

ViatyocauMU) vivipaMju!>

CcqfVLZocaaltu capKQ.oti

MazZleAivu capltloAli,

Buno^&tomum 4 p .

HaemonchuA ^p.

CoopoAia. 4 p .

TKicho-UAOtiggluuS, 4 p .

WZCAAtOCAAJuX^

Sheep

Sheep

C a t t l e

Goats

Mice

P i g l e t s

P i g l e t s

P i g l e t s

Roe deer & Mufflon

- do -

- do -

- do -

Catt le

Catt le

Catt le

Catt le

Catt le

99.6

90.7

99

90

99

100

80

60

75-100

100

70.9

70.9

99.4

96.4

98.3

99.9

100

152

152

153

154

81

155

155

1.55

156

156

156

156

157

157

157

157

157

89

1

4 . 5 mg/kg

- do -

1.6 mg/kg (S ing le )

10 mg/kg

3 mg/kg

2.5 mg/kg

2.5 mg/kg

1.5-9.0 mg/kg (Mul t ip le )

9.0 mg/kg

(Mul t ip le )

2.8 mg/kg

- do -

PARBENDAZOLE


P-CC-tyocoUt tLi iiZoAia. sheep 94-100

MorUez^ sheep 96-100

Tn.ickin<Llia ^p-uiatLU Mice 99

Ponies > 96 OxyaA<y{> tqtxl, St/iongyiwi, vaZgaA-Ut, S. zdzntaiwi, TxlodontjophoKubti, TKicho^tAonglix^ axai

HyoAôngyZiu Aubidiu Pigs 100

VictyocauMjLi vivipajia^ Calves 100

O^Wltag-i/l OÔAtagi, Calves 97-100 CoopÂia oncophoAd, C. mo^mcu>tZAl, C. pe-ctinata, C. panctaXa, Hdjnoitodvujud h(ilv(Ltioinu^, T. longi^picuZoA^^, OeAophogo^tomxm KoAiatum

A^COAIA -6UUin Pigs 99.2-100

MztcutAongyiuu apfii S Pigs 99.5 M. pudzndotzctijUi

HaemoyickLU contoAtwii Goats 82

CoopeAia CUAticeJ,, Goats 91 .3 BuA04>tomm tfiigonocz-phaJtum

Ha£jnondiuu!> contoAtwi, Calves 100 TAicho^tAongytuA colub- Calves 100 AlioAiruA, Buno-itomum Goats 100 tAigonoczphalum, Sheep 100 StAongyloid(Li> paplUo^(ju> Dog 100 ToxocoAa cants

111

111

158

159

160

161

162

163

163

164

164

165 165


UixAjJ/wdioL iaZcA-iiAn Elephant 100 165

90

20 mg/kg (Multiple)

30 mg/Ig active ingradient


41 mg/kg (single)

30 mg/kg

K-iCOAAJt VAXuZoAam Calves Ayicylo^&toma CjOAinum Dog TAldkuALi globaZoMi Goat

O^teAtagicL o^tvitagi Catt le CoopcAia. oncophoAo. Cat t le

A4CaA>C6 4tunm Boar GlobocZfhaMu uAoûbaiataiBoav

A'&caAidUa. numidaz Guinea-fowls

Q^ZAto-qia. CAAcumcinctn, 0. txliiiAcata., Gcu.Q<iAia. Sheep pachy^cztii, ChahiAtia. ovina, hlejnaZodoLu^ •bpathigoji, Ozôphago^ttormm coiwrbianum

98.5-100 66.1-88.7 51.6

96 100

100 59-100

99

94.1-100

165

166

167

168

169

PEBENDAZOLE

1.5 mg/kg (Mult iple)

VictyocjOJuZuA vivlpoAiu, HaijmonchLLi, contoKtiu,

Roe deer 100 & mufflons

- do -

- do -


- do -


- do -

- do -

- do -

- do -

Capi'iofffufi'A f^p^^fff-i, MueMeAAjii caplUoAii,

MoniizlcL

TOKOC/VLOL ca.ni/i

AncuflostomoL canimm

0. lyAdta

HouLmonchiu placzi

NemotodcAoA hzlvztianu^

TîchoAtAongyluA coiubAiioAjniA

- d o -

-do -

Puppies

Puppies

Calves

Calves

Calves

Calves

Calves

60.

75

64

88

71.

64,

85,

96

<99

6

3

.9

.9

.7

.9

170

170

170

171

172

173

173

173

173

173

91

1


200-500 mg/kg (Mul t ip le )


10 mg/kg

100 mg/kg (Mul t ip le)

7.5 mg/kg

- do -

- do -

2.5 mg/kg ( s i n g l e )


LEVAMISOLE



3 mg/kg

6 mg/kg


10 mg/kg

- do -

- do -

2

TcLZrUa ^âg-ouita

StKongyloideJi,

Ki)CjaA-ii> ûaun

KbCjaAidi/i galti

SyngamuA tAocJiZjii, CoupWLaAloi ob^gnaXoL,

CoopQJiia punctata

HaejnoncJuu

Oiôphagoômum JiadUatum

Hztaî>tA.ongyluu> apî

MoifUzzia ^coticz^

A. matay^znî^

TAlcho^tAongyiuu!) axzl, CoopZAia cxpiticzi, NzmvtodUnjUt ^pattigzA,

A.&coA'U

NzcatoA amzJiicanui^

0^6tejLtagla oÛeAtagl

PaAoAcat-cA zqaoAum

TKichonzma 4p.

StAongytiu dzntatwi>

3

Calves

Horses

Pigs

Chicken

Pheasants

Calves

Calves

Calves

Pigs

Calves

Rats

Sheep

Chi ldren

Chi ldren

Calves

Horses

Horses

Horses

4

100

98.5-100

100

100

< 90

75.4

100

100

97.8

91.7

90.77

97 .9 -99 .9

99.99

99.99

99.7

100

9

61

5

174

175

176

177

178

179

179

179

180

181

182

183

184

184

185

186

186

186

92

1



- do -

- do -

- do -


- do -

- do -

0 .6 -2 .4 mg/kg (Mul t ip le )

TETRAMISOLE

10 mg/kg (Multiple)

- do -

- do -


15 mg/kg

10-100 mg/kg

- do -

- do -

- do -

2 3

O^tteAtagia cltcuMuncta., Lambs 0. tJiiiuJicata, TtiadoA^a-gia davtlcuu., TAidio-!>tA.o-ngyiiU) vitAinwi,, T, cotijd)AA.^oKmli, hl^mcutodVLuUD battai

HaimondiuA pZxKKU.

8uno4^mu/n phZzbotamum

Oojiophago^tormm xadiatim

Victyocauitubi vivipa/m^

KdcaX'ii) 4uuff»

JjiickuJiLd 61x16

MeXoitnongyluui, 6pp.

[micJioiiZoAlxz]

VlgoMAvz 6tJLongyi(U>

Victyocaatiu

PA. oto6tAongylu6

K6caA'i6 6aum J Oe4 ophzgo6tomum de.ntatum

HaemoKidiuA, chabzKiMi

Calves

Calves

Calves

Calves

Pigs

Pigs

Pigs

Man

Sheep

Sheep

Sheep

P i g l e t s

Lambs

GanguZzWcakli 6pumo6tcc/uneX£a 6pAAatU -do-

HymeMolzpi^ nana - d o -

4

100

100

98.5

99.6

90.9

100

91

99.5

98.75

85.1

97.1

100

90-100

100

97

90.3

61

Ineffective

5

187

188

188

188

188

189

189

189

190

191

191

191

155

192

193

193

193

193

93

20 mg/Kg Anc£/-£o4^ma caninum, Toxocxfia fixvfiu>, Toxocxvil6 tzorUna

Dogs 100 194

- do TAichuA-Li valpi^, Vipytidium cauoAJUM, Eciru-Yiococcuud gAamiZo4>a^

Dogs Infec- 194 t ive

FEBANTEL




- do -

- do -


10 mg/kg

- do -

- do -


PoAcuccuiAyi, zqaoAwn

TAA.chin.zZta. ipiAOLtUt

HaanoncJuu plauczl, TAichO'itAongylvui axzi, OeMjphagO'&tatim AodlaXum, CoopzAlu pzctinata, C. oncophoACL, NejKVtodiAu^f ^patki.Q(iA.

O-iteAtcLg-ia o^ZAtagl

MonJjz.zia. hzmdini

Hyo-itAongyZtu Aubidiu

VictyocauliLit vivlpoAu^

V. oAndiaMl

ToxocoAa VAJtaloAum

Haemonc/iu4 contoAttJu, O^it&Atagia ciAcumcMicta,

H o r s e s

R a t s

C a l v e s

C a l v e s

C a l v e s

P i g s

Camel

Z e b r a s

B u f f a l o e s

Sheep

100

9 4 . 4

97-100

93 -98

N i l

99

100

100

; 100

99

195

196

197

197

197

198

199

199

199

200


JAinho^btAongyluuii coZixb-AlioAjnli, Oz^pha.gO'btomum vznulo4>um

ContoAtiu oncophoAa Calves 100 201

PRAZIQDAMTEL

30 mg/kg (single)

HzteAophy^^ hztzAophye^ Rats 100 202

http://TAA.chin.zZta

94

30 mg/kg (Double)

40 mg/kg (single)

40 mg/kg (single)

50 mg/kg (Multiple)

25 mg/kg (Multiple)


10 mg/kg (Multiple)

15 mg/kg (Multiple)

25 mg/kg (Multiple)

5 mg/kg (Multiple)

5 or 10 mg/kg (Multiple)

40 mg/kg (Multiple)

25 mg/kg (Multiple)

1-0 mg/kg (single)

1.0-2.5 mg/kg (single)

2.0-2.5 mg/kg (single)

Schi6to4>oma joponlcum Man 94.2

Op<yiÔA^dili> \)l\JViKinl Man 95.8

FcuciolofyiAJt ba6ki

SchLitoômcL mcLruoYu.

CtonoKok-Ld înznM^

Ta£.nia. M.ginata

T. ôLLum

Hyrmnole.pZ& nana

Chi ldren lOO

Man

Man

93.3

100

Calves loo

Man

Man

100

88.5

V. pac-Liicum Man 100

Ecft-inococcoA g>ianuZoâ^, Dogs loo TaejfUa nuttice.p4), T, hydatigzna

SchUto4oma haejmtob<im Children »97

ScJvL&to/iOma bou-u

TazrUa ptu.i(Vimi^, VipytidMun caiumm

Dog 100

203

204

205

206

207

174

208

208

VlphytlobotifiKiim iactum Man 95.3 208

208

209

210

Calves 98.9 2 I I

ViphytZobotlnAium ZfiAjnac<u. Cats loo 212

EdvinococcuA muZtAJtocataA-U Dog 99.9 212

212

95

1




- do -

- do -

- do -




2

Taen>ui tazyUciaioAnuu

Uonizzia. e-xpayua

C. iJa4cco-£a-4^

C. t/LnuicoUjU,

HytnznoZe.pi^ micAo-itoma.

VicAocozLLum dzridKltiam

ToLZYUjx hydoutLge.na

3

Cat

Goat

C a t t l e

Rabbit

Mice

Sheep

Mice

Sheep

Dogs

4

100

100

100

100

100

100

100

92.2

100

5

213

213

n 4

214

214

214

215

216

217

96

anthe lmint ic agent . In the l ight of the p r e s e n t l y mapped endemic

a r ea s of t h i s coun t ry , a compound capable of evoking an t i ces toda l

and antinematodal act ions would be an adequate coverage for achieving

the de s i r ed o b j e c t i v e ,

A ra t ional approach for the development of a wide - spec t rum

anthelmint ic i s , t h e r e f o r e , e s s e n t i a l . In p r i n c i p l e , a b r o a d - s p e c t r u m

anthelmint ic agent should possess the a b i l i t y to en ter ces tode and

nematode p a r a s i t e s i r r e s p e c t i v e of t h e i r t i s sue locat ions and should

be capable of binding e f fec t ive ly wi th p a r a s i t e t u b u l i n s . Yet another

approach to ach ieve the same ob jec t ive is concerned with the s imula

tion of pharmacophores in the molecular a r c h i t e c t u r e of compounds

for provid ing c a p a b i l i t y to in t e r f e re wi th more than one t a rge t s i t e s

of the p a r a s i t e s . Besides microtubule po lymer i sa t ion , energy and

l i p i d metabolism of the p a r a s i t e s have been iden t i f i ed by WHO as

effect ive a l t e rna te t a rge t s i t e s for the development of an the lmint ic

agen t s . It i s , t h e r e f o r e , obvious tha t a knowledge of pharmacophores

is of pr ime impor tance . The e a r l i e r work of t h i s l a b o r a t o r y concern

ing the ident i f ica t ion of p r o p e r pharmacophores r e l a t e s to changes

c a r r i e d out in five different a r e a s (F ig .3a) of the Mebendazole mole

cule and the r e s u l t s ob ta ined have suggested tha t subs t i tuen t s at

pos i t ion 5(6) of methyl benz imidazo le -2-carbamates g r e a t l y influence

218

antinematodal and an t i ces toda l p r o p e r t i e s . Minor s t r u c t u r a l v a r i a

t ions in the subs t i t uen t s at pos i t ion 5(6) has been found to influence

the t i s sue nematocidal a c t i v i t y . In continuation of t h i s work i t was

cons idered of i n t e r e s t to s y n t h e s i z e compounds s t r u c t u r a l l y r e l a t e d

to p ro to types IX-XXII and to evalua te them as b r o a d - s p e c t r u m an the l

mintic agen t s . The a c t i v e compounds could then be eva lua ted for

t h e i r a b i l i t y to b ind p a r a s i t e tubulin and as i n h i b i t o r s of p a r a s i t e

energy metabol ism. The p re sen t s tudy has a l so been ex tended to

obta in compounds s t r u c t u r a l l y r e l a t e d to p r o t o t y p e s XVI & XVII. The

design of t he se two p r o t o t y p e molecules is based on the concept

t ha t d i h y d r o p y r i m i d i n e d e r i v a t i v e s may in te r fe re s e l e c t i v e l y wi th

the r edox reac t ions of the p a r a s i t e energy metabolism and if found

ac t i ve may s e r v e as lead compounds for drug development work .

9.7

(F ig . 3a)

98

^

N ^ N - - C 0 2 M G

MGO2C/ \ N ^ < ^

.CO2H

X

NOH

N

H ^

XII

M G 0 2 C ^ ^ N H

XI

MGO2C'" \ H

XIII

.CO2MG

H

XIV

N

N - ^ N H C 0 2 M G

N ^COpMG H

XV

99

N-Me

XVI XVfl

NH

N ^ N H C 0 2 M e

MGO2C

NHCO2MG

CO2MG

XVIII

H

XIX

R

" ^ ^ C H ^ N H ^ / NCO2MG

NHCO2MG

XX

M G

^ N H C 0 2 M G

M G 0 2 C - •N / CO2MG

-H

/ ; •N :

Ar- -N CO2MG

XXII

CHAPTER 5

5.1 : Syntheses of 4-Benzoyl-N-[ 5( 6)-(2-methoxycarbonylamino)-benzimidazolyl ]pyrrolidin-2-one

5.2 : Syntheses of |3-[ (2-Methoxycarbonylamino)-5(6 )-benzirnida-zolyl ]-acrylic acid. Methyl 5( 6)-[ 5-ethoxycarbonyI-6-methyl-3 ,4-dihydropyrimidin-4-yl lbenziinidazole-2-carba-mate and 2-Methoxycarbonylaniino benzimidazole-5( 6 )-acetoxime

5.3 : Methyl-5(6)-[2 ,5-dimethyl-3-methox year bony 1-N-substituted pyrrolo ]-benzimidazole-2-carbaraates and Methyl-5(6)-N-[2 ,5-dimethyl-3-methoxycarbonyl-4-substitutedphenyl ] -benzi mi dazole-2-carbamates

5.4 : Comparative C-NMR study of tetrasubsti tuted furans and tetra/pentasubsti tuted pyrroles

5.5 : 2 ,2'-Dicarbomethoxyamino-5 ,5'-dibenzimidazol-yl-methanol

5.6 : 5-Ethoxycarbonyl-(2-( l-methylpiperazino )-6-phenyl-4( IH)-pyriraidine and 5-Aryl-6-ethoxycarbonyl-7-methyl-3-oxo-2,3-dihydro-5H-thiazoIo [3,2-a] pyrimidine

5.7 : Methyl 5(6 )-N-[ (2-methylmercapto-6-methyl-4-phenyl pyrimi-din-5-yl )methyl lamino benzimidazole-2-carbamate

5.8 : Methyl-5(6)- l4-(2,6-dimethyl-3,5-dimethoxycarbonyl- l ,4-dihydropyridino) 1-N-substituted benzimidazole-2-carbamate and N ,N-Dimethoxycarbonyl-N-(substitutedbenzyl )quanidine

5.9 : Methyl-4(5 )-methyl-5(4)-[ 3 ,4-methylenedioxyphenyl ]-4 , 5 -dihyd^oiraidazole-2-carbamate

5.9a : 1,6-Bis carbomethoxy-2-methoxycarbonylamino-7-methyl-5-(2^-methoxyphenyl)-4,5 ,6 ,7- te t rahydro- l ,3-diazepine

100

CHAPTER - 5

The synthesis of prototype IX required appropriately substi

pyrrolidine-2-one der i -

o Ph

5.1

tuted

vative as the starting material

and for which a model study

was considered desirable because

substituted butyrolactones after

ring opening with primary aro

matic amines, in pr inciple ,

can recyclize to give compounds structurally related to either 99a

or 100.

100

Benzoyl and (£-toluoyl)- / -butyrolactones (103, 104) , required

for the model s tudies , have been prepared by reacting appropriate

aroyl propionic acid (101, 102) with formaldehyde and hydrochloric

acid. Reactions of 103 and 104 with £-anisidine yielded compounds

which were devoid of IR signals for lactone carbonyls. This suggested

that the lactone ring had part icipated in the reaction and the mass

spectra of the compounds suggested that the opening of lactone ring

must have been followed by a ring closure reaction. In order to

eliminate one of the two possible structures (99a and 100), studies

on the C-nmr spectra of two compounds (105, 106) were undertaken.

The difference in the chemical shifts of methylene carbon attached

to the hetero atom in 104 and 105 (Scheme 19) was of a diagnostic

value for the structural assignment of the reaction product. The

appearance of the methylene carbon attached to the hetero atom in

104 at 69.07 6 ppm and in 105 at 50.58 6 PPm indicated that the

methylene carbon in the reaction products was not attached to oxygen.

This lent support for structure 105 and further support for the ass i

gned structure was obtained from the N signal. The appearance

101

/ « ,

1-Q7 (O)

erx}^' H /H

1Q9

-COOH

1Q1.R=H

102 -R^Me

a". 69.07 103 •.R=H

1 0 4 •R = CH3

"a 5056 b:3fr02 c-35.68 d 171.21 e:5547 f:2165 g:19737

o:5121 b 38A8 c:35J6 d:173A0 e: 55.39 f.2106 g 75.41

a,HCHO/HCi, b,g-Anisidine, c,H2/Pd,C; d,2-Nitro-p-phenytenedianiine,e.H2/Roney-Ni, f,HN=C-NH2/ClC02Me.»)^C-NMR data

S'*'* SCHEME 19

102

of the N signal at 33.689 <5 pprn indicated the amide character of 218a the nitrogen . Catalytic hydrogenation of 105 gave 106 and the

methylene carbon of this compound appeared at 51.27 £ ppm providing

further evidence for the attachment of the methylene carbon with

the nitrogen atom.

These model studies prompted the synthesis of the ninth

prototype molecule namely 4-Benzoyl-N-[ 5( 6 )-(2-methoxycarbonylamino )

benzimidazolyl ] pyrrolidin-2-one (109). Reaction of ^-benzoyl-y-buty-

rolactone (103) with 2~nitro p^-phenylene diamine in presence of t r i -

ethylamine yielded 4-benzoyl-N-( 4-amino-3-nitrophenyl )-pyrrol idin-

2-one (108; Scheme 19). Catalytic hydrogenation of 108 in presence

of Pd/C followed by the ring closure of the resulting diamine with

S-methylisothiouroniumsulphate and methylchloroformate furnished

the desired prototype molecule (109; Scheme 19).

5.2 Unlike the synthesis of IX, the preparation of compounds

relating to prototypes X-XII required the 0-nitroaniline derivat ives

(114, D & 123) as the starting materials, which after catalytic

hydrogenation and ring closure with N ,N-dimethoxycarbonyl-S-methyl-

isothiourea could be expected to furnish the desired compound. Reac

tion of 4-acetamido-3-nitro benzaldehyde (112) with malonic acid

o =c^N-^ OM<2

.C02M<2

X xn

H2N'

NOH

o . ^ M(2

Dl

XI

103

in the presence of c a t a l y t i c amount of p y r i d i n e gave 4-acetarnido-

3-ni t ro cinnamic ac id (113; Scheme 20 ) . Alkaline h y d r o l y s i s of t h i s

compound y i e lded the d e s i r e d s t a r t ing mater ia l (114; Scheme 20 ) .

Pre fe ren t ia l hydrogenat ion of n i t ro group in 114 was p o s s i b l e in

p resence of Raney nickel but i t fa i led for the s t a r t ing mater ia l (D ) .

o

112

H-o=9

OMe

N-</

115 114

a , malonic a c i d , pyridine/MeOH; b , 10% a q . NaOH/MeOH;

c , H , Raney-Ni/MeOH; d , HN=C-NH,/ClCO,CH, SMe

(Scheme 20)

The reac t ion of 4 -ace tamido-3-n i t ro benza ldehyde (116) wi th e t h y l -

ace toace ta te and th iourea under ac id i c condit ion gave 5 - e t h o x y c a r b o -

ny l -6 - r ae thy l -2 -oxo -4 - ( 4 -ace tamido-3-n i t ropheny l ) py r imid ine (117;

o . . - ^

AcHN

a , CHjCOCH^CO^Me, Urea; b , 10% aq.NaOH;

c , H-, Raney-Ni/MeOH; d , MeS-C=NCO,Me '2* NHCO^Me

,N-

OM(2 H NH

M<2 118

(Scheme 21)

104

Scheme 21) which on a lka l ine h y d r o l y s i s y i e lded 118. Hydrogenation

of t h i s compound in p resence of Raney-Ni furn ished the d e s i r e d d i

amine. The diamines p r e p a r e d from 114 and 118 were cyc l i zed by

react ing them with N ,N-d ime thoxyca rbony l -S -me thy l i so th iou rea to

obta in the d e s i r e d p r o t o t y p e compounds 115 and 119 r e s p e c t i v e l y .

Since the p r epa ra t i on of diamine D was not p o s s i b l e , 5 ( 6 ) - a c e t y l

benz imidazole-2-carbamate (124) , p r e p a r e d by the method r e p o r t e d

e a r l i e r , was reac ted wi th hydroxy lamine under cont ro l led condi t ions

to furnish 125 (Scheme 22 ) ,

NOH

AcHN

H 2 N - Y j ^ - ^ ^ M e

D2

Q

AcHN

120

122 121

O2N

123

^ Me02C^

O

124

N

MeOpC^ ^N H

NOH

Me

^

< -

125

a , AC^O; B, Fuming HNO^; c , 10% a q . NaOH/MeOH; d , H , Raney-Ni/MeOH;

e , HN=C-NH2/ClC02CHj; f, NH^OH/MeOH

SMe (Scheme 22)

105

5.3 In o r d e r to s tudy the effect of NH and NR in p lace of the

oxygen atom in the furan r ing of a promising anthelmint ic compound

83/148 (CDRI Code n u m b e r ) , t he syn thes i s of XIII was under taken

and as an extens ion of t h i s s t u d y compounds r ep resen t ing p r o t o t y p e

XIV were a lso s y n t h e s i s e d .

O M G H

CO2MC

83 /148 X=0

X111X=NH,NR XIV N'' f>IH

o=c OMe

The syn the s i s of XIII (X=NH) owes i t s or ig in to the o b s e r v a

t ion made during the upscal ing s tud ies of the compound 83/148. It

was o b s e r v e d tha t during the p r e p a r a t i o n of 2 , 5 - d i m e t h y I - 3 - m e t h o x y -

ca rbony l -4 - ( 4 -amino-3-n i t rophenyl ) furan, ( the r e q u i r e d in t e rmed ia te

for the p repa ra t i on of 83/148) was i n v a r i a b l y a s soc i a t ed wi th a

small amount of a r e d compound formed in the reac t ion mix tu re .

This was ident i f ied as 2 , 5 - d i m e t h y l - 3 - n e t h o x y c a r b o n y l - 4 - ( 4-amino-

3 -n i t ropheny l ) p y r r o l e . The formation of t h i s p y r r o l e d e r i v a t i v e

along wi th the cor responding furan in the react ion of 126 in p resence

of methylace toaceta te and p i p e r i d i n e suggested tha t a careful s t udy

of t h i s reac t ion may furnish a reac t ion condition in which the p y r r o l e

could be obta ined as the predominent p roduc t . This prompted to

s tudy t h e Nef react ion of 126 in presence of p r i m a r y , secondary 219

and t e r t i a r y amines and the r e s u l t s were monitored by HPLC

The reac t ion of 126 wi th methylacetoaceta te in p resence of

p r i m a r y amines gave N-subs t l t u t ed p y r r o l e d e r i v a t i v e s (132-137;

Scheme 23) and the cor responding furan d e r i v a t i v e s could not be

de tec ted in the reac t ion m i x t u r e . This method has a p r e p a r a t i v e

value and g ives be t t e r y i e l d s of the p y r r o l e d e r i v a t i v e s as compared

to the method r e p o r t e d in l i t e r a t u r e which i n v a r i a b l y gave poor

y i e ld s of 2 , 5 - d i m e t h y I - 3 - m e t h o x y c a r b o n y l - 4 - s u b s t i t u t e d phenyl p y r r o

les in our hands . The r eac t ion of 126 and methylace toace ta te In p r e

sence of secondary amines gave different percentage r a t i o s of 128

and 130.

106

129 : R = CI

a 131 : R = CI

CH3

CO I — CH2C02Me

126 : R = NH.

127 R = CI

W

128-129 Nl/

H2N

O2N CONHNH2

CH3 CO2CH3

148 :

148a

^ = NHNH.

R = OMe

a , P i p e r i d l n e ;

132 : R = n - B u

133 ; R = CH^Ph

134 : R = 4 - a n i s y l

135 : R = 3 , 4 , 5 - t r i m e t h o x y p h e n y l

136 : R = 4 - a m i n o - 3 - n i t r o p h e n y l

237 : R = ( C H 2 ) j - N ( C 2 H ^ ) 2

b . P r i m a r y a m i n e ; c , T r i e t h y l a m i n e ;

d . H y d r a z i n e h y d r a t e

(Scheme 2 3 )

107

o IJJ U z UJ in UJ cc GL

z o t-u < UJ

a: UJ

z u. o z z < X u UJ

UJ

z

> •

<

cc U)

CI o u

c o u

z x

i ,0 o

W^ o

@ 2 - 0 4 ^

Q::

O

u X

l O

H cc

X u

W [ / 2 O

o

X

r» X

o u X U

cr o u X

u w

K»

X

109

CO

c o U

110

LL)

z < >-

<

cc a.

O B «)

u

I l l

For example in presence of piperidine the percentage ratio

of 128 and 130 was 96:4, in morpholine it was 80:20, in N-methyl-

piperazine i t was 89:11 and in presence of N-phenylpiperazine it

was 87:13. Replacement of piperidine with triethylamine in the reac

tion of 126 and methylacetoacetate gave only 128 and the presence

of 130 in the reaction mixture was not detected.

Recent studies on the mechanism of Nef reaction indicate

the formation of an intermediate 138 (Scheme 24). It is l ikely that

reaction of 126 and methylacetoacetate in presence of triethylamine

also yielded an intermediate structurally similar to 138, which after 220 elimination of H O and HNO as suggested by Boberg , yielded the

tetrasubsti tuted furan. In the presence of piperidine the reaction

of 126 and methylacetoacetate possibly proceeds in three direct ions. 221 The formation of intermediates 138 and enamine (143) possibly

c6ntribute towards the formation of tetrasubstituted furan. The th i rd

pathway might involve the addition of water in 140 which leads

to the ring opening and recyclization of the open product to give

tetrasubsti tuted pyrrole (Scheme 24). This pathway possibly becomes

more facile in presence of primary amines and therefore leads to

exclusively N-substituted pyrrole der ivat ives . Despite a successful

synthesis of N-substituted pyrrole derivatives (132-137) , the synthe

sis of 130 by Nef reaction of 126 in presence of methylacetoacetate

and ammonia was not at all promising due to extremely poor yield

( —10%). An alternate preparation was, therefore, sought and this

prompted the reaction of 126 with 144 (Scheme 25) in different sol

vents. The yield of the compound 130 has been improved upto 90%

by using pyridine as a catalyst and methanol as a solvent.

Me-'^NHR 126 M G ^ ^ N R

1 4 4 : R = H 130 : R = H 1*5 ! R = CH3 146 ; R = CH^

(Scheme 25)

112

Based on t h i s o b s e r v a t i o n , a t t empt s were made to p r e p a r e 2 , 5 - d i m e -

t h y l - 3 - a c e t y l - 4 - ( 4 -amino-3-n i t ropheny l ) p y r r o l e by reac t ing 126 wi th

enamine 147. However, i t fa i led to give the d e s i r e d p roduc t . It

w a s , t h e r e f o r e , cons ide red of i n t e r e s t to a sce r t a in the reason which

led to the fa i lure of t h i s r e ac t i on . One of the logical explana t ions

for the fa i lure of 147 to r eac t wi th 126, would be the lack of a d e

quate nuc l eoph i l i c i t y of the carbon csC to the ace ty l g roup . In p r i n c i

p l e , exper imenta l ev idence for the same may be obta ined from the

chemical shif t of carbon atoms s ince subs tan t ia l difference in nucleo

p h i l i c i t y of the carbon atom c< to methoxycarbonyl and ace ty l groups

in enamines 144 and 147 i s bound to make a s ignif icant difference

b ^0CH3

H3C

• a = b = c = d = e =

^ \r

49.48 170.72 81.56 162.59 18.76

a b c d e

= 28.97 = 196.17 = 95.33 = 162.68 = 21,93

13 C-NMR data

(Fig . 4)

in t h e i r chemical s h i f t s . Th i s prompted a compara t ive s tudy of the 13

C-NMR s p e c t r a of 144 and 147. As would be e x p e c t e d , the carbon

o< to the ace ty l group in 147 was d e s h i e l d e d by almost 14 ppm. This

suggested t h a t the enamine carbon in 147 d id not pos ses s adequa te

nuc l eoph i l i c i t y to r eac t w i th 126. The p r e l im ina ry data of the C-

chemical sh i f t of carbon in enamines 144 and 147 tempted to s tudy

the chemical sh i f t s of carbon

atoms in t e t r a and pen t a subs t i t u t ed

p y r r o l e s (154-158 and 158a-158b).

