OF E Mottor of PliUogopIip · stereochemistry an biologicad assal (anticancey anrd antimicrobia...

DESIGW AND SYNTHESIS OF BIOLOGICALLY ACTIVE COMPOUNDS WITH SULPHUR AND

NITROGEN CONTAINING HETEROCYCLES AND RELATED COMPOUNDS

RESUME

T H E S I S SUBMITTED FOR THE DEGREE OF

Mottor of PliUogopIip IN

C H E M I S T R Y

BY

MUJEEBUR RAHMAN V. P

DEPARTMENT OF CHEMISTRY FACULTY OF SCIENCE

ALIGARH MUSLIM UNIVERSITY ALIGARH (INDIA)

2002

RESUME

The thesis describes the work related to synthesis, structure elucidation

stereochemistry and biological assay (anticancer and antimicrobial screening)

grouped in three parts Wdti six chapters as long alkyl chain substituted-j'-

tr iazolo-thiadiazepines, long alkyl chain substi tuted-benzopyrano-^-tr iazolo-

thiadiazepines, long alkyl chain substi tuted-^-triazolo-thiadiazoles, 2-aryl-3-

7V-long alkyl chain substi tuted-thiazolidin-4-ones/thiazan-4-one, 2-aryl-2-

methyi-3-A^-long alkyl chain- •substituted-thiazolidin-4-one and biological

screening: study for anticancer and antimicrobial activity.

The techniques used for structure elucidation are 'H-NMR, '^C-NMR,

APT,NOE, 2DNMR (COSY, HETCOR) including DCI-MS, IR and FTIR.

PART ONE

l,2,4-TRIAZOLOf3,4-b|fl,3,4]THIADIAZEPINES/THIDIAZOLES

CHAPTER I. Long alkyl chain substituted-s-Triazolo-Thiadiazepines

4-Amino-5-mercapto- l ,2 ,4- t r iazoles and their A^-bridged heterocyclic

derivatives have received much attention during recent years on account of

their wide spectrum biological potentialities. The 1,2,4-triazole nucleus has

been incorporated into a wide variety of therapeutically interesting drugs

including Hi/H2-Histamine receptor blockers, cholinesterase active agents,

CNS stimulants, antianxiety agents and sedatives. Also 1,3,4-thiadiazepine

derivatives are associated with a varied range of biological activities such as

anticancer, anti-HIV, anticholinergic, antidepressant, anaesthetic, calcium

antagonist choleretics and ACE inhibitory activity. Prompted by these

observations and as part of our continuing programme in the search of

biologically active compounds with sulphur and ni t rogen containing

heterocycles, we have undertaken the synthesis of two novel compounds,

2-(4-chlorophenyl)-4-[2-(4-chlorophenyl)ethenyl]-7-(9-decenyl)-2,3-dihydro

-l,2,4-triazolo[3,4-b][l,3,4]thiadiazepine (4) and 2,4-bis(4-chlorophenyl)-7-

(9-decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4]thiadiazepine (7) by the

reaction of 4-amino-3-(9-decenyl)-5-mercapto- l ,2 ,4- t r iazole (1) with ( i ) l ,5-

bis(4-chlorophenyl)pent- l ,4-dien-3-one (2) and (ii) 4 ,4 ' -dichlorochalcone (5)

.In each of the above synthesis, a Michael adduct of (2)/(5) and the s-triazole

(1) were also obtained. The reaction of (1) with l ,3 -b is (2- th ienyl )propen- l -

one (8) furnished only the Michael adduct (9) as the sole product . (Scheme 1)

The structures of these compounds were established on the basis of

spectral studies such as IR, DCI-MS, EI-MS, 'H-NMR, and '^C-NMR spectra.

LOT > K " ^ H H Y

(4) CI

(7)

(N X D u

o

dj c >o • — -

m c o

-C

:5 W t/1 15 fo

w

K U CZ)

c <u a . o

c D

CHAPTER II. Long alkyl chain substituted-Benzopyrano-s-Triazolo-Thiadiazepines

As part of our programme in search of biologically active compounds and in

continuation of the work described in previous chapter, we extended our work on the

reaction of the ^-triazole (1) with exocyclic a;y^unsaturated enone systems and

synthesised two novel compounds, 2,3-bis(4-chlorophenyl)-ll-(9-decenyl)-2a,3-

dihydro[l]benzopyrano[3,4-e]-2,2a-dihydro-l,2t4-triazolo[3,4-b][l,3,4]thiadiazep-

ine (12) and 2,3-bis(phenyl)-ll-(9-decenyl)-2a,3-dihydro[l]benzopyrano[3,4-e}-

2,2a-dihydro-l,2,4-triaz olo[3,4-b][l,3,4]thiadiazepine (15) by the reaction of the i--

triazole (1) with £'-3-(4-chlorobenzylidene)-4'-chloro flavanone (10) and E-3-

benzyiidene flavanone (13) in dry benzene using j:?-TsOH as catalyst. An adduct, 2-

(4-chIorophenyl)-3-[3-(9-decenyl)-4-amino-5-(4-chlorobenzylthio)-l,2,4-triazolyl]-

2,3-dihydro-4-benzopyrone (11) in the former and 2-(phenyl)-3-[3-(9-decenyl)-4-

amino-5-(benzylthio)-l,2,4-triazolyl]-2,3-dihydro-4-benzopyrone (14) in the latter

was also obtained in good yields(Scheme 2).

(11), (14)

( 1 1 ) , ( 1 2 ) : R = R ' = C I

( 1 4 ) , ( 1 5 ) : R = R ' = H

( 1 2 ) , ( 1 5 )

The structures of these compounds were confirmed by IR, DCI-MS, 'H-NMR.

'^C-NMR, COSY and HETCOR spectral studies. The configurations at the three

stereogenic centres C-2, C-3 and C-l '" in compounds (11) and (14) were established

on the basis of the inspection of models and the values of coupling constants.

N N

SH +

O H

(10) R = R' = C1 (13) R = R' = H

Molar ratio (1:1) Diy benzene Reflux 38/32 h

(11) R = R' = C1 (14) R = R' = H

(12) R = R' = C1 (15) R = R' = H

SCHEME 2

6

CHAPTER HI. Long alkvl chain substituted-5-Triazolo-Thiadiazoles

1,2,4-Triazole and its derivat ives have been found to play vital role in

medic ine , agricul ture and industry. The fused r ing sys tems such as

5- t r iazolo[3 ,4-b] [ l ,3 ,4] th iad iazoles have varied range of b io logica l activities

as ant ibacter ia l , ant i fungal , antiinflammatory, antihypertensive, analgesic and

an the lmin t ic agents. These findings have tempted us to under take this

p rob lem on the synthesis of three new compounds, (i) 2-(4-methoxyphenyl)-

5-(9-decenyl)-2,3-dihydro-l,2,4-tnazolo[3,4-b] [l,3,4]thiadiazole (17) (ii) 2-

(4-chlorophenyl)-5-(9-decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4]thia-

diazole (19) and (in) 2-(2-thienyl)-5-(9-decenyl)-2,3-dihydro-l,2,4-

triazolo[3,4-b][l,3,4]thiadiazole (21) f rom (i) p - a n i s a l d e h y d e ^ (ii)

/7-chlorobenzaldehyde and (iii) th iophen-2-a ldehyde respec t ive ly using

4-amino-3- (9-decenyl ) -5 -mercap to- l ,2 ,4 - t r i azo le (1) in the presence of a

catalyt ic amount of / j -TsOH. In each react ion, the co r re spond ing Schi f f bases

(16), (18), and (20) have also been obtained. The react ion of fu r fu ra ldehyde

with the t r iazole (1) yielded only the Schif f base (22) ( S c h e m e 3).

The structure of these compounds were es tabl ished on the basis of

spectral s tudies of IR, DCI-MS, ' H - N M R and '^C-NMR spectra .

N N

jl '1

I NH2 (1)

-CHO O - CHO

N N

I N = C H -

(16) R = 0CH3 ( 1 8 ) R = C1

N — N

( 2 0 ) X = S

( 2 2 ) X = O

+

N N

\ H N -

Q ( 1 7 ) R = 0 C H 3

( 1 9 ) R = CI

( 2 1 ) X = S

SCHEME 3

PART TWO

2,3-DISUBSTITUTEDTHIAZOLIDIN-4-ONES/THIAZAN-4-QNES

CHAPTER IV. 2-Arvl-3-iV-Long alkvl chain substituted-Thiazolidin-4-ones/Thiazan-4-one,

The interest in substituted thiazolidin-4-ones and thiazan-4-ones in

particular for medical application is increasing strongly. The thiazolidin-4-

ones, their derivatives and analogues have been found to exhibit usually high

in vitro activity against Mycobacterium tuberculosis, highly potent and

selective anti-Platelet Activating Factor activity and broad spectrum activity

against various diseases. Because 4-thiazolidinones substi tuted in the

2-posit ion were proven to be biologically very potent and selective and in

anticipation that by incorporating a long chain alkyl thioether moiety to

thiazolidin-4-one ring, the biological activity may be enhanced due to

lipophilization of the resulting molecule, we have under taken this problem. It

consists of the synthesis of the following four novel compounds , (i) 2-(3-

nitrophenyl)-3-[ll-(carboxymethylthio)undecanamido]thiazolidin-4-one (26),

(ii) 2-(3-nitrophenyl)-5-(methyl)-3-(10-undecenamido)thiazolidin-4-one (27)

and 2-(3-nitrophenyl)-5-(methyl)-3-[ll-{(l-carboxy)ethylthio}undecan-

amidoJthiazolidin-4-one (28), (Hi) 2-(3-nitrophenyl)-3-[ll-(2-carboxyethyl-

thio)undecanamido]tltiazan-4-one (30) f rom m-ni trobenzaldehyde-10-

undecenohydrazone (24) using mercaptoacetic acid in (i), 2-mercapto-

propionic acid in (ii) and 3-mercaptopropionic acid in (iii) in dry benzene. An

adduct, (25) in the former and (29) in the latter was also obtained in good

yields. (Scheme 1)

The compounds (24), (25) and (29) were found to exist in to be in its

two forms, synperiplanar (major) and antiperiplanar (minor) as represented

in F ig . 1. The s tructure and s tereochemistry of these compounds were

es tabl ished on the basis of IR, DCI-MS, 'H-NMR, '^C-NMR, NOE

experiments,COS Y and HETCOR spectral studies.

synperiplanar

H H

/ He ( 2 4 ) R = — C H = < ; ^

HZ

( 2 5 ) R = — ( C H 2 ) 2 — S — C F ^ - C O O H

( 2 9 ) R = — ( C H 2 ) 2 — S — C H J — C H 2 - C O O H

H R'

NO2

(26) R=

(27) R =

(28) R= -

-(CH2)2—S— CF^-COOH R' = H

— C H = < ( Hz

(CH2)2—S—CH-COOH

CH3

R = CH3

R = CH3

10

o z

O

(N JC

- I X' X , 5= 3 <U JJ i | i u § " c ^ fi I ^ ^ "o

.2 — S d « S O I-E iS ^ o I E

f s i O I ^ o u

i u

(N

I X C/i C/i 00 XXX

o fO

^

0 a c s -^ .o

1 s s s e C/5

S faJ I U c/D

11

CHAPTER V. 2-ArvI-2-Methvl-3-7V-Long alkyl chain substituted-Thiazolidin-4-one

As part of our programme in search of biologically active compounds with

sulphur and nitrogen containing heterocycles and in continuation of the above

mentioned work, we have undertaken this problem on the synthesis of 2-(4-

chlorophenyl-2-methyl-3-[l l-(carboxymethyl/2-carboxyethylthio)undecanamido]thi-

azolidin-4-ones/thiazan-4-one from /7-chloroacetophenone-10-undeceno-hydrazone

(31) using (i) mercaptoacetic acid and (ii) 3-mercapto propionic acid. An adduct,

(32) in the former and (35) as the sole product in the latter was also obtained

in good yields. (Scheme 1)

The structures of these compounds were established by IR, DCI-MS.

'H-NMR, '^C-NMR, NOE experiments, COSY and HETCOR spectral studies. Here

again the compounds (31), (32) and (35) were found to exist in its two forms,

synperiplanar (major) and antiperiplanar (minor) as represented in Fig. 2

( 3 2 ) R = — ( C H 2 ) 2 — S — C H 2 - C O O H

( 3 5 ) R = — ( C H 2 ) 2 - S - C H 2 - C H 2 - C 0 0 H

Fig. 2

12

O I

O::

to fO

. oo ^ <N

X

§ C g ^

.9 6 "tS

W S w W u CAJ

fO

o X

IT)

r<

01 c

- E O O

O ^

oi n: a:

fO

PART THREE

SCREENING FOR BIOLOGICAL ACTIVITY

CHAPTER VI. Study for Anticancer and Antimicrobial Activities

(A) A N T I C A N C E R A C T I V I T Y

The compounds (I) , (4), (7), (9), (12), (16), (17), (20), (21), (22), (24),

(26), (27), (30), (31) and (32) have been evaluated for ant icancer activity first

in the 3-cell lines of three types of human cancers: breast (MCF7), Lung

(NCI-H460) and CNS (SF-268) at one concentration as the one dose primary

anticancer assay. Out of the sixteen compounds screened, only one

compound, (27) was found to be active. This compound was then screened for

cytotoxic activity at five different concentrations against 60 cell lines of nine

types of human cancers: leukaemia, lung, colon, CNS, melanoma, ovarian,

renal, prostate and breast. This compound showed anticancer activity only at

higher concentration, Ix 10"^. Noteworthy results was obtained in the case of

non-small lung cancer, melanoma and renal where the reduct ion in growth is

75, 74 and 70 and 77% respectively .

(B) A N T I M I C R O B I A L ACTIVITY

A total of 10 compounds were tested against two Gram +ve bacteria

{Staphylococcus aureus, Bacillus subtilis) and two Gram -ve bacteria {Escherichia

coli, Pseudomonas aeruginosa) and a yeast, Candida albicans. Varying level of anti-

microbial activity was detected against one or more test organisms. Solvent control

(DMSO) showed non-significant inhibition to test microorganisms. Antimicrobial

activity against Gram -ve bacteria was deduced in all compounds. However such

activity could be detected only in four compounds against Gram +ve bacteria. The

14

compounds (25), (26), (30) and (31) demonstrated activity against both Gram +ve

and Gram -ve bacteria.

The compounds exhibited antimicrobial activity in the concentration range

200 ng to 500 fig/100 |il. Two compounds, (30) and (31) also demonstrated

antifungal (anticandidal) activity. Overall broad spectrum antimicrobial activity

i.e. active against Gram +ve and Gram -ve was deduced in (25), (26), (30) and (31).

These compounds may be directly explored in the preparation of topical antiinfective

agents. However, further exploration requires detailed study on exact MIC values and

least toxicity to host cell and system.

DESIGN AND SYNTHESIS OF BIOLOGICALLY ACTIVE COMPOUNDS WITH SULPHUR AND

NITROGEN CONTAINING HETEROCYCLES AND RELATED COMPOUNDS

T H E S I S SUBMITTED FOR THE DEGREE OF

B o t t o r o f $ l ) i l Q ^ o p t ) p

IN

C H E M I S T R Y

BY

MUJEEBUR RAHMAN V. P

DEPARTMENT OF C H E M I S T R Y F A C U L T Y OF S C I E N C E

A L I G A R H MUSLIM U N I V E R S I T Y ALIGARH (INDIA)

2002

THESIS

DEPARTMENT OF CHEMISTRY A L I G A R H M U S L I M U N I V E R S I T Y

A L I O A R H — 2 0 2 0 0 2

P H O N E S Int. 3 1 7 3 1 8

Certtticate

I certify that the work presented in this thesis entitled "Design and

synthesis of biologically active compounds with sulphur and nitrogen

containing heterocycles and related compounds" has been carried out by

Mr. Mujeebur Rahman V.P. under my supervision. It is original in nature

and has not been submitted for any other degree.

(Prof. W.H. Ansari)

I bow my head with all humility and reverence to the Almighty

for the completion of this task.

Words fail to express my indebtedness that I owe to my erudite

research guide, Prof. Wajid Husain Ansari, who inspired to take up

this task. I will cherish throughout my life the valuable suggestions,

constructive criticism and unending support that 1 received from him

throughout the course of this study.

I wish to place on record the kind help received from Prof.

Shafiullah and Prof. M. Ilyas, who as Chairmen of the Department

provided all the necessary facilities for undertaking this research

problem. It is my privilege to have the guidance of Late Prof. M.S.

Ahmad, who as my teacher was generous enough in showing me the

right path. I would like to thank all the faculty members of the

department, specially, Dr. A. Rauf for their constant support and

encouragement.

I M!ish to express my veneration to Prof. G. Lemiere, University

of AntM'erp (RUCA), Belgium, who as our collaborator carried out the

spectral studies of our compounds. Dr. S.K. Raza and Dr.V.V. Singh,

Defence Research Development Organization, Gwalior, India are also

acknowledged deeply for their kind cooperation in providing some

spectra. The help received from National Cancer Institute, Maryland,

Bethesda, USA in carrying out the anticancer activity of our

compounds is also truly acknowledged. I wish to express my

benevolence to Dr. I. Ahmad, Department of Agricultural

Microbiology, Faculty of Agricultural Studies, A.M.U., Aligarh, India

for his help in the determination of antimicrobial properties of some

novel compounds.

/ M'ish to record my debt of gratitude to many people who directly

or indirectly extended their support and encouragement especially my

lab colleagues, Sayeed Mukhtar and S.P. Singh for their all out help

and manifold cooperation throughout the course of this work.

I would like to express my profound sense of gratitude and

felicitation to Mr. K. Alavi, Mr. Naushad, Mr L.K. Peter and Mustafa

Kausar for all the help received from them.

I find no words to express my feelings for my dear parents whose

blessings and prayers was always a source of inspiration to me. I will

always cherish the valuable advice of my Late father-in-law. Last but

not the least 1 thank my wife without whose magnanimous support

nothing could have been done and my son Waquid vt'ho waited patiently

and stayed without me for days. I also thank my all family members for

their concern and suggestions at various stages.

J

(Mujeebur Rahman V.P.)

CONTENTS

Synthesis, Structure Elucidation and Biological Screening

CHAPTERS Contents Pages

General Introduction (i-ii)

PART ONE: l,2,4-TRIAZOLO[3,4-b][l,3,4]THIADIAZEPINES/ 1-164 THIADIAZOLES

THEORETICAL 1-19

CHAPTER I : Long alkyl chain substituted-5-Triazolo-Thiadiazepines 20-68

DISCUSSION 20-58

EXPERIMENTAL 59-68

CHAPTER II: Long alkyl chain substituted-Benzopyrano-^-Triazolo- 69-115 Thiadiazepines

DISCUSSION 69-107

EXPERIMENTAL 108-115

CHAPTER III: Long alkyl chain substituted-5-Triazolo-Thiadiazoles 116-164

DISCUSSION 116-148


REFERENCES 157-164

PART TWO : 2,3-DISUBSTITUTED THIAZ0LIDIN-4-0NES/ 165-286 THIAZAN-4-0NES

THEORETICAL 165-178

CHAPTER IV : 2-Aryl-3-A^-Long alkyl chain substituted-Thiazolidin- 179-243 4-ones/Thiazan-4-one

DISCUSSION 179-232


CHAPTER V: 2-Aryl-2-Methyl-3-A^-Long alkyl chain substituted- 244-286 Thiazolidin-4-one

DISCUSSION 244-270


REFERENCES 277-286

PART THREE: SCREENING FOR BIOLOGICAL ACTIVITY 287-300

CHAPTER VI : Study for Anticancer and Antimicrobial Activities 287-299

(A) ANTICANCER ACTIVITY 287-295

(B) ANTIMICROBIAL ACTIVITY 296-298

REFERENCES 299

CONCLUSION 300

LIST OF PUBLICATIONS 301

GENERAL INTRODUCTION

Heterocyclic compounds are very widely distributed in nature and are

essential to life as they play a vital role in the metabol ism of all living cells.

Thus, for example, the fol lowing are heterocyclic compounds: the pyrimidine

and purine bases of the genetic material DNA; the essential amino acids

proline, histidine and tryptophan; the vitamins and coenzyme precursors

thiamine, r iboflavine, pyridoxine, folic acid and biotin; the B12 and E families

of vitamin; the photosynthesizing pigment chlorophyll ; the oxygen

transport ing pigment hemoglobin, and its breakdown products the bile

pigments; the hormones kinetin, heteroauxin, serotonin and histamine,

together with most of the sugars. There are a vast number of

pharmacological ly active heterocyclic compounds, many of which are in

regular clinical use. Some of these are natural products , for example

antibiotics such as penicillin and cephalosporin, alkaloids such as vinblastine,

ellipticine, morphine and reserpine, and cardiac glycosides such as those of

digitalis. However, the large majori ty are synthetic heterocycl ics which have

found widespread use, for example as anticancer agents, analeptics,

analgesics, hypnotics and vasopressor modif iers , and as pesticides,

insecticides, weedkillers and rodenticides.

There is also a large number of synthetic heterocycl ic compounds with

other important practical applications, as dyestuffs , copolymers , solvents,

photographic sensitizers and developers, as ant ioxidants and vulcanization

accelerators in the rubber industry, and many are valuable intermediates in

synthesis.

The broad spectrum of biological activities associated with these

compounds and the multiplicity of action displayed by certain individual

members make them one of most interesting class of compounds . The current

research in organic chemistry and closely allied branches of biology is

characterized by extensive investigation of physiologically active compounds

encountered in the plant and animal world. The search for these active

constituents which control the biological process of various systems have

acted as a powerful st imulus to the further development of the chemistry of

heterocyclic compounds. Synthesising any useful organic compound is easier

than to isolate the same from natural resources. Therefore , in recent years

much effor t are being applied by chemists in synthesising such organic

compounds which possess high degree of biological activities. Innumerable

heterocyclic compounds have been prepared across the world for better

therapeutics, part icularly the compounds having hetero atoms such as O, S

and N.

A study on the inf luence of structure on activity shows that sometimes

a minor change in heterocyclic nuclei enhances the pharmacological profi le

many folds than its parent nuclei. The search for new eff ic ient and safe nuclei

lead to an improvement in the existing drugs by increasing their potency,

duration of action and decreasing side effects as well as creating new

biologically active agents by molecular modificat ion.

The successful application of heterocyclic compounds in these and

many other ways, and their appeal as materials in applied chemistry and in

more and fundamental and theoretical studies ensures virtually limitless

series of structurally novel heterocyclic compounds with wide range of

physical, chemical and biological properties.

Keeping in view of the importance of heterocyclic compounds , we

have undertaken this problem on 'Design and Synthesis of Biologically

Active Compounds with Sulphur and Nitrogen Containing Heterocycles ' -

A H

1,2,4-TRIAZ0L0[3,4- b]ll,3,4J THIADIAZEPINES/THIADIAZOLES

I jy "'J »I ^^

J J, J a U J J r .

THEORETICAL

Thiadiazepines and thiadiazoles are two important classes of

biologically active seven and five membered heterocycles containing one

sulfur and two nitrogen atoms. The isomeric structures are also possible for

thiadiazepine and thiadiazole rings depending upon the relat ive posit ions of

the three heteroatoms and the degree of oxidation of the ring systems.

Thiadiazepines and thiadiazoles having sulfur and ni t rogen atoms at 1,3,4-

posit ions are called 1,3,4-thiadiazepines and 1,3,4-thiadiazoles (I and II) .

N N

R^ III

N N

R' "N-/

N

R •R'

V R^

4 N - N 3

II

R'

N N

•N

N:

IV R'

R = Alkyl/Aryl/Heterocyclic r ' = Alkyl/Aryl/Heterocyclic

R^ R^ = H/Alkyl/Aryl

R'

N N

\ H N -

VI

R'

l ,2 ,4-Triazolo[3,4-b][ l ,3 ,4] thiadiazepine/ thiadiazole ( I I I / IV) are the

fused rings system of 1,2,4-triazole with l ,3 ,4- th iadiazepine/ l ,3 ,4- th iadiazole

and their dihydro derivatives are represented by the s t ructures (V) and (VI)

respectively.

2

1,3,4-Thiadiazepine derivatives are associated witli a varied range of

biological activities such as antimicrobial'"^, anticancer^, he rb i c ida^ ' ^ anti-

viral^, anti-HIV'®"'"', anticholinergic'^, antidepressant'^"'^, anaes thet ic '^

cholere t ics '^ calcium antagonist '^ and A C E inhibitory'^ activity. They have

also gained industrial imiportance as charge generators for

electrophotographic photoconductors^"®-'' and photoreceptors^^^

The 5-Triazoio-thiadiazole derivatives have been extensively studied in

the recent past. Numerous reports are available in the literature which

highlight their chemistry and use. l ,2 ,4-Tr iazolo[3 ,4-b][ l ,3 ,4] th iadiazole

derivatives have been found to be associated with diverse biological activities

such as antibacteriaP'"^^ antifungaP'"^^'^^"^®, anti-inflammatory^"''^'"^-',

anthelmintic^'•^^ ana lges ic" , antihypertensive^^-^^'"''', herbicidal' '" and

pesticidal activity'^'.

Several methods are reported for the synthesis of 1,2,4-triazolo[3,4-

b][ l ,3 ,4] thiadiazepines and l ,2 ,4~tr iazolo[3,4-b][ l ,3 ,4] thiadiazoles .

l , 2 ,4 -Tr iazo lo f3 ,4 -b] f l , 3^4] th iad iazep ines

The substituted and unsubst i tu ted-4-amino-3-mercapto- l ,2 ,4- t r iazoles

on cyclocondensat ion with ^./^-unsaturated ketones in presence of acidic or

basic catalyst yield 6 ,8-disubst i tu ted- l ,2 ,4- t r iazolo[3 ,4-b][ l ,3 ,4] th iadia-

zepines.

N.N. Koles et al.''^" have reported the s tereoselect ive addition of

4-amino-3-mercapto- l ,2 ,4- t r iazole with acetylenic ketone to give the Michael

adduct (VII) which on cyclocondensation with D M F yielded 6,8-diphenyl-

1,2,4-triazolo [3,4-bJ [1,3,4] thiadiazepines (VIII). Similar reaction of the

4-amino-3 -mercap to - l , 2 ,4 - t r i azo le with 4-n i t rocha lcone a f fo rded only the

Michae l adduct (IX) and all a t tempts fai led to undergo cyclocondensation'*^' ' .

N N

I NH2

O N N Ph

Ph SH

O2N.

NO2 0

N N

I NH2

0

•Ph

IX

N N

VIII Ph

J.E. Glo tova et a l / ^ have shown that the pheny l subs t i tu ted acetyl inic

ke tone with 4 -amino-3 -mercap to - l , 2 ,4 - t r i azo le in acet ic acid g ives6 ,8-d iphe-

nylsubs t i tu ted- l ,2 ,4- t r iazolo[3 ,4- -b] [ l ,3 ,4] th iad iazepine (X ) whi le unsubst i -

tu tedacety l in ic ke tone yieldsonly the Michael adduct (XI).

N N

N ^SH

O

'Ph

AcOH

NH2

AcOH

0

Ph

N N O

NH^ XI

-Ph

N N

I N

Recent ly , U.V.Laddi et al."^'' have synthes ised 3-subs t i tu ted-6 ,8-

d iphenyl -d ihydro- l ,2 ,4 - t r i azo lo [3 ,4 -b] [ l ,3 ,4 ] th iad iazep ine (XII) by the

cyc locondensa t ion of 3 - subs t i tu ted-4-amino-5-mercap to- l ,2 ,4 - t r i azo les with

cha lcone in presence of catalytic amount of acetic acid.

N N N N O

+ R"

I NH2

Ph AcOH^

MeOH

R = Ph PI

=N-0-(CH2)n-

=N-0-(CH2)n-

R' N ' I

N

Ph

Ph

XII

B. Kal luraya et al.'*''^ reported that the ace ty len ic ke tones on

condensa t ion with 3 - subs t i tu ted-4 -amino-5-mercap to - l ,2 ,4 - t r i azo les in

ethanol f irst gave the Michael adduct , l - a ry l -3 - (5 -n i t ro -2- th ieny l ) -3 - [3 -

sus t i tu ted-4-amino-5-mercap to- l ,2 ,4 - t r i azo le (XIII) wh ich on fur ther

t rea tment wi th conc. H2SO4 yielded 6 -a ry l -8 - (5 -n i t ro -2- th ieny l ) - l ,2 ,4 - t r i azo lo

[3 ,4 -b] [ l ,3 ,4 ] th iad iazep ines (XIV).

NO-

N N

Ph'

XIV

Ph

O

N N

/^N-^SH ^ I

NH2

EtOH, A

r

N N

N' EtOH/conc. H2SO4

Ph'

O C-

X I I I

N02

R = H, CH3, C2H5, C3H7, Phenyl, jPtolyloxymethyl

In a similar reaction'*''®''', a - b r o m o c h a l c o n e on condensa t ion with

3 - subs t i tu ted-4-amino-5-mercap to- l ,2 ,4 - t r i azo les us ing sodium acetate as

catalyst a f fo rded the Michael adducts (XVa-d) which on cycl i sa t ion furnished

the cor responding t r iazolo- thiadiazepines (XVIa-d).

Ar'

Br N N

,Ar + NaOAc / EtOH.

0 R / ^ N - - S H

N H ,

N N

A

•N-^S -HBr

Ar XVI a d

a: Ar = Phenyl Ar' = 2-Nitrothienyl b: Ar = Phenyl Ar' = /?-anisyl c: Ar = p-chiorophenyl, Ar' =p-Tolyl. d: Ar= Phenyl

N N

A r -

N N H , O X- Ar'

Br

XVa-d

i-H20

N

R' I

N

\ O h Br

Ar'

When 3-ary l -2-bromo-1 - [4- (n i t ro-2- th ienyl ) ]propen-1 -one condensed

with 3 - subs t i tu ted-4-amino-5-mercap to - l ,2 ,4 - t r i azo les , the produc t obtained

was 3-subs t i tu ted-7-a ry l idene-6- (4-n i t ro -2- th ienyl ) - l ,2 ,4 - t r i azo lo[3 ,4-b]

[l,3,4]thiadiazines'*'*'^ (XVII) instead of the expec ted t r iazo lo- th iad iazepines .

NO2 N- -N

R' ' N ^ SH I

NH2 NaOAc/EtOH, A

N-

R" "N I

N

-N N-

R'

Ar

'N' I N,

• N

O2N Ar = Phenyl,/?- anisyl, p-chlorophenyl, /p-methylphenyl XVll

^CH-Ar

O2N

They'*'''' have also synthesised novel 3-subs t i tu ted-6- (3-ary l

sydnony l ) -8 -a ry l - l ,2 ,4 - t r i azo lo [3 ,4 -b ] [ l ,3 ,4 ] th iad iazep ines ( X V I I I ) by

reac t ing 2 -b romo- l - (3 - a ry l sydnony l ) -3 - a ry l -2 -p ropen - l -one wi th 3-subst i -

tu t ed -4 -amino-5-mercap to - l ,2 ,4 - t r i azo les in ethanol in p resence of sodium

acetate.

N - • N

R ' "N SH

N H , O O

Ar = Ph, Ph-0CH3 R = CH3_ Ph, o-cresyloxymethyl,

p-cresyloxymethyl R'= H, CH3, OCH3.

N -

N - N E t O H , N a O A c 0 N / K , ^ Ar

R' 0

O

•N' /

• N

O,® N.

XVIII ^R'

G.J. Reddy et al.'^^ have synthesised 7 - (2 ' -hydroxybenzo ly l ) -3 -phenyl -

l ,2 ,4 - t r i azo lo [3 ,4 -b ] [ l ,3 ,4 ] th iad iazep ines (XIX) by the react ion of

3 - f o r m y l c h r o m o n e s with 3 -pheny l -4 -amino-5 -mercap to - l , 2 ,4 - t r i azo le s in

presence o fBu4N^HS04~ catalyst .

N N

P h - ^ N ^ S H O H C

N H ,

r ' = H, Me, CI, Br, R^ = H, Me

XIX

A.M. El -Sayed et al.'^^ have repor ted the synthes is of 3-( t r i f luro-

methyl ) -6-amino-7 ,8-d isubs t i tu ted-1 ,2 ,4- t r iazo lo[3 ,4-b] [ 1 ,3 ,4] th iadiazepines

(XX) by the react ion of 4 -amino-5 -mercap to - (3 - t r i f l u rome thy l ) - l , 2 ,4 - t r i azo le

with yl id ine malononi t r i les .

N N N C 33 IM N

F3C Y SH ^ NHj

, 1

R' F r- "N F3C I N

R = H, Ph, 4-CIC6H4, 4-NO2C6H4,2-Pyrrolyl, HjN 2-Pyridyl

R^ = CN, C02Et

R"

X X

The react ion of 3 - subs t i tu ted -4 -amino-5-mercap to - l ,2 ,4 - t r i azo les with

2 ,4 -pen tand ione yielded 3-subs t i tu ted-6 ,8-d imethyl - l ,2 ,4- t r iazolo[3 ,4-

b][13,4] th iadiazepines ( X X l / ' - ' ^

N N H" R

R = 3-Me,3-Me, 4-Me, 4-NO2, 4 - C l , 2—Br, Me

; "Me

XXI

N i z a m u d d i n et al.^ have prepared some 3 -a ry loxyme thy l -8 -n i t ro - l , 2 , 4 -

t r i azo lo[3 ,4 -b] [ l ,3 ,4 ]benzoth iad iazep ines ( X X I I I ) as po ten t herb ic ida l agent

by the cycl isa t ion of 5 -a ry loxymethy l -4 - (2 -ch loro-5-n i t robenzy l ideneamine) -

3 -mercap to - l , 2 ,4 - t r i azo le s (XXI I ) with pyridine.

NO. N N

OCH2- + OHC-'N SH

I NH2 CI'

O MeOH

R N N

-OCH2- -^SH CI

N = C H

XXII

R

Pyridine

NO2

R = 2 - C 1 , 4 -C i ,2 ,4-Ci2 . 2-CH3, 4 - C H 3

N N

-OCH2

/ V

xxni

s 3

NO2

B Prabhuswami et al have synthesised s- tr iazolo th iadiazepino-

quinol ines (XXIV) by the condensat ion of ch loroformyl quinol ines with

3 - subs t i t u t ed -4 -amino -5 -mercap to - l , 2 ,4 - tnazo l e s The base catalysed faci le

in t ramolecular rearrangement of (XXIV) yielded 5- t r iazolothiadiazino-

qumol ines (XXV) by the rearrangement involving N - N bond scission

CHO N N

NH,

XXIV XXV

R = H Mc CI R' = Me Ph X = C1

A Brukstus et al ^ have shown that the cyc locondensa t ion ol

pyr imidine carboxaldehyde with benz imidazole th iones gave benzimidazo

[2 , l -b ]pyr imido[5 ,4 - f ] [ l , 3 ,4 ] th i ad iazep ine ( X X V I ) and 5H-benz imidazo

[ 2 ' , r 2 ,3 ] [ l ,3 ] th iaz ino[6 ,5 -d]pyr imid ine (XXVII )

MeS. ^ N .