These p y r r o l e d e r i v a t i v e s may

be cons idered as the E-configura-

t ion (F ig . 5) of enamanine in a | r i g i d geometry . Since p y r r o l e s E-Configuration of

enamine e x h i b i t he te roaromat ic c h a r a c t e r (Fig . 5)

13 wh i l e furans fai led to do s o , t he compara t ive C-NMR s tudy of

..'CO2CH3

H^C

113

tetra or pentasubstituted pyrroles (154-158, 158a, 158b) and tetra

substituted furans (a-q ) was considered interesting. The te t ra-

substituted pyrroles (154-158) were prepared by reacting enamine

144 with appropriate nitrostyrene derivatives (149-153; Scheme 26).

The details of th is study are presented in sec. 5,4.

149

150

151

152

153

144

= R = H

R = 3,4-dimethoxy

R = 3,4-methylenedioxy

R = 4-benzyloxy-3-methoxy

R = 4-acetamido

->

154-158 Substituent R correspondence to 149-153

(Scheme 26)

The next phase of the studies on Nef reaction was concerned

with the reaction of 126 with methylacetoacetate in presence of hy

drazine hydra te . Mechanistic considerations of this reaction suggest

the formation of 148a but the excess of hydrazine hydrate possibly

gave 148 (Scheme 23; page 106 ). Preparation of compounds belonging

to prototype XIV required N-substituted pyrrole derivat ives (171-

175) as the starting material. These were obtained by the reaction

of 166-170 with methylacetoacetate in presence of 2-nitro-£-phenylene

diamine (Scheme 27). Hydrogenation of 130, 132-139 and 171-175 in

presence of Raney-Ni followed by their cyclisation with N,N-dimethoxy

carbonyl-S-methylisothiourea yielded the compounds structurally rela

ted to prototypes XIH and XIV respectively (Scheme 26 & Scheme

27).

5.4 Comparative C-NMR study of 144 and pyrrole derivat ives

154-158 revealed two interesting features. Despite the fact that pyr ro

le has a heteroaromatic character , the C-2 in compounds 154-158,

which corresponds to carbon d of 144 (Fig. 4; page 112 ) , appears

as a shielded carbon while C-3 in 154-158 which corresponds to

114

130.132-137 qb

Me02C

a , H , Raney-Nt/MeOH

h, MeO,CN=C-NHCO-Me 2 / 2

SMe

159 ; R = H

160 : R = CH^

161 : R = n-Bu

162 : R = CH Ph

163 : R = 4-OCH^-Phenyl

164 : R = 3 , 4 , 5 - t r i m e t h o x y p h e n y l

i •" = tty 166

H

NHC02Me

R = (CH^)3-N(C2H^)2

(Scheme 26)

166a

167 !

168 ;

169

170

Mo- HMe? Lv_y.

R = H

R = 3 ,4 -d ime thoxy

R = 4-methyl

R = 4-acetamido

R = 4 -benzy loxy-3 -methoxy

a , 2-Nitro-£^-phenylene diamine

b , H , Raney-Nl/MeOH c , MeO-CN=C-NHCO,Me

2 / 2 SMe

Subst i tuent R co r r e sponds to 166a-170

Subst i tuent R co r r e sponds to 166a-170

N ^ N H C 0 2 M G

(Scheme 27) 176-180

115

carbon C in 144 (Fig.5; page ) appears as deshielded carbon.

This may be explained by assuming that 144 in solution possibly

exists as 144a. This is why the

methyl carbon in 144 appears K-) t ^ "

downfield than the methyl carbon

attached to position 2 in comp-

ounds 154-158. 1 4 4 Q

The comparative study of the C-NMR spectra of te t rasubst i -

tuted furans and tetra or pentasuhstituted pyrroles (Tables A.B)-/as

concerned with the chemical shift of C-Z, C-3, C-4 and C-5 in these

class of compounds. As would be expected C-2 and C-5 in furans

appeared downfield than the corresi^onding pyrrole der ivat ives . How

ever , C-3 and C-4 may be expected to show different chemical shift

in these two class of compounds, because furan lacks heteroaromaticity

while pyrrole possesses the same. Surprisingly the chemical shift

of C-3 and C-4 in furans are marginally different from those of py

r ro les . This would also explain that effect of substituents on the

phenyl ring on C-4 in these class of compounds remains unaltered.

The effect of substituents on C-3, the chemical shifts of C-2 in furans

(a-q) indicated that a methoxycarbonyl or an acetyl residue predomi

nantly deshielded C-2 while an amide function led to a shielding

of almost 9-10 ppm. In continuation of this study the observed

chemical shifts of C-2, C-3, C-4 \ C-5 in furans and pyrroles were

compared with the theorit ical values computed by a computer for

representative compounds (Table C).

5.5 The synthesis of next prototype molecule (XV) namely 2,1'-

Dicarbomethoxyamino-5,5'-dibenzimidaaolylmethanol is based on the

presumption that the carbinol of 82/437 (CDRI Code number) in bio-

phase may slowly get oxidized to the parent compound which has

exhibited significant broad-spectrum anthelmintic ac t iv i ty . In a situa

tion such as this the carbinol may exhibi t longer duration of action.

MG02C-"' \ N ^ L ^ l ^ ^ y ^ N ^ C02M<2

82/437: X = CO XV:X= CHOH

116

to

c Id u b

•a

m 3

1-4

o H

« o

(0 U

tn 3 O • Id >

o

Id

e

•a H

(0 C o X) t-4 (d U )-• (U J3

X u

I i n

•X,

u

in

U

U

I O

I U

o;

i n t~

. - ; rH -O i n

i-t O

o- NO U l IM <NI I « ,

f\J ro

>o Cs] i -H

i n

o • t -

o f - H

* o CO

• fo >£)

^ ^H

• o i n

• N O

^ • o

o

<3D m °o •"• * ^ O fVJ O

• m o - r o • _ . vO in in ^ in

CO • *

i n

00 00

po m X X

u ul (M fM

o,o u lu

r-4

^ «o ^

I -H

p j <M

^ I—t

i n <=> U l •J-

X X O u Ul r>J o o u ulu o

m u l •-• (J _ _ ,->

X X X u u lu

<M (NO O O X u l u rvj

I-H O

r - .-H

• ^ <N1

o o («1 fM

.O 00

i n

00 i n

ic a z E o u

o

o

i n

X U o u

o i\j

in 00

o 00

X u o u

I V c 0)

r—* >» •M 0)

E

oo

o fM

00

i n

X u

fv o u

»< o J3 •J V E

t--

o (M

PO P ^

i n

X u o u

O

po "O

X u o u X 2 I

117

O '^

u I o

•4->

E o u

o I i n

X U

81 2

O^ I—I t— sO ^ r o NO (^ li^ i n

• • • • • o t ^ t ^ f* ^ m f M f M <-! vO

I

O f M f M >-l sO c\ n-t 1-1 i-H r H ^

m 0 0 ^ t ^

t ~ t ~ >o "* N fvj

CO

0.00

X ul o u X

- <=> o — i n ^

ol I o

^ X

, O

"a" o >o vO ( M ^

i n (va • * rsj i-H vO

(\r~ o O MM U == J

rsj i-H vO ( - ) .-H i-H i - l " - '

o

^ Ul X U

o ^ S O

i n (vj rs) r j o * pn psj <M

0 0 0 0 t ^ fvj < U l -o 2 TT -v] r j 04 - o t - <M tr ^ CO Q

• • • U m i n <*i . . ^ m fM vO

,— NO 00

<M O

m r o i n f—

Ul I o +-•

• nj

E . O

l-i

<

X u

X X """ u u U UlU m — >-- i n

0 0 CT-vO CO

1 " - O f l ( M

o o I - l 0 0

Ul I .

0 0

X U O "^

o i n i n

o (M

o o

CO CO 0

o

f M

i n

X u o u

° S 1 — O

a V 0) E

£> I I en

• * I

i n

o ( M

O <M

i n

X U

PS o u

1

>. X 0

0) 1

>. X 0 J3 • » - •

E 1

I M

i n i n

vO ( M

i n

X u (VJ o u

o xi + j «) E •a

H)

0 0 i n

<M

00 i n

X u o u

o u o

o I

1 <

118

^ ^ 00 m <-i t~

p l r^ f O r g 1—4 r-H

O 0 0 in o^

(M t^ m (M I - l 1—1

ry oo

O^ 0 0 1 " (VJ

f o

• 1 0 0 ( M m O

^O •—(

O^ (NJ

Ul 1

u ^ 4 j c n

(4 IC E U O ^ ( V ^ iH O . f ^

< UlK,

«k

<M i n

• 1—1

m r—1

O^ 0 0

t—*

INI 0 0

PO

"*

i n i n

« <7> O

• 0 0 i - H i -H

t>-0 0

1—«

r g t—(

o t n

-O Psl

•. 1 i n f\4

1" O

^ • ^ vO f - H

t^ 0 0

. o

Ul 1 ^ o .^

4J f >

B U O ^rvr^ »H O f ^

< u|3:,

* <?• f o

• 0 0 ( M F-H

cr

• — 1

o

m CO

<M O

i k

t ^ O -

• >o r j r—i

f~

( M 1—1

OO (NJ

» ^^ 0 0 f n

a- K • Ul

28 • — •

i-H 1—1

U l ^ 1 0 .

••H

0 y u 0 , <: ul

* 0 0 0 0

• vO ( M 1—1

i n > 0

1—1

• f ~ i *

pn 0 ^

« i n 0 0

• r-0 r -H

a> 0 0

0-0 1 — *

i n 0 0

I M <N]

* *^ ( ^ <*^ 00 X

• Ul Su

-

0 . 0 •^H ^ - . •

n» -Tin 0 y • ^. 0 , 0 < U 0

* 0 0 0 ^

• i n <M i -H

Csl

0 0

<-H

f - t

« 0 0 'a*

vO I -H

•

o» .-H

• f—1

I -H

»-H

PO ( M

^H I -H

i n 0

• f M f M

•k - - ^ . —

0 0 r o ro • 0 ^

r g 1—(

po 0

0 p n I -H

• t— PO

0 0 fVJ

* r~ i <

" o^ ^H I -H

t -

1—t

I-H

i n

• 0 0 (Nl

•• 1 -H 0 <=> u • " >o ^^

<j -i -H

N O

' T

. 0 - - ^ P^

Ul 1 0 .

4-1

•**» r^

a; u 0 ul X ^

• U 0^ 3C

:2 2 (0 f ^ ~

H f ; - in 0 y u 0 0 ^^ ^j4 '^r ^ ^ ^ ^ ^^

ij* t— m I-H in o in •<a< v O vO c- r-f~ o^ (\] (M rg (M i-i I-H (\j <vj rg <\i

t^ pn in (T-

fvj 0 0 o o ft (M e g <M pg

00 * rg I—I o~ ^ " t^ sO in in o c^ " in in in >o in in in in in in

pn m X X CO pn pi pii

U U X X X X

o' o' 8 8 8 8 u u u u u u

o § 0 H -^ a e X -o u S E ^ I O I X I I I

pg ^ po

V c: 0)

I -H

^ +J

2 >• e X 1 0

* - w ' T3

pp^ 1

S>s

X 0

J3 4->

0

E "O

1

^ p n

I

119

• • • • r< -^^ i n O* m C^ >0 GO r»T5^ O^ fNj "q* 00 • nC •

• • • • l A f ^ l i T i vO t^ • « f n O X ' ' m (VI (M >-< i-H J-^

• i n

o^ 00 t~ >o o o to • o o

• • ^ • , CO - O N O C^ ^

fo o o in i n I—I 00 (\j

»o oo "T fs] m fM O %0 ^ (M

o o

in r-i o o ,^ r - t ^ 0 0 <\j . "

«' Jri ;d -' ^ ' ° ^ (\J (M •—4 I I—I I—I I—I I—-H

C^ OO NO rsj

u o

u

O^ 1-1

00 r- c j l fV) (M I ^ - • (J

. r-i ^j rr

. (»i 00 U I 0) c

M (M <N 0 00 Ql ^r o

in o

I—I ^-^ >-^ f s j ^

E * o • • O ';^ rj<

^ (\] < ( J i n 1-4 <-H ^-^ •—' (M

I

c

a o

(M

o 00 O

in in

i n

o in

i n

(M

m

o

in CO

00 o^

m in

00 00

X U o u

0 O 0 = 0

I ono I

(N) >- o c •<-'

^ E I I

I <D C 0)

4-1 4)

E >. I X

•V O • 1 ^

120

w (U

I—*

o u u a,

in

3 (0 ta + j

c a. T3 C n]

(0

H

V) C o

(d u (fl 3 O

>

o (0

X!

<4 O

E

CQ

«

ztr

C o t-i

u u

x:

X u

I i n

U I

i n I

U

I U

I

u

(M I

u

oc

o a E o U

. . . I t— vO vO OJ

(\j vo 'J' o • • • n

r- I" vo 3 , (NJ rH \ 0

.-H O

O i n ( j |

•O CO O^ CT^ rg fO

0» t -

O fM u a: — u

o

»o >o o* O <?• (M

o o >o IN O >0

O f

in

e u

t^ fvj 0 m o <

•-I '^ O .^ ^^

o in c

• * <v < U\^, ^ ^ ^ — Ul

t^

^

o

• ^

00

00

(M <M

O

TT r-

ivj

I ' 00

CO

i > 00

in CO

CO in

M

o

CO

PO

• i n

• vn *—^ - ^ -s

m po I K U * U

fM— O o. ' x B'gi::i

i n ro 1—1

*

fNJ O 1—1

o o

fM

o

fM

00

fM

o

o

PO

Z

^r^\

X 2

I V a

>>

E >• ^ o

X 2

O J5 •»->

V

E

X 2

to

U X 2

I

00 lo

121

•k «

(M (\J O ' T

O O r1 O r-i >-H

» 0 0 ro • (J- (J

-• i 4^

I 1 1

fvc: o O 2 u —

0 0

" .« . in o f^

00

o 1—1

m

3 E U -• o O >H

m CJ|E,0 K, — o lu l u l

rsl in

r - l

» i n c«-

r-l

0 0

, 0 0 <sr f - i

o

0 0

r-

0 0

»—1

« o rr,

r H

OO

O^ »M 1—1

1—1

^ ( M r H

« ^ o

1—1

0 0 ( M 1—1

m 0 0

f t

• - H

* m rg

i n

r H

o

rg rg 1-1

^ fvr^ pg O "^

:2 — u Tf ^ ^ 1—I

• . O fvj

1 »•"< <J ^ " ^ .4^ rn

O f M ^

U o o^ O N

^ o, ' • < ulfc ^ ^ u

m i n

* ro 0 ~

0 0

1—1

I k

0 0 pg

1—1

00 i n

o^ m 1—1

r-l pg

* in I—1

o

1—1

« m o

1—1

1—1

m CO 1—1

« O rg r>-vO •

rg rH r H

* ^ m

i H " ^

00 00 00 CT-

• • 00 Tl-rg rg i-i 1-1

O cu U r j — X.

ul rg

o in o

. . « . . I ^- o^ in r- a* rg t— o "a* t~ o^ O 00 o^ >o r~ in i £ ^ ro rg rg iH >o _ , , , , , f

X u o

P*^ i H

«k »

rg ^ i n f-i

00 rsi

. o

CJI

•H _ -

E U

0 r^

00 in rg rg

Ul I U

—i •M

(0 E o u <

o

CO

00 i n

r-

i n i n

r-> rg

i n rg rg

o 00

rg rg

i n rg

rg rg PO

o

rg

o

P<1 pn

po

PO

2

rg

a: X!

I

2 = ^ o

rg

>» X o >. ta d » XI

^

X o X!

4->

t) tl

m

.5 ^ E;:; « c I I

^ pn

•H IH

E -^ Id c

4. J,

^ • 1 in s CO

122

J 3 u

•a u -»^ E 4)

u

V

+1

u

X

a . o z u

o +1

ul o

o o

tn o vO +1

00

• o rt +1

i n (vj

o o j ro +1

Ul I U

<

(M t ^ rg r -

00 O 00 O fVJ +1 (M +1

N O 1-t

00 O (M +1 pg +1

oo o r j +1

M

02 Ul

o +1

ro • ^ rH

in •

00 'T

O •

o +1

""^ - m •

•X) +1

* tn t

ro FH

pg eo • <o *

0 \ o^ M +1

in o ^ +1

pg

m

^ o«

oo o m +1 i n

t~- 00

pQ +1

00 ^ O^ +1

in +1

in fM (M t~-sO +1 r<i ^^

• • O (M m +1

oo o (M +1

Ul 1 u <

t-^ UU

OO O rvj +1 »—i —

(M

PO • m 1—4

in • 00

Ti<

in •

(M +1 —

_ in •

o +1

* 00 •

r-l t-l

in in t

m *

t o

w

eo

eo

<N

X o

123

r\j t»

00 O oj +1

CO ir> CO r -

O O CO O • * +1 <M +1

CO O CM +1

(M in (M r~

O O 00 O "* +i rvj +1

oo o rs] +1

(M +1

- < 00 ^

00 O "^1

r-H - ^ O

Ul I

<:

O^ r-4

m +1

^ 0 i-H

(M +1

<y- ^

m +1 u o

ul 1 l-l

<

O^ r H

PO r-1 f O +1

N O 1-4

r~ i-i (M +1

o -^

m t—1

m +1

. - t o *

o o >o +1

« «

— oo rH O 0»

• • I v O 1-1 Vf» •O +1 >0

o

• i n r^ +1 m +1

^a

0 0

• m r-l

«o • ( M

(Vl

O

• o +1

o « o

<M

+1

^ ««" • en

1-4

eo r* • f O

rg

(NJ

« 1—1

1—1

( M

* 0 0

(*>

1—1

• o +1

o • I *

1—1

+1

oo m * i-t

i - i

ro tn

f

\r\ fM

1—1

• o fM

00 •

<NJ I—1

+1

flO OJ

• «NJ C4

I - I t

o f M

0 0

• f M i - «

+1

eo 0* • *

fSJ

00 m

^ +1

o • 0 0

o

.*-> m • o

+1

M m • e

i H

( M O O

ro O •^ +1 fO

0» f l

O O CO +1

2 u 2

S

8 8

124

It was a lso cons idered d e s i r a b l e to eva lua te the ca rb ino l as a chemo-

p r o p h y l a c t i c . The s y n t h e s i s of 183, r e p o r t e d e a r l i e r in t h i s l abo ra

to ry was s l i g h t l y modified to obta in a be t t e r y i e l d and the main

sequence of reac t ions a r e d e s c r i b e d in Scheme 28 . The reduc t ion

of the carbonyl group in 183 was diff icul t but large e x c e s s of sodium

b o r o h y d r i d e he lped in obta ining . 184. Hydrogenation of t h i s compound

followed by ring c losure wi th N , N - d i m e t h o x y c a r b o n y l - S - m e t h y l i s o t h i o -

urea furnished 185 (Scheme 2 8 ) .

a O2N

• ^

NO2

181 182

184

M G 0 2 C - ^ ^ N H

183

^C02M(2

185

a . Cone. HNO ; b , a q . NH ; c , NaBH4/MeOH; d , H^, Raney-Ni/MeOH e , l/IeO,CN=C-NHCO,Me

2 I SMtt

(Scheme 28)

125

5.6 As an extension of our exploratory research act ivi t ies for

identifying new molecular structures associated with broad-spectrum

anthelmintic ac t iv i ty , the syntheses of prototype XVI-XVII were consi

dered of interest . The starting compounds 192-197 were obtained

by Biginelli reaction.

Et02C

H / \ Nv^/N N-Mc

II "^^ N

Me

XVI xvn

Acid catalysed reaction of aromatic aldehyde with ethylaceto-

acetate in presence of urea to yield 5-ethoxycarbonyl-6-methyl-4-

phenyl-3,4-dihydropyrimidin-2-one (202) , commonly known as Biginelli

reaction , was reinvestigated by Hinkle and Hey and it was

observed that the replacement of urea with thiourea led to the forma

tion of the corresponding 3,4-dihydropyrimidin-2-thione derivatives

(192-197; Scheme 29).

Before undertaking the synthesis of dihydropyrimidine proto

types XVI and XVII, model studies on the alkylation of 5-ethoxycar-

bonyl-6-methyl-4-phenyl-3 ,4-dihydropyrimidin-2-thione (192) , were

considered as essential prerequis i tes . Reaction of 192 with methyl

iodide under various experimental conditions was studied. This reac

tion in presence of sodium hydroxide in methanol yielded 198 (Scheme

29). Earlier workers have prepared th is compound by reacting

5-ethoxycarbonyl-2-methylmercapto-4-methylpyrimidine with phenyl

magnesium halide and the spectroscopic data of the reaction product

reported by them agreed well with that of 198. Reaction of 192 with

methyl iodide in presence of soidum hydroxide in DMSO gave 190

(Scheme 29). Deraercaptation of 199 with strong acid gave 201 (Scheme 225

27). The U.V. absorption maxima at 290 nm supported the assigned

structure. Finally the reaction of 198 with N-methylpiperazine yielded

the prototype molecule 200 (Scheme 29).

126

Q

186

187

188

189

190

191

:

:

:

:

:

R

R

R

R

R

R

=

=

=

=

=

H

3 ,4 -d imethoxy

4-niethoxy

3-niethyl

4-acetaniido

4-n i t ro

\l/

M G

,NH2

K ^NHo

Subst i tuent R co r re sponds to 186-191

\l/

H r-\ N x ^ N N M G

11 ^ N

M G

201

M G

202 a, CH^I, NaOH/MeOH; b , CH I , NaOH/DMSO; c , N - m e t h y l p i p e r a z i n e ;

d , H,SO^/MeOH 2 4

(Scheme 29)

127

In the l ight of t h e s e obse rva t ions a l k y l a t i o n of (192-195)

with monochloroacetic ac id in presence of potassium h y d r o x i d e in

methanol could be expec t ed to y ie ld the p ro to type (XVII) molecules

202-205 but the reac t ion of 192, 194, 195 wi th monochloroacetic ac id

in presence of potass ium h y d r o x i d e y ie lded 205-208 (Scheme 30)

whi le the reac t ion of 192-195 wi th monochloroacetic ac id in presence

192-195

rile COOH

202 :

203 :

204 :

20'5 :

R = H

R = 3 ,4 -d imethoxy

R = 4-OCH^

R = 3-CH,

206

207

R = H

R = 4-OCH.

208 : R = 4-NHCOCH.

a , CICH CO H, BF .Et^O/MeOH; b , CICH^CO^H, NaOH/MeOH

(Scheme 30)

BF e t h e r a t e y ie lded 202-205 (Scheme 30) . The cyclocondensat ion

of 192-196 wi th monochloroacetic ac id i s expec ted to y ie ld two i so

meric b i c y c l i c s , 5 - A r y l - 6 - e t h o x y c a r b o n y l - 7 - m e t h y l - 3 - o x o - 2 , 3 - d i h y d r o -

5H-thiazolo [ 3 , 2 - a ] p y r i m i d i n e s (202-205) and 7 -Ary l -6 -e thoxyca rbony l

5 - m e t h y l - 3 - o x o - 2 , 3 - d i h y d r o - 7 H - t h i a z o l o l 3 , 2 - a l p y r i m i d i n e s . Of t he se

the former could be i so l a t ed from the react ion mix tu re . The s t r u c t u r e

of 202-205 was suppor t ed by t h e i r PMR data (Table 24 , page n ^ )

which r evea l ed a downfield shi f t {—0.70 S ppm) of the methine proton

at posi t ion 6. Th i s can only be exp la ined if 3-N i s invo lved in r ing

c losure to form 202-205.

As a next s t ep towards the s y n t h e s i s of yet another 1,4-

d i h y d r o p y r i m i d i n e d e r i v a t i v e , benza ldehyde was r eac t ed wi th e t h y l

128

t r i f luoromethy lace toace ta te in p resence of t h i o u r e a . It was in t e re s t ing

to note tha t ins tead of 1 , 4 - d i h y d r o p y r i m i d i n e d e r i v a t i v e , t he i n t e r

mediate 209, 210 (Scheme 31) was obta ined as a mixture of d i a s t e r e o -

mer s . Isola t ion of t h e s e compounds has a s ignif icance because no

in te rmedia te a p p e a r s to have been i so la t ed from the Biginel l i r e a c -223 t ion . Ring c losure of 209 wi th p o l y p h o s p h o r i c ac id gave 211

(Scheme 31) .

R = H, 4-NHCOCH.

+ CF3COCH2C02Et

S ^

a

211 H a . Thiourea ; b , Po lyphos

pho r i c ac id

209 H

210 : R = 4-NHCOCH,

MeO

(Scheme 31)

5.7 In o rde r to eva lua te the cont r ibut ion of p y r i m i d i n e moiety

appended with benz imidazo le -2-carbamate nuc leus , the s y n t h e s i s of

p ro to type XVIII namely methyl 5 (6)-N-[ ( 2 - m e t h y l m e r c a p t o - 6 - m e t h y l -

4-phenyl p y r i m i d i n - 5 - y l )raethyl ] amino benz imidazo le -2 -ca rbamate

was under taken . The syn the t i c

s t r a t egy of p ro to type XVIII invo lves

oxida t ion of 198 wi th manganic

ace ta te to obta in 212 (Scheme 32)

which on reduc t ion wi th sodium

b o r o h y d r i d e y ie lded a mixture

of 213 and 214 (Scheme 32) which XVIII

were s epa ra t ed through column

chromatography . However, LAH reduct ion of 212 d id not lead to the

129

Ph>YXN>Y'SCH3

CH3

198

SCH3

Phv,,;;^Nv>^SCH3 II

SCH3

R, , '"Yi^' E t 0 2 C ^ ^ X ^ ^

CH3

213 R:SCH3

214 R=0CH3

IV

216

Ph> -N.

CH3

217

SCH3

N>^SCH3 ;i7 NH CH3

VII,VIII P h \ ^ N ^ S C H 3

NH MU I

VI

H N ~ ^

CH3

NO2 NH2

NHCO2CH3

220 219

Ph. -N.

X- ^ S C H 3

Br CH3

218

I.Mangnic acetate/Benzene. II NaBH^/MeOH.III LAH/Ether,IV Pyridimum

chlorochromate,V.PBr3,Et3N/Benz€ne;VI.H2N-0-NH2,Et3N/ Benzene

VII H2/RQneyNi.VIlI.Me5-C=NC02Me ^^^ NHC02Me

Scheme 32

130

reduction of pyrimidine ring but gave a mixture of 215 and 216 (Sch

eme 32) which were also separated through column chromatography.

The spectroscopic data of these compounds supported the assigned

structures and as an additional evidence 216 was oxidized with pyr i -

dinium chlorochromate to yield 217 (Scheme 32). Reaction of 216

with PBr gave 218 which in turn was reacted with 2-ni tro-p-phenyl-

enediaraine to get 219. Finally hydrogenation of 219 in presence of

Raney-Ni as a Catalyst and cyclisation of the resulting diamine with

N,N-dimethoxycarbonyl-S-methylisothiourea furnished the prototype

molecule 220 (Scheme 32).

5.8 Synthesis of molecules belonging to prototype XIX required

87, 88 and ^ as the starting materials and their synthesis have

already been reported in section

2 .5 . Hydrogenation of 87^ or 88^

gave the desired diamines which

were cyclized with N,N-dimethoxy-

carbonyl-S-methylisothiourea to

obtain the desired benzimidazole

carbamate derivatives (Scheme

33). The compounds representing

prototype XX (227-229) were pre

pared by reacting appropriate

benzylamines with N .N-dimethoxy-

carbonyl-S-methylisothiourea (Sch

eme 33).

/NHCO2MG CHpNHA

\ N C 0 2 M ( 2

XX

5.9 The synthetic strategy employed for the prototype XXI invol

ved the reaction of 2-n i t ro- l -

(3 ,4-methylenedioxyphenyl )-propene

with hydroxylamine hydrochloride

and as would be expected, a

mixture of theo isomers of 1-

hydroxylamino-l-(3 ,4-methylene

dioxyphenyl )-2-nitro propane XX i

was obtained (230; Scheme 34). Separation of these two isomers by

fractional crystallisation or preparat ive TLC was unsuccessful but

the column chromatography of the isomeric mixture over silica gel

revealed an interesting feature. It was observed that only one of

131

M e 0 2 C COoMe

H

90

-^ MGO2C

NHC02Me

CO2MG

H

221

R

.-^^^^2

MGO2C CC^Me

H

87. R = NHM2 88R=NHCH2Ph

a,b

RN'

Q -> MGO2C

NHCOpMe

i N

CO2M2

H

2 2 2 . R = M G

223 R=CH2Ph

^ '

CH2NH2 R-

2 2 4 : R = H

225: R=3,4-m(2thylGnGdioxy 226.R=3.4-dimgthoxy

R- 0 CH2NH^ NCO2MG

NH CO2M2

227-229 SubstituGnt R as in

224-226

a = H2,RaneyNi/MeOH, b= MeS-^'^J^J^'^® ^NHC02M2

Scheme 33

132

Me^ ^N02

a • >

^

CO2MG

NHOH

W

NH2

231

a , NH OH/MeOH; b , H /Raney-Ni; c , MeS-C=NCO Me

NHCO Me

HOHN

N02

230a(FfF^:Thrgo) NHOH

230b(Frs*:Ervthro)

(Scheme 34)

133

the two p o s s i b l e d ia s t e reomers was obta ined in pure s ta te whi le

the second isomer was pos s ib ly involved in a r e t ro r eac t i on leading

to the formation of s t a r t ing n i t r o p r o p e n e . Two p o s s i b l e conclusions

can be drawn from t h i s obse rva t i on . F i r s t l y , t he or ien ta t ion of the

NHOH and H in one of the d ias te reomers must be anti to each o the r

o the rwise the e l iminat ion of NH^OH would not have occured . Secondly,

the isomer which was obta ined in a pure s t a te must possess NHOH

and H groups in o r ien ta t ion which would not allow the e l iminat ion

of NH OH. In a s i tua t ion such as t h i s , two pos s ib l e d i a s t e reomers

(230a and 230b) can be env i saged . The dias tereoraer 230b {R*S*;

e r y t h r o ) would be e x p e c t e d to be ene rge t i ca l ly favoured since the

bu lky groups (Ph & NO^) a re t r ans to each o the r t h e r e b y minimising

the gauche i n t e r a c t i o n s . The s t e r eochemis t ry would a lso permit h y d r o

gen-bonding between NHOH and NO and thus becomes the favoured

d ia s t e reomer , Hydrogenation of 230 wi th Raney-Ni led to the formation

of 231 which was cyc l i zed with N ,N-d ime thoxyca rbony l -S -me thy l i so -

th iourea to ob ta in the r e q u i r e d p ro to type molecule 232 . The i n t e r

mediate diamine 231 was not ve ry s t ab l e and was the re fo re c h a r a c

t e r i z e d as i t s d i h y d r o c h l o r i d e . The r e l a t i v e s t e r e o c h e m i s t r y in 232

was asce r t a ined from the decoupled PMR spec t rum. A doublet (J =

7Hz) at 3.63 - 3.71 d ppm for the methine proton adjacent to methyl

group and a doublet (J = 7 Hz) at 4.23 - 4.31 ef ppm for CH adjacent

to a r y l group suppor t ed t rans s t e r e o c h e m i s t r y for two p r o t o n s .