N,

CI

H

^CHO H2N' Q

NaOAc

MeS^ N. ^ S

N;

Ci :N

XIII

-N. .N

OH CI

XXVIl

A.Kakeh i et al.^' have carried out the react ion of unsymmetr ica l ly

subst i tu ted 1-pyr id in ioamidates (XXVIIIa-e) wi th d imethylace ty lene

d icarboxyla te ( D M A D ) which a f fo rded the corresponding d imethyl -2-ary l -6-

me thy l -5aH-pyr ido [ l , 2 -d ] [ l , 3 ,4 ] th i ad iazep in -4 ,5 -d ica rboxy la t e s (XXIXa-e)

along wi th the rearranged produc ts (XXXa-e) in minor amount . Similar

react ion of l - (2 -methy lqu ino l ino)amida tes (XXVIII f ,g ) wi th D M A D gave

only d ime thy l -2 -a ry l -5a -methy l -5aH- l ,3 ,4 - thad iazep ino [4,5-a] quinol ine-

4 ,5-d icarboxyla tes (XXXm,n)^'.

COjMe CHCI3, 50-60°C

COiMe 8h

R', R2 = (CH=CH)2-

R4 = Me

r ' = R'' = H, R = Me CHCI3, 50° - 60°C 4h

Me COjMe

Ar XXIX a - e

R '

COjMe

Ar

'Me

COOMe COOMe

n : Ar = 2-thienyl

XXX a - e COiMe

R' R^ Ar

Me H H Ph

Me H H ;?-MeC6H4

Me H H p- M e O C e H

Me H H P-CIC6H4

Me H H 2- th ienyl

H Me Ph

H Me 2- th ienyl

a

b

c

d

e

f

H

H

H

H

H

-(CH=CH2)-

-(CH=CH2)-

1 0

J Bolos et al have reported the multistep asymmetr ic synthesis of

(3R,rS)-3-[ l ( -carboxy-3-phenylpropyl)amino]-4-oxo-2,3 ,4 ,5- te t rahydropyrrolo-

[2 , ] -b ] [ ] ,3 ,4 ] th iad iazep ine-5-ace t ic acid (XXXIId ) , a potent A C E inhibition

active agent f rom A/^-carbobenzoxy-L-senne through a sequence involving

pyrrole th iocyanat ion, fo l lowed by the reduct ive c leavage of the th iocyanate

group, and cyclisat ion of the result ing halo-thiol m the key synthet ic steps

Thus ( /^ ) -2- [ (benzyloxycarbonyl )amino] -3- iodo- / / - (2 - th iocyana to- l -pyr ro ly l )

p ropanamide (XXXI) (prepared f rom A^-carbo-benzoxy-L-serine) , on

reduct ive t reatment with NaBH4 afforded ( /^ ) -3- [ (benzyloxycarbonyl )amino]-

2 .3 ,4 ,5- te t rahydropyr ro lo[2 , l -b] [1 .3 ,4] th iad iazepin-4-one ( X X X I I )

NHZ HO. ^

HO i) DDC/N-H^drovs'succinimidc ^COOH ^ ^

») N-NH2

N

Z N H ^ \ / N H

o I

Z N H — - NH

O

XXXI

Thiadiazepin-4-one (XXXII ) on t reatment with e thy lb romoace ta t e in

the presence of sodiumethoxide af forded ( X X X I I a ) wh ich on hydrolyt ic

c leavage with lodo tnmethy l s i l ane (ISiMes) a f fo rded ( X X X I I b ) Alkylat ion of

( X X X I I b ) with racemic e thy l -2-bromo-4-phenylbu tanoa te gave ( X X X I I c )

which on hydrolysis a f forded the target molecule ( X X X I I d )

1 1

Z N H '

/ N ' I BrCHzCOOEt

N H NaOEt *

o

(XXXII)

. N

0 COOEt

(XXXIIb)

ZNH

/ ^ N - ^

V S o C O O E t

(XXXIIa)

i) IS iMe j

ii) M e O H

R = Et R = H

(XXXIId)

Recently, V.A. Bakulev and co-worker s . " have reported the first

synthesis of some novel bis t r iazolo[ l ,3 ,6] thiadiazepine derivatives by the

base catalysed condensation of aromatic and al iphatic 1,2-diamine with

5-chloro- l ,2 ,3- th iadiazoles via a multistep process which is thought to

involve two Dimroth rearrangements and subsequent loss of hydrogen-

sulphide by intramolecular nucleophilic subst i tut ion. Thus, when

5-chlorothiadiazole ester (2 equiv) on reaction with 1,2-phenylenediamine

( l equ iv ) in presence of EtsN undergo Dimroth rear rangement to yield a

mixture of 5-( l ,2 ,3- t r iazol -5-ylsul fanyl) - l ,2 ,3- th iadiazoles (XXXIIIa-e) and

novel b is t r iazolo[ l ,5-b;5 ' , r - f ] [ l ,3 ,6] thiadiazepine derivatives (XXXIVa,d-e).

Prolonged heating of the reaction mixture gave (XXXIVa,d-e ) as the pure

products. Saponificat ion of (XXXIVa,d-e) gave the diacid (XXXIVf) which

on decarboxylat ion by heating to melt at 180 °C gave the bistriazolo

[ l ,3 ,6] thiadiazepine (XXXIVg)

1 2

N // \ • ci

H2N

H2N' Q EtjN, EtOH Reflux

a: r ' = R^ = COjEt b: r ' = R = CONH2 c: r ' = R^ = CONHMe d: r ' = CONH2, R^ = C02Et e; R ' = CONHMe, R^ = C02Et

N -

J \ NH

. N H ,

N -

N // w R2 ,NH2

XXXIII a-e

N -

N '/ \ N SH

,NH2

N W / N-N N-

XXXIV a, d-e

n - n n - n 180 °C

iNaOH, thenHCl ^ (Saponification)

COOH COOH

N , \\ I

(Decarboxylation) N - N N - n '

/ ' ^ \

XXXIV g XXXIV f

l ,2 ,4-TriazoIof3 ,4-b][ l ,3 ,41thiadiazoles .

3,6-Disubsti tuted-2,3-dihydro-l ,2,4-tr iazolo[3,4-b][l ,3,4]thiadiazoles

can be conveniently synthesised by reacting 3-substi tuted-4-amino-5-

mercapto-l ,2,4-tr iazoles with aldehydes in presence of an acid catalyst like p-

TsOH, HCl etc.

S. Sangeetha et al.^^'^^ have prepared a number of 3-substituted-6-

(2 ' ,4 ' -dialkoxy-5-alkyl)phenyl-5,6-dihydro-l ,2,4-tr iazolo[3,4-b][l ,3,4]thiadi-

azoles (XXXV) by the reaction of 4-amino-3-phenyl substi tuted-5-mercapto-

1 3

1,2,4-tr iazoles with 2 ,4-d ia lkoxy-5-a lky lbenza ldehydes in presence of

/ j - T s O H as catalyst .

N- -N

NHz

OHC-

R O'

R'

OR /j-TsOH

Benzene R

R = 2-Me.C6H4, 2—OH.C6H4,2—CLC6H4 r ' = Et, Pr R^ R = Me, Et

XXXV

S. Hai jan et al^^^'^^ have synthesised var ious 3 ,6-disubs t i tu ted-2 ,3-

d ihydro - l , 2 ,4 - t r i azo lo [3 ,4 -b ] [ l , 3 ,4 ] th iadiazoies ( X X X V I ) by the react ion of

3-ary lsubs t i tu ted-4-amino-5-mercapto-1 ,2 ,4- t r iazo les wi th d i f fe ren t a ldehydes

in the presence of catalytic amount of HCl.

N- -N N-

Ar I NH2

+ R—CHO HCl

SH Ar

-N

HN—( R

XXXVI

Ar = 4-NO2.C6H4, 3-NO2.C6H4, 2—CLC6H4, 2-MeO ,2-Furfuryl, CH3.CH=CH

R = 4-MeO.C6H4, 4-Me2N.C6H4, 4—CLCeH,, 2—OH.C6H4, 3-MeO.C6H4, C6H5-CH=CH

A number of 5 ,6 -d ihydro-6- (subs t i tu ted-phenyl ) -5- (2 ,3-epoxypropyl ) -

3 -methy l - l , 2 ,4 - t r i azo lo [3 ,4 -b ] [ l , 3 ,4 ] th iad iazo les^^ ' (XXXVI) have been

prepared by first condens ing 3 -methy l subs t i tu ted-4-amino-5-mercap to-1 ,2 ,4 -

t r iazo leswi th var ious subst i tuted aromat ic a ldehydes in b e n z e n e using Dean

Stark appara tus in presence of /7-TsOH as catalyst f o l l owed by alkylat ion

with l - ch lo ro -2 ,3 -epoxypropane in ace tone and K2CO3

1 4

N N + R—CHO

- CH3 SH NH,

p-TsOH

Benzene

N N

HN-R

K2CO3 C H 2 - C H - C H 2 C I

O N N

C H s '

H2C-CH —CH2

0

A-n-N-

R XXXVII

R = 4-MeO.C6H4, 3,4—(MeO)2.C6H3. 4—CLC6H4, 3, 4—(OCH20).C6H3

In a s imilar react ion, they have also prepared some 2-subs t i tu ted

phenyl -3- (3-a lky l /a ry l ) -5 ,6 -d ihydro-1 .2 ,4 - t r i azo lo[3 ,4 -b] [ l ,3 ,4 ] th iad iazo l -6-

y l indoles (XXXVIII ) by the condensat ion of 2 -a ry l indo le -3 -a ldehydes with

3-subs t i tu ted-4-amino-5-mecapto-1 .2 .4- t r iazo les in p resence of /?-TsOH as

catalyst .

N N

C H 3 ' SH

NH2

Q H

D M F

N N

C H / ^S

H N -

Q

Q 'N'

R = CH3, C2H5, C3H7, CfiHj, CH2.C6H5, 2 - M e CeHj , 4 - M e O . C 6 H 4

r ' = CH3, CI

XXXVIIl

G. A. E l - S a r a r ° has syn thes i sed s o m e 3 - ( 2 - b e n z o f u r a n y l ) - 5 , 6 -

d i h y d r o - 6 - s u b s t i t u t e d - l , 2 , 4 - t r i a z o l o [ 3 , 4 - b ] [ l , 3 , 4 ] t h i a d i a z o l e s ( X X X I X ) by

r e a c t i n g 4 - a m i n o - 3 - ( 2 - b e n z o f u r a n y l ) - 5 - m e r c a p t o - 1 , 2 , 4 - t r i a z o l e wi th

1 5

d i f f e r e n t a l d e h y d e s such as jO-anisaldehyde, s a l i c y l a l d e h y d e , /?-chloro-

b e n z a l d e h y d e and 2 - t h i o p h e n a l d e h y d e in p r e s e n c e of p y r i d i n e as ca ta lys t .

Q N N

+ R - C H O P i p e r i d e n e ^ ^ ^ |

N N

I NH,

a b s . E t O H S H

H N -

X X X I X

R = 2 - O H . C 6 H 5 , 4 - M e O . C 6 H 4 , 4—CI .C6H4, 2 — t h i o p h e n y l

Z.Wang et al.^'^ have carried out the s te reose lec t ive synthes is of a

number of new chiral 3 ,6 -d ia ry l -5 ,6 -d ihydro t r i azo lo [3 ,4 -b] [ l ,3 ,4 ] th iadiazoles

( X L ) by the Mannich react ion of 3 -a ry l -4 -amino-5 -mercap to - l , 2 ,4 - t r i azo le s

with var ious alkyl or aryla ldehydes in the presence of cata lyt ic amount of

L-(+)- tar tar ic acid.

N- -N

^ r + R—CHO L-(+)-Tartaric acid

C H 3 ^ Y S H

NH2 H "R HN

H XL

Ar = 2-MeO.C6H4, 4-MeO.C6H4 R = C H = C H - C H 3 , 2 - M E O . C 6 H 4 , CGHJ, 2 - O H . C 6 H 4 , 4 - O H . C 6 H 4 ,

4-NO2.C6H4, 4-Me2N.C6H4, 4-CI.C6H4

In a s imilar react ion, they have prepared 3 - (3 -pyr idy l ) -6 - (pheny l /

2 - fu ry l ) subs t i tu ted - l ,2 ,4 - t r i azoIo [3 ,4 -b ] [ l ,3 ,4 ] th iad iazoIes 5 4 ( X L I ) by

reac t ing 3 - (3 -pyr idy l ) -4 -amino-5-mercap to - l ,2 ,4 - t r i azo le wi th benza ldehyde

or 2 - f u r f u r a l d e h y d e in the presence of L-(+)- tar tar ic acid . But , in this case,

the absolu te conf igura t ion was not detected.

1 6

CPJO L-(+) -Tartaric acid N N

R = C6H5, ^ O O XLI

R. Gupta et al .^^have also synthesised a series of novel 7/9-subst i tuted-

4 - (3 -a lky l / a ry l ) -5 ,6 -d ihydro- l ,2 ,4 - t r i azo lo [3 ,4 -b ] [ l , 3 ,4 ] th iad ia zol-6-yl) te t ra-

zo lo [ l , 5 -a ]qu ino l ines ( X L I I ) by the condensa t ion of 7 ,9-disubs t i tu ted-4-

fo rmyl t e t r azo lo [ l , 5 -a ]qu ino l ines with 3 -subs t i tu ted-4-amino-3-n ie rcap to-

1,2,4-tr iazoles under mic rowave irradiat ion us ing basic a lumina as solid

support .

N N

R^n-^sh Q NH2 r

C H O

' N I

fiv/A

R = CH3, C2H5, C3H7, CH2.C6H5, 4-OMe.C6H4, 2-Me.C6H4, 4-OMe.C6H4 1 1 N = N

R' = H, CH3, OCH3

R = H, CH3

M. Kidwai et al.^^ have shown that the react ion of 4 -amino-5 -mercap to -

3- (4 ' -methylquinol iny l -2 ' -oxymethyl ) / ( te t razo ly l -1 ' -methy l ) -1 ,2 ,4 - t r i azo les

with benza ldehyde in D M F under mic rowave i r radia t ion y ie lded the

cor responding 3 - subs t i t u t ed -6 -pheny l -5 ,6 -d ihydro - l , 2 ,4 - t r i azo lo [3 ,4 -b ] [ l , 3 ,4 ]

th iadiazoles ( X L I I I ) .

N N

R-^n-^sh

N- -N

+ R — C H O [iv/A DMF" K

NH2 \

HN

R = C6H5, O 0 N ' "OCHr

n . n XLIII R

N - C H 2 -

1 7

The react ion of 3 -subs t i tu ted-4-an i l ino-5-mercapto- l ,2 ,4 - t r iazo les with

oo-bromoacetophenones yielded 4-an i l ino-5-phenacylmercap to-3- th iophe-

noxyme thy l - l , 2 ,4 - t r i a zo l e s which on brominat ion fo l lowed by cyciisat ion

a f fo rded 3-subs t i tu ted-5-phenyl -6-benzoylsubs t i tu ted- l ,2 ,4 - t r i azo lo[3 ,4-b]

[ 1 ,3 ,4 ] th i ad iazo l id ines" (XLIV) .

N N

N I

P h N H SH

O II

BrCH-C-I R2

Ri jxvM DMr'

N N I I

N

P h N H

Br j AcOH UV-iig)it

N N

N \

P h N

N N

,Q R

EtOH

Pyridine

N

P h N H S I

BrC

0

XLIV

R = CjHj 0 CHj, 4-CH3 CaH) OCH2,4-CLC6H4 SCH,. 2.4-CI2 C6H3OCH2 C f i H j SCH2, 4-CH3 C a R , SCH2, 4-CLC6H4 SCH2

R= H, CI, CH3 R = H, 4-CLC6H4S, 4-CLC6H4O, CfiHj

N.D. Heinde l et al.^^ have reported the synthesis of 3 ,6-disubs t i tu ted-

l ,2 ,4- t r iazo lo[3 ,4-b] [1 .3 ,4] th iad iazoles (XLV) by the condensa t ion of 1,2,4-

t r iazoles wi th c innamaldehyde in ethanol. Dur ing this reac t ion , the format ion

of 3 -R-8-a ry l - l , 2 ,4 - t r i azo lo [3 ,4 -b ] [ l , 3 ,4 ] th iad iazep ines in mino r amount was

also observed .

N- -N

R^n-^sh +

^SH NH2

R = H , AJkyl, Aryl

Ph EtOH

XLV

1 8

R. Gupta et al.^"' have prepared several 3-alkyl /aryl-6-(2-chloro-2-

subst i tu ted-phenylethenyl)-5 ,6-dihydro- l ,2 ,4- t r iazolo[3,4-b][ l ,3 ,4] thiadiazo-

les ( X L V I ) by the condensation of 3-subst i tuted-4-amino-5-mercapto-1,2,4-

triazoles with /7-chlorocinnamaldehyde in benzene conta in ing catalytic

quantity of p - T s O H using Dean Stark apparatus.

N - - N

SH

)3-TSOH

E t O H

N H .

X L V I

R = CH3, C2H5, C3H7, CH2.C6H5,4-OMe.C6H4, 2-Me.C6H5 R, = CI, Br. OMe

Z. Y. Zhang and co-workers^^ have shown that the react ion of 4-amino-

3-( l - jC-chlorophenyl-5-methyl- l ,2 ,3- t r iazol-4-yl) -5-mercapto- l ,2 ,4- t r iazole

with p-chlorobenzaldehyde/benzaldehyde in presence of catalytic amount of

p - T s O H in ethanol afforded 4-benzyl ideneamino-3- ( l -p-chlorophenyl -5-

methyl- l ,2 ,3- t r iazol-4-yl- ) - l ,2 ,4- t r iazol-5- thiones ( X L V I I ) instead of the

cyclised product .

N - - N H

N N H

N - ^ N A , . O H C ^ p-TsOH

EtOH

N N

( X N = C H -N " ^ C H 3

N C H 3

O

Q R = H, CI

X L V I I

CI

CI

B. Kalluraya et a l . " have reported the condensat ion of 5-nitro-2-

(diacetoxymethyl) thiophene with 3-substi tuted 4-amino-5-mercapto-1 ,2 ,4-

triazole yielded the corresponding Sch i f f ' s base ( X L V I I I )

N NH

~ I ^ S NH2

R = H, Aikyl Ph, Ph.0CH2, Ph-CHj

EtOH

1 9

N-

R

-NH

N = CH

XLVIII

-NO2

Similar condensat ion of 3 -subs t i tu ted-4-amino-5-mercap to-1 ,2 ,4 -

t r iazoles with 5 - (2 ,4 -d ich lo ropheny l ) -2 - fu r fu ra ldehyde y ie lded 3 -subs t i tu ted-

4 - [5 - (2 ,4 -d i ch lo ropheny l ) -2 - fu r fu ry l iden^^ ( X L I X )

N N H

^ N ^ , + O H C -

I ^ N H .

Q cv

CI EtOH

N N H

A N -C I ^

N = C H -

XLIX

O cr

CI

R=HAlkyl

brl

Long alkyl chain substituted-s- Triazolo- Th iadiazepin es

2 0

DISCUSSION

SUMMARY

The M'ork described in this chapter includes the reaction of 3-(9-decenyl)-4-

amino-5-mercapto-1,2,4-triazole (1) with:

i) l,5-Bis(4-chlorophenyl)pent-l,4-dien-3-one (2) which afforded an

adduct, 1,5-bis(4-chlorophenyl)-5-[3-(9-decenyl)-4-amino-5-thio-1,2,4-

triazolyljpent-]-en-3-one (3) and a novel compound, 2-(4-

chlorophenyl)-4-[2-(4-chlorophenyl)ethenyl]-7-(9-decenyl)-2,3-dihydro-

1,2,4-triazolo [3,4-b][1,3,4]thiadiazepine (4).

ii) 4,4'-Dichlorochalcone (5) which yielded an adduct, 1,3-bis(4-chloro-

phenyl)-3-[3-(9'decenyl)-4-amino-5-thio-1,2.4-triazolyl]propan-l-one (6)

and a novel compound, 2,4-bis(4-chlorophenyl)-7-(9-decenyl)-2.3-

dihydro-1,2,4-lriazolo[3A-b][l,3,4]thiadiazepine (7).

Hi) 1,3-Bis(2-thienyl)propen-l-one (8) which furnished an adduct. f3-his(2-

thienyl)-3-f3-(9-decenyl)-4-amino-5-lhio-J,2.4-lriazolyljpropan-I-one (9)

as the sole product.

INTRODUCTION

4-Amino-5-mercapto- l ,2 ,4- t r iazoles and their /V-bridged heterocyclic

derivatives have received much attention during recent years on account of

their wide spectrum biological potentialities as anthelmintic^' ',

antihypertensive^^ antitubercular''^'', antibacterial"' ' . ant i fungal" ' ,

antiinflammatory''®, anti-HIV^', antiviral^^ inhibition of cholinesterase ' ' ' .

analgesic^'' and anticonvuisant^^'^^. The 1,2,4-triazole nucleus has been

incorporated into a wide variety of therapeutically interest ing drugs' ' '

including Hi/H2-Histamine receptor blockers, cholinesterase active agents,

CNS stimulants, antianxiety agents and sedatives. Also 1,3,4-thiadiazepine

derivatives are associated with a varied range of biological activities such as

2 1

antimicrobial '"^, anticancer^, herbicidal^"^, antiviral^, anti-HIV'*^"''',

ant icholinergic"^ antidepressant'^"'^, anaes thet ic '^ calcium antagonist '^

choleretics'® and ACE inhibitory'^ activity. Prompted by these observations

and as part of our continuing programme in the search of biological ly active

compounds with sulphur and nitrogen containing heterocycles , we have

undertaken this problem. It consists of the reaction of 3-(9-decenyl) -4-amino-

5-mercapto- l ,2 ,4- t r iazole (1) with ( i ) l ,5 -b is (4-chlorophenyl )pent - l ,4-d ien-3-

one (2) which afforded an adduct, l ,5-bis(4-chlorophenyl)-5-[3-(9-decenyl)-

4-amino-5- th io- l ,2 .4- t r iazoly l ]pent - l -en-3-one (3) and a novel compound, 2-

(4-chlorophenyl)-4-[2-(4-chlorophenyl)ethenyl]-7-(9-decenyl)-2,3-dihydro-

l ,2 ,4- t r iazolo[3,4-b][ l ,3 ,4] thiadiazepine (4). (ii) 4 ,4 ' -d ichlorochalcone (5)

which yielded an adduct, l ,3-bis(4-chlorophenyl)-3-[3-(9-decenyl)-4-amino-

5- th io- l ,2 ,4- t r iazoly l ]propan- l -one (6) and a novel compound, 2,4-bis(4-

chlorophenyl) -7-(9-decenyl) -2 ,3-dihydro- l ,2 ,4- t r iazolo[3 ,4-b][ l ,3 ,4] th iadia-

zepine (7) and (iii) l ,3-bis(2- thienyI)propen- l -one (8) which furnished an

adduct, l ,3-bis(2- thienyl)-3-[3-(9-decenyl)-4-amino-5- thio- l ,2 ,4- t r iazolyl]

p ropan- l -one (9) as the sole product.

Structural assignment and biological assay are discussed. Screening

results of the compounds (4), (7) and (9) are summar ised for anticancer

activity against 3-cell lines of three types of human cancers : lung, breast and

CNS. Screening results of the compounds (4) and (7) are also summarised for

antimicrobial activity against four bacteria, Escherichia coli, Bacillus

subtilis, Staphylococcus aureus, Pseudomonas aeruginosa and one fungi

Candida albicans (details in chapter 6).

2 2

(1)

Hz

(3)

2 3

2' 4' 6 8 io'

(8)

(9)

2 4

R E S U L T S A N D DISCUSSION

The syntheses of compounds (3), (4); (6), (7) and (9) have been

performed by the reaction between a triazole (1) and <3r,;^-unsaturated

carbonyl compounds. The triazole, 3-(9-decenyl)-4-amino-5-mercapto-1,2,4-

triazole FTZ (1) was first prepared following the method adopted by Peng-

Fei Xu et al.^^ by fusing 10-undecenoic acid with thiocarbohydrazide (molar

ratio, 1:1) at 170 °C, as white crystalline needles in 85 % yield. It was then

treated with (i) l ,5-bis(4-chlorophenyl)pent-l ,4-dien-3-one (2) (ii) 4.4'-

dichlorochalcone (5) and (iii) l ,3-bis(2-thienyl)propen-l-one (8) (molar ratio,

1:1) in dry benzene in presence of /?-TsOH. (Scheme 1). The products on

chromatography over a silica gel column using pet. ether-EtOAc (8:2 v/v) as

eluent yielded compounds FBCA (3) and FBCB (4) in (i), F T Z X 4 A (6) and

F T Z X 4 B (7) in (ii) and FTCA (9) in (iii). The reaction of the triazole with

l ,3-bis(2-thienyl)propen-l-one (8) didn't yield the desired product, triazolo-

thiadiazepine, but yielded only the adduct (9) as the sole product.

2 5

2. •a c TO

ro X! 3

!=:

c (U N c 

T 3 ^

o d X 3 (D

<D c 0 1 ^ c u '-3

I

I S 1 ^

o N ^ C —' U ^ JD

2 -a td

o d B

c Q.

_r C o

>1

cx I 0 1

ra Hi

I I s i s s o o. _o O "c3 ^ o " S o V CJ C/3 •3 ^

iri -T T /

S

X u C/)

2 6

3-(9-Decenyl)-4-amino-5-mercapto-l,2,4-triazole F T Z (1)

The outl ine of its synthesis is as fol lows:

. r „ Vr II S

1 7 0 ° C , 1 . 5 h ( O H 10-Undecenoic acid ^

' -H2O

h o ) I NH2

-H20

N N I I i l

I

NH2

FTZ (1)

H N — N H

( > P h n HN I

NH2

Structure Elucidat ion of FTZ (1)

It was a whi te crystal l ine solid, m.p. 82 °C. The s t ruc ture of F T Z was

fu l ly es tabl ished by IR, DCI-MS, ' H - N M R and ' ^ C - N M R spectra . IR-

spec t rum (KBr) gave character is t ic peaks at 3250, 3100 ( N H ), 2900(CH)

2350 (SH) , 1640 (C=C), and 1610 (C=N) cm"'. The D C I - M S spec t rum of the

sample F T Z showed a [M+1]"^ signal at m/z 255 c o n f i r m i n g its molecular

weight 254 which corresponds with the ion [C12H22N4S] . Th i s is equal to the

sum of the molecular weights of 10-undecenoic acid (184) and th iocarbo-

hydraz ide (106) minus two molecules of water. This sugges ted the format ion

of 5- tr iazole r ing by the condensat ion of t h i o c a r b o h y d r a z i d e with

2 7

10-undecenoic acid. It also showed two prominent f ragment ion peaks at

m/z 130 [(M+O-CgHn]^- and m/z 143 [M-CgHu]^ due to the (5- and y-

cleavage to the triazole ring on the long alkyl side chain.

All the signals in 'H-NMR and '^C-NMR spectra (acetone-c/g) (Fig. l a

and l b ) have been assigned to specific H- and C-atoms (Table 1) f rom their

typical chemical shift values, multiplicity and relative integrations. The

' H - N M R spectrum showed a weak broad singlet at 5 12.25 and a sharp

intense singlet at 5 5.18 which were attributed to the thiol and amino

protons. The two double doublets at 5 4.90 (J=10.26, 2.20, He) and 6 4.98

(J=17.21, 2.20, Hz) coupled with each other and with the ad jacent methine

proton were assigned to the two terminal o lefmic protons. HE -10 ' and H ^ - I O '

respectively. The signals appeared as triplet double doublet at 5 5.81 (J=6.78.

10.26, 17.03) were due to the coupling of the methine proton ( - C H = , H - 9 ' )

with the protons at H-8' and HE-10', HZ-10'. Also in '^C-NMR spectrum, the

characterist ic signals appeared at 5 153.34 and 5 168.26 were assigned to the

triazole ring carbons at C-3 and C-5. The other signals were at tr ibuted to the

9-decenyl side chain carbons.

On the basis of above facts, F T Z was characterized as 3-(9-decenyl)-4-

amino-5-mercapto-l,2,4-triazole (1).

FTZ (1)

2 8

J

3

< :

2 9

L •-'tfV »o •> I b W « M > « O C V N «

c f u f f i x i et < > W > > « « * MJ I O « O I tJ • >*

— < • • © «

C tf <D E i n c PC < « x — 4 o »- a . 4. h- z C itl ® £ s:«* oc 3 > — < — o - o e

4. z a u — • c o o « o > o

ao \ t oo 5 t

O o D w C 0

s o w e .

C2. C

E

J w — « i p I

« - «

e Q . Q .

• PJ fO « c

I I I M C O t o e i i «or> . I

a . •> » « •H o a

CM W «g •H .

UJ

W)

3 0

Table 1. 'H-NMR and ^^C-NMR spectra] data of FTZ (1),

H-nr 5(ppm) Integ-ration

Multi-plicity

J (Hz) C-nr S(ppm)

SH 12.25 IH brs 3 153.34

NH, 5.18 2H s 5 168.26

] ' 2.73 2H t ] ' 25.40

2' 1.75 2H br p J « 7.46 2' 26.68

3'-7' 1.33 5x2H br s 3'-7' 29.40-30.20

8' 2.03 2H m 8' 34.40

9' 5.81 IH tdd J9',8' 6.78, Jq' he 10.26. 9' 139.82

J9',Hz 17.03 114.61 HH-IO' 4.90 IH dd JHE,9' 10.26, JHL, Hz 2.20 10' 114.61

Hz-10' 4.98 IH dd JHZ„9' 17.21, JHE, Hz 2.20

l,5-Bis(4-chlorophenyl)-5-[3-(9-decenyl)-4'amino-5-thio-l,2,4-triazolyl]

pent-l-en-3-one FBCA (3) and 2-(4-chlorophenyl)-4-[2-(4-chlorophenyl)

ethenyl]-7-(9-decenyl)]-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4]thiadiazepine

FBCB (4). [Reaction of the ^-triazole (1) with l ,5-bis(4-chlorophenyl)

pent-l,4-dien-3-one (2)J,

A solution of the 5-triazole (1) and l ,5-bis(4-chIorophenyl)penl-1.4-

dien-3-one (2) (molar ratio, 1:1) in dry benzene was ref luxed with stirring for

32 h in presence of a catalytic amount of /7-TsOH. The products on usual

work up and column chromatography (silica gel, pet. e ther-EtOAc, 8:2 v/v)

yielded a colourless solid FBCA (3) (38%) and a yellow solid, FBCB (4) in

47.5% yield. The reaction sequence is as follows.

3 1

N N CI

FTZ (1) i) p-TsOH /dry benzene Reflux 32 h

N N

CI

Q

N ^ S '

NH2

N N N N

CI

FBCA (3)

Structure Eluc idat ion of F B C A (3)

It was a colour less glassy f lakes, m.p. 84 °C. The s t ruc ture of F B C A

was es tabl i shed by DCI-MS, ' H - N M R and '^C-NMR spectra . The DCI -MS

spec t rum (Fig. 2a) of FBCA showed a set of [M+I]"^ peaks at m/z

557/559/561 showing its molecular weight 556 /558 /560 [C29H34N40C1^S],

which is equal to the sum of the molecular weights of ^- t r iazole (1) (254) and

the d ienone (2) (302). This indicated that an addi t ion p roduc t has been

formed by the nucleophi l ic addi t ion of the thiol g roup to the enone func t ion

of the d ienone (2) and showing that the one ene g roup r ema ins intact . The

prominen t f r agment ion peaks were observed at m/z 255 [ (M+l ) -C i iH80S2]^

as the base peak and at m/z 303/305/307 [(M+1)-C,2H22N4S]^ fo rmed by the

3 2

I f t i l - wlOv-'lli ' U I HL i

t ' TDH '-i -TCMD = r - ' T i f " " r T r c

~fi 'kPi OLD »C<C> ' < Ci ' " L AI< " T;; ' I,,:, ' ~F-I I ~ C' I ON "CILFI "1 " "J'CI "''iCt ~ dCi 'X^Ci

llll I I I'" 1 1 1—'—I" 'I" "I'—I—1—r— 'fct '--Ti "-LI -'nc- •--C' ' T i '•.aft 'Aft ""'Pi ' " f i AC<fi <11 a AOO ii'-tci

• SI.' M A J .1 1 ^ .111 _ V «-R, 1 * 6 546 556 '5?^ 5£S . • '7- ' 553

1 62S

'-L' IT- .N 21 1: Y = N /IP S JIRI/GN =0X100 I NNV= 3325?S2 ~I~LE PPR A

1 'J 1 0 0 A 1 - ^ ^ FIN w - 00 5 306 an W W 4 N RF. W A < 02 00 'j^Ui-R c. 30 I 0 0 C- i 307 0 0 R 3

GO C C 10 302 0 0 1 5 55? 00 35 4 00 i. P 257. 01' 10 C FJ 3GZ OC 73 T n ft N N 1 ^ W 1/ I 00 ?

W 5 y £ U C J- ZO": N 0 1 / T V

t h V A =

1 ::5 ''' oc c r - tl 7 fi

V \ k

^C1 3Z5c. 5

''•fZS'' I • ?£3C"-

iP L

'^CC'L'ISI~ION PR.RAIIETER "' .E'LE

MASS "X'IMGES 7'.' 70C JislTFflPiiTTnw Tjwrc TYP- nr DiiM MdN 1-'-.

Fig. 2a

3 3

re t ro-Michael addit ion of F B C A , in addit ion to other peaks at m/z 165, 143

and m/z 130 as shown in Chart 1.

CH..

[M +1] m/z 557/559/561( 35.4 / 19.6/3.4)

retro-Michael addition

< m/z 255 (100.0) m/z 303/305/307 {12,3IAim3)

N N H

P-cieavage

N ^ S H I

NHj m/z 143(9.2)

N N

i [[ ^ N ^ S H

NH2 m/z 130(15.7)

CHART 1 m/z 165 /167 (28.8/9.2)

' H - N M R and '^C-NMR spectra (Fig. 2b and 2c) showed the s ignals as

assigned (Table 2). ' H - N M R spect rum showed a sharp singlet for amino

protons at 6 5.21. The signals cor responding to the methy lene p ro tons at C-4

appeared as two double doublets at 5 3.47 (J=17.25, 5.50, H-4up) and S 3.97

(J=17.25, 9.31, H-4dn). The signal for the meth ine proton at C-5 appeared as a

double doublet at 5 6.43 (J=9.31, 5.50, H-5) . The signals for the t rans-olef in ic

protons at 5 6.90 (J=16.33, H-2) and 5 7.67 (J=16.33, H-1) indica ted that the

Michael addit ion of thiol has occurred to only one ca rbon-ca rbon double bond

of the dienone (2). It was fur ther conf i rmed by the ' ^C-NMR spec t rum which

3 4

} :

X!