A r e t r o s p e c t i v e a n a l y s i s of the s t e r eochemis t ry of 230b, on the bas i s

of obse rved r e l a t i v e s t e r eochemis t ry in 232 a l so suppor t s the ass igned

s t e r eochemis t ry of 230b.

5.9a A retro-synthet ic approach for obtaining prototype XXII re

vealed methyl-2-substituted-3-amlno crotonates (233-236) as starting

materials. These were prepared M G p p ^ i .

by reacting l -ary l -2 -n i t ro ethylene M c 0 2 C \ / ' "N 227

with methyl-3-amino crotonate

Nucleophilic displacement of NH Ap

with NHOH group in 233-235 gave ^..

237-239 (Scheme 35) , The structural and stereochemical assignments

of these compounds were concerned with three problems. F i r s t ly ,

it was essential to ascertain whether the compounds ex i s ted as oximes

or as the tautomeric hydroxylamines. In case the compounds were

oximes , the stereochemistry across the -C=N, the second problem

.l^^<" CO2MC

1 J 4

R-NO2

.CO2M2

M G ^ ^ N H 2

a

.C02M<2

'^ MG

233 : R = H

234 : R = 3,4-Dimethoxy

235 ; R = 4-Methoxy

236 ; R = 4-Nitro

b V

. 9^C02M2

Me02Cv^X^N-C02M<2 / ^ N H C 0 2 M e N

MsO 249

\|/

248 : R = 4-Methoxy

a , MeOH; b , NH OH/MeOH; c , d i l .H^SO^/MeOH;

d . Mono p e r p h t h a l l i c ac id or m-CPBA; e , H , , Raney-Ni/MeOH;

f, Me§?C=NC02CH2 NHCO^CHj

(Scheme 35)

135

in the present context, had to be ascertained. Thi rd ly , the relat ive

stereochemistry across the two assyrametric carbon centres (in case

the compounds were oximes) had to be worked out. Indirect evidence

for the structure of compounds 237-239 was obtained from the following

chemical reactions. Acid hydrolysis of 233-235 furnished the ketones

240-242 which could then be converted into 237-239 by reacting them

with hydroxylamine hydrochlor ide. The direct evidence for the assign

ed structures of the compounds was obtained from the C-NMR spec-

trum of one of the representative compound (239). The C signal

for -Cj=N at 158.51 i ppm in the NMR spectrum of 239 and the appear

ance of the carbon of methyl group at 12.05 6 ppm indicated it to

be anti oxime . In order to ascertain the stereochemical purity

of compounds 237-239, the multiplicity of the methine proton (Ctl-

CO Me) was studied. However, this methine proton in these compounds

appeared along with the signals of other methine and methylene pro

tons in a 90 MHz PMR spectrum. This prompted to record a 400 MHz

PMR spectrum of one of the representative compound (239). The app

earance of a neat doublet at 3.72 6 ppm in the PMR spectrum sugges

ted stereochemical homogenity and indicated stereoselective formation

of only one diastereomer. The other minor diastereomer could not

be isolated from the reaction mixture and in the absence of this

minor diastereomer, the relat ive stereochemistry of 239 could not

be ascertained. Reaction of 233-236 with ei ther monoperphthalic acid

or m-chloro perbenzoic acid yielded 243-246. It was interesting to

note that the te r t ia ry hydroxyl group in these compounds was not

prone to easy elimination reaction and the reaction of 243-244 with

hydroxylamine hydrochloride yielded oximes 247-248 without eliminat

ing a molecule of water. The presence of a ter t iary hydroxyl group

in these compounds was evident from the PMR signal (DO exchangea

ble) at 4.0 6 ppm. The s tabi l i ty of the te r t ia ry hydroxy groups

in compounds 247-248 suggested that the hydroxyl group was not

placed anti to vicinal CH[. In the light of this observation the Dreid-

ing models of various rotamers of the two possible diastereomers

were studied to ascertain the s ter ic crowding in the molecule. On

the basis of Dreiding model the stereochemically most favoured dia-

stereomer (I ; R R , threo) is described below:

136

OH

HON.

O2NH2C

^ ^ / ^ ^ ./NOH

OMe' 3

I II In order to obtain a representative compound of the desired

prototype XXII, compound 239 was hydrogenated over Raney-Ni and

the resulting solution was reacted with N ,N-dimethoxycarbonyl-S-me-

thyl-isothiourea to yield 249. The appearance of one broad singlet

and a sharp doublet for the methyl protons in the PMR spectrum

(400 MHz) indicated it to be a mixture of stereomers. This was fur

ther supported by the appearance of two extremely close peaks in

the HPLC spectrum. Preparative HPLC to separate the isomers was

not attempted. The possible mechanism of formation of compound 249

is described in scheme 36.

137

H H

O2N-

,C02Me ^^ H H

A r ^ 1 A ^COaMG

• ^

NOH HoN H rt NHOH/NH2

Mc Mc

M G S H ^ NCO2MC

-NH2OH/NH3

Ar CO2MG /^

HN / /V -NHC02Me

N C02Me

NHC02M(2 V

A r C02M<2

H 2 N - ^ ^)~^

M<2

Me

M < 2 0 2 C ^ X N , / .C02Me

-N -H

A r ^ \ ^N / / ' ^ C 0 2 M e

Scheme 36

CHAPTER 6

6 , 1 : EXPERIMENTAL

6 .2 : T a b l e s

6 .3 : BIOLOGICAL ACTIVITY

13TB

CHAPTER - 6

EXPERIMENTAL

6.1 The melting points were determined on a sulphuric acid

bath or an electrically heated block and are uncorrected. All the

reactions were checked by thin layer chromatography over silica

gel G or alumina (basic or neutral) plates using iodine vapours.

KMnO. spray or Dragendorf's spray as the developing reagent. The

structures of all the compounds were routinely checked by IR and

PMR Spectroscopy-IR spectra were recorded on Perkin-Elmerl57 infra-

cord and Beckman Acculab-1 grating instruments and values are expre

ssed in cm . PMR spectra were recorded on varian EM 360L or

Perkin-Elmer R-32 using TMS as internal reference (Chemical shift

in S ppm). C-NMR spectra were recorded on CFT-20 spectrometer

operating at 20 MHz, 8192 data points were collected and a sweep

width of 5000 Hz was used. Pulse delay of 0.5 sec. was essential

for recording the signals of carbonyl carbons. C-NMR spectra were

also recorded on a Bruker WM-400 FT NMR spectrometer. Mass spectra

of these compounds were recorded on Jeol-D 3000 instrument. The

second place after the decimal point in the required value of C,H,N

analyses have been approximated.

P-Aroyl propionic acid (101-102)

These compounds were essentially prepared by the method

reported earlier

/3-Aroyl-/-butyrolactones (103,104)

These compounds were prepared in better yields by modifying 41a

the method reported earlier . The details are as follows:

Formaldehyde (37.41%; 0.30 mole) was added to a stirred

solution of /3-aroyl propionic acid (103, 104; 0.27 mole) in NaOH

solution (0.5N, 60 ml, 0.30 mole). After stirring at room temperature

(25') for 1 hr. the mixture was acidified with cone. HCl (^15 ml)

and stirred at room temperature (25°) for additional 12 hr. The

separated semi solid was triturated with ether to obtain the required

compound. The mother liquor was again stirred at room temperature

(25°) for 4 hr. and the separated solid was filtered. The combined

solid product was recrystallised from hot benzene.

139

41a +

103 : Y i e l d 82%; m . p . 62 -64° ( L i t . m . p , 6 3 - 6 5 ° ) ; M

a t m/z 190

I R ( K B r ) : 1670 & 1775 (CO) PMR ( C D C l , ) : 2 . 7 1 - 2 . 8 8 ( t , 2H, C H , ) , 4 . 2 5 - 4 . 5 3 ( m , 3H, OCH )

3 i ^ & C H ) , 7 . 2 6 - 7 . 5 8 ( m , 3H, 3 , 4 i 5Ar -H) , 7 . 7 2 -7.90 ( m , 2H, 2 & 6 A r - H ) .

41d +

104 : Y i e l d 80%; m . p . 84-85° ( L i t . m . p . 8 5 - 8 7 ° ) ; M

a t m/z 204

IR (KBr) : 1670 & 1760 (CO) PMR(CCK+CDC1,) : 2 . 4 0 ( s , 3H, C H , ) , 2 . 6 9 - 2 . 8 2 ( { t . 2H, CH ) ,

4 3 ~i '^

4 . 1 2 - 4 . 5 8 ( m , 3H, OCH^ & C H ) , 7 . 2 0 - 7 . 2 9 ( d ,

2H, 3 & 5 A r - H ) , Jo = 9 H z ) , 7 . 7 3 - 7 . 8 3 ( d , 2H,

2 & 6 A r - H , Jo = 9 H z ) .

4-Aroyl-N-(£-methoxyphenyl)-pyrrolidin-2-ones (105 & 107; Table 15)

General Procedure:

To a mixture of 103 or 104 (0.1 mole) and £-anisidine (0.1

mole) in alcohol (6 ml) was added triethylamine (0.2 mole) and ref-

luxed for 8 hr. Water was added to it and extracted with ethyl aceta

te. Usual work up of organic layer yielded a residue which was trea

ted with ether to obtain a solid. It was recrystallised from a mixture

of chloroform-hexane.

4-( 0(-Hydroxy-4-methyl) benzyl-N-(£-methoxyphenyl) pyrrolidin-2-one

(106)

A solution of 105 (0.01 mole, 3.0 gm) in methanol (35 ml)

was hydrogenated in presence of Pd/c (10%, 0.3 gm) at 2,5 kg/cm

for 2 hr. The catalyst was filtered off and the filterate was concen

trated under reduced pressure. The residue on trituration with CCl

furnished a crystalline solid which was recrystallised from hot CCl .

106 : Y i e l d 90%; m . p . 1 1 5 - 1 6 ° ; M"^ a t m/z 311

PMR (CDCl^) : 2 . 3 0 ( s , 3H, C H ^ ) , 2 . 5 4 - 2 . 7 2 ( m , 2H, C H - C H ^ ) ,

3 . 4 0 - 3 . 8 0 ( m , 6H, OCH ) , NCH^ & C H ) , 4 . 4 0 -

4 .60 ( m , I H , CHOH), 6 . 7 0 - 7 . 4 5 ( m , 8H, A r - H ) .

4-Benzoyl-N-(4-amino-3-nitrophenyl)-pyrrolidln-2-one (108; Table 15)

The experimental procedure was essentially same as described

for 105 but instead of 2.-anisidine, 2-nitro-£-phenylene diamine was

140

used . The compound 108 was r e c r y s t a l l i s e d from e thy l a c e t a t e - e t h e r

mix tu re .

Catalytic hydrogenation of 108

A solution of 108 (0 .65 gm, 0.02 mole) in methanol (30 ml)

was hydrogena ted a t 2.5 kg/cm in the presence of Raney-Ni ( 0.3

gm) io r 30 minutes . Ca ta lys t was f i l t e red off and the solvent was

removed under reduced p r e s s u r e . The r e s idue without fu r ther pu r i f i ca

tion was u t i l i s ed for the next s t e p .

4-Benzoyl-N-I5(6)-(2-methoxycarbonylainino) benzimidazolyl 1 pyrro l id in-

2-one (109; Table 15)

Aqueous NaOH solution (25%, 3 ml) was added under s t i r r i n g

to a precooled (10-15°) mixture of S-methyl isothiouronium su lpha te

(0 .01 mole) and methylchloroformate (0.02 mole) in water (4 ml)

and the pH of the reac t ion mixture was maintained between 8.0 - 9 .0 .

Glacial ace t i c ac id was then carefu l ly added for readjus t ing the pH

to about 5.0 and to t h i s mix tu re , was then added the r e q u i r e d diamine

(mentioned a b o v e , 0.01 mole) in methanol (15 m l ) . This reac t ion mix

tu re was re f luxed for 3 h r , cooled to room tempera tu re and f inal ly

d i lu ted wi th w a t e r , the so l id so obta ined was f i l t e r e d , washed wi th

water and d r i e d . I t was r e c r y s t a l l i s e d from e thano l .

4-Acetanildo-3-nitro benzaldehyde (112)

This was p r e p a r e d by the method r e p o r t e d e a r l i e r

4-Acetamido-3-nltro cinnamic acid (113)

To a mixture of 112 (2 gm, 0.01 mole) and malonic ac id

(1 gm, 0.01 mole) in ethanol (5 ml) was added p y r i d i n e (0 .1 ml)

and re f luxed for 2 h r . The s e p a r a t e d so l id was f i l t e r e d , washed

wi th hot ethanol (5 m l ) , d r i e d and r e c r y s t a l l i s e d from DMF-H O mix-

Yield 88%; m . p . 254-5°; M* at m/z 250

1620, 1690 (CO)

2.08 ( s , 3H, NHCOCH^), 6 .50-6.58 ( s , IH, =CH,

J = 16Hz), 7 .40-8 .20 (m, 5H, Ar-H, =CH. & NH)

Requires : C, 52.80; H, 4 .00; N, 11.20

Found: C, 52.60; H, 3 .80; N, 11.50.

t u r e .

113

IR(KBr)

PMR (DMSO-

Sl"l0^2°5

•'^6^

141

4-Ani lno-3 -n i t ro c innamic a c i d (114)

To a s u s p e n s i o n of 113 ( 2 . 5 gm, 0 . 0 1 mole ) in m e t h a n o l (8

ml) w a s s l o w l y a d d e d a q u e o u s NaOH s o l u t i o n (10%, 2 . 5 ml) a n d s t i r r e d

a t room t e m p e r a t u r e ( 2 8 ° ) for 30 m i n u t e s . A m i x t u r e of w a t e r (10

ml) a n d a c e t i c a c i d ( 0 . 1 ml) w a s a d d e d to t h e r e a c t i o n m i x t u r e a n d

t h e s e p a r a t e d s o l i d w a s f i l t e r e d , w a s h e d w i t h w a t e r , d r i e d a n d

r e c r y s t a l l i s e d from DMF-H O m i x t u r e .

114

IR (KBr )

PMR (DMSO-d, ) o

S«8^2°4

Y i e l d 9 0 1 ; m . p . 2 0 6 - 7 ° ; M a t m/z 208

1690 ( C O ) , 3350 (NH)

6 . 1 0 - 6 . 4 0 ( d , I H , =CH, J = 1 6 H z ) , 6 . 9 0 - 7 . 0 8

( d , I H , =CH-CO H, J = 1 2 H z ) , 7 . 3 0 - 8 . 0 0 ( m ,

5H, Ar-H & NH^)

R e q u i r e s : C , 5 1 . 9 2 ; H, 3 . 8 4 ; N, 1 3 . 4 6

Found : C , 5 2 . 2 0 ; H, 3 . 5 0 ; N, 1 3 . 1 1

C a t a l y t i c h y d r o g e n a t i o n of 114

T h e e x p e r i m e n t a l p r o c e d u r e w a s e s s e n t i a l l y s ame a s d e s c r i b e d

for t h e r e d u c t i o n of 108 a n d t h e r e s u l t i n g d i a m i n e w a s u s e d for t h e

n e x t s t e p w i t h o u t f u r t h e r p u r i f i c a t i o n .

/ 3 - I ( 2 - M e t h o x y c a r b o n y l a m i n o ) - 5 ( 6 ) - b e n z i m i d a z o l y l ] - a c r y l i c a c i d (115)

T h i s w a s p r e p a r e d b y t h e m e t h o d d e s c r i b e d for 109 a n d

r e c r y s t a l l i s e d f rom DMF-H O m i x t u r e .

115

I R ( K B r )

PMR ( D M S O - d . ) 6

S2«llN3°4

Y i e l d 63%; m . p . > 3 0 0 ° ; M a t m/z 261

1665 ( C O ^ H ) , 1720 (CO)

3 .73 ( s , 3H, CO^CH^) , 6 . 2 4 - 6 . 4 1 ( d , I H ,

J = 16Hz, 7 . 1 0 - 7 . 7 8 ( m , 4H, =CH & A r - H )

R e q u i r e s : C , 5 5 . 1 7 ; H, 4 . 2 1 ; N, 16 .09

Found : C , 5 5 . 1 0 ; H, 4 . 2 6 ; N, 15 .80

=CH,

5 - E t h o x y c a r b o n y l - 5 - m e t h y l - 2 - o x o - 4 - ( 4 - a c e t a i i i i d o - 3 - i i i t r o p h e n y l ) p y r i m i -

d ine (117; T a b l e 16)

T h i s w a s p r e p a r e d b y t h e m e t h o d r e p o r t e d in t h e l i t e r a

t u r e 223

5 - E t h o x y c a r b o n y l - 6 - m e t h y l - 2 - o x o - 4 - ( 4 - a i i i i n o - 3 - n l t r o p h e n y l )

d ine (118; Table 16)

p y r i m l -

T h i s w a s e s s e n t i a l l y p r e p a r e d b y t h e m e t h o d d e s c r i b e d for

114 a n d r e c r y s t a l l i s e d f rom DMF-H O m i x t u r e .

142

Methy l -5 (6 ) - t5 -e thoxycarbony l -5 -methy l -3 ,4 -d ihydropyr imid in-4 -y l ]

benzimidazole-Z-carbamate (119; Table 16)

The method for p r epa ra t i on of 119 was the one r e p o r t e d

for 115.

4-Acetaniido acetophenone (121)

231 This was p r e p a r e d by the method r e p o r t e d e a r l i e r

4-Acetainido-3-iiitro acetophenone (122)

This compound was p r e p a r e d by the method r e p o r t e d e a r -

, . 231

l i e r

4-Amino-3-nitro acetophenone (123)

This was p r e p a r e d by the method d e s c r i b e d for 114.

Methyl -5(6) -acy l benzimidazole-Z-carbamate (124)

This was a lso p r e p a r e d by the method r e p o r t e d in the l i t -232

e r a t u r e 2-Methoxycarbonylamino benzimidazole-5-(6)-acetoxime (125)

A mixture of hydroxy lamine h y d r o c h l o r i d e (0.96 gra, 0.015

mole) and sodium ace ta te (1.36 gm, 0.015 mole) was added to a solu

t ion of 124 (2 .3 gm, 0.01 mole) in methanol (12 ml ) . The mixture

was re f luxed for 5 h r . cooled and d i lu ted wi th water (10 ml ) . The

so l id so obta ined was f i l t e r e d , washed wi th w a t e r , d r i e d and r e c r y s -

t a l l i s e d from DMF-H O mix tu re .

125

IR(KBr)

PMR (DMSO-d, ) 6

Yield 80%; m . p . 281-2°; M" at m/z 248

1640 (C = N) , 1720 (CO), 3300 (NH)

2.22 ( s , 3H, CH^), 3.80 ( s , 3H. CO^CH^) ,

7 .40-7.70 (m, 3H, Ar-H)

*^ll"l2 '^4°3 ' Requires : C, 53.22; H, 4 .83 ; N, 22.58

Found ; C, 53.15; H, 5 .31 ; N, 22.20

2-Ni tro - l - (4 - subs t i tu ted phenyl) propene (^26, 127)

These compounds were p r e p a r e d by the method r e p o r t e d

e a r l i e r

2,5-Dimethyl-3-methoxycarbonyl-4-subst i tuted phenyl fiiran (128, 129;

Table 17) and 2 ,5-dimethyl -3-methoxycarbonyl-4-subst i tuted phenyl

pyrrole (130, 131; Table 17)

145

Method A:

To a suspension of 126, 127 (0 .001 mole) in methanol (10

ml) was a d d e d p i p e r i d i n e (0.002 mole) and methylacetoaceta te (0.002

mole) under cooling (5-10°) and s t i r r i n g . I t was s t i r r e d at room

tempera tu re (25°) for 24 h r . Water was added to the reac t ion mixture

and e x t r a c t e d wi th e t h y l a c e t a t e . Usual work up of organic l aye r

y ie lded an oil containing a mixture of 126, 129 and 130, 131 which

were s e p a r a t e d through column chromatography over s i l i ca gel using

hexane : Chloroform (80:20) as an e luent . These compounds were r e c -

r y s t a l l i s e d from chloroform - hexane mix tu re .

Method B for 128

To a suspension of 126 ( 1 .1 gm, 0.005 mole) in methanol

(8 ml) was added t r i e thy lamine (1 .0 ml, 0.01 mole) and methy lace to

ace ta te ( 1 . 1 . ml, 0.01 mole) under cooling (5-10°) and s t i r r i n g .

I t was then s t i r r e d at room tempera ture (20°) for 24 h r . Water (20

ml) was added to the reac t ion mix tu re , the s epa ra t ed so l id was

f i l t e r e d , washed wi th water and d r i e d . I t was pur i f ied through socc i -

lat ion using benzene as a so lven t .

Method B for 130

A mixture of 126 (1 .1 gm 0.005 mole) , methyl-3-amino c ro to -

nate (1.15 gm, 0.01 mole) and p y r i d i n e (0 .1 ml) in methanol (10

ml) was re f luxed for 36 h r . Water was added to the reac t ion mixture

and af ter 4 hr the s e p a r a t e d so l id was f i l t e r e d , washed wi th water

and d r i e d .

2,5-Dimethyl-3-methoxycarbonyl-4-substituted,_^ phenyl-N-subst i tuted

pyrroles (132-137; Table 17)

General Procedure:

A mixture of 126 (0 .01 mole ) , methylacetoaceta te (0.02 mole)

and p r i m a r y amines (0.02 mole) in methanol (15 ml) was re f luxed

for 12-14 h r . Water was added to the reac t ion mix tu re , the s e p a r a t e d

so l id was f i l t e r e d , washed wi th water and d r i e d . These compounds

were r e c r y s t a l l i s e d from chloroform-pet ro leum benzene mix tu re .

2,5-Diinethyl-3-methoxycarbonyl-4-(4-anilno-3-i i l trophenyl )-N-methyl p y

rrole (146; Table 17)

A mixture of 126 (2 .2 gm, 0.01 mole) and 145 (2 .58 gm.

144

0.02 mole) in methanol (15 ml) was re f luxed for 36 h r . Water was

added to the react ion m i x t u r e , t he s e p a r a t e d sol id was f i l t e r e d ,

washed wi th water and d r i e d . I t was r e c r y s t a l l i s e d from aqueous

methanol .

3,6-Dimethyl-4-hydrazinocarbonyl-5-(4-amino-3-nitrophenyl) pyridazine

(148)

A mixture of 126 (0.002 mole) , methylace toace ta te (0.004

mole) and hydraz ine h y d r a t e (0.004 mole) in ethanol (10 ml) was

re f luxed on a wa te rba th for 3 h r . The reac t ion mixture was d i lu ted

with w a t e r , the s e p a r a t e d so l id was f i l t e r e d , d r i e d and r e c r y s t a

l l i s e d from e t h y l a c e t a t e - h e x a n e .

148

IR(KBr)

PMR (TFA)

Yield 70%; m . p . 170-71°; M+lat m/z 303

1650 (CO)

2.18 ( s , 6H, 2xCH ) , 7 .47-7.61 (m, 3H, Ar-

H)

C, H, N O, : Requires : C, 51 .65; H, 4 .63 ; N, 27.81 13 14 6 J

Found : C , 5 1 . 4 5 ; H, 4 . 3 1 ; N, 2 7 . 9 6

2-Nitro- l - subst i tuted phenyl propene (149-153)

These compoxinds were p r e p a r e d by the method r e p o r t e d ,. 43 e a r l i e r

2 ,5-Dimethyl-3-methoxycarbonyl-4-subst i tuted' '^phenyl pyrro les (154-

158; Table 18)

The exper imenta l p rocedure was e s s e n t i a l l y same as d e s c r i b e d

for 146.

Catolytlc hydrogenation of ^30^, 132-137

The exper imenta l p rocedure was e s s e n t i a l l y same as d e s c r i b e d for

108.

Methyl 5 (6) - I2 ,5 -d imethyl -3-methoxycarbonyl -N-subst i tuted'~ ' pyrrolo]

benzlniidazole-2-carbaiiiates (159-166; Table 19)

To a solution of diamine (mentioned a b o v e , 0,01 mole) in

methanol (10 ml) was added N ,N-d ime thoxyca rbony l -S -me thy l i so th iou rea

(0 .01 mole) and ca t a ly t i c amount of 2.-TSA and ref luxed for 5 h r .

Water (20 ml) was added to the reac t ion mix tu re , the so l id so o b

ta ined was f i l t e r e d , washed wi th water and d r i e d . These were r e c r y s

t a l l i s e d from aqueous methanol .

145

2-Nitro- l -subst i tuted' 'phenyl propene (166-170)

43 These were p r e p a r e d by the method r e p o r t e d e a r l i e r

2,5-Dimethyl-3-methoxycarbonyl-4-subst i tuted ^ ^ phenyl-N-(4-anuno-3-

nitrophenyl) pyrrole (171-175; Table 20)

These compounds were e s s e n t i a l l y p r e p a r e d by the method

d e s c r i b e d for 132-137.

Methyl-5(6)-N-[2 ,5-dimethyl-3-inethoxycarbonyl-4-substituted'^~^ phenyl 1

pyrrolo ben2imidazole-2-carbamates (176-180; Table 21)

The exper imenta l p rocedure was same as d e s c r i b e d for 159-

166.

4 , 4 ' -D ich loro -3 ,3 ' -d in i t ro benzophenone (182)

232a This was p r e p a r e d by the method r e p o r t e d in l i t e r a t u r e

4 ,4 ' -Diamino-3 ,3 ' -d in i tro benzophenone (183)

This compound was also p r e p a r e d by the method r e p o r t e d

2,2 • -1 B i s - (4-ainino-3-nitrophenyl) ] methanol (184)

To a suspension of 183 (3.04 gm, 0.01 mole) in methanol

(10 ml) was a d d e d sodium b o r o h y d r i d e (4.56 gm, 0.12 mole) and

ref luxed for 4.5 h r . Water was then added to the reac t ion mixture

and pH was ad jus ted to 7 by adding glacia l acet ic ac id d r o p w i s e .

The so l id so obta ined was f i l t e r e d , washed with w a t e r , d r i e d and

r e c r y s t a l l i s e d from aqueous DMF.

184 : Yield 85%; m . p . 230-32°; M" at m/z 304

PMR (DMSO-d^) : 5.50 ( s , IH, CH), 6 .88-6 .98 and 7 .23-7.31

(m, 6H, Ar -H) , 7 .70-8.01 ( b s , 4H, 2xNH )

^ 1 3 " l 2 ^ 4 ° 5 ' Requires : C, 51 .31 ; H, 3.94; N, 18.42

Found : C, 51.80; H, 4 .24; N, 18.51

2,2'-Dicarbomethoxyaniino-5,5'-dibenziniidazolylmethanol (185)

This compound was p r e p a r e d by following the method d e s

c r i b e d for 159-166. I t was r e c r y s t a l l i s e d from aqueous DMF.

185 s Yield 78%; m . p . > 300"; M" at m/z 410

IR (KBr) : 1725 (CO)

146

PMR (TFA) : 3.90 ( s . 6H, 2xNHC02CH^), 5.28 ( s , IH. CH)

7.38-7.70 (m, 6H, Ar-H)

C,„H,„N^O^ : Requires : C, 55.60; H, 4 .39; N, 20.48 19 18 6 5

Found : C, 55 .41 ; H, 4 .78; N, 19.94

5-Ethoxycarbonyl-6-methyl-4-subst i tuted'^phenyl-3 ,4-dihydrophyrini idin-

2-thione (192-197; Table 22)

These compounds were p r e p a r e d by the method r e p o r t e d ,. 223

e a r l i e r

5-Ethoxycarbonyl-2-methylmercapto-6-methyl-4-phenyl-4( lH)-pyrimidine

(198; Table 23)

A mixture of 192 (0 .01 mole ) , Potassium h y d r o x i d e (0 .01

mole) and methyl iod ide (0.012 mole) in methanol (5 ml) was s t i r r e d

at room tempera ture (SO'C) for 1 h r . The s e p a r a t e d so l id was f i l t e r e d ,

washed with water (50 ml) and r e c r y s t a l l i s e d from aqueous methanol .

5-Ethoxycarbonyl -2- ( l -methyl p iperaz lno) -6 -methy l -4 -pheny l -4{ lH) -

pyrimidine (200; Table 23)

A mixture of 192 (2 .9 g, 0.01 mole) and N-methyl p i p e r a z i n e

(2 ml , 0,02 mole) was hea ted a t 70-75°C for 5 h r . Water was added

to the reac t ion mixture and e x t r a c t e d wi th e thy l a c e t a t e . Organic

l aye r was washed f ive t imes wi th water and d r i ed on anhydrous

sodium s u l p h a t e . Removal of solvent y i e lded the compound 200 as

an o i l .

l , 6 -Dimethyl -5 -Ethoxycarbonyl -2 -methylmercapto-4-phenyl -4 - [ lHl -pyr i -

mldine (199)

A mixture of 192 (0 .01 mole ) , potassium h y d r o x i d e (0.02

mole) and methy l iod ide (0.022 mole) in N.N-dimethyl su l foxide (3

ml) was s t i r r e d at room tempera tu re (30°C) for 1 h r . Water (5 ml)

was a d d e d to the reac t ion m i x t u r e , the s epa ra t ed so l id was f i l t e r e d ,

washed wi th water (50 ml) and r e c r y s t a l l i s e d from aqueous methanol .

199 : Yield 70%; m . p . 160"; M" at m/z 304

IR (KBr) : 1680 (CO)

1.03-1.19 ( t , 3H, CH^), 2.30 ( s , 3H, CH ) , PMR (CDCl )

2.43 ( s , 3H, SCH^), 2.90 ( s , 3H, NCH ) , 3 .88-

4.10 ( q , 2H, CH^) , 5.11 ( s , IH, CH) , 7.18

( s , 5H, Ar-H)

147

C , , H - - N , 0 , S : Requires : C, 63 .15 ; H, 6 .57; N, 9.21

Found : C, 63.00; H, 6 .25; N, 8.80

1,6-Dimethyl -5-ethoxycarbonyl -4-phenyl -3 ,4-dihydropyrimidin-2-one (201)

To a solution of 16^ (0 .5 g) in methanol (2 ml) was added

cone. H SO (2 d rops ) and ref luxed on steam ba th for 4 h r . Water

was added to i t and e x t r a c t e d with e t h y l a ce t a t e . Usual work up

y ie lded a so l id which was r e c r y s t a l l i s e d from aqueous methanol .

201

IR (KBr)

PMR (CDCl )

Yield 55%; m . p . 160-61°; M"*" at m/z 274

1660, 1690 (CO)

1.03-1.20 ( t , 3H, CH^), 2.27 ( s , 3H, CH^).

2.77 ( s , 3H, N-CH ) , 3 .79-4 .11 ( q , 2H, CH^) -

5.12 ( b s , IH, CH), 7.21 (m, 5H, Ar-H) , 8.50

( b s , IH, NH)

C ,^H, -N-0- : Requires : C, 65 .69; H, 6.56; N, 10.21 15 l o & J

Found : C, 65 .73 ; H, 6 .81 ; N, 9.84

5-Aryl -6-ethoxycarbonyl -7-methyl -3-oxo-2 ,3-dihydro-5H-thiazolo [ 3 , 2 -

a ] pyrimidine (202-205; Table 24)

General Procedure:

A mixture of 192-195 (0 .01 mole ) , monochloroacetic ac id

(0.05 mole) and BF e t h e r a t e (0 .1 ml) in methanol (5 ml) was s t i r r e d

at room tempera tu re (30°C) for 3.5 h r . Water (10 ml) was then added

to the reac t ion mix tu re . Extrac t ion wi th e t h y l ace ta te (10 ml) followed

by usual work-up y ie lded an oil which on t r i t u r a t i on wi th petroleum

benzene furn ished a so l id 202 which was r e c r y s t a l l i s e d from c h l o r o -

form-hexane mix tu re , wh i l e o the r compounds 203-205 were ob ta ined

as o i l .

l -Carboxymethyl-5-ethoxycarbonyl-6-methyl-4-subst i tuted^^~^phenyl-3 ,4-

dih7dropyrlmldin-2-thione (206-208; Table 25)

General Proce dure :

A mixture of ^92, 194-195 (0 .01 mole) , monochloroacetic

ac id (0 .05 mole) and potassium h y d r o x i d e (0 .01 mole) in methanol

(5 ml) was re f luxed on steam ba th for 2 h r . The s e p a r a t e d so l id

was f i l t e r e d , washed wi th water (40 m l ) , d r i e d and r e c r y s t a l l i s e d

from methanol .

148

Ethyl ( c<- tr i f luoroacety l -^- th iour ido) subs t i tu ted^phenyl propionate

(209. 210)

General Procedure;

A mixture of a p p r o p r i a t e benza ldehyde (0.01 mole ) , E t h y l -

t r i f luoromethyl ace toace ta te (0 .01 mole ) , th iourea (0.02 mole) in

HCl (3 ,5 ml , 3N in 1:1 methanol : wa t e r ) was s t i r r e d at room tempe

r a t u r e (20°C) for 48 h r . The s e p a r a t e d so l id was f i l t e r e d , washed

wi th water and d r i e d . I t was r e c r y s t a l l i s e d from chloroform-hexane

mix tu re .

5-Ethoxycarbonyl-6- tr i f luoromethyl -4-phenyl-3 ,4-dihydropyrimidin-

2-thione (211)

0.1 g of 209 was taken in 2 gm of p o l y p h o s p h o r i c ac id and

hea ted on water ba th for 8 h r . water was added and on t r i t u r a t i o n

a so l id was obta ined which was r e c r y s t a l l i s e d from benzene.

5-Ethoxycarbonyl-2-methylmercapto-6-methyl-4-phenyl pyrimidine (212;

Table 26)

A mixture of 198 (5 .8 g, 0.02 mole) and manganic ace ta te

(11.8 g, 0.04 mole) in d ry benzene (50 ml) was re f luxed for 2.5

h r . Manganous ace ta te was f i l t e r ed off. Water was added to the f i l t e -

r a t e and i t was e x t r a c t e d wi th benzene. Usual work up y i e lded an

oil which was pur i f i ed through a shor t band of s i l i ca gel using h e -

xane as e luent .

5-Ethoxycarbonyl-2-methylmercapto-6-methyl-4-phenyl-2-( lH)-pyri inidine

(213) and 5-Ethoxycarbonyl -2-methoxy-5-methyl -4-phenyl -2- ( lH) p y r i

midine (214)

To a solution of 212 (5 .6 g, 0.02 mole) in methanol (8 ml)

was added NaBH (1 .5 g, 0.04 mole) under cooling (5-10°C) and s t i r r

ing . It was s t i r r e d at room tempera ture (20°C) for 1 h r . Water was

added to the react ion mixture and e x t r a c t e d wi th e t h y l a c e t a t e . Usual

work up y ie lded a mixture of two compounds 213 and 214 which were

s e p a r a t e d through a column of s i l i ca gel using hexane , chloroform

as e luent .

5,6-Dimethyl-2-inethylinercapto-4-phenyl-pyrimidine (215; Table 26)

and 5-HydToxymethyl-2-methylmercapto-6-methyl-4-phenyl pyrimidine

(216; Table 26)

149

To a suspension of LAH (1.14 g, 0.03 mole) in d r y e t h e r

(5 ml) was added a solution of 212 (8 .7 g, 0.03 mole) in d r y e t h e r

(20 ml) under cooling (S-IO'C) and s t i r r i n g . I t was s t i r r e d at the

same tempera tu re for 0.5 h r . Ord ina ry benzene was added followed

by add i t ion of w a t e r . It was e x t r a c t e d wi th benzene. Usual work

up y ie lded a mixture of 215 and 216 which were s epa ra t ed through

a column of s i l i ca gel using hexane , chloroform as e luent .

5-Formyl-2-methylmercapto-6-methyl-4-phenyl-pyrini idine (217; Table

26)

To a solution of 216 (2 .4 g, 0.01 mole) in d r y CH CI (15

ml) was added pyr id in ium chlorochromate (2.2 g, 0.01 mole) . I t

was s t i r r e d at room tempera tu re (25''C) for 40 minutes. Solvent e t h e r

was added and eluted wi th e the r through a shor t band of s i l i ca

ge l . Solvent was removed and on t r i t u r a t i o n with hexane a c r y s t a l l i n e

so l id obta ined was r e c r y s t a l l i s e d from chloroform-hexane mix tu re .

5-Bromomethyl-2-methylmercapto-6-methyl-4-phenyl pyrimidine (218;

Table 26)

To a solution of 216 (2 .4 g, 0.01 mole) in d ry benzene (10

ml) was added a mixture of PBr (0.95 ml, 0.01 mole) and t r i e t h y l a -

mine (1 .0 ml , 0.01 mole) in d r y benzene (5 ml) under cooling (5°C)

and s t i r r i n g . It was s t i r r e d at room tempera tu re (25°C) for 1 h r .

Water was added and e x t r a c t e d wi th e t h y l ace t a t e . Usual work up

y i e lded a so l id which was r e c r y s t a l l i s e d from chloroform-hexane

mix tu re .

5-IN-( 4-Amino-3-nitrophenyl) ] aminomethyl-2-methylmercapto-6-methyl-

4-phenyl pyrimidine (219; Table 26)

To a solution of 218 (0 .3 gm, 0.001 mole) in d r y benzene

(5 ml) was added t r i e t hy l amine (0 .1 ml, 0.001 mole) and O-nitro-

£-phenylened iamine (0.15 gm, 0.001 mole) . It was s t i r r e d a t room

tempera tu re (25°C) for 1 h r . Water was added to i t and e x t r a c t e d

wi th e t h y l a c e t a t e . Usual work up y ie lded an oil which on t r i t u r a t i o n

wi th p e t . benzene y i e lded a so l id which was r e c r y s t a l l i s e d from

chloroform-hexane m i x t u r e .

Methyl-5(6)-N-I (2-methyImercapto-6-methyl-4-phenyl pyrimidine-S-

y l ) methyl] amino benzimidazole-2-carbamate (220; Table 26)

To a solution of 219 (0.38 gm, 0.001 mole) in methanol (20

150

ml) was added Raney-Ni (0 .1 gm) and hydrogena ted at 2.5 kg/cm

for 1 h r . The ca t a ly s t was f i l t e r ed off and the solvent was r educed

to 5 ml. N ,N-d imethoxycarbony l -S-methy l i so th iourea (0.2 gm, 0.001

mole) was added to i t and re f luxed for 3.5 h r . Water (20 ml) was

added to i t , the s epa ra t ed so l id was f i l t e r e d , washed wi th water

and d r i e d . It was r e c r y s t a l l i s e d from chloroform-hexane mix tu re .

M e t h y l - 5 { 6 ) - [ 4 - { 2 , 6 - d i m e t h y l - 3 , 5 - d l m e t h o x y c a r b o n y l - l , 4 - d i h y d r o p y r i -

d y l ) ] benzimidiazole-2-carbamate (221; Table Zl)

A mixture of ^ (0.331 gm, 0.001 m o l e ) , N ,N-d ime thoxyca rbo -

ny l -S -me thy l i so th iourea (0.206 gm, 0.001 mole) and c a t a l y t i c amount

of £-TSA in methanol (5 ml) was ref luxed tor 4 h r . The reac t ion

mixture was d i lu ted with w a t e r , t he so l id so obta ined was f i l t e r e d ,

washed with water and d r i e d . I t was r ec r v s t a l l i s e d from aqueous

methanol .

Methy l -5 (6 ) - (4 - (2 ,6 -d imethy l -3 ,5 -d imethoxycarb«myl - l , 4 -d ihydropyr i -

dy l ) I-N-substltuted benziinidazole-2-carbainate (222-223; Table 27)


for 159-166.

N,N-Dimethoxycarbonyl-N-( subst i tuted benzyDguAnldine (227-229; Table 28)


for 221.

l -Hydroxyla in ine- l - (3 ,4 -methylenedloxyphenyl ) -2-n i tro propane (230)

To a solution of l - ( 3 ,4 -me thy lened ioxypheny l ) -2 -n i t ro propene

(2.07 g, 0.01 mole) in methanol (10 ml) was added hyd roxy lamine

h y d r o c h l o r i d e (1.39 g, 0.02 mole) and sodium aceta te (1 .64 g, 0.02

mole) . The react ion mixture was s t i r r e d at room tempera tu re (20°C)

for 5 h r . Water (20 ml) was a d d e d to i t , the p r e c i p i t a t e d so l id

was f i l t e r e d , washed wi th water and d r i e d . It was pur i f i ed through

column chromatography over s i l i ca gel using hexane : chloroform (20

: 80) as e luent . The compound was r e c r y s t a l l i s e d from chloroform-

hexane mix tu re .

230 : Yield 82%; m . p . 123-24 'C; M* a t m/z 240

IR(KBr) : 3000 (NH) & 3320 (OH)

1.18-1.25 ( d . 3H, CIJ^, J = 7Hz), 4 .20-4 .30 PMR(CDCl,+DMSO-d^) i o

151

( d , IH, CH, J = 9Hz) , 4 .79-5.02 (m, IH,'

CH), 5.89 ( s , 2H, OCHÔ), 6 .66-6 .87 (m,

3H, Ar-H)

'^ lo"l2 '^2°5 • Requires : C. 50.00; H. 5.00; N. 11.66

Found : C, 50 .41 ; H, 4 ,82 , N, 11.40

1 ,2-Diamino- l - (3 ,4-methylenedioxyphenyl)propane (231)

To a suspension of 230 (1.2 g , 0.005 mole) in methanol (20

ml) was added Raney-Ni {'^0.5 g) and hydrogenated at 2.5 mg/cm

for 2 h r . After removal of the c a t a l y s t solvent was r educed to

--lO ml and t h i s was used for the next s tep as such .

However, for c h a r a c t e r i s a t i o n 231 was i so la ted as i t s d i h y d r o -

c h l o r i d e . 231 : m . p . l O l ' C ; IR (KBr) : 3000 (NH) PMR(D20) : 1.31-1.39 ( d , 3H, CH^), 3 .60-3.85 (m, IH,

CH), 4 .05-4 .17 (m, IH, CH), 6.19 ( s . 2H.

OCHÔ), 7 .08-7 .21 (m, 3H, Ar-H)

C ^ Q H ^ ^ C I N ^ O ^ : Requires : C, 52.06; H, 6.50; N, 12.14

Found : C, 52.00; H, 5 .95; N, 11.94

Methy l -4{5 ) -methy l -5 (4 ) - [3 ,4 -methy lened ioxypheny l ] -4 ,5 -d ihydro imida-

zole-2-carbaniate (232)

To a solution of 231 ob ta ined from above in methanol (5

ml) was a d d e d , N ,N-d ime thy lxyca rbony l -S -me thy l i so th iou rea and £ -

TSA in c a t a l y t i c amount. The reac t ion mixture was re f luxed for 5

h r . I t was cooled to room tempera tu re (20''C) water was a d d e d to

i t , the s e p a r a t e d so l id was f i l t e r e d , washed wi th water and d r i e d .

It was r e c r y s t a l l i s e d from aqueous methanol .

232 : Yield 68%; m . p . l 7 1 ° C ; M" at m/z 277

IR(KBr) : 1700 (CO)

PMR(CDCl2) : 1.26-1.32 ( d , 3H, CH^, J = 6Hz), 3.42 ( s .

3H, NHCO^CH^), 3 .58-3 .74 (m, IH, CH),

4 .23-4 .31 ( d , IH, CH, J = 7Hz), 5.89 ( s ,

2H, OCHÔ), 6 .65-6.85 (m, 3H, Ar-H)

^13"lfc^3°4 ' Requires : C, 56 .31 ; H, 5 .4 t ; N, 15.16

Found : C, 56.12; H, 5.59; N, 15.06

Methyl-3-ainino-2I j3 - subst i tutedphenyDnitroethyl ] crotonate (233-236; Table 29)

152

General Proce dure :

To a solut ion of l - a r y l - 2 - n i t r o e thy lene (0 .01 mole) in metha

nol (10 ml) was added methyl -3-aminocrotonate (0.02 mole) and s t i

r r e d at room tempera tu re (20°C) for 8 h r . Water (20 ml) was a d d e d ,

the p r e c i p i t a t e d so l id was f i l t e r e d , washed with w a t e r , d r i e d and

r e c r y s t a l l i s e d from chloroform, hexane mix tu re .

3-Carbomethyoxy-5-i i i tro-2-oxiinlno-4-substituted'^ phenyl pentane (237-

239; Tab le 30)

General Procedure:

To a suspension of 233-236 (0 .01 mole) in methanol (10 ml)

was added hydroxy lamine h y d r o c h l o r i d e (0.02 mole) and sodium ace

ta te (0.02 mole) . It was s t i r r e d a t room tempera tu re (20°C) for 5

h r . Water (20 ml) was added to the reac t ion mixture the s e p a r a t e d

so l id was f i l t e r e d , v/ashed wi th w a t e r , d r i ed and r e c r y s t a l l i s e d

from aqueous methanol .

3-Carbomethoxy-5-nitro-4-substituted _ phenyl pentan-2-one (240-242;

Table 30)

General Procedure:

A mixture of the compoiind 233-235 (1 g) and sulfur ic ac id

(2 d r o p s ) in methanol (10ml) was re f luxed on water ba th for 6 h r .

Water was added to the reac t ion mixture and e x t r a c t e d wi th e t h y l

a c e t a t e . Usual work up of organic l aye r y i e lded an o i l .

3-Carbomethoxy-3-hydroxy-5-nitro-4-substituted^ phenyl pentan-2-one

(243-246; Table 30)

Method A:

To a solution of 233-236 (1 g) in d r y CH CI (5 ml) was

added e t h e r i a l solut ion of monophorphtha l l i c ac id (15 ml) and s t i r r e d

at 20''C for 20 h r . The s e p a r a t e d p h t h a l l i c ac id was f i l t e r e d , so lvent

was removed and on t r i t u r a t i o n with pet ro leum benzene a so l id was

ob t a ined . I t wa r e c r y s t a l l i s e d from chloroform, , petroleum benzene

mix tu re .

Method B:

To a solut ion of 233-236 (0 .01 mole) in d r y CH CI (10 ml)

• was added m-ch loroperbenzoic ac id (0.02 mole) and s t i r r e d a t 20°C

for 24 h r . Work up was same as in method A.

153

3-Carbomethoxy-3-hydroxy-5- iutro-2-oxini ino-4-subst i tuted ^ phenyl pen-

tane (247-248; Table 30)

The exper imenta l p rocedure was same as d e s c r i b e d for 237-

239.

1,6-Bis carbomethoxy-2-methoxycarbonylaniino-7-methyl-5-p-methoxyphe-

n y l ) - 4 , 5 , 6 , 7 - t e t r a h y d r o - l , 3 - d i a z e p i n e (249)

This compound was p r e p a r e d e s s e n t i a l l y by following the

method as for 232 but he re ref luxing time was 36 h r . The compound

was pur i f i ed through column chromatography over s i l i ca gel using

hexane : Chloroform (1 : 1) as e luent . It was r e c r y s t a l l i s e d from

ch loroform, hexane mix tu re .

249

I R ( K B r )

PMR(400 MHz, CDCl )

13 C-NMR{DMSO-d, )

6

S9»25^3S

Y i e l d 20%; m . p , 139 ' 'C; M a t m/z 407

1730 (CO)

1 . 5 4 - 1 . 5 5 ( d , 3H, C H ^ ) , 1.59 ( b s , 3H, C H ^ ) ,

2 . 9 0 - 3 . 0 2 ( m , I H , CHG ) , 3 , 50-3 .60 ( m , I H ,

C H ) , 3 . 4 5 ( s , 3H, CO^CH^) , 3 .75 ( s . 6H,

OCH^ & NCO^CH^) , 3 .82 ( s , 3H, NHCO^CH^),

4 . 5 9 - 4 . 7 0 ( m , 2H, CH ) , 6 . 8 6 - 6 . 8 7 ( d , 2H,

3 & 5 A r - H , Jo = 9 H z ) , 7 . 1 9 - 7 . 2 1 ( d , 2H,

2&6 A r - H , Jo = 9 H z ) , 7 .27 ( s , I H , NH)

1 8 . 6 1 ( q , C H ^ ) , 4 5 . 6 4 ( d , A r - C H ) , 51 .78

( q , CO^CH^) , 54, 65 ( t , C H ^ ) , 54 .92 ( q .

NCO^CH^ , NHCO^CH

C=N), 160.95 ( s ,

& OCH^) ,

NCO^CH^ &

1 5 0 . 0 1 ( s ,

NHCO^CH^),

171 .84 ( s , CO^CH^) , 1 1 3 . 8 4 , 1 2 8 . 4 1 , 129 .44

& 158 .39 ( A r - C )

R e q u i r e s : C , 5 6 . 0 1 ; H, 6 . 1 4 ; N , 1 0 . 3 1

Found : C , 5 6 . 5 1 ; H, 6 . 3 5 ; N , 10 .65

6.2 TABLES

Physical and Analytical data of Compounds, which have not

been included in Experimental section, are described

under vertical columns and their spectral

characteristics have been recorded in

horizontal sequence.

154

iT

a 3 o fa

>.

c <

T3

U •wi 3 a (U

3 •

o _: f o " S o S

e.^

T3 . O. O S 2 o u

U

U

00

i n

oo o

i n

OO

O

z — rt O

•—I U

u

e

XI u o

X

t -

X

u X .

o • m

T)

o "i

I —I +

X

XI I

<

^ X 00

o

-t E ( \ j — •

.. o — <?•

U Q U

2

o oo

XI u

o •» XI

•= si-f - Z

X

o 00

o

Ui o

0:1 •

<M r - t

i n

m O 2 '-

r- in X (M

00 ~ ^ 1—1

U

i n

f - H rg

^

0 (T-

rr\

^^ PO

X I u 0

. X pn

•« w

0 f -

•

in M «J S

-

X 1

) H

<:

• X ( M

• t

E -^

0 0

• 00

1 f - H

0 0

t~-

00 s

U] M

-~" ITJ

XI ^ u ..

i n

r X u

* X <NJ

i n 00

I <?•

I l-i

<

X i n

o

I

o

J ^ 3:1 U Q U

S a.

i-i <

X

J: E o ^ u

00

i n

o I i n

o 00 ^^ S XI

00 00

i n

C3 in

fM

IM

O 2 —

in in

X ' ' I

u

u CQ

XI u

SI 2

in o

(M <M

155

1 O CO

IN = 1 u

I 2

XI u

I

2 "a"

XI u o u

X

in •«a<

o I " I

o o

XI ^

X IN

to

00 ^

o X •O 2

I O OS w S XI Q I u a. <

U Q U cc:

CO

' J '

(N

o 2 —

CO CO

X f^ o -^ fM

U

o u

3: o i -H

CO o • CO

1

co "

o 2 w

o I O •v ^

z ^

Z'^ Z I

Hi

CO in

2 « • m m

N

XI * .

A-

x S in

• ^ s o •-f - . - .

• XI IN 2 o . X

6S S CO

e

-::xi

u <

o X IN CO

B

^ '

o • • CO

« - ; K|X|

156

5

in

c <

•o a 3 o

•a 0) u 3 cr

nj

9 • Eg o _• ^ O

o

- o

a:

a o E Z o o

u

a:

u

00

00

i n

o

CO

in

2 '-00 (M

U

o 00

= a •4-«

o

< ^ ro

CO "E

in ta

<M

^ . m

XI u o u E 2

* I t n

s „ Ul Ul «J 2

• » — =1 2

•s

• 1 1

1

o o

32 i n

=:l u

X

M

-M

—' I—1 1—1

• 1—1

CO

f~ 1

00

t~-

^ X u

a,

1—1

lf> I

o w

O u

Kl U

X o o , ^ cr

o o

00

o 1"

fM

O o

o in (M in

in O

' J ' 2

i-H O

X '^

u

o

I o

C O V c ^ E o o I -H

•• *- u CQ M ^1 H H |

(0 ^ 00 o

• i n

• ^ X I u

o

OO

o

i n

o

'3' in

in

O in

2 — X <

u

o o CO

A w

O X O

^ : z x

o

I o u 2

157

UJ

• en

. -—

U)

s • •

J»-s.

XI 2

m X u ^

a: ro

^ to

INI rg

• (NJ

^ ^

o8

XI 1

<

X i n

• E

r—i

•Ji f n .

I u ,

X ro

+ j

•—1 tvi

• 1

>o CT>

• o

•• ^

t-1

o t--

>o

^ XI 2

* X i -H

• n X) ^ CO

T3 1

O W

S Q

OS S cu

•* O U

o PS)

r-1—(

•a

o m vO 1—1

,- u 03 : ! < : 2 • 1

i n 1

m i -H

i n

* (M

XI u

X (NJ

* cr *^ INJ o •

^ 1 o 00

f n

• •• , X I r>.

u f o

158

A &

in

>• c <

C 3 O

XI 0) l-l

• 1 -4

3 o* (U

OS

o • ^. o - . S o S

5H

* O

X

a o B Z o u

u

u

i n

o -

<M CO

• 'a*

o > o

r -i n

i n N O

• c r

e g CO

^

m o ^

t -i n

i n

O ( M

2 — ' T O rt O^

X " J TT ^ -

1 i n

_

u (V)

o u *

a:

o sO

r o

— r<^

El U

( M

. X

• in

^-^ o >

•* ( M

* .-

CO

OCi u

1 i n

• X m

Ul

1 u

<

X i-H

•a " • ^

0 0

t>-1

00

« ^.^ ti

X

II

E "-)

N

X

II

0 >->

„ S I

1

< vO

^ X <—1

, •a T3

CO

r~

^

• ^

oo

" m

( M o ^

• m i n

INJ i n

• Tj<

f -oo

m

0 0 fV]

• ^ i n

i n

O 2

U fvj t-l

X ^

—, i n

• o -o m

^

1 m

,-,

X l u ( M

o u • X

in

i -H

^ o

1*1

• — »

f i

X l U

1 <M

• X

* in

i—t

•>* (M

« ^ • 1 ,

P^

3=1 U

1 i n

• X r»1

in

a^ o r o

N

~E

-M (Q

•1-

s in

0 = ;

• N

I <N]

II

E 1-)

•

0 0

f—1

" T ( \ ]

• i n

f M i n

• r -i n

CO i n

• •«r ^H

CT-

!—( • i n

m I -H

• 0 0 i n

I H

< OJ

* X r-*

I "

0 r o

2 — i n ! > i-H oo

X "M I " "--

u

i n

X 2

' ^ rU

CM

^'^ X

II

U Q U

a.

O U

o

o

331 I l-i <

U

IM

O I

in

o

^ - <

OQ

0:1 • I-H O

(M

II

E u

in ^

- * I m o

U Q U

S a,

O o

XI I

<

in o o * - X

i-H psj

- E

CQ tali 00

^ ^ 0:1 • i-i t -

U

o

(M I

m

2

X 2

159

r o

3=1 U

I N

O u

• X m

•» V)

•^ o i n

• r o

* m

XI u

1

« K r o

en

—.^^ o •«j<

• ( N

,-» m

XI u

1 i n

. X r o

• CO

m o

« »M

„ ^

sO

T J

6 Q

+

X nH

K

T 3

O oo

• p -

1 oo r -

f ~

«, ^-^ N

X (NJ

II

E I D

^ N

X <T-

II

o I - )

« XI

1

u . «

•« < > — t .

O

o o t>» t-i

..

u m Ui < t a ^

K -<

o •-J

* X i u N O

• s o

+ S

• •

(0

S

X ( M

II

E <-i

x" 1-1

< p j

i n <T-

1

m^

2

U

M

w i - i

I i-H

m

XI u o u

X CO

i n

oo o

XI u 1 <M

^ X CO

. w

-' i n

•^ •

(VJ

A

-- m

XI u 1 i n

. X m

+ s M (0 (3

2

•« -—* N

X f M

II

E 1-1

» X

1 u

< r g

vO o

( M

•• ^ - s

, ^

o

• r~ 1

m 00 r-H

u Q U . B:

S a.

• « "—^ O u ^ i ^

o 1—1 p -1—I

^ ^1

0 Ui ^.M"

« HH

>o r~

* -~* X u <

-o • 8

i n

• X f M

^ E ^ <M

^ •

<-

00

m oo

i -H

o

tM >o

O ro

2 — m in f M • ^

X f^ 00 ^-'

U

I o

3 03

rg X 2

«M

•. ^^ m

XI u i n

* X m

* V)

•

o

• e g

• ' • " *

XI u X

f M

a.

X "<1<

( M r -

• •£> 1

fM >o O

.— ' M

XI 2

X <M

^ M

J 3

— •

oo f-H

. > o

1 i n tr -

E >->

^ XI

1

< f M

> X <—t

• • 0

i n CO

h -1

oo

. P -

« - N

X f M

i n XI U

fM fM

N X

— o

xTs u

I

XI I u < ^0

m

o

O 1

i n oo

• o

••

m

XI f M

o u

X m

ro * ,.H

u Q U OH S Ok

•* O

o

r-H

a. ^•s

tH

CQ M >»>• OS M

u

pn i n

p->

x"i t i-i

< iS\

X r-i

* •o *—•

a

i n

•^ PO

N

"E •M (d

+ S

•• (0 (0

S • at

^ N

X r g

II

00

oo

fM OS

• sO

(M OO

• o 1—t

f-sO

l4 u T3 >s

x; • E (U

x M ta

XI (U (0

>> ^ - 4

10

c <

OS

sO

2 — •-I O S

(M t~-X <^

I-H s—

(M u

o sO

I fM

a.

u I

2

(M

X 2

160

o

XI U

X i n

o

I o in

:i u o

X

o

i n

i n t~-

• ro

• ^

ro x\ u

00

O U

X

* X 1—1

• •o T3

^^ 00

t^ 1

o

i n

o 00

X\ u o o

X

o i n

• f^

_ po

XI u

1 Ps]

X m

<:£

•• M (0

(0 Si •• N

T ^

(NJ

II

E <->

i n l-i < rv4

• X '~' 1—I

ppi

XI . U T3

X

* n ^ o o f M

*-

1 1—1

a> • t ^

-X

1

u <

J^x" u Q U

« S a. ••

« * H

o u ^^ o 0 0 sO 1—1

•• rf-^

u 0 :-1

f - H

o

. ^

r X u

» X INJ

m in

in O 2 --

i-H i n

X f^

i-H * - ^

U

o in

o in 1

in in

0 - 2

B:|

X 2

M

0 0

XI u 1 pg

» X pn

, i n t ^ PO

N

~ E

o

PO

XI

<M

X

i n

XI u I

X PO

X

XI I u <

Pvl PvJ

• Pg

» j . , ^

P*!

XI u

1 i n

. •nH

PPl

. M

•o —' (\J

o~ • t— 1

o

r-^

^ XI

1

u <

X

00

oil

•• pn

o " * I - '

f—1

u u >—^ a: S a. —

O u ' -' o o r-1—1

„ ^ • ^

4->

(0 0) 2

«

f

o i n

a

vO

* I M

XI 2

<M

* X ^

* w

( M

*—' pn pn

• vO

_

161

1—t

ro i -H

p - i

PsJ

t ^

CJ> ^H

( M • O

CO O^

• m I -H

>£>

co

<M vO

rr O

^ 2

o X

f—1

CM

U

CO

O 1—1 1—t

1

m

fVJ

Z u 2

<M

2

f~l fo P H

^» o M"

(NJ

•M U

2

1 1 U

1 1

u 2

CO X I u

1 i n

( M

1—t

• e ...

oo i n

" ( N 1

1—(

O

( M

• -•-

I M O

N

E • M

nj

+

s V) in

S

.• /—V

X I 1 U H X

(M

. X

• 1 - '

o *-H

»—1 1

0 0 0 0

o

„

<

* X m

E ' • ^

i n

m r-

1 i-H

o

^ f

^ X

U Q u S s a, ,„^

o u o

1—1

^

Ui >»« K HH

r

o u • 8

( X

1 2

X i n

E*

1 m •««•

1 • m

ro I -H

r-0 0

i n

CO

o CO i n

•o oo

• ro I -H

1—1

i n

o

<T-i n

O i n

__-

" H i - H |

U 1

2

•-T H-t

in

o • ^

m

__-

X <M

II

E

N

X o

II

o >-l

X I 1

) H

<

m X I U

1 I M

A

H H

f<l

. M

i n

^ •

( M

• " ^

* X I -H

^ •o T3 ~—

CO I -H

1

o •

t -

^^ m N

X I U

1 i n

TJ«

O X ro (*1

2 '—

X i n ^ ^ i-H

o

n 1 o V)

m vO

1

1 ^

ro

••

X o

II

0 <-t

, X I

1 u —

o o • -H

(v j • •

X 2

O * r H

••

tH

Uj ^.M^

cd HH

^ O^ 0 0

•

>o

* c

X I

u fM

O

u

x"

» <o

—

m O CO

N

"E

(0

+ 2

in in

S

N

X ( M

II

E >->

x"| 1 u <

• X I -H

•o

f M

1 o >o

r-

162

Z X

Xi

u

(0 c <

•V

o

3 CT 0)

3 .

u •> o • t . o ^ S o 2

(U

E o "

X) . a o E 2 o u

u

u

o i n

o

o "S"

N O

CO I -H

• r^ r-

r-H 1—1

vO

m i n

, >o

CO

• ro r-

CM

O z i n

1—1

ac '»< I -H

u

~ o <NJ <VJ

^

1

o

,., m

u o u »

I -H

i n

m

X I u 1

X CO

* U)

o

rvj

• ^^ XI o

1 i n

fc

X p n

(0 • - ^ OS

( M

f M O

^

0 0 I -H

-o

o I -H

• sO vO

^ oo

"J"

r-i n

o

• sC N O

f

O 2

o> r H

X >o I -H

u

—. o CO fvj

—'

CO

XI u o X (VI

X o

w (M

o3

fm^

ro X I U

X m

* w

i n

<NJ

* XI U

1 i n

X* pn

» V) • ^ - ^

o o

0 0 (\J

N

~E

Id

+ S

(0

.. — X

1

u < • X

I -H

0 0

^

i n I -H

i n

o ro • vO

vO

<NJ i -H

i n

CT~ ^r

i n

i n ^O

^ o 2

i n i -H

X i n I -H

u

—^ ro r-<M

—'

CO in

o

m

Q S U

O u

o

— E IH . ^

OQ ^ O

TT OSI • I-H t ^

i n

M (0 Id

2 • •k

**•**

X 1

I H

< « X

i n

r?