Dio

} :

J

3 5

Q . Q .

W o o i / i i - m o •• ee. a i«) E « « i > < « « ( r i u « > ^ »» U o fi o I H - O •

C O E < >t UJ ^

or - r c s.

•» c tf oi . . -n — ^ n O ' - O C i-H o 9 z le o . £ . • - >,-a w ccj o

o ©• X'W «<» SM-OiA « • ) 0 0 . 0 3 E V > « f « . o « 0 o o ** t

** ui^ C t e c A N o v £ 0 > ( L C — fSiCL — • O CL Z} •]

B T - c e u o > c - j < c «

o <N

• t V •« cn

) O) M « O •

36

fO «n in Tf- 1—H '—1 ro

in

o

o d cn 1 o on (n

>n o <w rf

u-) o< cn

vo

in oo a\ n

(n o<

m (N —< fn in cs 1

ro 00 on

+ 2 ^

o m m

c .

O in in

in (N ^ i> CO —'

a.

m o<

in (N

d

0 O

(N

m (tl 00 V) rn no no no m r—(

— <N fs « 1—> 1—> 1—5 •—5

w a:

On (N (N

u X

(N (N

On CNl (N

N 3: a n: I—!

on" O

in in CO 00

OS ro 00

in S £ ^

<

<

On ro 00

<

13 Td -O -a -o V -o

i u

•o c c5

z a

fs 0)

3 «

h

c « »-( iM

s A a to"

T) ! ^

in NO o m

r-on o On r-no

m vo o (n

o r-' »n (n o o o

00 o on

00 on (S in

no C<) o fs| on rt t^ rt K t^ r-

u C 1 «

in D.

rj- -o T— ^ fS rn 00

W Kl X X

z o on

cn t i r-f ro" I I

^ ^ ^ ^

3 7

exhibited peaks for the saturated carbon-carbon bond at 5 57.69 (C-5) and

5 45.05 (C-4) and the carbonyl carbon 5 196.04 (C-3). It also showed the

signals corresponding to the ^./^-unsaturated olefenic carbons at 5 127.64

(C-2) and 6 142.08 (C-1), in addition to the usual peaks of i '-triazole moiety

and 9-decenyl side chain carbons as shown in Table 2.

On the basis of the above facts, the structure of compound FBCA was

conf i rmed as 1,5-bis(4-chlorophenyl)-5-[3-(9-decenyl)-4-amino-5-thio-1,2.4-

triazolyljpent-l-en-3-one (3).

FBCA (3)

Structure Elucidation of FBCB (4)

It was a yellow globular shaped crystalline solid, m.p 161 °C. The

structure of FBCB has been established by DCI-MS, 'H-NMR and '^C-NMR

spectra. The DCI-MS spectrum of the sample FBCB (Fig. 3a) showed a set of

[M+l]"^ peaks as the base peak at m/z 539/541/543^ showing its molecular

weight 538/540/542 [C29H32N4SCI2] which is equal to the molecular weight

of (3) (556) minus one molecule of water. This indicated the formation of the

thiadiazepine ring by the intramolecular cyclocondensat ion between the NH2

and CO groups of (3). The formation of the molecular ion as the base peak

indicated the stability of the molecule. The peak at m/z 415/417/419

3 8

1 H M " . - 1 ' ' 4 1 1 - • ' 1 . - > « .M

I 1 f t l i OL. 1 f, S I U I H L :

-L4 CCRN 'TTi r""rT ' r r "" "=r-i

r

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—.—i ^ — I — — — r — -f- 'Ti -u' <r-,fi • < r. 1'f' •'-f' <ar-

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i Hi i I I v I III <11 !i;i

.iiiiiii •'fl J "J 0 dACj -1 " l" ^ " f-l J M L" SK" 52? 528 54?

-i_ .il '"CAN iC ST = ij u .irk;r.T)= 0X10 0 « w w 0^2560 - J ">•[_£ FI'CP

10? 0 0 0 - 30 C 00 - M 0 0 6 . 542 15 n f •7 7Cl 01 V 53t GO lO 2 543

l2? 0 0 Q W V k. n n 30 £ M n w w i nn * V • 0 544 240 0 P < -Kfl-W V w fl u V V w 540 Ci 45 1 2^:5 n n 3 304 0 0 Ic: 541 (1 fl w w 1

28 ? 1 6 . ?

Q

^T/iPT = TDTP.L

240 0 302 0 jj^c; n 505 i •-7Q ' fi

1 rwri = "c;

12i 21s

- g72

•^'•"•J AA

iPi.

^CC'UISniON PARftflElLR 1

M;iC5 Pii|WG£;5 ?C'-?OC' ' TL.GRftTIC.' TIrtES

TYPr nc" CllN Miliv Ti-'c; Fig. 3a

3 9

[(M+O-CgHij]"^ was arised by /7-cleavage of the long alkyl chain to the

t r iazole ring. The peaks at m/z 302/304/306 [(M+l)-C,2H2iN3S]^ and an

another peak at m/z 240 [ (M+O-CiyHj jNCla]^ arised by the c leavage of 1-7

and 3-4 bonds are the diagnost ic peaks for the t r iazolo- th iadiazepine ring

system as shown in Chart 2.

CH7

N N H

\

6 Q

N N

^ N f S s McLaffe r ty rear rangement 3 ^ 6

XI Q "a

m/z 539/541/543 (100.0/75.7/18.7)

m/z 415 /417 /419 (7.6/5 8)

N N

m/z 240(25.1) n ^ o /

P-cleavage

m/z 302 /304/306 (30.6/16.5/4 .2)

H

^ S H N N H

miz 128 (5 9) N ^ S H H

m/z 115 (6.3)

N N

X X H ^ N - ^ S H

H

m/z 102 (9.3)

C H A R T 2

'H -NMR and '^C-NMR signals (Fig. 3b and 3c) were assigned to individual

H-and C-atoms (Table 3) from their typical chemical shift values, multiplicity,

relative integrations and by the comparison with the spectra of (3) (Table 2). The

signal corresponding to NH2 protons at 5 5.12 of (3) has disappeared in the

'H-NMR spectrum of FBCB, showing that NH2 group was involved in

4 0

3

h n I ^

WD

3 3 j j :

4 1

/

ID

Q . Q .

E C

c> V I v>

C C - O f v c w >,

fA u o r ot ^ Oia> — o UJ ft) 4-»

>>-c — • -UJ o o « j f n -H T. — in o 4 » - - x t ;

or - c o 6 u <r V — p

UJ X «>

a . O — O t/ e V

c r> «

x; in « c «o -

V O O O UJ ^ 1- <

- ^ e ,

C CW > Q. _

c — 0 " 0 UJ o >

o o o

t: N «) -J

v> V — C _ I C c <5 O m u> rj CM r» in "C cn cn CM ^ - o in as CM in Ol CO CM tn CO rv- eo <0 O <st <o 00 ui op «r <o W eo e> n <M en m U) m CO in lO cn CO tn rv o in cn cn in 10 m eo CM w lO ^

CM CM n r«. M rg CM rj «D C0 <0 fv 10 in 10 lO cn B «0 tn TT 10 (O in ^ ee o CO

r "

tn in ^ <0 N. «> •-4 fM cn CM cn in ^ (0 «> CM CM in 03 at «e ^ o ^ „ CSJ tn 10 ^ o o cn o ^ us - V r. GO fn iO Op CM n n (0 10 m in cn «o cn w> <0 10 10 CM o to rv cn in in cn o tn <n e

C. te \n tn in tn n N e wa in IT O et in n (C (0 CM o o CI cn eo in in tn o o ^ o o o o e in *r n <0 cn CM cv CM cn P- tn <n CM <v CM r>. CM CM CM fv w **

> o o O e\ lO ^ Q s> Q « in o cn _ «6 CM <n <n «> *0 tn «e in lO ^

w « €9 CSJ (0 0t o fs. o in in CM •> in (O CM cn CNi o eo eo CO cn CM n cn CM ^ 10 CM o in o cn '-I oi o O if <0 <e e <0 «c rs. in •n to CM in *M e (O o V, in ir (n •o rv «0 eo M in tn tn o o so tM o UJ rw O 09 «B <S( *) in CO 0» o O o *r en CM CO 10 fs. in cn w> c CD in tn 0) eo in 10 10 CM in cn a: r- 40 <0 (O CO <0 in m o o o <0 in cn 10 c 9> CO rs. in cn CI eo *y U. O o O O n n « CO CO CM in CO <n tn cn tn cn CM <M CM CM CJ

(M N

X O in CD e cn 10 eo Wi o CM 10 CO <9 CM cn eo O) o Csl cn IT 40 2 in tM CM CM CM CM CM rM cn cn tn cn tn n cn tn tn •c ry rr

CO " T

I c

4 2

E a. o. lO

NO ON ( N 00 CM On r - NO »—1 NO

i r i CO ON v n NO d i n CO

NO CO CM l O l > m CM CM

o 'Cl-O m o <N ON (N

ON

o cn 1—I ON

CM oo On ON

XI- CO t—( CM CO

la C

c3 ON (Ti CN) rn 00

O OS - -

+

03 U CO b 0 C3 «5

•o

u O)

a. PS 1

u

•o c «

Z I

fo O)

CS H

N

>o ON 00 On

00 <M (N o. c D •o •o

1—1 CNT >—J fO ro irf r- 00 00 CO CO irf r- ^ rt

NO NO -o T3 D. ' — '

d 3 U a

3 cs —^ CM ro m <N CM 1—> 1—1 1—J I—J 1—1

On O

o\ I—> r i (N

On <N (N

O E

» io ON

(N <N O

ON w I

ON (N (N

N a:

ON o

ON l-l a:

. i 3 '.ii

T3 •T3 -a t3 -a T3 T3 T3 13 T3 13

a « rc ^ X X X y — (N (N K n: X CnI —< <N

E Cu a to'

r--o <N in <N (N (N CO 00

ON On (N O O 00 00

00 On rn ro r^ <N >r>

vo r--I <N

u a c. -a m (N (N

I rs) CO

o o ^ td PC "t

5o ON ffi nc <;

4 3

the formation of the ring. The absence of NH2 protons signal (§ 5.12) in

' H - N M R spectrum and CO signal (5 196.04) in '^C-NMR spectrum of F B C B

confirmed the formation of thiadiazepine ring. The format ion of the ring has

also been supported by the shifts of two diastereotopic hydrogens to higher

fields, f rom 5 3.47 (Hup-4) to 5 3.07 and from 5 3.97 (Hdn-4) to 6 3.52 and also

of the methine proton from 5 6.43 (H-5) to 5 5.22. The exocycl ic t rans-olefinic

protons signals appeared as doublets at 6 7.27 (J=16.48, H - l " ) and 5 7.66

(J=16.48, H-2"). In '^C-NMR spectrum, in place of CO signal at 6 196.04

(C-3), a signal appeared at higher field at 5 143.16 (C-4) in the formation of

ring. The other thiadiazepine carbons appeared at 5 53.59 (C-2) and 5 36.92

(C-3). The exocyclic olefinic carbon signals appeared at 6 126.76 (C-1") and

5 140.69 (C-2"). The terminal o lefmic carbons appeared at 5 139.81 (C-9')

and 5 114.59 (C-10').

Based on the above discussion, compound F B C B was characterised as

2-(4-chlorophenyl)-4-[2-(4-chlorophenyl)ethenyl]-7-(9-decenyl)]-2,3-dihydro

-1,2,4-triazolo[S,4-b][ 1,3,4]thiadiazepine (4).

4 4

l,3-Bis(4-cltlorophenyl)-3-[3-(9-decenyl)-4-amino-5-tltio-l,2,4-triazolylJpro-

pan-l-one FTZX4A (6) and 2,4-bis(4-chlorophenyl)-7-(9-decenyl)-2,3-

dihydro-l,2,4-triazolo[3,4-b]ll,3,4]thiadiazepine, FTZX4B (7) [Reaction of

the 5-triazole (1) with 4 ,4 ' -dichlorochalcone (5)]

A solut ion of the ^-tr iazole (1) and 4 ,4 ' -d ichIorocha lcone (5) (molar

ratio. 1:1) in dry benzene was re f luxed with st i rr ing for 26 h in presence of a

catalytic amount o f p - T s O H . . The products on ch romatography over silica gel

column using pet. e ther -EtOAc (8:2 v/v) as an eluent a f fo rded two

compounds , a colourless oily liquid FTZX4A (6) (37%) and a yel low

cry stall ine solid FTZX4B (7) (51.9%). The reaction sequence is as fol lows:

N N

FTZ (1)

N N

CI

Q

I NH2

o

(5)

(i) p-TsOH/dry benzene Reflux 26 h

N N

Q

FrZX4A (6) FTZX4B (7)

4 5

Structure Elucidat ion of FTZX4A ( 6)

It was a colour less oily liquid. The s tructure of FTZX4A has been

es tabl ished by FTIR, EI -MS and ' H - N M R spectra. FTIR spect rum gave

character is t ic bands at 3178.55, 3074.26 (NH2), 2927.0 (CH) , 1689.17 ( C = 0 ) ,

1639.66 ( C - N ) and 1590.04 (C=C) cm"'. In EI -MS spec t rum, the molecular

ion peak was absent , but two prominent peaks, one at m/z 254 [ (M+I) -

C i s H n O C b ] ^ ' and another at m/z 277/279/281 [(M+1)-C,2H,4N4S]^

cor respond ing with the tr iazole f ragment and 4 ,4 ' -d ich lo rocha lcone f ragment

ions, appeared possibly by the re t ro-Michael addi t ion of (6) The

f ragmenta t ion pat tern is shown in Chart 3.

CHo.

N N

NH. m/z 531/533 (absent)


N N

I

. NH, m/z 254 (89.0)

t N N

N" ^SH I

NH2

m/z 143 (28.3)

H

m/z 277/279/281 (16.0/9.2) P-cleavage m/z 255 (21.2)

N N Cl-

I SH

o -c=o

NH2

m/z 130 (67.8)

m/z 139/141 (100.0/37.8)

CHART 3

4 6

The 'H-NMR (acetone-cf^) signals were assigned to individual

hydrogen atoms (Table 4) on the basis of typical 5-values , multiplicity,

relative integrations and by comparison with the spectral data of (1) (Table 1)

and the previously established compound (3) (Table 2). The spectrum showed

no signal corresponding to thiol proton at 5 12.25 and in place additional

signals appeared due to the addition of the SH group to a,y5-unsaturated

funct ion of chalcone forming an adduct. A double doublet at 5 6.40 (J=9.08,

5.58) was assigned to H-3 and the two double doublets at 6 3.60 (J=17.10,

5.58) and 5 4.20 (J=17.10, 9.08) were attributed to the two diastereotopic

protons, Hup-2 and Hdn-2. The aromatic protons signals appeared as A2B2

pattern at 5 7.20 (d, J=8.55) and 6 7.42 (d, J=8.55) for one benzene ring and

at S 7.84 (d, J=8.42) and 6 7.70 (d, J=8.42) for another benzene ring. The

other signals were assigned to the 9-decenyl side chain protons.

Based on the above evidence, FTZX4A was character ised as 1,3-bis

(4-chlorophenyl)-3-[3-(9-decenyl)-4-amino-5-thio-l ,2,4-triazolylJpropan-l-

one (6).

' S ^ N

FTZX4A (6)

4 7

Table 4. 'H-NMR spectral data of FTZX4A (6).

H-ni 6(ppm) Integra-tion

Multi-plicity J (Hz)

N H , 5.14 2 H s

3 6.40 I H dd J3.2dn 9.08, J3.2UP 5.58

2up 3.60 I H dd J2up.2dn 17.10, J2UP.3 5.58

2dii 4.20 I H dd J2up,2dn 17.10, J2dn. 3 9.08

1" 2.68 2 H t J,..2" 7.46

1.64 2 H br p J « 7.33 1 1 H "711 J - / 1.23 5x2H br s

8" 1

1.98 2 H m

! 9" 1 5.70 i 1 I H tdd 1

J9..8.. 6.71. V „ r , 10.22. J9". Hz 17.08 |

1 Ht-10" 4.88 I H dd .IHC 9" 10.22. Jn, , , 1 .2 .28 ;

1H/-IO" 4.97 i ! i I H dd J„,,9" 17.08. 2.28 '

Ar-2.6 7.20 2 H d JAr-2.6. Ar-3,5 8 . 5 5

1 1 Ar-3.5 7.42 2 H d JAr-2.6, Ar-3.5 8 . 5 5

Ar'-2.6 7.84 2 H d JAr'-2.6,Ar'-3.5 8 . 4 2

Ar'-3,5 7.70 2 H d JAr'-2.6, Ar'-3.5 8 . 4 2

Structure Elucidation of FTZX^B (7)

It was a pale yellow globule shaped crystalline solid, m.p 148 °C. The

structure of FTZX4B has been fully established by DCI-MS. 'H-NMR and

'^C-NMR spectra. The DCI-MS spectrum displayed a set of peaks

corresponding to [M+l]"^ as the base peak at m/z 513/515/517 showing its

molecular weight to be 512/514/516 [C27H30N4SCI2] which is equal to the

molecular weight of (6) (530) minus one molecule of water. This

4 8

indicated that tlie format ion of thiadiazepine ring has occurred by the

in t ramolecular cyclocondensat ion of (6). Moreover , the appearance of the

molecular ion peak as the base peak highl ighted the stabil i ty of the molecule .