I-H 0 0 I -H

>» X o x • J

0) E • Q

1

^ « m

<M

'U

o u K 2 a.

• at

^ o u ^"^ o

i - H

vO

( • vO

ro X I o

PM

O U

X

0 0

V C <0

ID 2

1 0

—' M U w

09 M rH »— i n

Kl •

po "O

o 00

Xi U

IN O u

i n

i n

X\ u I

<M

I M

XI u 1 i n

. X m

^ U)

^-'

o o

• M

+ S ,, V)

w <o S

•» X

1 l->

J <

Q E o —

O U

o

CQ

i n

O XI

X

i n

o 00

m 00

o

INJ

O 2 --

CO in (VJ - O

X f

U

I O ^0

° X >< 0 N 42 C •>-'

I I

XI t^ u s

(M O " Cj in

XI

i n

U

<

m XI u

1

. X ro

E

o

t ^ 1

i n

o

0 0

( M

^

XI 1 u

<

rr\ X X u 1 i n

a k

X pn

V)

^ 0 0

o» i - H

•• ^

^

* G *— i n 0 0

>o 1

i n

o

— I M

XI u r<-i

f - H

o Q U tim^

cn s a.

"* o o

o o o »-H

^ V4

CQ ^ 1-^'

OS

* X PM

• U]

'" m o

i n

* ^ f i

XI u o

X m

. • i n

W ^0

i n

o fM

<M

O 2 —

00 NO <-l 00

X (N

I-H <-- m

U

i n

O

B

V o <

eo to

1 f \ l

» X m

• w '* o^ r^

rva

i k

, f ^

I M

. X fV]

n TD

'— i n r r

1

r o

• rn t ^ X o

1 i n

X m

^ (A

• O^ CT-

r~i

• ^~-

- N

X

II

E •-> ^

N

II

O

XI u o u X 2

^ X m

• in

— t ~ o^

• 1—(

._

^ XI

1

u o I-H

°0

o ^ o I - l

, - M

n :^ "*«•*

oc -•

0 0

vO

PO

r~ •

•o

^ ,^ po

XI u

IM o u

X PPI

• w

^ • ^

0 0 i n

• m

• '"

m X u

vO 0 0 I M

N

^ E

•>-•

1^

+ S

,, (0 in

*i,

.-" • ^

N

X I M

II

E l->

* X

n

o —> «

XI 1

u <

•o • 0

163

164

C 3 O

in

c <

^ z c r

•V

)H

3 cr (U oi

n) I—t

3 .

o •

o

0)

u

O. O

e 2 o u

u

u

i n

o

<M

«o 00 i n

u-i CT-

• i n i—\

o >o

(fl IH

•V

>. J3 - r ^

E (U x: w m

T3

<a in >> ^ nJ C <

00 in

Z ' -00 (M •- I »)<

t~- — r H

u

o

<M INI

I 1—1

<M

K

m XI u

CM o u

XI U

X

XI u

I i n

X

H

a.

O u

XI I u <

X

o

I

00

XI U

2 o * X

OQ t>i 00 ^ ^ 00 Kl .

(M

O o

o

o

i n

o

2 ' -o o fsj in

00 ^ -1—1

U

X

u

XI u

I 2

X

P-r i ^

u 1 rg

, 1

X m

^ in

• (M

« ^-^ XI

u 1 in

X* PO

XI 1

<

X PO

s *'"' o

1

o

00

<

H

cu

O u

CO

XI u O U X 2

X po

oo

o "

^ X u

o "

M OQ

m o

oo

O

P O

2 — >0 00 (M O^

X rn 1—1 —

(M U

OO

I 00 o

3 03 I C

165 ir>

OS

* VI

o

(Na

« -—»

r\ X\ U X

<\J

« ^

• E ^^ vO r-

• 1

rri I -H

1—1

« —

0 0

vO

m Kl U

o u 2

• "X m

* tn

oo

m

^ rg

= 1 U

1 2

^ X

• f ^ - ^

0=1 u

• X rr,

» +J

>o o*

O

O 0 0

o

^

^^

• m 1

sO

* m

-m

acl u

<M O u

X f o

U Q U ^ 1 -

•• O

o <*>

«a

o o r-i-H

,. ** u CQ !<::

OS l - l

• (0

^ • ^

i n

CO

^ m

SI u PM

^ X

. w ^ o i n

• f M

* ^

1 X I 1 o

0 0

N

"e • J

+ s •. in

"^^ x\ 2

fvi

o8

<

* X i n

• E

m m

t ^

m o

o 00

in in

O 2 —

<\] ro X ^

Pvl

U

i n 00

00 I

o

a. X

u

m Kl U

CM

O u

« X m

Ul

•- m vO

« PO

• ,-—«,

,_ =1

1

<

• 3: 0 0

* ro E X\ U

1 <M

a: CO

•k

u ~—'

r~ m

• rg

« i

- - V

»_ o i n

f -1

1—1

o • t—

. 1 1 2 X

<VJ

* X

m (\j 331 U

1 i n

• X rr,

-

o c»

• 1—1

•• o 1

o

N O

* "

X I

u •

X pg

* in

o o

• ^ »

w *

PO

XI

o u X 2

ik

X

«k

W

_ o* 0 0

• ro

r->o

1—1

o ty-

i n

o rr\

• •"T >£>

O i n

( M 1—(

i n m

i n

CO (VJ

i n

O

2 — •'J' 0 0 PM T T

X '*' PM

U

(—1

o

1—1

1—1

1 o 1—1

PM

t—4

>. c 0) JS &

m X u o

1

fn o F H

XI u o u

X

i n

XI I u <

X

u 1

« X m

i—i

rr t--

1

pg

o a-o

pg o

1—1

pg

PM

^

X I

I H

< «

X f ^ ' '

>Tll h—1

u 1 i n

X pr,

. to

o 0 0

^ E

^ o 1—1

t^

1 o 0 0

vO

XI u - - , OM

CO O 1—4

u Q U -

o: S O i

• a .

^ o u ^ o PM r~

u X 2

^ X m

^ U}

""•^

0 0

• p n

M

«-

1—(

i n

i n

I—t

•

vO

t~-

O

2 — 0 0 « > PM O

X i^ vO — PM

U

ro 0 0

o o-

I

0 0 I—1

PO

X I U

PM

o u

«, X f ^

. w '— >o i n

^ xl

1

<

X PM

XI u 1 PM

«

o

1 o i n

i n I M

• CM

* r^

Xl u

1 i n

* T ^

f O

l-l

<:

X m

• E ^"^ 1—)

i—i

p -

1 o i n

o u

. X I I U

PM

. O - u m X

2 X

00

o -^ f- X I ' - U

o o ^ X

' ^ in u ^

CQ

a:| •

166

o 1—(

, 0 0 1—1

0 0 i n

• •XT

O -fSI

00 i n

i n

^ oo

u ( M

O u

w

i-H vO

m

.

u 1

* X pn

i-H

^ i n r—1

o m • r-

1—1

i n

0 0 PO

i n

S | U - 2

3:1

i n

»a

( M

a: ^ ^H

e

o vO

• 1

~* 11 1 l-l <

* X

i n f*^

r-

fVJ

o r-" j *

i n

co i n

vO

O

2 i n e g

X vO INI

U

0 0

>o

>o 1-1

1 i n i—t I-H

_^ I -H

m i n

—

6»

2 O o X z

M X r 0—•'

\ ^

ml •« F H |

Z I

)f 'yi

o o

( M

-r o

u 1

i n

*

^ (0

^ • * '

t n

t * -

I -H

••

aci 1 u < ^

E

CO <V.'

• r-1

-o

u Q CJ ^ w

a.

,, O u "^ o

•s o

I -H

•• u

03 iiei I M ^

OSI I-.I

«o

•. • — •

m

U

o u X 2

^ as

M

u f M

" ^ 0 0

m

•s

i - i CO

• r^

i n I M

r-

m vO

^ —-X\ 2c

f M

. X

* •*-»

o CM

o r-^

i n 1

2 — 2 on i n <^ m i n •

X ^ o • ^ —

(VJ •• U

0 0 0 0

0 0 fVl

1

<M •-H

U (M

o u X 2

* X m

V)

^ CO

ro

^ .

rrt

'^r^a^l ^ 1

u a u

a,

^ „

O o '"^ o

fM r-^ l - < +J

U •a

2 on o -~~ pg fM t^

a: u -,_,-

o <« ^

I -H

•• u

CD i<

K\ H H !

u CM

o u •B

(M XI u 2 ^

S i n

E' ~ -m t ^

1

pn

• po

*—H

—" SI U

167

V)

c <

c 3 o

u

T3 0)

3

0)

cc:

u

0)

Id

3 E o

l-H

o

i n

o

i n

o in

c

i n

in 0

2 — o ir, i-l O

o (M

U

CC

•o , O. O E 2 o U

u

0 0 i n

CO I

i n 0 0

I—I

3:1 U O u X

m in

i n

f o

0=1 u rg

. X m

m

V) • * — '

t~ IM

, i n •£> CO

N

~-E

CO 0 0

^^^ m

x\ u 1 i n

. X CO

^ u ^ on CO

• M l

+ IS

•• in U] (4 S

^ n:

1

< ^

K CO

i n

( M

o 2

fM in X csj

p j ^ (M ^-^

U

u Q U

a: S ou

,

0 u "^ 0 0^ vO <—1

•• ^ w 03 Ui ^w**

OS 1—•

0^ Pvl

f -

1

r-•

>o

r X 2

. X (NJ

M

V) J3

0 en

1 • >o

in

o >«

o Xi +J 4)

E

I

«s

m = 1 U

(VJ o u

X m

00

0

CO i n

• ro

^

u

i n

XI u

I (VI

in o

00 o

i n

m XI u

I in

X

in 00

XI I

<

X

0 in •a 1

0 S Q +

o» r-

f—1 l-H

. r-

ro t - H

u Q U —' B: S Qu

,, -

0 U

0 Pvl t--1—t

.. ^ u CD Ui •^„^ 0: l-H

^ „-^ r

X u 0 X

PJ

• X >o

u> (M

0 CO

m

•a

r-t~-

1 • m

i n

O m

2 — (\j r-

I—( — ' (M

U

i n

o o (M

+->

( M

XI u

I

CO (NJ

XI u

I in

X

in 00

•a I

O w

Q +

u Q U a S

o u

o o

03

168

rg

X I 1

<

X

* • '

rsi i n

t ^ 1

1-^

m

h -

* I I U

<

X V

E

t -<M

• t

t ^ <T-

*

X I u

CM o u X rr\

* to

—•

r-4 O

f—1

i-H sO

• I f l

o

• ( M vO

• CO I-H

1—1 ( M

i n

i n i n

« 0

,_ CO

XI o o u 2

X*

* n

i-H

• ^ • ^

PO

X u

1 <N

^ X PPI

V]

0 0 I-H

(NJ

-.

XI 1 u < »

X m

E* r o — '

X u

1 i n

X* m

i n •

O U Tl< ^ ^

2 ^» pg <M i n INJ ( M t ^

X ^

u

t ^ • o

1—1

0 0 1

o CO <M

O T3

E (0

< 1 TT

* p -i H

.. ^^ o •

m

0 0 I -H

0 0

I -H

<M

• i n

I-H

• t ^

>o

oo PO

oo

0 0

m i n

vO

o

>o

>o O

pp

2 f -

(\ X

-~ r^ O i n

0 0 —' pvi

U

PM i n

^ H

0

o I -H

c CQ

1

ml t -i - i |

>, o

E po

pg p -

• ppi

* ^^ pn XI o

Pvl o u

f r ^ M ^

V)

o i n

. r o

e n

X u

J

X*

CO

o p j

•

p j

~ XI 1

<

x* I-H I-H

ppi

X u

1 i n

> X ppi

(0

o 0 0

• I -H

.,

E

i n ppi

p -1

I -H

PJ

>o

• ^^ PM

X I U

« — X ^ p>]

I -H u u • ^ - ^

cc s O i

> « t

, . ^ - N ,

o o

o I-H

i-H

_ •M <0 0) 2

K HH

(0

^^ ^ 0 ^

^ ^

X I

8 x* PPI

• k

CO

169

in

C <

c o

•a

3 O* V

OS

u

u

o

<d

3 .

o • ^. o ^ S o

0)

* u

T3 . a o B 2 o

INJ

i n

i n

CO

O

'J"

O 2 —

(M 00 <M 1—1

X "* m *^ IM

O

o in I

u rsj

O

U I

VI

(M i-H

XI u I

in

^ XI u

r H <

— X >0 00

O w S Q i n

'J '

t^ 1

1—1

00

O

XI u o u X 2

o

O

X

CQ

^ - 00 OSI .

in o

o

o in

(M

2 — IM t ^

X ^ in ' ^ (M

U

00 1 "

i n (M

>.

O JS • t - «

0) H

XI u o u

X

vO

X I 1

r g

^ X m

^ t/i

N M '

CO I -H

• fNJ

« .— m

X I u 1 i n

X r o

,^ X I

1 l-i

< * X

vO

• e

•o o • p -

t i n I M

o o p-

U O IM

X ^0

U ^ CQ

(Sj •

CO

<M

in in

o

o «>

u-i i n

o •O

CO

^ t -1—4

< t^ H

K •*r* «£

a,

••. "—^ o u

o

^ ^^ m

XI u fS3 o u X 2

• X pn

in

o-0 0

• ppi

^ pp)

•T i l

•<9<

O "* 2 p j

X pg

U

0 0 t ~

Pvl

1

pg

^ pg

S I

170

o t^

• m

• ^

u

X pn

* M ^ • ^

PM r^

(M

* ^"^ ro XI u 1 fVJ

. X m

• V)

"- ^ XI <M 1

U

T—t

T

o

i n

o i n

• m >o

m t~-

' T 1—1

LTl

• *"* ro X I u o u X 2

» X

. w

o TJ<

a

<N

•• '- P O >T>1 1 ^ 1

u 1 (M

X <*> •

Ul

'—

• <M

,— XI 1 u

< -

X m

m

E —'

o p-

p-1

o ' T

t~-

^ X

1 IH

< ^

X '»'

fM

'— p^

X I u 1 i n

> X PO

X r~

* E

Pvl 00

•

pn >0

•

O^ t ^

• i -H

•• "— < b H • * - *

OS S a. •• O O

o ^ p-f—1

•a

o o p-f—1

•• ^ u CQ Ui

r*

f - H

O^

O

• .«-s

m X I u

PJ

o u X z • X

m

^ U)

oo

• p^

« .»— pn X I U

rg O U

* X pn

i n O

i n

in in r\j r-

X -^ i n •—' fM

U

00 o^

o o pn

A

o •o •» E nl

•4-»

O o < 1 "*

XI U 1

in

* X pn

^ H

m

t~-1

pn I—«

o

1—*

- t *

r-

oS

o o

i -H

.. ^ )H

CQ :<:: . a;|

-•1

X I o

Csl

o u » X

pn

. to

^ ^ o • pn

171

>. I—»

«J c <

T3 C 3 O

U

(d H

T3 (U

"B zr v

u

.—* 3 .

o • •* o ^' S o

4J ^

J U

»

•a . Q. o E 2 o U

a: N

i n

i n

o

(M

CO

o

IM O 2 —

X <^

U

o 00

o o I

•>3<

1 O

o m

-• — •

m 3:1 u •> a:

po

. V)

• ^

m ro

M M <0 S

<* i : 2

K ^H

. Ui

JD

-^ r~ vO

• o

(*1 J2

w o

1 i n

o • • — 1

•• • • " ^

X I 1

I H

< •

X i n

sO TI

1

O w S Q +

. m

1—(

fSJ

• rr, r-I—»

U Q U -

o: 2 cu

••, «—« o u ^ o 0 0 vO

^ ^ 0=1 U

* X • — 1

* T3

-' o m i n

1

Vi

QQ M

c l HH

• •" . 331 t~ U (Nl

00

CO

in

(M

i n

^ O

i n

o 2 —

as f

U

o

C3 i n

0~

>» X O

<u E

Q I

00

Xl U o INI

X ^0

o

- XI

X f i m

O i n

^ ^ I

i n

o XI u

- X < ^ nH

u Q U « a,

o u

o

IM XI U

X rg

— cr l-i ^ .

ia in ^ - ^ 1—I

l - t •«»•

CO

i n

i n

CO CO

INJ 0 0

• CO i n

W pn

o I M

2 CO f - H

X i n

>-* vO

o m "—

u

CO ^ 0

I

X o

• • J

172

o 00

• m

• ^"^ m El U o

^ X m

(0

N O

•o •

m

^ *^

m I I U

•, X f ^

f S

•• (fl w (Q

««r<

<::

•• X 2 X

rvi

• X f M

. W

X i

—' o <M

• oo

•a

N O

sO

sO (NJ

• IVJ

__; m

XI u

XI 1

t-l <

. X ^

E

X

o

I .—I O

in

o I

sO

XI U

u Q U ^ ^ f

o: S cu ..

^ o u ^ • ^

i n

in X>

o <M

• i n

f

X u

X .-H < >

-^ cr 03

o

N

i n

o

oo

i n

o

o

CO <M

O 2 —

00 O •~^ <J-

X <M in —'

U

I

X

u I

' J '

X

o

I i n oo

o

rg

N

"E

m Xl U X

rg

^ X o

* m

(Nl

i n <M

• (M

<>a

(VJ rg

a

rg

^ -^

f«^

XI U

• t

X r^

^ 4->

•~^ •^ 1—1

• 1—1

1

in (0 nj S

.— X 2 >i

rg

X rg

^ U}

JD rg

'^ o •*

• 0 0

°9

o oo

t - -

• .*—N

"-n

1 U

< .

X ^

« B

U Q U ^ • c

o: ^ CU

• «

o u -

o N O

vO

vO

• X U

X 1—1

« u

J3 ^ • < '

1-4

0 ^ K j HH

• ^ pg

XI U

o

i n

i n

rg

o

i n

i n

c/J (*>

O

2 ' -O m r-i m

X f

U

CO

I o

o •a •^ E Id + j o o <

^0

1 —- rg

ro >-i

==' «:

X N

rg

m

o

II

E

rg N

. X - ^ ty

ro XI U O

u X 2

II

i n r j

XI I U < in

X rg

M X) XI X) U

X m

I o o

XI

u ^ X

vO (0

6 ""

»_- +

o in

' 5

N - X

rg

II

E

X - U

X

o O . ^ U X ^ ^ rg rg

o- X ^ ^ rg

OQ ,

^-' 00 rg

0 0

m

1—1

1—t

•«r

' T

^H r<-i

r j i n

OO O

m 1—t

t ^ vO

' T

ro m

rg i n

W

NX>

rg

i n

**"*

o 1-3

^ XI

1 u

XI ^ u

X rg

«,

o

r r 1

i n 0 0

m

—^

vO

°a

rg

X rg

•a X>

I -H

r g

CO 1

o ro o

XI u

•>

X ro

* cn

— o^ rg

rg

* • — *

• 0 0

* -^ X rg

II

E •-)

N

X o

m " X u

, X m

Tl<

O ^ >J^ -^ 2 ^ ^ • •

m r-H , - . r j ^

n-» CO * ' i -H

^ ^-^ rt

u

i n

«o

rg 1 f^ 1 ,

r-( r-1—1

1

o

o

^

II

0 •-)

» XI

u <

LTI

oQ

m

^ X rg

PO t—»

U Q U ">—^ o: r

&

• .

O rg U

O 2

1 I "

e* » P H

^ o vO

I -H

,, ^ i ^

^ QQ i<1

« -.

» • a

-a ^ 17-

1 0 0 ro

t«-

^ -_ XI u

* X I -H

» V}

X> -

I -H

rg PO

N

"E • M

Id

+ S

w Id S .. ** M

X rg

II

E >-) •.

N

X c-II

173

(0

r—• (0 a <

T3 G S O

• a <o u d a* o a:

u

u

X) (d H

n)

"3 • E -u " o •

« s o

0)

U

•a . o. o 6 2 o U

CT»

"* O^

C3 i-H

• O

1—(

(Nl sO

m vD

• o

o

• vO

N O

o

< 0

O

2 — CO O

^ o 3 3 fvj

i n " - ' I -H

u

m r~

t ^

1

>o I -H

u

ee o» r H |

1 o

• pn

•

X I u

1 w

•x. (*>

M • • • • ^

r-f ^

•

(VJ

. '•

o o

xT"^ u

X p ^

w

0 0 ( \ i

<V]

'-^

N

- ^ E

(0

+

'• CO "

SI ^ u

X m

•t->

o fVJ

( -H 1

O

I -H

u < .

X i n

6 "*—' o r g

• t ^

* X

f^ "

u Q U

a. • *

o u

o

^ N

u oq ui «*.' OS >-•

X I -H

M

'»' i n

• m

r X U

X fSJ

O* ^ p n I -H

• ^

o 0 0

• I -H

o i n • t ~

oo I-H ,

O vO

t -r o

• >o i - \

o

• f -

N O

^. >o

I N

o 2 —

vO r o fvj "a*

X f ^ a - •^ I -H

u

I -H

'a'

• M

O

. 1 r^^-^

1

o| o «Nj|

1

o I-H

• CO

*

•4-"

Id

+ S

I I 12 u 2 <a

s

X I ( ^ 1

X I U

^

I H

<

* CvJ ^

XI u x

( M

X o I-H

* E

o

1 O o PO

,-^ p

X U

X PO

*J

'*-' PO I -H

• I -H

1

o» o

,-,

i n

•

r - l

• t~-1

o o t - -

X I U

X I-H

'i t ^ I -H

i n

* f M

XI u «

X IVJ

cr

0 0 T f O-

1—1

U U

CU

.» ^ O U

o i n

I -H

4 . (

2

OS «

• po 1

PPI

» '— (NJ

XI u INI

X "a*

E CM

_ ^ pn

p n

m N . ^

r^ E

174

M

H

in >.

t—t (d c <

X) c 3 o

(U

3

v

u

<d

3 • o • fa ":; . o o S

r: u

a:

•o . o. o E 2 o u

CM

i n

0 0

o

0 0

o

i n

o

en

o 2 ' -

i - l i-H

1—1

U

i n

•vj

= 1 u

f M

o t^

m

* —

m X I u

X m

vO .—t fo

N

"E ••-»

(d

. s

^ XI

XI u X m

• • ->

^^

1—1

i -H

1—1

o

I-l <:

«. X i n

"•^

00

t--

00 <—(

nH t ^

U Q _ :

a: S cu

. . <= ^^ o O ^ u

o •43

o

XI u

X

CQ T Ui o

DSI .

i n

0 0

i n

m

in

^ ^

i n

i n O

2 — O vO

00 -^ I-H

U

<VJ

>« X! O

JC

O E

i n >o

m

.

X I u

^ X <M

at

w • o . ^

o i n

f

s crt w

. . — X

1

<

» •"r" ro

X | f-

X

<M XI u

XI w

X

00 I-H

I-H 1

(M O

(M XI u

^ X 00

cr

r -H

u u oc 2 Oi

•• **

o u .

o >o r -i - H

•0

o rv] t -1—1

.. -%

§ K 0WM»

OS HH

vO o I *

1 m 00

f n

a^

^^ pn

XI u o X (M

X

* in

eg ^^

f\

•»

. >o f -m

N

^

• M

<4

0 0

oo •<J"

oo i n

o

o

i n

oo in

I * O

2 - -00 vO

I - l ^

r- -^ I-H

u

o

2 O

«M

175

U II

rg

X o

00

m X u

• X m

* in

• ^

m

( M

» ^—^

-X

^ T3 T5 . . o vO

-o 1

0 0 i n

• o

fO -X

u •

X f» l

• k

• M

0 0 1—1

r H

o

c-H

•—

XI U

^ *~r^

•—)

« w

0 0 t^

i n

^ IM

XI u

•-• X u u OS

s a. .

O u

o i n i>-1—1

•a

o

1—1

••

Id 0) 2 i a _ i >

B: -"

rg

U

oo o

1 m 0 0

m

• -•

m XI

8 *

•-M ro

(0

O

<o •

CO

-o T

^ E • M

(d

+ S

•" (0 VI nt

s •* N

X I M

II

E • D

N

X (T-

II

o

» X u < o

•s (VJ

X ( M

» •o T3

^ i ^

f»i 1—1

• 1

rH O

t^

.—^ N

X (M II

B •^

m in

00

in 'I"

00

1—1

O (M 00 O 1-t CO

X o

• PO

* —* ro XI u X! ( \ j

• X

• n

( M

TJ" fNJ

, PsJ

°a

i -H

<M

• I M

^ —•

cr, XI u

x* n

^ m ro

N

~E -M Id

+ 2

•• U) (0 Id

S

•* X

1

u <

• X • ^

. E _

>o 1—1

« 1

p-t -

^ o

-. 5' r-l

I 00 X

e

^ in r—1

U U ^ M "

o: ^ H

^ cu •* . ^ O U ^ M l '

i n TT h-1—1

«B

O I M t«-r H

•• '^ (d (U 2 ^ j r -

OS - H

r-•

i n

* I M

XI u

a^

X I M

» tj

^•^ CO

o ^ ^ 0 0

• m

* , r ^

PM

XI u • X

pg

176

(0

I—I

c <

o

3

cr cc

(0 t—(

3 . 1-1 ^ O •

o S o

2

0)

u

T3 . CX O E 2 o u

u

O

<T-

in

in

CO

in

00

oo m

00

O 2 — 00 'J" i-H fO

a: <^

u

o

CO

I O

e

x CM

* (0

*- r-^H

ro a.

^ - K

CO

=1 u

« 3C f o

•« to —' 0 0 OJ

• Osj

* '—^ r<

X I u

* ac CO

-M

• .

0 0 r-i

1—4

1 1—1

o •

S

., w w nl

s

Xl 2

* X r ^

W ja —'

( N O^

• 0 0

« ^^ XI

1 u <

« X i n

K

E

m M

t - -

• * - » . X u

^ K o "

•o 1

O w ^ H

^ Q +

* V "-^

^ p j

m • «—1

U Q U

a s a,

,» ^ O u -

o rg r-i'^

i n 1

i n

^ ^-.

CM

XI u

. X ( M

, a-

o >o o r-t

•• u CQ Ui

• TC

1

r-0 0

ro

» <>>

a

')' rr\

ro

N

"" E

CO

in

in

o

in

in

O ro

2 — O 'T

X •—

^

CO 0 0

N O

1 r-r-

• ^

• ~

X | u

XI .,: u X m

^ + j

(M 1-1

I—1 1

CO C3

O

••

' • ' ^

^ T3

0 0 o

• i n

1 i n

o • i n

• t

<M

XI u o

T3 1

o to S Q — Q:

S a. .«

O u "haC

O P J

r~ 1—1

00

O

1—1

„ .—s

I H

OQ ilj

OS

* X rvj

« cr —' TT O

• 1

1—1

00

• <»1

. ^ m XI u o

^ X pn

. «]

'— 0 0 sO

•

-M

nl

4-

S

<n M

to

^

_ X 2

„

1—t

V)

X I

vO

o •

o

• , — V

N

X (M

II

s • -1

a.

N

X

II

o i - j

,> X

1 u <

>o °s

( M

a j

X (VJ

. •o T3 ' ' <y-f—1

1

CO o t>-

^-. N

X (M

II

E •n

* N

X <T-

177

•J3

i n

i n i n

1 "

in in

in

o m

2 — I—( I—I

X f^ QO - ^

u

i n

I O <V1

O •o

E Id •M u ->

» X I

1

o

* X m

+->

o 1—(

, • — 1

1 Ti<

o • o

••

U

. X 1—t

U] ^ H ^

<M O

• i n

« - - N

»M

X I

6 2 Q

o: s cu

.» ^-^ o u —' o pg t ^ 1—1

»0

o -o

X

cr • ^ ^

o

T r 1

o 0 0

f»1

* ^ CM

XI U

^ X

• ^H O^

r< ^g

"E

4-"

m +

<£

,, (0 U) fl 2 ,

X <\1

II

E • n

« N

X o II

o >-> ^

X 1

t-l

< vO

oO

(VJ

0t

X I M

* •0 •0 ^"^ t~ >* • t ~

i n

m h -

r^ f^

u CQ

Kl 00

X

II

E

178

X)

c o

6)

o N

Id

H

•V (0 u 3 o* (U

U

U

O • 1 o

- S o S

u

a:

T3 . o. o E 2 o u

o

i n

r-

o in

O 2 —

>o oo i-l oo

X IN in ^-^

u

i n

fM

u o o

1 " I

o 00

x\ u

I

= 1

i \ l

(M

m

(M CO

. 00 —^ fM

m

U ^

X

I

O U

(4 a> 2

XI I

<

i n

I

o

fN)

XI u X fNJ

i n

fM

OO

o •o

OO

o

in

o

X u

to CO

XI I

t-l

X in

o i n

t -I

fM

XI o r

10

X

t^ f

w fM

2 — «r o •- I m

X fM m - ^ .—t

U

XI u X m

• (0

^ o •^

m XI u

X

• o

•• fM

oo fM

o

O 2 --

rH ^ X fM

U

i n fM

o

X o

fM X u

ID f>4 «M

Q E U • . ^ * OS ^r< alS Ou

+ S

>o i H rg

179

i n vO

• 1

OJ ro t~

^ (M

11

u *

o m ^H 1—1

^ r-

• •

i n

o •

o

_ El

1 ^ 1

<

X in

^ E

^ 00

00

•<r ro

• I *

•• U) u ^ •e:

J! E

1

<

3 i n

-

"T 1—1

QO -H

1—1

fO

• i n

i—t

CO

~E -M

fl

ro (M

O^ r~t

vO 00

• T

O^

00

V) ^^ i n i n

1—1

IM

"<r vO

o o

• r-1

o

XI u

I

t ~

^ i -H

f—1

1—t O^

* -^^ Xl U

1 w

m

CT> O

o

(M (M

o

I

XI u

o

X\ I

<

X 00

i -H

i n

oo o

N

i n

aci

i n

Q

fM

w

X «» O S

CD

l-i <

X m

>0

O 2 —

I-H ^ <

X (M m — f—I

U

o

O X u

IM

XI

X rp>

i n

' J *

Q E

o in

C/3

2 VH

m 00 i-H O

X '^ I—t

U

i n i n

i " " ^

o u ^"^ o o f -i -H

•• ^ »4

oa i<j

(fl M (4 2

•• ^ o X u * X

I -H

ta (M

X u

XI u

I

X

(M

00

i n

o in

2 — o ^ I-H 00

X rn I-H

U

00

in

I

^0

IM XI u

(M

oo o

XI U X

(NJ

X

<SJ

in

U Q U

«3

cu

<Na 00

o

m O 2 - -

rg ' T <M CO

X "* rg — (M

U

i n

1

o o rg

CM l O z-o

z x

180

\o\

o

XI u o u

X m

oo XI u I

X

o in

XI U

fvj

PM

<M

(M

-i - H

•• ^ ^ CQ Ui

00

* E •-^

ID

• t~ 1

r-H

i n

o

. ^ • ^

<M

XI U

M

X fM

181

w K Z ^ P4 1 o ""V «A S ' ^ :

; ti •y 2 ^ eg

o V" JL-Z ^ 2

Id H

to

>< m c <

•o

o

T3

3

0)

u

u

cd

3 •

o •

o

a

«

•a . 0. o E 2 o u

i n i n

m 1—1

o u K 2

s" r

00 i n

i n

i n

o

I o

00

Kl u ( M

o u X rg

^ a vO

* w -'

oo i n

•

, ^ i -H •"S"

N

-*. E

t~-

o •

t ^ i n

O

O • ' I '

2 fVl f M

X C3 (SJ

u

* — N

^ t-\

TT

.•

rp+ XI u X fVl

•

tf]

o rg

fsi

,.

2 ,

VI U)

nJ 2

_, X

IH

< m,

X CO

S 04

O u

o

I

XI u

i n

(M

i n

CO

o

o

00

00 in

O "a*

2 — 'a* oo

X ' i * I—I ^ - ^

u

o 00

I m o

X

o (J«

>o 1—1

•• .0-^

u D3 id

«[ -HI

M N _ ^

O O

• i n

• ""^ XI u

4

«M

o u X (Nl

. X o

(M

( 1—t

r H

o u X 2

^ X PO

' . X I

X I X

X

I

00

^ X I < 2

X I

X X

X

o

IM

m X

U Q

U

2 a.

X I t\

o u X 2

"a" i n

<M i n

»«< N O

m

r-i

XI u o

"^^ 1

<

X 0 0

E"

00 INJ

X I '^ u 2 —

oo rg

f* - i - ac in

O u

-I X

•3

O M OS ^

i-H 0 0

• • •

—. f n

CQ

OSj X I

u

co I

00

X u

u X

X 00 00

in Hil • 2

^ X

u X IM

. X sO

en JD

o i n

• N O

o (\1

<\J

;; g' X

X I

u Q U C6 2 OH

O u

o

o o

u

QS| X I

o u u I 2 Z

V X

a

jQ

H

(0

C <

o CL4

T3 O

3

a:

u

u

n) r-H

3 •

o • . o ^ S o S

(1)

u

T3 . O. O E 2 o U

00

1" in

00

in

O m

2 — m in

U

o a-

fM

El U

in

I

SI

u (M

o u 2

Kl U (M

o u 2

== 4.

w CM

vO

•a

m in

SI 2

;: s

U 00

U 00

XI I

<

X in

O U

o

O

Ui o ~- o Kl •

oo in

o in

in

in 00

00

o in

O

2 — in o^ r-l O

U

fM 00

00

d 0)

0)

S >« ^ o

00

00

U

a:

I O

= 1 U fM O u

182

v> fM

O

•a

U Q U

o U

El

<

X

o 00

o m

•N

-^ 8 ^ 1

fM

O

in

<M

•a

ro

XI a^ U o fM (V,

O U N 2 ^

E

• ^

00

• in

O m

2 — cr in

•TT' fM

X m U

00

o

X o J3 • < - •

(U

E 'ft Q I

eg

U O

» m — 00

r-• m

--^ ro

XI U fM

o u -r«

2

oa

CO

XI u o u 2 * X vO

^ </] fM ^m^

OO NO

• ro oB

1" O

• r^

fM fM

N

"- E -M (d

• ( -

2

M « (0 2

•• .^-N

M «

2

Hrt

1—4

w XI N — ^

in

o 00 1 in

m 00

. >>-s

X 1 t-4

<

• X en

U Q U ^^ a: S cu •• O u • — •

o rr, r~ • — 1

., ^-^ u m « So.*

OS -•

o 00

>o in vO

vO

,-» fM

XI U

« X fM

. •a ^^

o in

• TT

183

C <

o o

CM T Z

S I z CM

o

•0

0)

0)

oc:

u

o

cr

(M

ns 1—4

o _! c o J S o S

T3

a E

•a . o. o e 2 o u

o

o

o o

in

o

o

o

in

O (M

2 -> >0 ' J '

I—I U

i n

I o-00

XI U

o

I

XI u

( M

o u .

X m

at

in

o T

ro

, ^

a T T

vO Csj

N

^ E

4-" 4

+ s m

X u

x" ff^

v>

<N4

o •

.. ^

.. U)

u R]

2 • ik

• XI 1 IH <

« t

X i n

^ (0

J ^ J3 u Q U K S cu •«

. ^ O u ^ M '

0 0 l«-

<o r - l

„ --

I H

OQ » * k ^

U HH

^ • * '

o

t^ 1

1—1

o • t~

. ^^ XI U

« X fSl

. E

OJ

o •

in

OO

i n

i n

in in

in in in in

o IM

2 — O ' i f IM og

X <^ i n ~- ' r-i

U

00

<M I

X o (U

E

XI U O X

X

O 00

<M m

'^ *.

m XI u (VJ

o u

« X rr\

-10

CO i n

s „ w tn Id

S

.. X

1

u <

—r*

CO

XI u

X

o

^0 I

O

XI U

X (\1

-H E U Q U +

in o

IN

f-H i n U I U 12

.. =1 o H X

o 00

cni •

00

o in

i n

i n

IM

r-H

vO

in

in O 2 —

00 (T-

X (M

u

CO

o I

00 rg

X u o

o

I 00 m

XI u o

o

(M

X

w

0 0 vO

• m

* "" m XI u o u b r ^

m

• tn ^~^ o i n

' m

« m

XI u

* X ro

* X CT-

II

o •-)

x"| 1

o

I M

N

^ -M «J

+ s n w nj SI

o IM

U Q U

2

O U

o

x l u

X IM

(M

o in I

o 00

XI U

X IM

E

X a-

O

XI 1 u < in

X

T3

1

oo

Tl"

*

X

1

u < sO

•a

<M

z -o o 00

184

o in

I CI in

in

in CO

u "

CO

N

II

o O in

O m

2 — in o^ rH O

X c> (*1 * —

U

XI U

I u < in

>8

m

X

o

N

T3 • *

o

m

in

U Q U

OS

S a,

CO

I

N

X

U XI E ' - ' U •">

o z

o X

^ E

^- o HH in

N X

II

o •-J

X*l

185-

(S

O

I z CM

o (X

c <:

T3

c 3 O

0)

3 cr 0) a;

u

u

f^ o

o S

4)

a u

X

OS

OS

T3 a . E 0 o 2 U

O"

o

i n

o o

i n

i n m

in O

(M 2 —

>o o >-l 0 0

m —' 1—1

U

in CO

CO i n

X o 2

CO

asl o

(M

I O

acl u O U

XI u X

o

CO

N

. XI

^ < 00

* X r-H i n

o — ^ XI o rr,

X

— in

i n

i n

C<1

00 00

a-

i n

O fM

2 - -o o (M M"

X '^ in ^^

^ U

O

Q U

ec 2 a<

o^ <M

r -1

i n o

p-

o r-l

X

o 2

I

e<\ E

(M

XI u o X

X o

N

+

X XI

I

<

J2 X

' . E r»1 ^—

. 00 ^ ^ P-

XI t

» • X ^

"N

U Q U

«

XI u x"

i n

o in

O XI

i n

X

- E u ^

CQ

05 .

t n

i n

00

i n

o

o CO

i n

O 2 —

CO O r-H 1—1

^ U

o

X o 2

S o

XI u o

XI I u <

fM

in in

a-

XI U o u

X (M

2 II

^ ui

u l

•a — ^^ 00

in m

oo • o j

vO ^ - ^ I rv

2 •• X ' . - _ Ul

n I

O w S Q

o

u X m

o 00

XI U

X IM 2

I U

E -

a-' I * fM

I

XI U

T3 X I ^ o S E Q -'

o <-<

00

X ul + O 2

cr " •Z. w

fd

in S 00

in U l I

^ u ^ <

X 2

in

rr\ I

(NJ 00

XI U

O X U X

o

•• * ^ QQ Ui •mm^

OSI l-<\

1*1 P-

1 1—1

P-

• m

XI I u <

•a

fM

X fM

•o

fM fM

f^

O^

X fM

X u

CM 0 U X

i n i n

0 i n 1—t

Ul

i n

i n

in fM

X

ul . fM >o

O >o U

m 0

• i -H

m

* "^^ X ul

fO X u fM 0 ul ^

Ul

186

i n

00

00 ir>

oo

oo

00 in

in O 2 -«

in in

X (^

U

00

o

I o o-

X\ U

O

X

V)

O

i n

>£> m

• CO

in Ul ni

s

S I

u

X

331 I

<

X in

I 00 O

o

00

.. x\

u -". U X

o u

o

in t-H

o

X\ U

X

^ E u .^ « o • » ^ i - i

«| •

o o

i n i n

o

00

oo

in in

O 2 —

o» i n •-H PsJ

i n —' 1—1

U

00 >0

I o

ro E

M

o •>-> 00 (d

Xi u o u •

X ro

* (fl (VJ

1—1

t -

f O

(fl (fl nj

s • «

^—« I

1

u <

. a: CO

^ E "-'

o i n

ro

1 oo i n

x\ u

a:

S I U X

(fl

Ci E

fM

I o

IM

on U

U Q U

2 04

X

o

o m

I 00

m X I U o X

(M

• in X <N >0 <^ OQ

M ^ E

00

o

i n

m

O 2 —

I—( cy>

X <N

U

X u o

X I u o

X I

u o u

X

(fl (NI

CO

1<

N X (N

II

E i-i

N •-t-t

CT>

II

O

X I I u

<

X (M

X I

u X

(fl (M

o8

t ^ O^

i n

(N

X I T3

•_

< 3 0 0

• vO

1 0 0

>o • o

• xl U X

<\j

(fl

+ "^T*

^ • • (fl (fl Ifl

s _ N

X m II

E

X . . (M

u Q U

2 04

O u

o

o (N

1

in X I I

<

<N X I U

X X (\J (N)

T3 T3

o

o

00 <3^

in in

O 2 ' -

i n 1-4 ^ 00

X (N

U

00

in (3^

I

X

o

^

rg <• (M

(d

2 > > H ^

tfl -"I

1 O 0 0

• PO

r~ vO <7^

• ^0

m V PSJ

X I

o

o

XI

u o u

X

o

oo (NJ

XI -J, U (fl

(fl

X I I u <

o

O 00

(M in

in

in

00

(NJ in

00 O 2 —

•-H ' * ' X ^

i n ^-' ^ U

1

o w s Q +

m

r-1

o

U Q U

*-• -< 0 4

• ^ ^

o u

o (NJ

l -H

^ u

CQ m •Om^

OS - 1

^ ( M

XI u 0 0

X I CJ

* X (*> ,

E —' ^ >c

1

^ rr,

• >*

1—«

00

t~-0 0

1

00 l -H

o

X o

-»4

Q I

T T

ro

* * <N

>< X 0 si <0 E

187

I i n

8 ^

X

^ m N

"E

CO

>o * in

>0

m

XI ; :

o L u <

X X

CO i n

XI u

o

X I o

XI o X en

CO

in

s a,

I

O ^ U X I — o o

— J3

h-i V

o in

i n

O 2 ' -

X

u

0 0

Q U

X o

X u o

I D

(M r ( M 0 0

. o

XI 4J U O -^

i n

i n

X ol

X m

CO

o

II

B

N X

o

XI u o u

X m

. in

XI

<

i n

•a

m

X I M

X ul O*

i n

X u 8i

in

o o

U

8i

0 0

o in

X I u

X

pg

f M

• 0 I

o w S Q

3 2 z

^ I

. m

-o X

w f n

in CO (fl

X I u e

. + (VJ

X U l w

V] . A

•M S

X I X

( M U l

I

o

00

pg

• X r j

•o T)

a>

U l o

CO

in

CO

i n

X pyj

U l -^ PM

O -

•w« • I—I * m

M ^ r- ^ rH

o in

0 0 in

Psl

X o ul .

I—1

i n CO

r-•"T

0 0

o PsJ

2 — l-H Ps]

X (^ 1—1

U

CO

m pvi

I p-i

X I I u <

•8

PM

^ X f ^ p^

X I

O "O

u — X o

0 0 I

p i

N

X pyj

PO

U E

X N X

o

T3 I "^

o ^ CO •«> S Q -^ +

X p j u

Q U

S

O U

o Psl

PSJ

2 "•

y. s i n

I Psl

X P J

.. X I — u u

CO M

« l S I S u

p g II

X a-

0 0

Psl

i n

i n

o

o

Psl i n

O Psl

2 —

r-i <y-X p>J

m — 1—I

U

in CO

E -"

X o 2

X o

X I o

X I u

PS o u

X >o

PSI

in

CO

X I u

X

o

"E

oo

oo i n

Psl i n

r-i i n

«J 2

^,.^ X

1 IH

< a.

X i n

i n

o" Psl

2 oo l-H

X TT

»-» vO Psl pn

— u

vO

1

O w S Q +

i n p j •

r 1

o o

m t~-f—1

U Q U

K 2 cu

^ O U

o Psl

r-t

,~^ u

« ^ > . ^ a: t-H

^ _»

•V" XI u •a

XI U

X* m ^

E ~-' l -H

i n

1 sO

. f ^

• V

i n o

r-m 1—(

X O 2

X

o

0) 2 o 1

eo * M

188

o-

<M

. 1

u o • t

X

(0

• . 0 0 i n

CO

•k

-

>o .

" • ^

O 2

1

11 U

^

•• >o

•0 00 1

O

Q

S 2

1

U

* T) ro "O

X l o

— f\] ^

o u ^

X m

^ V)

o

<*1

o i n

1 i n

a

' J '

^ S I u

^ ^ 1—1

SCI u

• t

X

* ^ M '

i n

• •M

^ o CO

1

>« • ^

•• o

•o 1

O w

Q

* X

o o

« S a,

• m

*- O u o

1—1

,, - u 00 !it:

_ B:

•-«

XI u X

• •o

i n po

^ 1

i n ( M

• V

X I o X i -H

, n •^^ o o • >*

^ • ^

9 X

0 0

0 0

11

o >-> X

u < •o

•8

* X M

•k

•o

o^ r H

r-

N

X

0 0

• oo

II

o <-> X

1

u < in

•0

m

X ( M

T

o» t>-

• >o

i n • — 1

• X

8i U)

—' CO

o CO

. — I M

X ul ^

+J

I -H

t ^

m X Ul o * O"

•<m^

f - H

CO

i n

, m

X Ul

( M o u

V >m^

GO ^H

• <NJ IT)

-^^ X ul

i . •a

O

•

-PO

X U l

* O"

^ -r o^ • o^

N

*" s • M

m f

:s

"• M M <d

•• Ul

1

u < o i n

f O i n 1—1

•a

o

o

I-H

1—t

O

m • CO

•—1 1—1

« ~ X U

f M

o ul " o CO

o

X o 2 II

U l • k

(0 % H ^

189

6.3 BIOLOGICAL ACTIVITY

The compounds syn thes i zed were t e s t ed for t h e i r in v-tvoantifila-

r i a l a c t i v i t y against LitomO'ioidZ'!^ CCUiinii in cotton r a t s , ces toc ida l

a c t i v i t y against Hymznoie-pi^ nana infection in mice and r a t s . Nematoci-

dal a c t i v i t y agains t AncyiO'itoma CZylanicum in hamsters and against

Ni]ppo6tXongLjtu^ bâ'Utiznî'i in r a t s in the d i v i s i o n of pa ras i to logy

of t h i s i n s t i t u t e .

METHOD OF ANTHELMINTIC SCREENING

1. Antifllarial screening of Compounds;

Litomoôidz^ caAiini infection in cotton r a t (Sigmodon hi'tpidu^j

was used as exper imenta l model for the de tec t ion of an t i f l l a r i a l a c t i

v i t y . The vector of t h i s infection namely LiponLj^'iu^ bacoti, ob ta ined

o r ig ina l ly from National Ins t i tu te for Medical Resea rch , Mill H i l l ,

London, was maintained in a room in which t empera tu re (26°) and

humidi ty (90%) were automat ica l ly con t ro l l ed . Culturing of the vector

and the propagat ion of infection to h e a l t h y cotton r a t s were c a r r i e d

as 230

229 out as per the methods of Hawking and Sewell and Sewell and Haw

king

(a) Preliminary tox i c i ty s tudies :

Before admin i s te r ing any compound to infected cotton r a t s ,

the p r e l i m i n a r y t o x i c i t y s tudy of each compound was conducted in 231

mice . This s tudy he lped in finding out the maximum to l e r a t ed dose

(MTD) of the t e s t compound in mice. One fifth of the MTD was used

against cotton r a t . (b) Selection of infected cotton rat:

Recently infected cotton r a t (3 months or so) wi th p r o g r e s s i v e

r i s e in p e r i p h e r a l microf i la r iae and having count of 250 mf or more

per 5 cmm of blood were used for sc reen ing .

(c ) Testing of Compotinds:

For p r i m a r y screening two infected cotton r a t s were used for

each compound a t any pa r t i cu l a r dose leve l s t u d y . Suspension of the

t e s t compounds were made wi th 0.1% Tween 80 af ter gr inding i t to

a fine powde r . The pH of a l l the solut ions were f ina l ly ad jus ted

to 7 .0 . Infected cotton r a t s were injected i n t r a p e r i t o n e a l ! y l / 5 t h MTD

190

in mice for six consecutive days. Tail blood of the animals were

examined just before the commencement of treatment and thereafter

at weekly interval upto 42nd day. Generally, 5 cmm of blood were

taken from the tail every time and smeared on a clean glass s l ide .

The smears were air d r ied , dehaemoglobinised, fixed in methanol

and stained with 5% Gilmsa for 1 hr in the usual way. These were

then examined under high power magnification of the microscope and

the number of microfilariae per cmm of blood counted. Compounds

capable of cleaning 90% of the pretreatment circulating microfilariae

were considered as the effective microfilaricidal. The animals were

sacrificed on 42nd day (counted from the day of commencement of

the treatment) and the adult filarial worms were removed from pleural

cavi ty , kept in a petry dish containing normal saline at 37° and incu

bated at this temperature for 15 minutes. Animals were sacrificed

and adult worms were examined for motility, cell adhesion on the

surface and changes in different developmental stages of mf in the

uterus. If the worms were found dead, the compound was considered

as an effective adult icidal .

2. Screening against Ancylo^toma ce.ytanicum

(a) Adulticidal activity:

This nematode was obtained from Sarabhai Research Centre,

Baroda and each hamsters (weighing 40-60 g) was administered 50

to 60 larvae oral ly . The larvae were harvested lO-iS day old faecal

cultures by the Baermann method. The screening technique was essen-232

t ia l ly that of Steward with slight modifications, to suit the local 2'i3

conditions . Animals in different groups were administered the comp

ounds (made to suspension with Tween 80) orally in varying doses

using three infected animals for each dose; the other three infected

and untreated served as controls. The administration of compounds

was initiated on day 18-20 of infection either in single or multiple

doses on successive days . The experimental and control animals were

sacrificed on day three after the last dose, their worms were counted

and compared with control group of animals. The efficacy has been

expressed in terms of percent worm reduction. (b) Larivicidal activity:

Golden hamster of ei ther sex weighing 40-60 g were inoculated

orally with 60 ± 5 infective larvae (L ) and divided into several

191

groups, depending on the number of doses, routes of the drug adminis

tration and the stages of parasites against which the action of the

drug was to be evaluated. L of this parasite moults to L on the 2:34

day 2 and L. to L on day 5 . The test compound and the reference

drug were administered on days 1,3 and 6 post infection ( p . i . ) so

that effect on L . , L. and L_ stages respectively could be evaluated.

Treated and untreated hamsters were necropsied on day 20 p . i . , when

worms reached maturity in the intest ine. . The efficacy was expressed

in terms of absolute worm expulsion from the host and per cent worm

reduction compared to untreated controls

3. Screening against Hyme.note.pi^ nana:

(a) Adulticidal activity:

The screening technique followed was essentially that of Ste

ward with modifications as described by Gupta et aj_ , The test

compounds were screened in male swiss mice (18-20 g) and male albino

rats (40-45 g) . The experimental animals were infected with 20 viable

eggs and 17-20 days after infection, the faeces of the individual ani

mal was examined for the presence of eggs and those found infected,

were used for screening. The animals were then colour marked, weighed

and divided into groups of three animals, each for different compounds

under tes t . The compounds synthesized were administered init ially

at a dose of 250 mg/kg x 3 (given for three consecutive days) . Inso

luble compounds were made into fine suspension with the help of

Tween 80 (the amoiont used just to moisten the compound), gumaccacia

or 0.1% agar. The experimental animals received the initial dose of

the test compounds, after they were starved for 6 hr. On day three

post-treatment, the animals did not receive any food for 5-6 hr before

they were sacrificed for assessing their worm burden. The intestine

from each animal was removed, washed with normal saline and examined

for worms and scolices under dissecting microscope. Absolute clearance

of parasite along with scolices from individual mice was taken as

the cri terian for assessing the act iv i ty . Even, if a single scolex

or worm remained in the intestine, the drug at that dose was consi

dered in-effective for that particular animal (because the range of

adult worms maturing from a particular inoculum is very wide) . The

percentage efficacy of the compound was obtained from the number

of treated animals freed from infection and those which did not r e s

pond to the drug treatment.

192

The compound showing act ivi ty at the initial dose level , were

persued further in lower doses t i l l a reasonable act ivi ty was retained

by the compound.

4. Screening against Nippo^st/LOngytuA bJiaAille.nAi6:

Young male rats of university of Freiburg strain weighing 25-

40 g were used as the host of N. bâ6itLzn6i6. This parasite was

originally obtained from Wellcome Laboratory, Bechenham, London.

Rats were infected by inoculating 500 infective larvae each (harvested

from 6-10 day old copro-cultures) sub-cutaneously.

The screening technique was essentially same as described

for A. cey£a»iicum(adulticidal) but in the case of W. bfiaîtidnî^, the ad

ministration of compounds was initiated on day 9 post-infection either

in single or multiple doses on successive days .

Parameters selected for identifying active compounds

The death of the adult worms was the primary parameter for

monitoring the antifllarial ac t iv i ty . The percentage of worm clearance

(PWC) was monitored for evaluating compounds against A. C£y£an-tcum in

fection. If PWC was less than 100 at a single dose of 250 mg/kg,

the compounds were considered as less active and were not evaluated

further. In cases where PWC was less than 50, the compounds were

considered as inactive against this infection. For evaluating compounds

against H. nana infection, the total clearance of scolex was taken into

consideration. In cases where scolex was not eliminated (SNE), the

compound was considered as inactive and the complete absence of

scolex was considered as 100% act iv i ty .

Profile of anthelmintic activity of the various compounds synthesized

Almost all the compounds were evaluated for their antifllarial

act ivi ty against Litomo^toidz^ caAA.nii in cotton r a t s , nematocidal act ivi ty

against Ancyio6toma ce-ijianicam in hamsters and against Nippo6tôngytu^

bA.a'SitLznî^ in rats and cestocidal act ivi ty against Ht/meno£ep-C'4 nana in

mice and r a t s . Only the active compounds have been described in

tables 31 and 34. Of these two compounds (159 and 185) had been

picked up for larvicidal act ivi ty and the results are described in

tables 32 and 33.

193

T a b l e 3 1 : A c t i v i t y of Compounds a g a i n s t A. ceyân-tcum ( i n h a m s t e r s ) a n d N. bâ'i>itLznM'i> ( i n r a t s ) i n f e c t i o n s .

Compound No.

A. czyianicum

Dose mg/kg

A c t i v i t y %

N. bâAitizn6i^

Dose m g / k g

A c t i v i t y %

125 250 X 1 100

50 X 1 0

250 X 1

159

160

177

185

221

232

250 X 1 100

100 X 1 100

50 X 1 97 .22

1 2 . 2 5 x 1 7 7 . 7 8

6 . 1 2 x 1 2 7 . 7 7

250 X 1 80.00

250 X 1 80.00

250 X

100 X

50 X

25 X

12.5x1

6.25x1

3.12x1

250 X

100 X

250 X

100 X

50 X

1

1

1

1

1

1

1

1

1

100

100

100

100

91

83.06

78.46

100

0

100

100

50

250 X 1

100 X 1

50 X 1

12.25x1

6.12x1

250 X 1

250 X 1

250 X 1

100 X 1

50 X 1

25 X 1

12.5x1

6.25x1

3.12x1

100

100

97.2

77.7

27.7

0.0

0.00

100

100

100

100

91

83

78

250 X 1

194

E 3 u C Id

u

in V 00

00 a a o

> •a

c n) 00 fl)

IT)

T3

c 3 O a a o u

u 10 U

U

^ ^

x: •M

• ^ I—< - ^ ^ ^ , _ ^

?is» c ? c o r o ^ u O 0) 5 M

V C ^ ft O n)

*t

a >H

^ 4

(d M O

ii ^ " - i H cu to

(0

E 0)

fi 3 

?? e M ^ (0 O

u d)

p > . o U u o a) a w.

c •-• 1) (d

0) c Hi Id

E ^

< o

o 00 n>

•» ->

w

,_ 00

A: « "~ W 00

s t:

1

3 ^ •o S

—, C JS 00 S * c ^ 1 2

V)

v • < - •

"0 in 0) E <d 0) O ^ tH 1-1 Id

^ XH D .

i O M

* J • ©

1

o 3 ^ •o S

^ i . ^

J3 0) 5 •^ 60 Id —< e «3: S E — c;

1

Id ^ °* « •a (u V C 4->

S g « >« £ <d

•o '^ £ O W S 0 a * j ^ fl)

>—1 i n

> 0

l-H CO

( M

O O

IN] I M

O

c H

I N

o o

l -H

l -H

l -H

o o

l -H

l -H

o o

i n

i n

m f M

OO rg

>0 f M

vO p l

vO (VJ

t~-i -H

f M P J

( M ( M

O r j

o> 0 0

^-^ 1

o < h ^

m • I M

>£> i n

l -H

CT»

^.^ i n

• f M l - H

l -H

I M l - H

0 0

*^^ o • o

l - H

I M i n

l-H 1

l-H l-H

*-^^ m • ( M

l - H

O o i -H

i - H

'M

2

' J * <T-

• ^

1 l - H

^ o > f - H

o o l -H

l - H

• -H

2

o o l -H

l - H

• 2

CO

i n l-H

1 o l -H

m m • ( M

l -H

o o o o o o o l-H i n l-H l-H

(M

O

*--f M

ro

CO

'^ o

•-H

f M

^ O

l -H

m

. o

f M

O

^ vO

f M

N O

^ ro

I M

o

* -* >o

I M

vO

- ^ vD

l -H

xO

-. o

' T C7«

^ o -^ f ^ f ^

• l - H

1 " r-

o

1 m ^^ • ^

• r-

^

CO 1

o ^ I M *

o '.fi • ^0

i n %0

o f - H

CO

. o o • o^

o o 1—1

f - H

• 2

r-CT»

I M 1

l -H

^ i n

• o

o o r - l

^ 1

• •H

2

o CD l - H

» - H

• ^4

2

l -H

r-

i-H l-H

1 •o ^ i n

• CO

o o o o >o o o l-H >0 l-H l-H

( M ^ — i - H t - t p g r v j f v i r y i i N j

• O ^ H r n f r i > 0 > 0 > 0 ' > 0 > 0

o o X X m o o m (M >-< i n pg

X o i n fV)

X o o l -H

X o i n f M

X o o l - H

f - H

X o i n

195

00

a o (U

> X)

00

CO

oo

T3

c O a E o o

u Id u

U

Id

w S (M w

[ o / ^

0' Z ^ Z I

Y z X

o eg

O w Z

^ o 00 d (0

— 1 ^

o >-< ^ X! +J C + j C fl) ~H

o « 5 U 6 --•

f - 4

« 0 e 0

••4 M » a Wi

e

o

C 1 <" p y

«) O 0) °

x: 1 ~4 0) %

E o c 00 i; > 5 c o 0 S m ^ u E M

1 4^ (/) Id (Q

c r a 0) * S TJ i ti E 5 s « « "S « o Id

W "0 ^ '"•

S " S o a cj 3 )H z 5 <C G - •^ «

* p y O t! 3 -0) O J, O & ^ IH H

c 'IT

. p «

— IH (-

o o > S o ^

C M 0 . n fl) Id •a o tJ e « E::; S •3 y o w CL 5 >-; t i Id tn Id >.-4 vn j ;

jo ^ —

'^ 0) irt • • '

•3 2 S 0 o-' ^ U 4-" ^ - 0)

V 00 rS 4->

w _^ 00

J<S © ~~-m op <s B

( M I M

0

1—1

CO

0

0 0

INJ

IN]

vO

0 1—1

-0

0 0

0

0 ? 0

0 0

0 0

r-7 I M

(V)

C3

' J "

0

i n ( M

0

0 0

i n i n

i n ( M

0 i n

0 0

0 0

i n m

1 ( M

0 0

I M

i n i n

i n

I I

IM I

i n rvj

I

i - l > 0

o I r-H r-

00

o in o

O ' -< M

00 (M

I

o

o in

o o

2 nH

o r o r- l r O

I M

0

• ^ 0

I M

^ •

( M 0 0

f>a

vO

^ 0

ro f ~

• »M

r-

i - H

ro —** 0

>o ^

• CT> i n

( M

sO

. m

0 0

<M

0

^ 0

t^ I M

• r-t^

1—1

m ** 0

a» I M

• 0

r-

(M

0

*»»^ v O

0 0

I M

vO

* ^ vO

0 0

( M

sO

^ ( M

I M

r-• r~

>o

^-* 0 0

( M

^^ m m

I M f—1

er\ ^^ 0 0

I " 1—1

1

^ H l '

0 0 i - H

0 r-H

1 i n ^ • i *

0

in

0 i -H

V ^ m ^

m m l - H f - H

I M

1 0

^ t^ vO

0

n^

0 ^

" - 0

0

I M

>o •* 0

0

1—1

m ***, 0

0 0 1-1

eg

>0 -"" >o

0

(M

N O

* 0

0

i -H

m *«»» 0

0 0 1—1

I M

sO

*•».«. >o

0 0 I-H

p g

0

* *»» N O

f O

I M

0

"" I M

X 0 0

X 0 i n

X i n <M

X 0 0

X 0 i n

X i n pg

X 0 0

X 0 i n

X i n fM

i V b

E 3 u C

H u

00

c a o

v—t

>

u C m 00

SI

o a E o u

u <d u

O

»-H

o u -1 4-. C C "1 O tt) U E

Id

o ••4

a M I H 4^ C

l - l

V N

l « | 4

o

c 4)

l i V

a.