A set of peaks at m/z 276/278/280 and an another peak at m/z 240 arised by

the c leavage of 3-4 and 1-7 bonds are the diagnost ic peaks for the tr iazolo-

th iadiazepine r ing system. The other f ragment ion peaks at m/z 102, 115, and

m/z 128 were obta ined f rom the ion m/z 240 by the c leavage on the long alkyl

side chain at a , P and y-posit ions to the tr iazole ring as shown in chart 4

N N a-cledvaae

N ^ S H H

m/7 102 (9 0)

H

N N

H

m/z 1 1 5 ( 7 9)

C H ,

N N

"CI

[M+H] m/z 513/515 /517 (100 0/72 1/18 3)

1 -7 and 3-4 -bond clca\ due

N- "SH H

m/z 240 (28 6)

y-cleavage

N N

H

m/z 128 (5.1)

CHART 4

m/z 276/278/280 (54 3/27 8/6 8)

Cl- - C = N O m/z 138/140 (5.9)

The ' H - N M R and '^C-NMR signals (Fig.4a and 4b) we re assigned to

individual H- and C-a toms (Table 5) on the basis of chemica l shif t values,

mult ipl ic i ty , re la t ive integrat ions and by compar ing the spectra l data with that

of (6) (Table 4). The absence of N H j pro tons signal (at 5 5 .14) in the

4 9

CL Q .

a

laD

} ^ J M

5 0

« M

w O E * « » > , « > 9 L> O « U

' C

3 Q. <

..com Vi i - w i X 0 0 « » C C 01 . QC oi«> — <n O'-Q c UJ m^a 3 t ^ - - o • - - CO ( u j 0 « 0 €>kn z r ^ o M 6 & ) x i t p « ^ c • «o au 3 iw eie> c a:<oE</)« f>.o«Oo • CL U J ^ C l a C A N «

C £ O > & . i - ^ M O , . o k 0 CM v^JK « «• « ^ e r - o o u J O ^ c ^ ^ C v U Z 3 U —NCOU O 4 <J D.O. O <-« SJIUO. U X .U. k > a o o Q

U J l f ) CSJ ^ «7

v> V <») rs.

rs. ^ in *> «s (M

rf

5 1

pa tT X

H ta 0 05 B

•V "a •4-1 a a O.

PJ S z 1 u m •a c a

Z I

X

\r) o

es H

E a a.

I. e I U

N

-M =

00

d

ON

00 o (N (N d.

On rW

13

ON O MD >n

q iri (N 00 m

cn

r-

rs

ON

r-

00 o (N o

CN i> ro

(N —

«

o Tt d

O-l (N

CO CO CO 1 ON •T) NO o (N 00 NO CO

n <N a< o< ON ro 1—' rN <N (N (N CO

(N I

CO CO o\ — +

<N T—H

N S OS

(N d

w E

00 r--

oc ON

o <N (N N X

tS X

<N

O Osl <N N X

C<l r-'

On ON

00 oo : M3 vo 00 00

i i o

oi <N o,' r^

< < <

-o T3 X) TZi 'X2 T3 T3 T3 -o -o -o

1 e J s n

^ u

s a a to rn

k. c a 3 m

0-) CN

X (N

ON 00 CN

X (N

00

(N X

o\ (N

X (N

O o — (N

X

OS r- ON 00

w X

c 13 en CNl ^ (N I

rn 00

ON

Kl X

I

d ON ^

>!Ih rC K ^ (N CS (N <N

ON '—1 NO — NO Tt IT) 00 t^ K

^^ NO in fo" (N I 1

5 2

' H - N M R spectrum of FTZX4B and shifts of 5 values of the two

diastereotopic methylene hydrogens to higher fields from 5 3.60 (Hup-2) and

S 4.20 (Hdn-2) to 5 3.27 and 5 3.66 respectively and also of the methine

proton from 6 6.40 (H-3) to 5 5.27 indicated the format ion of 1,3,4-

thiadiazepine ring. This was also supported by '^C-NMR spectrum. The signal

corresponding to CO group did not appear and in place other signals

corresponding to thiadiazepine ring carbons appeared which were assigned as

5 143.05 (C-4), 38.42 (C-3), 53.72 (C-2) and 170.18 (C-9a). In addition, the

signals corresponding to the 9-decenyl side chain and the 1,2,4-triazole ring

carbons appeared as shown in T a b l e 5.

Based on the above spectral evidence, compound FTZX4B was

characterised as 2,4-bis(4-chlorophenyl)-7-(9-decenyl)--2,3-dihydro-l,2,4-

triazolo[3,4-b][1,3,4]thiadiazepine (7).

FTZX4B (7)

5 3

l,3-Bis(2-thienyl)-3-[3-(9-decenyl)-4-amino-5-thio-l,2,4-triazolyl]propan-l-

one F T C A (9) [Reaction of the s - tr iazoie (1) wi th l ,3 -b i s (2- th ienyl )

p r o p e n - l - o n e (8)]*

*[This react ion fai led to yield the desired produc t 2,4-bis(2-thienyl)-7-(9-

decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-bJ[l,3,4Jthiadiazepine].

A solut ion of 5-tr iazole (1) and l , 3 -b i s (2 - t h i eny l )p ropen - l -one (8)

(molar ratio, 1:1) in dry benzene was re f luxed wi th s t i r r ing for 30 h in

presence of catalyt ic amount of /7-TsOH. TLC examina t ion of the reaction

mixture revealed only a single spot. The react ion was con t inued for further

10 h, but no other spot developed. The products on pur i f ica t ion by column

chromatography a f fo rded F T C A (9) as the sole produc t as a pale yel low oily

l iquid in 68 .6% yield.

N N

11 I I

NH2

(1)

"SH I I w

o

N N

(8)

i) /7-TSOH /dry benzene Reflux 30 h

F T C A (9)

5 4

Structure Elucidation of F T C A (9)

It was a pale yel low oily liquid. The structure of F T C A has been ful ly

es tabl ished by DCI-MS, ' H - N M R and '^C-NMR spectra . The DCI-MS

spectrum showed [M+l]"^ peak at m/z 475 showing its molecu la r weight to be

474 [C23H30N4OS3] which is equal to the sum of the molecu la r weights of (1)

(254) and l ,3 -b is (2- th ienyI )propen- I -one (8) (220). This sugges ted that an

adduct of (8) and (1) has been fo rmed by the nuc leophi l ic addi t ion (1,4) of

SH group to or.y^-unsaturated carbonyl funct ion. The m o d e of f ragmenta t ion

of the adduct FTCA is shown in Chart 5.

CH2v

N N

i 1 I

NH2

[M+1] m/z 475 (12.5)

/ O

H

r N N

A

# N N

NH2

m/z 255 (67.1)

P-cleavage H

N N W ^SH I

NH2

m/z 143 (3.7)

^SH NH2

m/z 130 (5.7)


// // w

o m/z 221 (100.0)

// 3

O

m/z 111 (32.0)

CHARTS

S 5

The signals in 'H-NMR and '^C-NMR spectra (Fig. 5a and 5b) have

been assigned to specific H- and C-atoms (Table. 6) f rom the chemical shift

values, multiplicity, relative integrations and by comparing with the spectral

data of a previously established structure of compound (6) (Table 4). The

absence of the thiol proton signal at 6 12.25 and <2,y9-unsaturated protons

signals at 6 7.48 and 5 7.93 of the substrate (8) (experimental) and appearance

of the diastereotopic proton signals at 5 3.82 (Hup-2) and 5 4.25 (Hdn-2) is the

clear evidence for the formation of the adduct. The signal of the methine

proton (H-3) appeared as a double doublet at 5 6.79 (J=9.01, 5.80) due to the

coupling with the two diastereotopic protons (Hup-2) and (Hd„-2). The structure

was further confirmed by '^C-NMR spectrum. The disappearance of the signals

corresponding to the a , (3-unsaturated carbons, C-2 (5 121.36) and C-3

(5 140.95) of (8) (see experimental) and the appearance of the signals of the

same positions at higher fields, 5 44.25 (C-3) and 5 54.03 (C-2) indicated the

formation of adduct. The carbon signals have been assigned as 6 189.06

(C-1). 5 152.51 (C-3') and 5 167.48 (C-5'). The signals of 9-decenyl side chain

and the aromatic ring carbons appeared as usual.

Based on the above spectral facts, the structure of the compound FTCA

was established as \,3-bis(2-thienyl)-3-[3-(9-decenyl)-4-amino-5-thio-],2,4-

triozolylJ propan- J -one (9).

^ - - - - «

>i s o s ^ i f

5 6

to • 1- V N z *» n o *>—• lA • ' v ttj wfs . c e

o o « « o o i H ' o o c o) . — ^ w i o « j i n

o- c tt^ae^Eo*' (J a> n wo^ •• (n — tov — t/) O V- O. • •» Ck.UJ N «> C t- X C lU yc 9

c r u i • « r « * • w w c

s a . 4 0 « « 0 . o £ & . 0 4 a c D « u . o " - O O h-00(0 l-> -o

O O (0 ^ t > ® ®

. €0 ® rv MS «) .

^ ^ CO r o o o o «> VI a .

CSi « «> ® «) M •

91 <0 « in u) I M ^ ^ o o ® tt ® (A in in to

• e T - Q . f . a J o

e •

. W)

V o wt e

5 7

e a. o.

« c « : u j . e.— J £ I . i •) e

o M « v> e V o « M «• . • « e e « N C e « > ««

C « oi «. w a : ^ o«»» — - ^ c i o - - O c £ UJ • ^ © • i V CL H . . • > . ' e M C < n O t n w « O M « v> -< r ^ i r t e w C M T i m • < « < » o a o 3 i t jcw^v

« T O « e U O ^^ E « V —Wt — O. - U J ^ C I O ^ D N * * ' UJ X V —NA. ^ i. x: « M « « » « • -

w O ) C - i s c Z « 3 U — V > « 0 U O

o o o o t -

3 l i

Xi in

t - fe

X M <M M

a . <o le o c _ _ -

>

5 8

a o. w to'

o d

00

NO 00 1 ON o CO >n o <r> OO o OO •vO m o (N) ON (N r-" NO fN 00 NO Tf" 00 >n NO >o vd ON Tf cn »—( (N

<N (N cn >—1

vo

c o u

0\ ^

U H b o «

T3

u a O) a,

0! S ^ u

•a c «

a

vo i i C3 H

I. c

m (N

N M

3 .a

Sf.2 C

s a o.

r^ In — (N r^ 00 ON +

2 ^

o 00 >ri

o CO >n

a

o On

Q. 3

Tt Tj-ON ON

O On'

c •T3 <N

C

ci 3 (S

r--Tt rs|

«

On P

N

CN

E

(N r-MD

00 r Ov

ON rs) <N N X

w K

<N (N

ON H

X w X

CN

w X

(N (N (N CN CN CN 00 (N (N rW —' rn —'

(TT o en" O r.

00 oo" 00 CTT 00 iri ))

lO II m II 11 11 II II )

II )

II II 1—) II 1—)

-o T3 -d T3 13 l-H Id 13 T3 -O 13 13 -a TS 13 13 "O 13

r-H ON cN r- (N

00 rW Tt (N —;

00 00 r-} o (N to

1—I I—i

00 ON ON rt-' r t

vo M CO Tf o m ON (N O CN ON r- oo r- K

s

cn o . 3 (N -a CN ^ CN 0-) 00 ON

U X

K1 in f? I

rt I >o I ^ ^ ^ ^ ^ ^

5 9

EXPERIMENTAL

Reagent and solvents were of commercial grade and were used without

further purification, 10-undecenoic acid was also of commercial grade (Fluka,

AG, Switzerland). All melting points were determined on a Koff le r hot-plate

apparatus and are uncorrected. IR spectra were determined on a Perkin-Elmer

621 spectrophotometer in KBr-pellets. FTIR were recorded on Perkin-Elmer

spectrophotometer RXj and wave numbers (v^ax ) were recorded in cm"'. 'H-

NMR and '^C-NMR spectra were recorded in Bruker Ac 300, Brucker DRX

300 and Varian Unity 400 in acetone-^/^ and DMSO-t/^ with TMS as the

internal standard. The abbreviations s, d, t, m, dd. ddd, br p, dt denote singlet,

doublet, triplet, multiplet, double doublet, triple doublet, broad pentet and

double triplet. ' X - N M R were recorded on a Varian unity 75 and 100

spectrometers. ElMS were recorded on V.G. Micromas model ZAB-IF

apparatus at 70 ev ionization voltage. DCI-MS spectra were recorded with a

Ribermag RIO-IOB quadrupole mass spectrometer, using ammonia as reagent

gas. Thin layer chromatography (TLC) was carried out on 0.25 mm layer

plates with silica gel G (Merck). Spots were visualised on exposure to iodine

vapour using iodine chamber. The column chromatography was performed on

silica gel [Merck. (60-120) mesh] using 25-30 g. per g of material to be

separated and purified. Dry solvents were prepared according to standard

procedures. The organic solvents used were of LR grade and were distilled

before use. The abbreviation used pet. ether for petroleum ether and EtOAc

for ethyl acetate,

6 0

4-Amino -3-(9-decenyl)-5-mercapto-l,2,4-triazole FTZ (1)

A mixture of 10-undecenoic acid (6.0 g, 0.0325 mol) and thiocarbo-

hydrazide (3.456 g, 0.0325 mol) taken in a 100 ml round bot tom flask was

fused by heating at 170 °C on an oil bath for 1V2 h, moni tor ing the progress

of the reaction (TLC, silica gel G, pet .ether-EtOAc, 8:2 v/v) which revealed

the formation of a single spot corresponding to the product . The reaction

mixture was then extracted with ethyl acetate, washed successively with

water (3x50 ml), 5% NaHCOs solution (2x50 ml), again with water (3x50

ml) and finally dried over anhydrous Na2S04. The solvent was removed under

reduced pressure and the solid residue thus obtained was crystall ised from

EtOAc-benzene as white crystalline needles, 7.078 g (85.6%), m.p. 82 °C,

R, 0.63 (pet. ether-EtOAc, 8:2 v/v)

Spectral data of FTZ (1)

IR (KBr): v 3250, 3100 (NHj) , 2900 (CH), 2350 (SH), 1630 (C=N), 1610

(C=C), 1550, 1470, 1410, 1370, 1280, 1230, 940, 890, 720 cm"'.

'H-NMR (400 MHz, ace tone-^) : 5h 12.25 ( I H , SH), 5.18 (2H, s, NH2), 2.73

( 2 H , t , J = 7 . 5 1 , H - 1 ' ) , 1 . 7 5 ( 2 H , b r p , J ~ 7 . 4 6 , H - 2 ' ) , 1 . 3 3 (LOH, b r s, H - 3 ' - 7 ' ) ,

2 . 0 3 ( 2 H , m , H - 8 ' ) , 5 . 8 1 ( I H , t d d , J = 6 . 7 8 , 1 0 . 2 6 , 1 7 . 0 3 H - 9 ' ) , 4 . 9 0 ( I H , d d ,

J = 1 0 . 2 6 , 2 . 2 0 , H E - 1 0 ' ) , 4 . 9 8 ( I H , d d , J = 1 7 . 2 1 , 2 . 2 0 , H z - 1 0 ' ) .

'^C-NMR (100 MHz, acetone-^/g): 5c 153.34 (C-3), 5 168.26 (C-5), 25.40

(C-1'), 26.68 (C-2'), 29.40-30.20 (C-3'-7'), 34.40 (C-8') , 139.82 (C-9'), 114.61

(C-10').

6 1

DCI-MS (NH3);; m/z (%) 254 ( M \ 7.1), 255 [ ( M + l ^ , 100.0], 256 (25.3), 257

(9.6), 143 (9.2), 144 (3.6), 130 (17.3), 131 (3.1).

l,5-Bis(4-chlorophenyl)pent-l,4-dien-3-one (2)

It was prepared following a literature procedure^^'^ by condensing

acetone with p-chlorobenzaldehyde (molar ratio, 1:2) in the presence of 2.5

equivalents of sodium hydroxide. To a solution of /?-chlorobenzaldehyde

(604 mg, 4.310 mmol) dissolved in ethanol (20 ml) was added acetone (250

mg, 2.155 mmol) dropwise keeping the solution in an ice bath. To this was

added sodium hydroxide solution (431 mg, 5.38 mmol) drop wise with

stirring for 2 h. until a yel low precipitate was formed. It was then worked up.

The ye l low solid thus obtained was recrystallised f rom benzene to give (2)

as bright yel low crystalline flakes, 710 mg (72.0%), m.p 176 °C, Rf 0.72

(pet. e ther-EtOAc, 8:2 v/v).

Spectra l data of (2)

' H - N M R (400 MHz, acetone-rfg): 5h 7.28 (2x lH , d, J=16.0, H-2/H-4), 7.70

( 2 x l H , d, J=16.0, H- l /H-5) , 7.73 (4H, d, J=8.50, 2xAr-2,6) , 7.45 (4H, d,

J=8.50, 2xAr-3,5) .