(U u

E o

c 0) o u v Cl<

«1

<« E • c < 4-»

c t) u u 0)

a.

1

u E u o

c u u V

04

V)

E - w

c <

V 00 <d •M W

,^ 00

X 4) -v . (0 «10

a B

^ o 00

c Id ) - i

J S • • - •

• *H

>

1

S 3 ^

° « .S ^ M 4->

- 4 - 1

> ! > . ^ ^ lU C 00 > n> C o g S u E IH

1

E 0) E —• •i « o <2 c IH V4 Id (d >4-l >M t-l

S S o a y * j ^ , fl)

1

^ 3 ^ * -o S i £ °

Id *

i-i

f X « -M > - o ?

1

^ a « Id -a S E V E :^ •^ 9 o n 5 V4 Wl 10 (d ^4H V4H >H

- . • « UH ' 7 "O 5 o 10

»H S O O" U 4-> ^ ^ 4)

1 fvj ( M

O O

vO I M

f O

' J ' 1—1

^^ vO 1—1

f O f S ]

M

^ 1—(

^ >o 1—1

CO INJ

1 "

1

r~ <—1

^^ • ^

o-IVI

C O

1 i n I M

^^ O o IJ^ IN]

1

•<)• PM

^ ro oo >o (Vl

T T)<

INI

•*^ CO 0 0

>o (NJ

•C 00 o o

o o o

00 1

IVJ

< o o • o

1—1

^ o-

1 • — 1

- i n

, I M

i - H

1 I M ' • W

i n

r-a

h *

1—t •w

2

0 0 1

•—1

^ »M t - -

• <M

,—1 •«H

2

f - t •f-t

2

o IM

O

o IM o o o

*—s ^ .^-v ^ - ^

»M , N H C

r g

- o

I M ^ M '

i n

-« 1—1

<M

«««' f M

"" O

m < h ^

0 0

*^ 0 0

^ i -H

• — 1

*^ m

fVJ

^ i n

^«.* i n

<M % H ^

i n - * N >

ir»

oo 00

IM in o o o

o o

r - i-H ' ^ - ^ ' ^ rt I [>- 00 <0 I I-H I I

I »0 l-H i-H i-H

^^ rg >o vO 00 O • • CO 0 0 t—I t—(

• (M i n • . . ^ ..H f o r-H i-i i-i pg 2 2

o in

m IM IM

O O

o o

• - I f \ j rg »— Tf rg <M

i n

o i n

o r - l

m I M

V U

i n

i n

i n

i n

i n

X o t n I M

X o o •-H

X o i n

X o o i - H

X o i n

X! o o 1-H

• — <

X o i n

197

u u

J l

•!-• M-l

T3 fl)

+ j U (1)

(0

u c o

o u

(fl T3

O

a e o u

u

^ ^

N E u o 5

3 •o

«]

c: n) 00 Id

• J

>

o u o

1

o

'1 Id

•O

c o

M

e Id BO Id

u Id

c o u « a.

00

m ^ 5 ^

o a E o o

o



u Id

i-i Id

Qu

o o

o o

o o

o r-H I

o o

o o

I

00

I

o CM

in

00

+

o o

o o

o o

m IM 1

1—1

PO

+

in n X . o ^ (f) « -»

^ .

• ^

" . ^ a o .

<^ ^

0) 3

i-H (d > 0) 00 Id

0)

> <

ID 00

198

CONCLUSION

The sensit ivi ty of pharmacophores in benzimidazole-2-carbamates

for evoking anthelmintic activi ty is evident from the results of the

biological screening listed in tables 31 and 34. For example, the

oxidised form of 1 ^ (CDRI Code No.82/437 page 115 ) is active as

a macrofilaricidal agent by oral route of administration while the

alcohol 185 fails to exhibit antifilarial act ivi ty by oral route. Yet

another striking example of the effect of minor change in the pharma

cophore on the profile of anthelmintic act ivi ty is obvious from the

comparative study of the results of biological screening of compounds

83/148 (prepared earlier in CDRI) and 159. Replacement of the oxygen

atom in the furan ring of 83/148 by a nitrogen atom (compound 159)

has resulted in the abolition of antifilarial ac t iv i ty . However, only

minor differences in the profile of biological act ivi ty against develop

ing stages of A, CS.yianicum of these compounds have been observed.

A comparative study of the anthelmintic act ivi ty of 159 and 177 indi

cates that the orientation of the ester carbonyl, with respect to benzi-

midazole nucleus and the linkage of the C-5 of the benzimidazole

ring significantly influence the profile of biological ac t iv i ty . Similarly

in compounds 221 the benzimidazole nucleus is not planar with the

1,4-dihydropyridine residue and this in turn makes the orientation

of the ester carbonyl different than the other compoiands. Possibly

th is is one of the contributing factor for the deminished anthelmintic

ac t iv i ty . The biological act ivi ty of compound 232 is interesting because

in th is compound the phenyl ring has been dissociated from the benzi

midazole ring system and the resulting substituted imidazole-2-carba-

mate is capable of exhibiting specificity of anthelmintic ac t iv i ty .

This molecule, therefore, provides a good base for undertaking lead

optimisation studies and should there be a need to develop a specific

antlhookworm compound, this exercise may prove to be beneficial.

The antifilarial act ivi ty of 185 is encouraging and calls for undertaking

special drug formulation studies to induce oral absorption. The proto

types XVI, XVII, XX and XXII have failed to yield a new lead for

developing an anthelmintic agent.

199

REFERENCES

1. S e g a l , S . J . , S y s t e m i c C o n t r a g e s t a t i o n a l A g e n t s , i n : C r e e p , R . O .

a n d K o b l i n s k y , M . A . , e d . F r o n t i e r s in R e p r o d u c t i o n a n d F e r t i l i t y

C o n t r o l , P t . I I , C a m b r i d g e a n d L o n d o n , MIT P r e s s , 170 ( 1 9 7 7 ) .

2 . O m o d e i - S a l e , Amedeo ; C o n s o n i , P . ; G a l l i a n i , C ; L e r n e r , L . ( G r u -

p p o L e p e t i t S . p . A . ) Ge r . Offen. 2 , 8 1 9 , 3 7 2 (C1.CO7D249/08) ,

16 Nov , 1 9 7 8 , B r i t . A p p l . 7 7 / 1 9 , 0 1 2 , 06 May 1977; 26 p p . ;

CA, 9 0 : P72206C.

3 . O m o d e i - S a l e , A . ; C o n s o n i , P . ; G a l l i a n i , G. , J . Med. C h e m . , 2 6 ( 8 ) ,

1187-92 (1983) E n g . ; CA^ 99 : 3 8 4 1 8 z .

4. T o j a , E. ; O m o d e i - S a l e , A . ; C a t t a n e o , C ; G a l l i a n i , G, , E u r . J .

Med . C h e m . , 1 7 ( 3 ) , 223 -7 (1982) E n g . ; CA, 9 8 ; 16614n.

5 . S a k s e n a , S . K . ; C h a u d h u r y , R . R . , I n d i a n J . Med. R e s . , 5 8 ( 3 ) ,

374 -6 (1970) E n g . , CA, 7 3 : 3 2 0 2 9 q .

6 . H o r a c e , A . D . a n d R o g e r , D . W . , U . S . 3 , 1 5 6 , 6 9 8 ( C I . 2 6 0 - 2 9 6 ) Nov,

1 0 , 1964 , A p p l . M a r c h , 1 1 , 1 9 6 3 , 4 p p . ; CA, 62 : P 5 2 5 8 d .

7 . S e t t y , B . S . ; Rao , K . V , B . ; I y e r , R , N , ; D h a r , J , D . (Counc i l of

S c i e n t i f i c a n d I n d u s t r i a l R e s e a r c h ( I n d i a ) ) I n d i a n 148 ,416 ( C I ,

C 0 7 D 5 / 4 0 ) , 14 F e b , 1 9 8 1 , A p p l . 78 /DE 3 8 4 , 20 May 1978; 6

p p . ; CA, 9 5 : P168979m.

8 . P a g e t , C h a r l e s , J . J r . ( L i l l y , El i a n d C o , ) U , S , 4 , 2 7 5 , 2 1 0 ( C I .

5 4 6 - 2 2 2 ; C 0 7 D 2 6 3 / 5 8 ) ; 23 June 1 9 8 1 , A p p l , 1 4 0 , 2 8 9 , 04 May

1 9 7 1 ; 15 p p . D i v i s i o n of U , S . 4 , 0 8 8 , 7 7 0 ; CA, 9 5 : 150641k .

9 . L i l l y , El i & C o . I s r a e l i 50 ,413 ( C I . C 0 7 D 3 3 3 / 5 6 ) , 30 May 1980 ,

US A p p l . 8 2 6 , 0 0 9 , 28 O c t , 1975; 56 p p , ; CA, 94 : P83931au ,

10 . J o n e s , C D , ; S a u r e z , T , ( L i l l y , E l i & C o . ) B e l g , 847 ,718 ( C I ,

C 0 7 D ) , 28 A p r , 1977 , US A p p l , 6 2 6 , 0 0 9 , 28 O c t , 1975; 43 p p ,

CA, 8 8 : P 3 7 6 0 2 r .

1 1 . B e l l , M.R, ( S t e r l i n g D r u g , I n c ) US P u b l , P a t . A p p l . B 402 ,162

( C I . 260-288 R; C 0 7 D ) , 02 Mar . 1976 , A p p l . 1 5 6 , 0 7 0 , 23 June

1 9 7 1 ; 18 p p . D i v i s i o n of U . S . 3 , 8 1 9 , 6 3 7 ; CA, 8 5 : P 46425a .

12 . V i n c e n t , M. ; Remond , G. ( S c i e n c e Union e t C i e . - S o c i e t e F r a n c a i s e

de R e c h e r c h e M e d l c a l e ) Ger Offen . 2 , 5 1 9 , 0 7 7 ( C 1 . C 0 7 D ) , 06 N o v .

200

1975 , B r i t . A p p l . 1 9 , 2 3 9 / 7 4 , 02 May 1974; 32 p p . ; CA, 84:

P 59451k .

1 3 . B e l l , M . R . ; Z a l a y , A.W. ( S t e r l i n g D r u g , I n c . ) U . S . 3 , 7 9 9 , 9 4 3 ( C I .

2 6 0 - 3 2 6 . 1 6 ; C 0 7 D ) , 26 May 1974, A p p l . 1 5 6 , 0 6 8 , 23 June 1971 ;

6 p p . ; CA, 80 : P 133246h .

14. K a m b o j , V . P . ; K a r , A . B . ; R a y , S. ; G r o v e r , P . K . a n d A n a n d , N. ,

I n d i a n J . E x p . B i o l . , 9 ( 1 ) , 103-4 (1971) Eng.

15 . C h a u d h u r y , R . R . , I n d i a n J . Med. R e s . . 5 6 ( 1 1 ) , 1720-7 (1968)

Eng.

16. Huang, W. ; Yang, Z. and Peng, S. , Nanjing Yaoxueyan Xuebao,

16(2), 75-6 (1985) Ch.

17. P a n d e y , V . K . ; R a j , N. ; a n d S a x e n a , Y. , B i o l . M e m . , 1 1 ( 5 ) , 2 0 1 -

4 (1985) Eng.

18 . J o n e s , C D . ; S a u r e z , T . ( L i l l y , El i & C o . ) Ger Offen . 2 , 6 4 7 , 9 0 7

( C 1 . C 0 7 D 3 3 3 / 5 6 ) , 12 May 1977, U . S . A p p l . 6 2 6 , 0 1 0 , 28 O c t .

1975; 81 p p . ; CA, 8 7 : P 84806z .

19 . O m o d e i - S a l e , A . ; T o j a , E . ; G a l l i a n i , G. ; L e r n e r , L . J . (Ger Offen.

2 , 5 5 1 , 8 6 8 ) , B r i t . A p p l . 7 4 / 5 0 , 8 5 5 , 23 Nov . 1974; 49 p p . ; CA,

8 5 : 192731p .

2 0 . Rao , B . V . ; a n d S o m a y a j u l u , V .V. , J . I n d i a n C h e m . S o c . , 5 7 ( 8 ) ,

837-40 (1980) Eng.

2 1 . C o o m b s , R .V . ( S a n d o z , I n c . ) U . S . 4 , 0 4 2 , 7 0 4 ( C I . 4 2 4 - 2 7 3 P ; C07D

2 3 1 / 5 6 ) , 16 Aug. 1977 , A p p l , 6 6 8 , 2 6 6 , 18 Mar . 1976; 11 p p . ;

CA. 87 : P 184493p .

22. H a b e c k , D . A . ; a n d H o u l i h a n , W . J . ( S a n d o z - W a n d e r , I n c . ) U . S .

3 , 9 3 2 , 4 3 0 ( C I . 260 -296 T ; C 0 7 D ) , 13 J a n . 1976 , A p p l . 1 8 8 , 9 3 8 ,

13 O c t . 1 9 7 1 ; 13 p p . ; CA, 84 : 121821m.

2 3 . G r u p p o L e p e t i t S . p . A . Ge r . Offen. 2 , 4 2 4 , 6 7 0 ( C I . C 0 7 D ) , 12 D e c .

1974 , B r i t . A p p l . 2 5 , 1 6 3 / 7 3 , 25 May 1973 ; 46 p p . ; CA, 8 3 :

P 206286V.

2 4 . H o u l i h a n , W . J . ( S a n d o z - W a n d e r , I n c . ) U . S . 3 , 8 1 6 , 4 3 7 (CI . 2 6 0 - 2 9 4 . 8 B ;

C07D) 11 June 1974 , A p p l . 3 1 7 , 5 1 9 , 22 Dec . 1972; 4 p p . ; CA.

8 1 : P 105516J.

201

2 5 . H o u l i h a n , W . J . ( S a n d o z - W a n d e r , I n c . ) U . S . 3 , 8 1 6 , 4 3 8 (CI . 2 6 0 - 2 9 4 . SB;

C 0 7 d ) , 11 J a n . 1974 . A p p l . 3 1 7 , 5 4 4 , 21 D e c . 1972; 5 p p . : CA,

8 1 : P 91519n.

2 6 . Rosen , M.H. ( C i b a - G e i g y C o r p . ) U . S . 3 , 7 8 1 , 2 9 3 ( C I . 2 6 0 - 2 9 3 . 5 7 ;

C 0 7 d ) , 25 Dec . 1 9 7 3 , A p p l . 1 7 1 , 7 2 0 , 13 Aug. 1 9 7 1 ; 3 p p . ; CA,

80 : P 82633X.

2 7 . Yamamoto , M. ; H i r o h a s h i , T . ; I n a b a , S. ; a n d Yamamoto , H. ( S u m i

tomo C h e m i c a l C o . L t d . ) J a p a n Kokai 7376 ,886 ( C 1 . 1 6 E 5 5 ) , 16

O c t . 1 9 7 3 , A p p l . 7 2 , 0 7 , 8 3 0 , 19 J a n . 1972; 3 p p . ; CA, 80 : P48050v .

2 8 . L o o z e n . H u b e r t J a n J o z e f , ( A k z o , N . V . ) E u r . P a t . A p p l . EP 50 ,387

(C1.C07D 4 9 1 / 0 4 ) 28 A p r . 1982 , NL A p p l . 8 0 / 5 , 7 5 4 , 18 O c t . 1980;

24 p p . ; CA, 9 7 : P 127622n.

2 9 . O m o d e i - S a l e , A . ; T o l a , E . ; G a l l i a n i , G. a n d L e r n e r , L . J . (Guppo

L e p e t i t S . p . A . ) P a t . S p e c i f . ( A u s t . ) AU 516 ,343 ( C I . CO7D471/04)

CA, 9 6 : P 162693n.

3 0 . Ho C h i h Yung ( M c N e i l a b , I n c . ) PCT I n t . A p p l . WO 8504,554 ( C I .

A01N43/84) 24 O c t . 1 9 8 5 , US A p p l . 5 9 9 , 0 9 5 , 11 A p r . 1984 , 35

p p . ; CA, 104: 224922f.

3 1 . M e h r o t r a , P . K . ; N e e l i m a ; B h a d u r i , A. P . a n d K a m b o j , V . P . , I n d i a n

J . Med . R e s . , 8 3 , 614-617 ( 1 9 8 6 ) .

32 . N e e l i m a ; M e h r o t r a , P . K . ; B h a d u r i , A . P . a n d K a m b o j , V . P . , I n d i a n

J . Med. R e s . , ( i n p r e s s ) .

3 3 . P i a n c a t t e l i , G. ; S c e t t r i , A. a n d D ' A u r i a , M. , S y n t h e s i s , 245 ( 1 9 8 2 ) .

34 . P i a n c a t t e l i , G. ; S c e t t r i , A. a n d B a r b a d o r o , S. , T e t . L e t t e r s , No. 3 9 ,

3555 ( 1 9 7 6 ) .

3 5 . A k h t a r , M . S . ; S e t h , M. a n d B h a d u r i , A . P . , J . H e t . C h e m . , 2 2 ,

1323 ( 1 9 8 5 ) .

3 6 . A k h t a r , M . S . ; S e t h , M. a n d B h a d u r i , A . P . , I n d i a n J . C h e m . ,

( i n p r e s s ) .

3 7 . M a d h u s u d a n a n , K . P . ; A k h t a r , M . S . ; S e t h , M. a n d B h a d u r i , A. P . ,

I n d i a n J . C h e m . , 2 5 B , 915 -21 ( 1 9 8 6 ) .

3 8 . A u d i e r , H . E . ; M i l l i e t , A. a n d M o u s t a p h a , C . , O r g . Mass S p e c t r o m . ,

1 8 , 132 ( 1 9 8 3 ) .

202

39 . H o l t z c l a w , J , R . ; W y a t t , J . R . a n d C a m p a n a , J . E . , O r g . Mass S p e c -

t r o m . , 2 0 , 90 ( 1 9 8 5 ) .

40 . C l a e y s , M . ; Van A n d e n h o v e , M. a n d V a n d e w a l l e , M. , B u l l . S o c .

C h e m . B e l g . . 8 8 , 799 ( 1 9 7 9 ) .

4 1 . M a s s , R. ; N i x o n , W . B . a n d B u r s e y , M . M . , A n a l . C h e m . , 4 9 , 1071

( 1 9 7 7 ) .

4 1 a . S o m e r v i l l e , L . F . a n d A l l e n , C . F . H . , O r g . S y n . C o l l . , V o l . 1 1 ,

81 ( 1 9 5 5 ) .

42 . G r i f f i t h s , J . S . ; B e a m , C . F . a n d H a u s e r , C . R . , J . C h e m . S o c . ,

C , 974 (1971) Eng.

4 3 . Ho, B . T . ; M c l s s a c , W . M . ; Rong, A . ; T e n s e y , VV. ; W a l k e r , K . E . ;

E n g l e r t , L . E . a n d N o e l , M . W . , J . Med . C h e m . , 1 3 , 26 ( 1 9 7 0 ) .

44 . A r n e t t , C D . ; W r i g h t , ' J . a n d J a n k e r , N. , J . Med . C h e m . , 2 1 ,

73 ( 1 9 7 8 ) .

4 5 . C o h n , M . O . ; N a r i n e , B . a n d T a r n o w s k i , B . , T e t . L e t t . , 3 3 , 3111

( 1 9 7 9 ) .

4 5 a . O t t o , M . ; B r a m h a , N. ; B r i a n , T . a n d R o y , H. , J . C h e m . S o c .

P e r k i n t r a n s . , 2509 ( 1 9 8 1 ) .

46 . P a t e l , J . C . ( 1 9 5 4 ) : A n c y l o s t o m i a s i s in I n d i a , A r e v i e w w i t h s u g g e s

t i o n s for f u t u r e r e s e a r c h w o r k , I n d i a n J . Med . R e s . , 4 2 , 2 7 9 -

304 ( 1 9 5 4 ) .

47 . A r o r a , R . R . ; B i s w a s , H. a n d M a t h u r , K.K. ( 1 9 7 6 ) : P r e s e n t s t a t u s

of Hookworm in I n d i a , J . Commun. P i s . , 8 , 6 6 - 7 6 .

4 8 . Yunus , H. , " P r e v a l e n c e of i n t e s t i n a l p a r a s i t e s (a r u r a l s t u d y ) ,

J . I n d i a n Med. A s s o c , 6 9 , 241 ( 1 9 7 7 ) .

49 . S e n , A . B . a n d B h a t t a c h a r y a , B . K . , I n d i a n J . H e l m i n t h o l . , 16 ,

142 ( 1 9 6 4 ) .

50. Goonertne, B.W.M. and Sen, A.B. , Trans Roy. Soc. Trop. Med.

Hyg., 64, 935 (1970).

5 1 . M u r t h y , P . K , ; T y a g i , K. ; C h o w d h u r y , R .K. a n d S e n , A . B . , I n d i a n

J . Med . R e s . , 7 7 , 623 ( 1 9 8 3 ) .

52 . M i s r a , S. ; C h a t t e r j e e , R .K. a n d S e n , A . B , , I n d i a n J . Med . R e s . ,

7 8 , 210 ( 1 9 8 3 ) .

203

5 3 . M u r t h y , P . K . ; Roy C h o w d h u r y , T , K . a n d S e n , A . B . , J . Communic .

P i s . . 1 5 , 100 ( 1 9 8 3 ) .

54 . C h a t t e r j e e , R . K . ; Sen , A . B , a n d B h a t t a c h a r y a , B . K . , Ann. T r o p .

Med. P a r a s i t o l . . 5 9 , 434 ( 1 9 6 5 ) .

55 . P a n d e y , V . ; K a t i y a r , J . C . ; G e o r g e , P . A . a n d S e n , A . B . , I n d i a n

J . P a r a s i t o l . , 1 , 41 ( 1 9 7 7 ) .

56 . M i s r a , A. a n d K a t i y a r , J . C . , I n d i a n J . P a r a s i t o l . , 4 , 195 ( 1 9 8 0 ) .

57 . M i s r a , A . ; K a t i y a r , J . C . a n d S e n , A . B . , I n d i a n J . E x p t l . B i o l . ,

1 8 , 906 ' ( 1 9 8 0 ) .

58 . V i s e n , P . K . S . ; K a t i y a r , J . C . a n d S e n , A . B . , J . H e l m i n t h o l . ,

5 8 , 159 ( 1 9 8 4 ) .

59 . G u p t a , S . ; K a t i y a r , J . C . a n d Sen , A . B . , J . H e l m i n t h o l . , 5 5 ,

101 ( 1 9 8 1 ) .

60 . K a t i y a r , J . C ; G u p t a , S. a n d Sen , A . B . , I n d i a n J . E x p t l . B i o l . ,

2 1 , 371 ( 1 9 8 3 ) .

6 1 . S r i v a s t a v a , V . M . L . a n d G h a t a k , S. , I n d i a n J . B i o c h e m . B i o p h y s . ,

8 , 108 ( 1 9 7 1 ) .

62 . S r i v a s t a v a , V . M . L . ; C h a t t e r j e e , R .K. a n d G h a t a k , S. , I n d i a n

J . B i o c h e m . B i o p h y s . . 8 , 97 ( 1 9 7 1 ) .

6 3 . R a t h a u r , S. ; A n w a r , N. a n d G h a t a k , S. , 2 . P a r a s i t e n k , 6 2 , 85

( 1 9 8 0 ) .

6 4 . S a x e n a , J . K . ; S r i v a s t a v a , A . K . ; M u r t h y , P . K . ; C h a t t e r j e e ' , R . K . ;

G h a t a k , S. a n d W a l t e r , R . D . , T r o p e n m e d . P a r a s i t o l . , 3 5 , 174

( 1 9 8 4 ) .

6 5 . A g a r w a l , A . ; S a x e n a , J . K . ; K a t i y a r , J . C ; G h a t a k , S. a n d W a l t e r ,

R . D . , Z. P a r a s i t e n k . , 7 1 , 259 ( 1 9 8 5 ) .

66 . S i n g h , G. a n d S r i v a s t a v a , V . M . L . , I n d i a n J . E x p t l . B i o l . , 2 2 ,

91 ( 1 9 8 4 ) .

6 7 . S a x e n a , J . K . ; B o s e , S . K . ; S e n , R. ; C h a t t e r j e e , R . K . ; G h a t a k ,

S. a n d S e n , A . B . , E x p t l . P a r a s i t o l . , 4 3 , 239 ( 1 9 7 7 ) .

6 8 . M i s r a , S . K . ; A g a r w a l , A . ; S e n , R. a n d G h a t a k , S. , J . H e l m i n t h o l . ,

5 9 , 101 ( 1 9 8 5 ) .

6 9 . S r i v a s t a v a , D . K . ; R o y , T . K . a n d S h u k l a , O . P . , I n d i a n J . P a r a s i

t o l . . 4 , 187 ( 1 9 8 0 ) .

204

70. Roy, T .K . ; Agarwal , S. and Agarwal , D.K. , Biochem. B i o p h y s .

Res. Comm. , 106, 888 (1982) .

71. Gibson, D.W.; Connor, D .H. ; Brown, H . L . ; Fuglsang, H. ; Anderson,

J . and Duke, B . O . L . , J . T r o p . Med. & Hyg. , 25 , 74 (1976) .

72. Rew, R.S. and Saz, H . J . , J . P a r a s i t o l . , 63 , 123 (1977).

73. Walter , R . D . , Mol. Biochem. P a r a s i t o l . , 1, 139 (1980a) .

74. Wal ter , R.D. and Schu lzkey , Tropenmed. P a r a s i t o l . , 3 1 , 55 (1980b) .

75. Pampor i , N .A. ; Singh, G. and S r i v a s t a v a , V .M.L . , J . Helminthol . ,

58, 39 (1984) .

76. Ottesen : Transact ions of the Royal Society of Tropica l Medicine

and Hygiene, 78, 3 , Supplement (1984) .

77. Sharma, 5 . ; Dubey, S.K.; and I y e r R .N. , Progress in Drug Res

e a r c h , 24, 217-266 (1980) .

78. (a) Sharma, S. ; and C h a r l e s , E . S . , Progress in Drug Resea rch ,

26 , 9-54 (1982); (b) Sharma, S. and Abuzar , S. , P rogress

in Drug Research , 27, 85-161 (1983) .

79> Bwibo, N.O. and Pamba, H.O. , Helminthological A b s t r a c t , 53 ,

Apr i l (1984) .

80, Chandiwana, S,K,; Makaza, D, and Makura, O. , Central African

Journal of Medicine, 29 (11) , 213 (1983) (En. 9 r e f . ) .

81 Chee tham, R . F . and Markus , M . B . , South African Journal of Scien-

c e , 79(11) , 470 (1983) (En. 4 r e f . ) ,

82. Dickson, B. , Helminthological A b s t r a c t , 53 , April (1984) .

83 . S u p p e r e r , R. and Pfei f fer , H. , Beliner und Munchener T i e r a r z t -

l i che Wolchenschr i f t , 96 (10) , 333-334, 337 (1983) (De, en ,

8 r e f . ) .

84.. Bha ibu laya , M. and Pun thup rapasa , P . , South-East Asian Journal

of Trop ica l Medicine and Public Heal th , 15 (3 ) , 389 (1984) ,

(En, 11 r e f . ) ,

85. Bunnag, D. ; Pungpark , S. ; Har inasu ta , T. ; Viravan, C ; Vanija-

nonta , S. ; Suntharasamai , P . ; Migasena, S . ; C h a r o e n l a r p , P . ;

Rigant i , M. and Looareesuwan, S. , Helminthological A b s t r a c t ,

54, May (1985) .

205

86. B a s s i l y , S . ; E l -Mas ry , N . A . ; T r a b o l s i , B. and F a r i d , Z. Annals

of Tropica l Medicine and Pa ra s i t o logy , 78 (1 ) , 81 (1984) (En,

4 r e f . ) .

87.- Sa th ianesan , V. and Sundaram, R.K. Kerala Journal of Ve te r inary

Science, 14(2 ) , 1 (1983) (En, 18 r e f . ) ,

88. Anaya, R.M.; Ga r ib i , D. ; Milian Suazo, F. and Alcibar Mera, P.

Tecnica Pecuaria en Mexico, 45, 95 (1984) ( E s , en , 11 r e f . ) .

89. Abia-Vega, P . ; Bolas , F . F . and Mart inez, F . A . R . , Ravista I be r i ca

de Pa ras i to log ia , Special Volume, (1982) , 509-518 (En, en ,

11 r e f . ) .

90. T h e o d o r i d e s , V . J . ; Freeman, J . F . and Georgi , J .R . , Ve te r ina ry

Medicine and Small Animal Cl in ic ian , 77 (4 ) , 569 (1982) , (En,

7 r e f . ) .

91 . Han, Y. ; Liu, D. ; L i , P.Y. and Huang, Z .G. , Liaoning Animal

Husbandry and Veter inary Medicine , N o . l , 1-3 (1982) (Ch,

3 r e f . ) .

92. Todd , K.S. J r . and Mansfield, M . E . , American Journal of Ve te r i

nary Research , 43 (3 ) , 551 (1982) , (En, 15 r e f . ) .

93. Van Scha lkwyk , P .C . ; Geyser , T . L . ; Recio, M. and Erasmus ,

F . P . G . , J . of South African Ve te r ina ry Associa t ion , 5 0 ( 1 ) ,

31 (1979) , ( E n . ) .

94. Spaldonova, R. , C h e r t s e y , S u r r e y , U.K. ; Reedbooks L t d . . (1981) ,

323 (En, 4 ref. ) .

95. Cobra , J . ; Dubinsky, P. and Stof fa , P . , Blue Book for the Ve te r i

nary Profess ion , 30, 458 (1981) (En, 12 r e f . ) .

96. Rober tson, E .L . and Burke , T . M . , Journal of American Ve te r ina ry

Medical Associa t ion. 180(1) , 53 (1982) (En, 18 r e f . ) .

97. Bal i , M.K. and Singh, R . P . , Haryana Agricul tural Un ive r s i ty

Journal of Resea rch . 10 (1 ) , (1980) , 110 (En, 4 r e f . ) .

98. Malan, F . S . and Reinecke, R .K. , Journal of South African Ve te r i

na ry Associa t ion . 5 1 ( 4 ) , 223 (1980) , (En. 7 r e f . ) .

99. Cal l inan , A . P . L . and Cummins, L . J . , Austra l ian Veter inary J o u r n a l ,

55 (8 ) , 370 (1979) (En) .

206

100. Cal l inan, A . P . L . and Cummins, L . J . Helminthological A b s t r a c t ,

49, 325 (1980) .

101. Cobra , J . ; L i e t ava , P . ; Duwel, D. and Re i sen le i t e r , R. , B r i t i s h

Vete r inary Jou rna l , 135(4) , 318 (1979) (En) .

102. Duncan, J . L . ; McBeath, D.G. and Pres ton , N .K . , Equine Ve te r ina ry

Jou rna l , 12 (2 ) , 78 (1980) (En, f r , d e , 7 r e f . ) .

103. Duwel, D. , Kleint ier P r a x i s , 23 , 237 (1978) (De, en , f r , i t ) .

104. Gur lap , N. and T inav , R. , Ankara Univers i t e s i Veter iner Fakul tes i

Derg i s i , 2 5 ( 3 ) , 440 (1978) , ( T r . , f r . ) ,

105. Kutzer , E. , T i e r a z t l i c h e P r a x i s , 6 ( 3 ) , 325 (1978) , ( D e . ) .

106. Gupta, C . S . , Geobios, I n d i a , 8 ( 2 ) , 54 (1981) (En, 8 r e f . ) .

107. V a s i l e v i c h , F . I . , Nauchnye Trudy Nil Pushnogo Zerovodstva i

Kro l ikovods tava , 26 , 71 (1981) (Ru) .

108. C h a n d r a s e k h a r a n , K. ; Ramdas, K. ; Cheeran , J . V . and P a i l y ,

E . P . , Kerala Journal of Vete r inary Science, 10 (1 ) , 159 (1979) ,

(En, malayalam, 9 r e f . ) .

109. Santamarina, F . M . T . ; Sanmartin Duran, M.L. ; Mart inez, F . A . R . ;

and L l o r e n t c , L . V . , Revista I be r i ca de Pa ra s i to log ia , 4 4 ( 1 ) ,

91 (1984) , ( E s , en . 17 r e f . ) .

n o , T ina r , R. , Bursa Univers i t es i Veter iner Fakul tes i De rg i s i , 1 ( 1 ) ,

19 (1982) , (T r . f r . ) .

111, Denev, I . and Rostov, R. , Nauchni Trudove Raionen Nauchnoiz-

s l e d o v a t e l s k i Veter inarnomedi t s insk i I n s t i t u t , Veliko Turnovo,

2 , 81 (1982) , (Eg, en , r u , 10 r e f . ) .

112, Matta, S.C. , Indian Journal of Poul t ry Science, 15 (3 ) , 207 (1980)

(En, 16 ref. ) .

113, London, C . E . ; Rober tson , E . L . ; McCall, J .W. ; Guer re ro , J . ;

Panca r i , G. ; Michael , B. and Newcomb, K. , Journal of Ve te r i

na ry Resea rch , 4 2 ( 7 ) , 1263 (1981) , (En, 14 r e f . ) .

114, Matta, S . C , Indian Journal of Poul t ry Science, 15 (4 ) , 288 (1980)

(En, 5 r e f . ) .

115, Merd ivenc i , A. ; Sengi i l , M. and Baydemir , M. , Ce r r aphasa Tip

Fakul tes i De rg i s i , 7 ( 3 ) , 339 (1976) , ( T r , e n ) .

207

116. Clayton, H.M, and Neave, R . M . S . , Vete r inary Record, 104(25) ,

571 (1979) , ( E n . ) .

117. Jaroonvesama, N . ; C h a r o e n l a r p , K. and Muangmanee, L . , Journal

of Medical Association of Tha i l and , 61(12) , 675 (1978) (En,

T h a i ) .

118. Oguz. T. , Ankara Univers i t es i Veter iner Fakul ter i De rg i s i , 2 3 ( 2 / 4 ) ,

385 (1976) . ( T r , d e ) .

119. Krotov, A . I . ; Kovalenko, F . P . ; Cha rnyaeva , A . I . ; Budanova,

I . S . ; Bayandina, D. G. and Kuznetsova, O . E . , Helminthological

A b s t r a c t , 47, Apri l (1978).

120. Co lg laz ie r , M.L . ; Enz i le , F .D. and Kates , K.C. , Journal of Pa ra

s i t o logy , 6 3 ( 4 ) , 724 (1977) , ( E n ) .

121. Ray, D.K. ; Bhopa le , E.K. and S r i v a s t a v a , V . B . , Annals of T r o p i

cal Medicine and Pa ra s i t o logy , 72 (1 ) , 55 (1978) (En) .

122. C h o n g s p h a j a i s i d d h i , T. ; At tana th , P . ; Panasoponkul , C. and Rado-

myos . P . , Annals of Trop ica l Medicine and Pa ra s i t o logy , 7 2 ( 1 ) ,

59 (1978) (En) .

123. Andrasko , H . ; Pacenovsky , J . ; Krup ice r , I . and B i r c a k , A . ,

V e t e r i n a r s t v i , 2 5 ( 9 ) , 416 (1975) .

124. Goldsmith , J . M . , South African Medical Journa l , 48 (54) , 2265

(1978) (En) .

125. Kan, S . P . , Transact ions of the Royal Society of Tropica l Medicine

and Hygiene. 7 7 ( 5 ) , 668 (1983) (En, 15 r e f . ) .

126. Maki, J . and Yanagisawa, T. , Pa ras i to logy , 8 7 ( 3 ) , 525 (1983) ,

(En, 12 r e f . ) .

127. B r a d l e y , R . E . ; Guer re ro , J . ; Becker , H.N. : Michael , B . F . and

Newcomb, K. , American Journal of Veter inary Resea rch , 44 (7 ) ,

1329 (1983) , (En, 4 r e f . ) .

128. Notteghem, M . J . ; Leger , N. and C a v i e r , R. , Annals Pharmaceu-

t iqnes F r a n c a i s e s , 3 7 ( 3 / 4 ) , 153 (1979) , ( F r , e n ) .

129. B a r a n s k i , M.C. ; S i l v a , A.F.DA, Kotaka, P . I . ; Gomes, N . R . ; Gio-

vannoni , M. and T e l l e s , J . E . Q . , Revista do Ins t i tu to de Medi-

cina Tropica l de Sao Paulo . 2 0 ( 4 ) , 213 (1978) , ( P t , En) .

208

130. Reddy, A . B . ; Rao, U.R. ; C h a n d r a s h e k h a r , R.; S h r i v a s t a v a . R.

and Subrahmanyam, D. , Tropenmedizin iind Pa ra s i t o log i e , 34 (4 ) ,

259 (1983) (En, d e , 12 r e f . ) .

131. Pecheur , M. , Annales de Medecine V e t e r i n a i r e , 127(3) , 203 (1983)

( F r , en , 4 ref. ) .

132. Drudge, J . H . ; Lyons , E . T . ; T o l l i v e r , S.C. and Kubis , J . E . ,

Modern Veter inary P r a c t i c e , 64 (5 ) , 414, 416, 417 (1983) (En,

12 r e f . ) .

133. Sa th ianesan , V. ; Mohan, M.C. and Sundaram, R .K. , Kerala Journal

of Vete r inary Science , 10 (2 ) , 193 (1979) (En, Malayalam, 13

r e f . ) .

134. Pecheur , M. ; D e t r y - P o u p l a r d , M. and Benakhla , A . , Annales de

Medicine V e t e r i n a i r e , 124(8) , 609 (1980) ( F r , en , 6 r e f . ) .

135. Wil l iams, J . C ; Sheehan , D. and F u s e l i e r , R, , Proceedings of

the Helminthological Society of Washington, 4 5 ( 1 ) , 129 (1978)

( E n ) .

136. T h e o d o r i d e s , V . J . ; Nawal insk i , T . ; Freeman, J . F . and Murphy,

J . R . , American Journal of Veter inary Resea rch , 37 (10) , 1207

(1976) (En) .

137. Splandonova, R.; Cobra , J . and Velebnj , S. , Helminthological

Abs t r ac t , 46, Sep . (1977) .

138. Nawal inski , T. and T h e o d o r i d e s , V . J . , Ve te r ina ry Medicine and

Small Animal C l in ic i an , 72 (1 ) , 96, 99, 101 (1977) (En) .

139. Yalc inkaya , F. , Turk Hijiyen ve Deneysel Biyoloji D e r g i s i . 35(2-

3 ) , 101 (1975) ( T r , f r ) .

140. Cal l inan, A . P . L . and Bar ton , N . J . , Austral ian Ve te r ina ry Jou rna l ,

55 (5 ) , 255 (1979) ( E n ) .

141. Drudge, J . H . ; Lyons , E .T . and T o l l i v e r , S . C , Ve te r ina ry Medi

cine and Small Animal Cl in ic ian , 72 (3 ) , 433, 437 (1977) (En) .

142. Bhargava , R.K. ; S h a h , P . K . D , ; Ujwal, J . S . and Maheshwar i ,

K . L . , Journal of Associat ion of Phys ic ians of I n d i a , 23 (11) ,

763 (1975) ( E n ) .

143. Ziegler , J . , Helminthological A b s t r a c t . 4 1 , January (1972) , 41 .

209

144, Del ic , S . ; Cankovic , M. and Rozman, M. , Ve t e r ina r i a , Sara jevo ,

2 0 ( 4 ) , 507 (1971) (Sh , e n ) .

145. Cobra , J . ; Legency, J . ; Stoffa, P . ; Krup ice r , I . and Pacenovsky ,

J . . V e t e r i n a r i s t v i , 2 8 ( 6 ) , 274 (1978) ( C s ) .

1 4 6 . S a l a z a r , D. ; H e r r e r a , R.D. and Quiroz, R . H . , Teenica Pecuaria

en Mexico, No.26, 28 (1974) ( E s , e n ) .

147. Ba ransk i , M.C. ; S i l v a , A.F .DA, ; Kotaka, P . I . ; Gomes, N . R . ;

Giovarnoni , M. and T e l l e s , J . E . Q , , Revista do Ins t i tu to de

Medicina Tropica l de Sao Paulo, 2 0 ( 4 ) , 213 (1978) ( P t , En) .

148. Fo ix , J . , Revue de Medicine Ve t e r i na i r e , 128 (8 /9 ) , 1111 (1977)

( F r , en , d e , e s ) .

149. Spaldonova, R. ; Cobra , J . and Biologia, B r a t i s l a v a , B. , Zoologia,

32(11) , 4 (1977) .

150 .Furmaga, S. ; Gundlach, J . L . and P a t y r a , J . , Medyeyna Wetery-

na ry jna , 32(12) , 734 (1976) , ( P I , en , r u ) .

151. Tager -kagan , P . , Revue d 'E levage et de Medecine Veter ina i re

des pays T rop lcaux , 2 9 ( 4 ) , 317 (1976) ( F r , en , e s ) .

152 .Pecheu r , N. and L a b r i q u e , Y . , Annales de Medicine V e t e r i n a i r e ,

118(6) , 367 (1974) ( F r ) .

153. Poup la rd , L. and Pecheur , M. , Annales de Medecine V e t e r i n a i r e ,

116(3) . 229 (1972) ( F r ) .

154. C a b a r e t , J . ; Anjorand, N. ; L e c l e r c , C. and Mangeon, N. , Helmin-

thological A b s t r a c t , 54, March (1985).

155.Ziomko, I . , Medyeyna Wet r y n a r y j n a , 39(11) , 665 (1983) (P I ,

en , r u , 21 r e f . ) .

156 .Chrous t , K. , Sbornik Vedeckych P r a c i , USVU P r a h a , No .11 , 141

(1981) , (Cs , en , e s , r u ) .

157. Manuel, M . F . ; C a n t i l l e r , D . B . ; Cambero, W.R. and Dominguez,

R . V . , Ph i l i p ine Journal of Vete r inary Medicine, 2 0 ( 1 ) , 80 (1981)

(En, 4 r e f ) .

158. Karunakaran, C .S . and Denham, D .A . , Journal of Pa ra s i t o logy ,

6 6 ( 6 ) , 929 (1980) (En, 9 r e f . ) .

159. Kingsbury , P.A. and Reid , J . F . S . , Vete r inary Record , 109(18) ,

404 (1981) (En, 3 r e f . ) .

210

160. Kingsbury , P. A. ; Rowlands, D. AP T. and Reid , J . F . S . , Ve te r i

na ry Record, 108(1) . 10 (1981) , (En, 3 r e f , ) .

161. Cha lmer s , K. , New Zealand Veter inary Jou rna l , 2 7 ( 1 / 2 ) , 8, 13

(1979) (En) .

162. Bake r , N . F . ; F i s k , R.A. and Mil le r , J . E . , Journal of Ve te r inary

Research , 3 9 ( 8 ) , 1258 (1978) (En) .

163. Corwin, R.M.; Kennedy, J . A. and P r a t t , S . E . , American Journal

of Vete r inary Resea rch , 40 (2 ) , 297 (1979) (En) .

164. Michael , S .A. ; Higgins, A . J . and El -Refa i , A . H . , Tropical Animal

Health and Product ion , 11 (2 ) , 63 (1979) , (En, F r , e s ) .

165. C h a n d r a s h e k a r a n , K.; P y t h a l , C ; Sundaram, P.K. and P e t e r ,

C . T . , Kerala Journal of Veter inary Science, 5 ( 1 ) , 26 (1974)

(En) .

166. Weissenburg, H. and Mlela, VV.S., T i e r a r z t l i c h e Umschau, 2 9 ( 5 ) ,

272 (1974) ( D e ) .

167. Kutzer, E. ; Po in te r , J . and P r o s l , H. , T i e r a r z t l i c h e Umschau,

2 9 ( 5 ) , 268-270-271 (1974) (De ) .

168. Res tani , R. ; Pavonce l l i , R. and T a m p i e r i , P . , Veter inar ia I t a l i a n a ,

24 (11 /12) , 477 (1973) ( I t , En) .

169. Har t , J . A . and Bos man, C . J . , F l s . Afr. Vet. Med. A s s . , 42 (3 ) ,

243 (1971) ( E n ) .

170. Ch rous t , K. , Sbornik Vedeckych Praci Ustredniho Statniho Ve te r i -

narniho Ustavu u P r a z e , No.13, 98 (1983, Publ . 1984) (Cs ,

en , e s , r u ) .

171. Bruke , T.M. and Roberson, E . L . , Journal of American Ve te r ina ry

Associa t ion . 183(9) , 987 (1983) (En, 17 r e f ) .

172. Bruke , T.M. and Roberson, E . L . . Helminthological A b s t r a c t . 53 ,

1511 (1984) .

173. Yazninski , T . A . ; Greenway, T . E . ; T i l l e y , W. and F e a t h e r s t o n e ,

H, Ve te r inary Medicine & Small Animal Cl in ic ian , 78(12) , 1899

1904 (1983) (En, 20 r e f . ) .

174. A r k h i p o v a , N . S . ; Bessonov. A . S . ; Ubi raeu . S . P . ; Upsensky , A.V.

and S h e k h o v t s o v , N . N , , Prague , Czechoslovakia Academia, P r a h a ,

263 (1983) (En, r u , 16 r e f ) .

211

175. C o b r a , J . ; S t o f f a , P . ; D u b i n s k y , P . a n d H o l a k o v s k y , P . , F o l i a

V e t e r i n a r i a . 2 4 ( 3 / 4 ) , 121 (1980) ( S k , r u , 10 r e f ) .

176. S t e w a r t , T . T . ; B i n d e r , T . D . ; S o u t h e r n , L . L . a n d S i m m o n s , L . A . ,

Am. J o u r n a l of V e t e r i n a r y R e s e a r c h , 4 5 ( 5 ) , 984 (1984) ( E n ,

11 r e f ) .

177. V e l i c h k i n , P . A . ; G o l u b k o v , V . F . a n d S o l o v ' y a n o v , V . V . , He lmin -

t h o l o g i c a l A b s t r c t , 5 3 , O c t o b e r 1 9 8 4 ) .

178 . K i r s c h , R. , Av ian D i s e a s e s , 2 8 ( 2 ) , 311 (1984) ( E n , e s , 21 r e f ) .

179. L i m a , W. ; D o s , S. ; C o s t a , H.M. DE A . ; C o s t a , J . O . ; G u i m a r a e s ,

M . P . a n d L e i t e , A . C . R . , A r q u i v o s da E s c o l a de V e t e r i n a r i a

da U n i v e r s i d a d e F e d e r a l de Minas G e r a i s , 3 4 ( 2 ) , 325 (1982)

( P t . e n , 13 r e f ) .

180 . F e r g u s o n , D . L . , J a p a n e s e J o u r n a l of P a r a s i t o l o g y , 3 1 ( 5 ) , 369

(1982) ( E n , J a , 8 r e f ) .

1 8 1 . C i o r d i a , H . ; S t u e d e m a n n , J . A . a n d M c C a m p b e l l , H . C . , A m e r i c a n

J o u r n a l of V e t e r i n a r y R e s e a r c h , 4 4 ( 6 ) , 1091 (1983) ( E n , 10

r e f ) .

182 . Ambu , S. ; Kwa , B . H . ; Mak , J . W . a n d S i n g h , K . I . , F o l i a P a r a s i t o -

l o g i c a , 2 9 ( 4 ) , 361 (1982) ( E n , r u , 17 r e f ) .

1 8 3 . L e g u i a , P . G . ; G u e r r e r o , D . C . a n d R o j a s , C M . , R e v i s t a de I n v e s -

t i g a c i o n e s P e c u a r i a s , 4 ( 1 ) , 25 (1979) ( E s , e n , 10 r e f ) .

184 . M o y r o u d , J . , B r e u i l , J . a n d C o u l a n g e s , P . , A r c h i v e s de L ' I n s t i t u t

P a s t e u r d e M a d a g a s c a r , 5 0 ( 1 ) , 125 ( 1 9 8 2 , P u b l . 1 9 8 3 ) , ( F r ,

2 r e f ) .

185 . B e r g e r , H . ; G a r c e s , T . R . ; Wang, G . T . ; G a l e , CO.; S t e l l e r ,

W.A. a n d S i m k i n s , K . L . , A m e r i c a n J o u r n a l of V e t e r i n a r y R e s

e a r c h , 4 5 ( 1 ) , 162 (1984) ( E n , 9 r e f ) .

186. D o r c h i e s , P . ; F r a n c , M. a n d L a h i t t e , J . D . DE, Revue de Medec ine

V e t e r i n a i r e , 1 3 0 ( 3 ) , 4 1 7 , 421 (1979) ( F r , e n , d e , e ) .

187 . D o w n e y , N . E . , V e t e r i n a r y R e c o r d , 1 0 1 ( 1 3 ) , 260 (1977) ( E n ) .

188 . R o w l a n d s , D . T . a n d B e r g e r , J . , J o u r n a l of South Af r i can V e t e r i

n a r y A s s o c i a t i o n , 4 8 ( 2 ) , 85 (1977) ( E n ) .

189 . F e r g u s o n , D . L . a n d W h i t e , R . G . , J o u r n a l of Animal S c i e n c e s ,

4 0 ( 5 ) , 838 (1975) ( E n ) .

212

190. Moreau, J . P . ; Radaniel ina, R. and B a r b i e r , P . , Medecine Trop ica le

35 (6 ) , 451 (1975) ( F r , e n ) .

191. Samigull in, R .N. , Helminthological A b s t r a c t , 53 , July (1984) .

192. Samizadeh-Yazd, A . , Journal of Ve te r ina ry Facu l ty , Un ive r s i ty

of Tehran , 3^(4) , 21 (1981) ( P c , en , 24 r e f . ) .

193. Che rnyaeva , A . I . ; Kuznetsova, O . E . ; Manenkova, G.M.; Bene-

d i k t o v , I . I . and Krotov, A . I . , Medskaya P a r a z i t , 4 0 ( 4 ) , 414

(1971) .

194. Calzet ta Resio, E. and Basso , N. , Revlsta de Medlcina V e t e r i n a r i a ,

Buenos A i r e s , 5 1 , 87, 92 (1970) ( E s ) .

195. Ambrosi , M. and Morettini , B . , Ob ie t t i v i e Documenti V e t r i n a r i ,

5 ( 1 ) , 59 (1984) ( I t , en , 24 r e f ) .

196. Alcaino, H.A.; Gorman, T.R. and E l o r r i e t a , M.B. , Ve te r ina ry

Pa ras i to logy , 14(2 ) , 153 (1984) (En, 23 r e f ) .

197. C iod ia , H. ; McCampbell, H.C. and Stuedemann, J . A . , American

Journal of Veter inary Resea rch , 43 (12) , 2248 (1982) (En, 5

r e f ) .

198. Cannan, R .M. , Vete r inary Medical Review, No.2 , 145 (1978) .

199. Engl ik , K. and Dey-Hazra , A . , Vete r inary Medical Review, No .2 ,

195 (1978).

200. Hopkins , T . J . and R a t t e r t y , M. , Vete r inary Medical Review,

No.2 , 160 (1978) (En) .

201. Gre lck , H. ; Horchner , F. and Wohrl , H. , Vete r inary Medical

Review, No.2, 154 (1978) , (En) .

202. Abo-Shady, A . F . , Abdel-Magied, S.A. and Hegazi, M.M., Journal

of Egyptian Society Pa ra s i t o logy , 13 (2 ) , 629 (1983) (En, 4

r e f . ) .

203- Santos , A.T. , B i a s , B . L . ; P o r t i l l o , G. ; Nosenas, J . S . ; Po l i qu i t ,

O. and Papas in , M. , Arzneimit te l Forschung, 3 4 ( 9 b ) , 1221 (1984)

(En, d e , f r , 5 r e f ) .

204. Pungpak, S. ; Bunnag, D. and Har inasu ta , T. , South-East Asian

Journal of Tropical Medecine and Public Heal th , 14 (3 ) , 363

(1983) (En, 7 r e f ) .

213

205. Bunnag, D, ; Radomyos, P . and Har inasuta , T. , South-East Asian

Journal of Tropica l Medicine and Public Heal th , 14 (2 ) , 216

(1983) (En, 6 r e f ) .

206. Katz, N . ; Rocha, R . S . ; Lamber tucc i , J . R . ; Greco, D . B . ; Ped roso ,

E . R . P . ; Rocha, M.O.C. and Flan, S. , Revista do Ins t i tu t e

de Medicina Tropica l de Sao Paulo, 2 5 ( 4 ) , 173 (1983) (En,

P t , 17 r e f ) .

207. Chen, C.Y, and Hsieh , W.C. , Arzneimit te l Forschung. 3 4 ( 9 b ) ,

1160 (1984) (En, d e , f r , 7 r e f ) .

208. Grol l , E. , Helminthological A b s t r a c t , 53 , June (1984).

209. T i shchenko , V.V. and T i shchenko , L . G . , Byul le ten ' Vsesoyuznogo

Ins t i tu ta Gel 'minto . logi i im, K . I . , S k r y a b i n a , No.34, 62 (1983)

(Ru, en , 5 r e f ) .

210. Schu t t e , C . H . J . ; Osraan, Y. ; Van Deventer , J .M.G. and Mosese,

G. , South African Medical Journa l , 64 (1 ) , 7 (1983) (En, 12

r e f ) .

211 . Busha ra , H.O. ; Hussein, M . F . ; Majid, M.A. and T a y l o r , M.G. ,

Research in Vete r inary Science, 3 3 ( 1 ) , 125 (1982) (En, 8 r e f ) .

212. Sakamoto, T. ; Kono, I . ; Yasuda, N. ; Kitano, Y . ; Togoe, T . ;

Yamamoto, Y. ; I w a s h i t a , M. and Aoyama, K. , Bullet in of the

Facul ty of Agr icu l tu re , Kagoshima, U n i v e r s i t y , No.29, 81 (1979)

( J a , e n ) .

213. Sakamoto, T. ; Kono, I . ; Yasuda, N. ; Kitano, Y. ; Togoe, T . ;

Yamamoto, Y. ; I w a s h i t a , M. and Aoyama, K. , Bullet in of the

Facul ty of Agr icu l tu re , Kagoshima U n i v e r s i t y , No.29, 81 (1979)

( J a , e n ) .

214. Thomas, H. , Boletin Chileno de Pa ra s i to log ia , 3 2 ( i ) . 2 (1977) . r

215. Thomas, H. and Gonnert, R. , Zei t schr i f t fur Pa ra s i t enkunde , 5 2 ( 2 ) ,

117 (1977) (En, d e ) .

216. Gura lp , N . ; Oguz, T. and Zeybek, H. Ankara , Univers i tes i Ve t e r i -

ner Fakul tes i Derg i s i , 2 4 ( 1 ) , 85 (1977) ( T r , e n ) .

217. Baldock , F . C . ; F lucke , W.J. and Hopkins , T . J . , Research in

Veter inary Science, 2 3 ( 2 ) , 237 (1977) (En) .

214

218. Kumar, S. ; Se th , M. ; B h a d u r i , A . P . ; Visen, P . K . S . ; Anuradha ,

M. ; Gupta, S..; Fa t ima , N . ; Ka t iya r , J . C . ; C h a t t e r j e e , R.K.

and Sen, A . B . , J . Med. Chem. , 27 , 1083 (1984) .

218a .Akh ta r , M.S . ; Se th , M. and Bhadur i , A. P . , Ind . J . Chem. , 25B,

395 (1986).

219. Akh ta r , M.S . ; Sharma, V.L. and B h a d u r i , A. P . , J . Het. Chem. ,

(In P r e s s ) .

220. Meyer, H. , Liebigs Ann. Chem. , 1534 (1981) .

221 . Boberg , F. ; Ruhr , M. and Garming, A . , Liebigs Ann. Chem. ,

223 (1984) .

222 Boberg , F . ; Garburg , K.; Gor l ich , K. ; P i p e r e i t , E. and Ruhr ,

M. , Liebigs Ann. Chem. , 911 (1984) .

223. B ig ine l l i , Ber dt Chem. Ges. , 24 , 131 (1891); Brown, D . J . , The

P y r i m i d i n e s , (Wil ley , New York, N.Y. ) 440 (1962) .

224.. Hinkel , H.E. and Hey, D . H . , Reel. T r a v . Chim. Days Bas , 48,

1280 (1929) .

225. Sweet, F . and F i s s e k i s , J . D . , J . Am. Chem. Soc. , 95 , 8741

(1973) .

226. Hassner , A. and Michelson, M . J . , J . Org. Chem. . 27 , 3974 (1962) .

227. Gomez, S.A.; Mancera, A . ; C a b a l l e r o , F . J . and Bel lana to , J . , An. Quim. S e r . C , 7 9 ( 2 ) , 175-85 (1983) Span . ; CA: 102: 95485y.

228. Wehr l i , F.W. and Wir th l in , T. , in " In te rp re ta t ion of Carbon-13

NMR S p e c t r a " , HEYDEN, London, p . 187.

229. Hawking, F. and Sewel l , P . , B r i t . J . Pharmacol . Chemothe rapy ,

3, 285 (1943) .

230., Sewel l , P. and Hawking, F. , B r i t . J . Pharmacol . Chemothe rapy ,

5 , 239 (1950) .

231 . C h a t t e r j e e , R .K. , P h . D . Thes i s submit ted to Agra U n i v e r s i t y ,

Agra (1970).

232. S teward , J . S . , P a r a s l t o l . , 45 , 235 (1955).

233. Gupta, S. ; Ka t iya r , J . C . and Sen, A . B . , Ind . J . P a r a s i t o l . . 3, 199 (1979) .

234.. Ray, D.K.; Bhapa l e , K.K. and S r i v a s t a v a , N . B . , J . He lmin th . ,

XLVI, 357 (1972) .

215

2 3 5 , K a t i y a r , J . C ; V i s e n , P . K . S . ; M i s r a , A , ; G u p t a , S. a n d B h a d u r i ,

A . P . , Acta T r o p . . 4 1 , 279 ( 1 9 8 4 ) .

LIST OF PUBLICATIONS

Syntheses of 4-Aryl-2 , 5 - d i m e t h y l - 3 - ( N , N - d i s u b s t i t u t e d - c a r b a m o y l )

furans , subs t i t u t ed Isoquinolino [ 1,2-b Iquinol ines & Methyl 5 ( 6 ) -

subs t i t u t ed benz_lmidazole-2-carbamates, S. Niwas, M.S. A k h t a r ,

S. Kumar & A. P. Bhadur i , Indian J . Chetn. , 24B, 754-60 (1985) .

Novel ox ida t ion of t e t r a s ubs t i t u t ed furans by pyr id in ium c h l o r o -

ch romate , M.S. A k h t a r , M. Seth & A. P. B h a d u r i , J . Het. Chem. ,

22, 1323 (1985) .

Syntheses of Methyl 5( 6 ) - s u b s t i t u t e d b e n z i m i d a z o l e - 2 - c a r b a m a t e s ,

M.S. A k h t a r , M. Seth & A. P. B h a d u r i , Indian J . Chem. , 2 5 3 ,

395 (1986) .

Rearrangement r eac t ions in the mass s p e c t r a of t e t r a s u b s t i t u t e d

e t h y l e n e s , K .P . Madhusudanan, M.S. A k h t a r , M. Seth & A. P.

B h a d u r i , Indian J . Chem. , 25B, 915-21 (1986) .

Studies on Nef Reaction Induced by organic b a s e s , M.S. A k h t a r ,

V.L. Sharma & A .P . B h a d u r i , J . Het. Chem. , (In p r e s s ) .

Reaction of cC , f t -unsa tura ted ketones wi th n u c l e o p h i l e s , M.S. A k h t a r ,

M. Seth & A. P. B h a d u r i , Indian J . Chem. , (In p r e s s ) .

Syntheses of T e t r a h y d r o t h i a z o l o [ 3,2-a Ipy r imid ines and t e t r a s u b s t i

tu ted d i h y d r o p y r i m i d i n e d e r i v a t i v e s as p o s s i b l e an the lmin t ic

agen t s , M.S. A k h t a r , M. Seth & A. P. B h a d u r i , Indian J . Chem. ,

(In p r e s s ) .

Studies on the Syntheses of Possible Antifertility and ...

Documents

Transcript of Studies on the Syntheses of Possible Antifertility and ...