4,4'-Dicholrochalcone (5)

It was prepared following a published but sl ightly modified

procedure^^® by condensing j?-chloroacetophenone with p-chlorobenzyl idene

diacetate (both prepared according to reported procedure)^^^ in equimolar

ratio in the presence of 2.5 equivalents of sodium hydroxide . A mixture of

p-chiorobenzyl idene diacetate (6.5 g, 0.0268 mol) dissolved in (21 ml)

ethanol and p-chloroacetophenone (4.13 g, 0.0268 mol) was kept on an ice

bath in a 500 ml flask. To this, sodium hydroxide (2.65g, 0.0671 mol) was

6 2

added drop wise with stirring. After complete addition, the reaction mixture

was fur ther stirred for 3 h, diluted with 300 ml of ice cold water and was then

kept in the refr igerator overnight. The precipitate formed was filtered and

washed successively with water to make it neutral. It was then washed with

10 ml of cold alcohol. The yellow solid thus obtained was recrystal l ised from

acetone-benzene to give (9) as light yellow crystalline needles, 6.29 g (85%),

m.p. 156 °C, Rf 0.72 (pet. ether-benzene, 1:1 v/v)

l,3-Bis(2-thienyl)propen-l-one (8)

It was prepared following a literature procedure^^'' by condensing

thiophen-2-aldehyde (prepared from thiophene according to reported

procedure^^'') with 2-acetylthiophene in equimolar ratio, in the presence of 2.5

equivalents of sodium hydroxide. A mixture of th iophen-2-a ldehyde (500 mg,

3.96 mmol) dissolved in (20 ml) ethanol and 2-acetyl thiophene (554 mg, 3.96

mmol) was kept on an ice bath in a 500 ml flask. To this sodium hydroxide

(396 mg, 9.9 mmol) was added dropwise with stirring. It was stirred further

4 h and then worked up as above. The yellow solid thus obtained was

recrystal l ised (acetone-benzene) to give (8) as light yel low crystal l ine needles,

765 mg (78%), m.p 89 °C, Rf 0.63 (pet .ether-EtOAc, 8:2 v/v).

Spectral data of (8)

' H - N M R ( 400 MHz, acetone-dfi): 5h 7.48 ( IH , d, J=15.41, H-2) , 7.93 ( IH,

d, J=15.41, H-3), 7.67 ( IH , d, J=5.04, Ar-3), 7.60 ( I H , d, J=4.73, Ar'-3), 7.27

( IH , d, J=3.51, Ar-4), 7.17 ( IH, d, J=3.67, Ar'-4), 8.11 ( I H , d, J=3.82, Ar-5) ,

7.91 ( IH , d, J=3.67, Ar'-5).

6 3

' ^C -NMR (100 MHz, acetone-dtf): 5c 181.80 (C-1), 121.36 (C-2), 140.95

(C-3). 146.68 (Ar-C-2), 136.58 (Ar-C-5), 133.06 (Ar'-C-5), 129.34-134.25

(Ar-C-3, Ar '-C-3, Ar-C-4, Ar'-C-4).

DCI-MS (NH3): m/z (%) 221 (100.0), 222 (20.4), 220 (14.0), 191 (3.7). 137

(3.8). I l l (7.2).

l,5-Bis(4-cltlorophenyl)-5-[3-(9-decenyl)-4-amino-5-thio-l,2,4-triazolyl]

pent-l-en-3-one FBCA (3) and 2-(4-chlorophenyl)-4-[2-(4-chlorophenyl)

ethenyl]-7-(9-decenyl)]-2,3-dihydro-l,2,4-tnazolo[3,4-b][l,3,4]thiadiazepine

FBCB (4). [Reaction of the 5-triazole (1) with l ,5-bis(4-

chiorophenyl)pent- l ,4-dien-3-one (2)]

A mixture of the ^-triazole (1) (50 mg, 1.96 mmol) and 1.5-bis(4-

chlorophenyl)pent- l ,4-dien-3-one (2) (590 mg, 1.96 mmol) in dry benzene

(20 ml) was refluxed for 33 h in presence of yC-TsOH (5 mg). The TLC

examination of the reaction mixture revealed the presence of two spots of

equal intensity. It was worked up and subjected to column chromatography

(silica gel, pet. ether-EtOAc, 8:2 v/v) which afforded FBCA (3) as glassy

flakes. 416 mg (38.2%), m.p 84 °C, Rf 0.71, pet. e ther-EtOAc (8:2 v/v).

Further elution of the column using pet. ether-EtOAc (6:4 v/v) followed b>

crystallisation (benzene-acetone) afforded FBCB (4) as yel low crystalline

globules, 460 mg (47.5%), m.p 161 °C, Rf0 .30 (pet. e ther-EtOAc, 8:2 v/v).

Spectral data of FBCA (3)

'H-NMR (400 MHz, acetone-^/g): 5h 5.21 (2H, s, NHj), 6.43 ( IH , dd, J=9.31.

5.50, H-5), 3.47 ( IH , dd, J=I7 .25 , 5.50, H-4up), 3.97 ( I H , dd, J=I7 .25 , 9.31,

H-4dn), 6.90 ( IH , d, J=I6 .33 , H-2), 7.67 ( IH, d, J= 16.33, H - I ) , 2.73 (2H, t,

J=7.48, H- l " ) , 1.70 (2H, br p, J~7.36, H-2"), 1.25 (lOH, br s, H-3"-7") , 2.00

64

( 2 H , m , H - 8 " ) , 5 . 8 0 ( I H , t d d , J = 6 . 7 1 , 1 0 . 2 2 , 1 7 . 0 9 , H - 9 " ) 4 . 9 0 ( I H , d d ,

J = 1 0 . 2 2 , 2 . 2 9 , H E - 1 0 " ) 4 . 9 8 ( I H , d d , J = 1 7 . 0 9 , 2 . 2 9 , H Z - 1 0 " ) , 7 . 3 6 ( I H , d,

J = 8 . 5 4 , H - A r - 2 , 6 ) , 7 . 4 6 ( I H , d , J = 8 . 5 4 , H - A r - 3 , 5 ) , 7 . 7 2 ( I H , d , J = 8 . 3 9 , H -

A r ' - 2 , 6 ) , 7 . 4 9 ( 2 H , d , J = 8 . 3 9 , H - A r ' - 3 , 5 ) .

'^C-NMR (100 MHz, acetone-rf^): 5c 57.69 (C-5), 45.05 (C-4), 196.04 (C-3),

127.64 (C-2), 142.08 (C-1), 152.46 (C-3'), 167.62 (C-5'), 25.21 (C-1"), 26.70

(C-2"), 29.20-30.40 (C-3"-7"), 45.05 (C-8"), 139.35 (C-9"), 114.64 (C-10"),

139.85 (Ar-C-1), 129.29 (Ar-C-2,6), 129.99 (Ar-C-3,5), 134.59 (Ar-C-4),

134.04 (Ar ' -C- l ) , 130.81 (Ar'-C-2,6), 130.09 (Ar'-C-3,5), 136.63 (Ar'-C-4)

DCI-MS (NHs): m/z (%): 557[(M+1)^ 35.4], 558 (10.3), 559 (19.6), 560

(8.7), 561 (7.8), 301 (5.1), 302 (15.5), 303 (73.3), 304 (22.9), 305 (47.0), 306

(10.4), 307 (8.3), 267 (5.5), 254 (20.8), 255 (100.0), 256 (17.6), 257 (6.6),

204 (4.3), 165 (28.8), 167 (9.2), 143 (9.2), 137 (4.8), 130 (15.7), 101 (4.1),

102 (11.0).

Spectral data of F B C B (4)

'H-NMR (400 MHz, acetone-rfg): 5h 3.07 ( I H , dd, J=13.73, 11.75, H-3„p),

3.52 ( I H , dd, J=13.73, 4.89, H-3dn), 5.22 ( IH, dd, J=11.75, 4.89, H-2), 7.27

( IH , d, J=16.48, H-1"), 7.66 ( IH , d, J=16.48, H-2"), 2.83 (2H, t, J=7.47, H-

1'), 1.79 (2H, br p, J~7.44, H-2') , 1.29 (lOH, br s, H-3'-7 ') , 2.00 (2H, m, H-

8'), 5.78 ( I H , tdd, J= 6.7,10.22, 17.09, H-9') , 4.88 ( IH , dd, J=10.22, 2.29, HE-

10'), 4.96 ( I H , dd, J=17.09, 2.29, Hz-10'), 7.42 (2H, d, J=8.54, H-Ar-2,6) ,

7.49 (2H, d, J=8.54, H-Ar-3,5), 7.76 (2H, d, J=8.39, H-Ar '-2,6) , 7.57 (2H, d,

J=8.39, H-Ar'-3,5).

^^C-NMR (100 MHz, acetone-dg): 8c 171.27 (C-9a), 155.84 (C-7), 143.16

(C-4), 36.92 (C-3), 53.59 (C-2), 126.76 (C-1"), 140.69 (C-2"), 25.51 (C-T),

6 5

27.36 (C-2'), 29.20-30.40 (C-3'-7'), 34.42 (C-8'), 139.81 (C-9'), 114.59 (C-

10'), 142.37 (Ar-C-1), 129.47 (Ar-C-2,6), 129.61 (Ar-C-3,5) ,135.23 (Ar-C-4),

134.32 (Ar ' -C- l ) , 130.39 (Ar'-C-2,6), 129.95 (Ar ' -C-3,5) , 136.12 (Ar'-C-4).

DCI-MS (NH3): m/z (%): 538 ( M \ 10.2), 539 [ (M+1)^ 100.0], 540 (45.1),

541 (75.7), 542 (28.9), 543 (18.7), 544 (5.9), 415 (7.6), 416 (5.9), 417 (5.8),

300 (16.5), 301 (5.9), 302 (30.6), 303 (8.5), 304 (16.5), 255 (6.8), 240 (25.1),

128 (5.9), 1 15 (6.3), 102 (9.3).

l,3-Bis(4-chlorophenyl)-3-[3-(9-decenyl)-4-amino-5-thio-l,2,4-tnazolyl]pro -

pan-l-one FTZX4A (6) and 2,4-bis(4-chlorophenyl)-7-(9-decenyl)-2,3-

dihydro-l,2,4-triazolo[3,4-b][l,3,4]thiadiazepine, FTZX4B (7) [Reaction of

the 5-triazole (1) with 4,4'-dichlorochaIcone (5)]

To a solution of ^-triazole (1) (500 mg, 1.96 mmol ) in dry benzene (20

ml) was added 4,4'-diclilorochalcone (5) (540 mg, 1.96 mmol) and refluxed

for 28 h in presence of /?-TsOH (5 mg). The progress of the reaction was

monitored by TLC (silica gel G, pet .ether-EtOAc, 8:2 v/v) which showed the

presence of two new spots, labelled as FTZX4A (6) and FTZX4B (7). The

solvent was removed under reduced pressure and the residue left was

dissolved in ethyl acetate, washed with water and the organic phase was dried

over anhydrous Na2S04 The products on column chromatography over silica

gel using pet. ether-EtOAc, (8:2 v/v) as eluent y ie lded FTZX4A (6) as

colourless oily liquid, 384 mg (37.0%), Rf 0.63 (pet. e ther-EtOAc, 8:2 v/v).

Further elution of the column using pet. e ther-EtOAc (4:6 v/v) as eluent

yielded FTZX4B (7) as a yellow solid which was recrystal l ised f rom benzene

as pale yellow crystalline globules, 540 mg (51.9%), m.p 148 °C, Rf 0.32

(pet. e the r -E tOAc, 8:2 v/v)

6 6

S p e c t r a l da ta of FTZX^A (6)

FTIR (Neat): v ^^^ 3178.55, 3074.26 (NH2), 2927.0 (CH) , 1689.17 (C=0),

1639.66 (C=N) , 1590.04 (C=C) , 1571.40, 1491.79, 1426.78, 1402.11,

1360.60, 1308.68, 1260.38, 1205.25, 1174.16, 1091.68, 1014.26, 991.38,

961.79, 910.10, 827.98, 783.54, 723.97, 629.04, 531.33, 462 .77 cm"'

^H-NMR ( 300 MHz, acetone-flfg): 5h 5.14 (2H, s, NH2), 6 .40 ( I H , dd,

J = 9 . 0 8 , 5 . 5 8 , H - 3 ) , 3 . 6 0 ( I H , d d , J - 1 7 . 1 0 , 5 . 5 8 , H-2„P) , 4 . 2 0 ( I H , d d , J = 1 7 . 1 0 ,

9 . 0 8 , H-2DN), 2 . 6 8 ( 2 H , t , J = 7 . 4 6 , H - 1 " ) , 1 . 6 4 ( 2 H , b r p , J ~ 7 . 3 3 , H - 2 " ) , 1 .23

(LOH, b r s , H - 3 " - 7 " ) , 1 . 9 8 ( 2 H , m , H - 8 " ) , 5 . 7 0 ( I H , t d d , J = 6 . 2 8 , 1 0 . 2 2 , 1 7 . 0 8 ,

H - 9 " ) , 4 . 8 8 ( I H , d d , J = 1 0 . 2 2 , 2 . 2 8 , H E - 1 0 " ) , 4 . 9 7 ( I H , d d , J = 1 7 . 0 8 , 2 . 2 8 , HZ-

1 0 " ) , 7 . 2 0 ( 2 H , d d , J = 8 . 5 5 , H - A r - 2 , 6 ) , 7 . 4 2 ( 2 H , d d , J = 8 . 5 5 , H - A r - 3 , 5 ) , 7 . 8 4

( 2 H , d d , J = 8 . 4 2 , H - A r - 2 , 6 ) , 7 . 7 0 ( 2 H , d d , J = 8 . 4 2 , H - A r ' - 3 , 5 ) .

E I - M S : m / z ( % ) 2 7 7 ( 1 6 . 0 ) , 2 7 8 ( 4 . 2 ) , 2 7 9 ( 9 . 2 ) , 2 8 0 ( 3 . 1 ) , 2 7 6 ( 1 3 . 3 ) , 2 7 5

( 1 0 . 0 ) , 2 5 5 ( 2 1 . 2 ) , 2 5 6 ( 7 . 0 ) , 2 5 7 ( 3 . 1 ) , 2 4 2 ( 2 6 . 0 ) , 2 4 3 ( 8 . 9 ) , 2 2 1 ( 8 . 0 ) , 2 0 0

( 7 . 2 ) , 1 9 6 ( 1 4 . 0 ) , 1 5 7 ( 1 0 . 0 ) , 1 4 3 ( 2 8 . 3 ) , 130 ( 6 7 . 8 ) , 1 2 8 ( 2 4 . 0 ) , 130 ( 6 7 . 8 ) ,

128 ( 2 4 . 0 ) , 1 0 3 ( 4 . 8 ) , 8 3 ( 9 . 0 ) .

S p e c t r a l da ta of FTZX4B (7)

' H - N M R ( 4 0 0 M H z , acetone-^/<5): 5 h 3 . 2 7 ( I H , d d , J = 1 3 . 9 1 , 12.08, H-3,p),

3 . 6 6 ( I H , d d , J = 1 3 . 9 1 , 4 . 7 6 , H-3DN), 5 . 2 7 ( I H , d d , J = 1 2 . 0 8 , 4 . 7 6 , H - 2 ) , 2 . 8 9

( 2 H , t , J = 7 . 5 1 , H - 1 ' ) , 1 . 8 2 ( 2 H , b r p , J ~ 7 . 4 6 , H - 2 ' ) , 1 . 2 9 ( L O H , b r s , H - 3 ' - 7 ' ) ,

2 . 0 0 ( 2 H , m , H - 8 ' ) , 5 . 7 9 ( I H , t d d , J = 6 . 7 8 , 1 0 . 2 9 , 1 7 . 2 1 , H - 9 ' ) , 4 . 8 9 ( I H , d d ,

J = 1 0 . 2 6 , 2 . 2 0 , H E - 1 0 ' ) , 4 . 9 7 ( I H , d d , J = 1 7 . 2 1 , 2 . 2 0 , H z - 1 0 ' ) , 8 . 1 9 ( 2 H , d,

J = 8 . 7 9 , H - A r ' - 2 , 6 ) , 7 . 6 1 ( 2 H , d , J = 8 . 7 9 , H - A r ' - 3 , 5 ) , 7 . 4 1 ( 2 H , d , J = 8 . 6 1 , H -

A r " - 2 , 6 ) , 7 . 5 6 ( 2 H , d, J = 8 . 6 1 , H - A r " - 3 , 5 ) .

6 7

' ^C -NMR (100 MHz, acetone-rfe): 6c 156.09 (C-9), 170.18 (C-7), 143.05 (C-

4), 38.42 (C-3), 53.72 (C-2), 25.55 (C-T), 27.36 (C-2'), 29.20-30.40 (C-3'-7'),

34.42 (C-8'), 139.83 (C-9'), 114.63 (C-10'), 130.44 (C-Ar '-2,6) , 129.67 (C-

Ar'-3. 5), 130.03 (C-Ar"-2,6), 129.39 (C-Ar"-3,5), 142.31 (C-Ar"-1), 134.24

(C-Ar ' - l ) , 138.81 (C-Ar'-4), 134.39 (C-Ar"-4).

D C I - M S (NH3): m/z (%) 512 ( M ^ 6.2), 513 [ ( M + l ) ^ 100.0], 514 (40.7), 515

(72.1), 516 (25.2), 517 (18.2), 518 (5.4), 273 (3.9), 274 (23.7). 275 (7.4). 276

(54.3), 277 (12.6), 278 (27.8), 279 (6.7), 280 (6.8), 238 (3.9). 239 (3.7). 240

(28.6), 241 (7.4), 242 (5.7), 128 (5.1), 1 15 (7.9), 1 17 (5.1), 102 (9.0), 93

(6.4).

l,3-Bis(2-thienyl)-3-l3-(9-decenyl)-4-amino-5-thio-l,2,4-triazolyl]propan-l-

one FTCA (9) [Reaction of the s-triazole (1) with l ,3-bis(2-thienyl)

propen- l -one (8)]*

*[This reaction failed to yield the desired product 2,4-bis(2-thienyl)-7-(9-

decenyl)-2,3-dihydro-l ,2,4-triazolo[3,4-b] [1,3,4]thiadiazepine].

A mixture of the 1,2,4-triazole (1) (500 mg, 1.96 mmol) and l ,3-bis(2-

th ienyl )propen- l -one (20) (432 mg, 1.96 mmol) in dry benzene (20 ml) was

ref luxed for 28 h in presence of catalytic amount of p - T s O H (5 mg). The

reaction mixture on TLC examination (silica gel G, pet .e ther-EtOAc, 8:2 v/v)

showed the presence of only one spot even by prolonging the reaction. The

products on column chromatography (silica gel, pet. e ther-EtOAc, 8:2 v/v)

afforded FTCA aa pale yellow oily liquid, 640 mg (68.6%), Rt 0.69 (pet.

ether-EtOAc, 8:2 v/v).

6 8

Spectral data of FTCA (9)

^H-NMR (400 MHz, acetone-^/^): 5h 5.21 (2H, s, NH2), 6 .79 ( I H , dd, J=9.01,

5 . 8 0 , H - 3 ) , 3 . 8 2 ( I H , d d , J = 1 6 . 9 4 , 5 . 8 0 , H - 2 ) , 4 . 2 5 ( I H , d d , J = 1 6 . 9 4 , 9 . 0 1 , H -

2) , 2 . 7 1 ( I H , t, J = 7 . 4 7 , H - L " ) , 1 . 6 8 ( 2 H , b r p , J ~ 7 . 3 2 , H - 2 " ) , 1 . 2 8 (LOH, b r s,

H - 3 " - 7 " ) , 2 . 0 2 ( 2 H , m , H - 8 " ) , 5 . 8 1 ( I H , t d d , J - 6 . 7 2 , 1 0 . 2 3 , 1 7 . 0 9 , H - 9 " ) , 4 . 9 1

( I H , d d , J = 1 0 . 2 2 , 2 . 2 9 . H E - 1 0 " ) , 4 . 9 9 ( I H , d d , J = 1 7 . 2 4 , 2 . 2 9 , H ^ - I O " ) , 7 . 9 0

( I H , d d , J = 4 . 8 9 , 1 . 2 2 , H - A r ' - 5 ) , 7 . 2 4 ( I H , d d , J = 4 . 8 8 , 3 . 8 1 H - A r ' - 3 ) . 8 . 0 3

( I H , d d , J = 3 . 8 1 . 1 . 2 2 , H - A r ' - 3 ) 7 . 2 2 ( I H , d d , J = 3 . 5 1 , 1 . 2 2 . H - A r - 5 ) 6 . 9 7 ( I H .

d d , J = 5 . 0 3 , 3 . 5 1 , H - A r - 4 ) , 7 . 3 6 ( I H , d d , J = 5 . 0 3 , 1 . 2 2 , H - A r - 3 ) .

'^C-NMR (100 MHz, acetone-rf^): 5c 152.51 (C-3'), 167.48 (C-5'), 54.03 (C-3).

44.25 (C-2), 189.06 (C-1), 25.18 (C- l" ) , 26.6 (C-2"), 29.20-30.40 (C-3"-7"),

34.45 (C-8"), 139.89 (C-9") 114.64 (C-10"), 126.30-144.78 (Ar + Ar') .

D C I - M S (NH3): m/z (%) 475 ( M ^ 12.5), 476 (4.4), 254 (6.8), 255 (67.1), 256

(11.9). 257 (4.1), 220 (11.5), 221 (100.0), 222 (15.3), 223 (1 1.3), 137 (3.6), 130

(5.7). I l l (32.0).

1,11 11

Long alkyl chain substituted-Benzopyrano-s-Triazolo-Thiadiazepines

6 9

DISCUSSION

SUMMARY

The work included in this chapter deals with the reaction of 4-amino-3-(9-

decenyl)-5-mercapto-l,2,4-triazole (1 ) with :

i) E-3-(4-Chlorobenzylidene)-4'-chloroflavanone (10) which yielded an

adduct, 2-(4-chlorophenyl)-3-[4-amino-2-(9-decenyl)-5-(4-chlorobenzyl-

thio)-! ,2,4-triazolyl]-2,3-dihydro-4-benzopyrone (11) and a novel

compoundj2,3-bis(4-chlorophenyl)-1 l-(9-decenyl)-2a, 3-dihydro[l]benzo-

pyranof3,4-e]-2,2a-dihydro-l,2,4-triazolo[3,4-b][1,3.4Jthiadiazepine (12).

ii) E-3-Benzylidene flavanone (13) which afforded an adduct 2-(phenyl)-3-

[4-amino-3-(9-decenyl)-5-(benzylthio)-1,2,4-triazolyl]-2.3-dihydr0'4-hen-

zopyrone (14) and an analogous new compound, 2.3-bis(phenyl)-J I-(9-

decenyl)-2a,3-dihydro[I]benzopyrano[3,4-e]-2.2a-dihydro-l.2.4-lriazuh)

[3,4-b][},3.4]thiadiazepine (15)

I N T R O D U C T I O N

As part of our programme in search of biologically active compounds

with sulphur and nitrogen containing heterocycles, we have synthesised two

novel compounds, 2-(4-chlorophenyl)-4-[2-(4-chlorophenyl)ethenyl]-7-(9-

decenyl)-2 ,3-dihydro- l ,2 ,4- t r iazolo[3,4-b][ l ,3 ,4] thiadiazepine (4) and 2.4-

bis(4-chlorophenyl)-7-(9-decenyl)-2,3-dihydro-l ,2 ,4-tr iazolo[3.4-bJ[1.3.4]thi-

adiazepine (7) by the reaction of 3-(9-decenyl)-4-amino-5-mercapto-1,2,4-

triazole (1) with l ,5-bis(4-chlorophenyl)pent- l ,4-dien-3-one and 4,4'-

dichlorochalcone (described in Chapter 1). In continuation of the above

mentioned work, we extended our work on the reaction with exocyclic

^./^-unsaturated enone systems and synthesised two novel compounds, (i) 2,3-

bis(4-chlorophenyl)- l l - (9-decenyl) -2a ,3-dihydro[ l ]benzopyrano[3,4-e]-2 ,2a-

7 0

dihydro-l ,2 ,4- t r iazolo[3,4-b][ l ,3 ,4] thiadiazepine (12) and (ii) 2,3-bis(phenyl)

-1 l - (9-decenyl)-2a,3-dihydro[ l ]benzopyrano[3,4-e]-2 ,2a-dihydro- l ,2 ,4- t r iaz-

olo[3,4-b][ l ,3 ,4] thiadiazepine (15) by the reaction of 4-amino-3-(9-decenyl)-

5-mercapto- l ,2 ,4- t r iazole (1) with (i) £ '-3-(4-chlorobenzylidene)-4 '-chloro-

f lavanone (10) and (ii) £-3-benzyl idene f lavanone (13) in dry benzene using

p - T s O H as catalyst. An adduct, 2-(4-chIorophenyI)-3-[3-(9-decenyl)-4-amino-

5-(4-chlorobenzyl thio)- l ,2 ,4- t r iazolyl]-2,3-dihydro-4-benzopyrone (11) in the

former and 2-(phenyi)-3-[3-(9-decenyl)-4-amino-5-(benzylthio)-1,2,4-

tr iazolyl]-2,3-dihydro-4-benzopyrone (14) in the latter was also obtained in

good yields. £'-3-Benzylidene flavanones have been prepared by the reported

procedure^^ and their stereochemistry discussed.

3-Benzyl idene flavanones, often formed as co-products during the

preparat ion of flavanones or chalcones^° have structural similarities with

natural homo isoflavonoids, Eucomin and Eucomol isolated f rom Eucomis

bicolor (Liliaceae)^' and they have been the focuss of attention for their

methods of preparation and stereochemical implications during the past few

years^^. Although the reactions of various substi tuted/unsubsti tuted

benzyl idene f lavanones with 2-aminobenzenethiols to form the corresponding

1,5-benzothiazepine have been described^^'^'', no report on the reaction of

benzyl idene f lavanones with 4-amino-5-mercapto- l ,2 ,4- t r iazoles has

appeared in the literature.

Structural assignment, stereochemistry and biological assay are

discussed. Screening results of the compound (12) is summarised for

anticancer activity against 3-cell lines of three types of human cancers: lung,

breast and CNS (details in Chapter 6).

7 1

4 .R '

O H

(10) , (13)

(11), (14)

(12) , (15)

(10) , (11) , ( 1 2 ) : R = R ' = CI

(13) , (14) , ( 1 5 ) : R = R ' = H

7 2

R E S U L T S A N D D I S C U S S I O N

£-3-Benzyl idene flavanones, AF2 (10) and AFj (13) were first prepared

fol lowing the literature procedure '^ and their structures alongwith the

stereochemistry determined by IR, 'H-NMR and '^C-NMR spectra. IR spectra

(KBr) showed characterist ic bands corresponding to ( C = 0 ) , (C=C, phenyl)

and (C-O-C). The ' H - N M R and '^C-NMR spectral signals were assigned on

the basis of chemical shif t values, coupling constants and by comparison with

the spectral data reported in the literature^^. The absorpt ion of the vinyl

proton (H-y^ (H-1'") at very low field, 5 8.09/8.03 indicated that it is due to

deshielding effect arising from the diamagnetic anisotropy of the carbonyl

group (being in close proximity) which is consistent with the trans

configuration^^ (£ - fo rm) (H cis and Ph trans to the carbonyl group) (in the

cis isomer, the vinyl proton appears at 5 6.65^^). Thus, benzylidene

f lavanones here exhibit ^-conf igurat ion. The conf igura t ion at C-2 was

decided by the inspection of models. Models showed that ^ -S-benzyl idene

f lavanones adopt strainless stable conformation only, when the 2-Ph is axial

and 2-H equatorial as the 2-Ph equatorial in another possible conformer

nearly overlaps with ^ - P - P h causing severe non-bonded interaction. Such an

interaction is not present in the former conformer. It is well known that in

cyclohexanes and other related compounds, a large group normal ly adopts the

equatorial position to avoid 1,3-diaxial interaction. However , in our case,

there are no such axial groups to interact with an axial 2-Ph group. The

preferred conformat ion of £'-3 benzylidene f lavanones (10a and 13a) would

therefore, be (10b and 13b).

7 3

lOa. R=R' = C1 13a. R = R' = H

10b. R=R' = C1 13b. R=R' = H

The title compounds FF2C (12) and FTZX5 (15) were then synthesised

by ref luxing 4-amino-3-(9-decenyl)-5-mercapto- l ,2 ,4- t r iazole FTZ (1) with

£ '-3-(4-chIorobenzylidene)-4 '-chloroflavanone AF2 (10) and £-3-benzyl idene

f lavanone AFi (13) in dry benzene using />-TsOH as catalyst. In above

reaction, a Michael adduct FF2A (11)/ FFjA (14) was also obtained in good

yields (Scheme 2)

7 4

o .OH

+ OHC-,CH3 o -R

O molar ratio (1:3) EtOH/KOH

O H

N N

^SH

NH2

(1)

(10) R = R' = C1 (13) R = R' = H

Molar ratio (1:1) Dry benzene Reflux 38/32 h

(11) R = R' = C1 (14) R = R' = H

N N

(12) R = R ' - C 1 (15) R = R' = H

SCHEME 2

7 5

2,3-Bis(4-chlorophenyl)-ll-(9-decenyl)-2a,3-dihydroll]benzopyrano[3,4-e]-

2,2a-dihydro-l,2,4-triazolo[3,4-b][l,3,4]thiadiazepine FF2C (12) alongwith

the adduct, 2-(4-chlorophenyl)-3-[4-amino-3-(9-decenyl)-5-(4-chlorobenzyl-

thio)-l,2,4-triazolyl]-2,3-dihydro-4-benzopyrone FF2A (11). [Reaction of the

5-triazole FTZ (1) with £'-3-(4-chlorobenzyIidene)-4'-chloroflavanone AF2 (10)]

A solut ion of the 1,2,4-triazole (1) and £ ' -3- (4-chlorobenzyl idene)-4 ' -

ch lo ro f lavanone (10) (molar ratio, 1:1) in dry benzene was re f luxed for 38 h

in the presence of a catalyt ic amount o f /7 -TsOH, moni to r ing the progress of

the react ion by TLC. The react ion mixture revealed the p resence of two spots,

labelled as FF2A (11) and FF2C (12). The res idue left a f te r usual work up

was chromatographed over a silica gel column using pet. e ther -EtOAc, (7:3

v/v) as eluent . Elut ion first y ie lded a colourless glassy f lakes , FF2A (40.5%).

Further e lut ion of the column using pet. e ther -EtOAc, (1:1 v/v) y ie lded a pale

yel low solid, FF2C which was recrystal l ised (pe t . e the r -E tOAc) as white

crystal l ine needles in 44 .72% yield.

N N

N H 2

F T Z (1) Dry benzene//?-TsOH AF2(10) Reflux 38h

F F 2 A ( 1 1 ) F F 2 C ( 1 2 )

7 6

Structure Eluc idat ion of FF7A (11)

It was a colourless glassy flakes, m.p 93 °C. The structure of FF2A has

been established by DCI-MS, 'H-NMR, and '^C-NMR spectra. The DCI-MS

spectrum (Fig. 6a) displayed a set of [M+1]"^ peaks at m/z 635/637/639

showing its molecular weight 634/636/638 [C34H36N4O2SCI2], which is equal

to the sum of the molecular weights of the 1,2,4-triazole (1) (254) and £-3-(4-

ch]orobenzylidene)-4 ' -chloroflavanone (10) (380). This suggested that FF2A

is an adduct of the two compounds. The retro-Michael addit ion fragment ion

peaks were observed at m/z 255 [(M+l)-C22Hi4Cl202]^ as the base peak and

at m/z 381/383/385 [(M+l)-Ci2H22N4S]^. The retro-Diels Alder f ragment ions

were formed at m/z 121 and m/z 259/261/263. The f ragment ion peaks at m/z

130 and 143 were arised by the cleavage on the long alkyl side chain at and

/-posit ionsto the triazole ring as shown in Chart 6.

All the signals in 'H-NMR and '^C-NMR spectra (Fig. 6b and 6c) were

assigned to individual H and C-atoms (Table 7) on the basis of 5 values,

mult ipl ici ty and by comparison with the previously established structures of

similar adducts. The absence of vinylic proton (H- l ' " ) signal at 5 8.03 and

thiol proton signal at 5 12.25 and the appearance of new signals at 5 5.65 as

doublet for H - l ' " and 5 6.42 also as doublet for H-2 in the ' H - N M R spectrum

of FF2A suggested the addition of thiol group to C=C double bond of the

enone systems of £-3-benzyl idene flavanone. This is also supported by the

shifts of the vinylic carbons at 5 138.28 (C- l" ' ) and 133.28 (C-3) to higher

fields 5 60.01 and 5 79.04. The other 'H- and '^C-NMR signals were found

almost identical to that of £'-3-benzylidene f lavanone and tr iazole moieties.

• - . M

7 7

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8 1

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N N o O

NH2

CI' Q -O' Q m/z 635 / 637/639(5.4/4.1) [M+1]


-Km

N N

N- "SH

NH2

m/z 143(6.0)

m/z 255 (100.0)

P-cleavage H

N N • i 1]

^ N - ^ S H NH2

m/z 130 (9.4)

m/z 381/383/385 (22.6/14.7/3.2)

retro-Diels Alder fission

m/z 121 (12.4)

m/z 259/261/263 (14.3/6.8)

CHART 6

The large coupl ing constants of C-2 and C-3 pro tons of f l avanone

moiety (JH-2»H-3 11-14) conf i rmed that the two ad jacen t hydrogen a toms are in

axial disposi t ion. This showed that the proton of thiol g roup has been added

s tereoselect ively to C-3 of C=C bond f rom the axial s ide. A plausible

explanat ion may be of fe red for this s tereoselect ivi ty . The addi t ion of thiol

group of (1) to the exocycl ic a,y^-unsaturated carbonyl sys tem of E-3-{4-

chlorobenzyl idene) -4 ' -ch lorof lavanone (10) fo l lowing nuc leoph i l i c path '^

( S c h e m e 3) leads to the fo rmat ion of a s tereoselect ive t rans adduct probably

8 2

by in t ramolecular proton t ransfer of the 1,4-addition moie ty f rom the axial

side at C-3, assuming the bigger group adopts equatorial pos i t ion . Thus, the

conf igura t ions , of all the three s tereocentres (C-2, C-3 and C - l ' " ) could be

settled f rom the coupl ing constants of protons at C-2, 6 6.42 ( I H , d,

J2ax,3axl l . l4) , C-3, 5 4.37 ( I H , dd, J3ax,23x11-14, J3ax ,1'" 2 .75) and C - l ' " , 5

5 . 6 5 ( 1 H , d, J r . . 3 a x 2 . 7 5 ) .

N N

I I

I

NH;, F T Z (1)

N N

AF2(10)

FFjA (11) S C H E M E 3

8 3

Based on the above spectral evidence, s tereostructure of FF2A was

formulated as 2-(4-chlorophenyl)-3-[S-(9-decenyl)-4-amino-5-(4-chloro-

benzylthio)-!,2,4-triazolyl]-2,3-dihydro-4-benzopyrone (11).

CI

0 1 N N Ha (ft

1 U' , 1 4 ? ^ (ft

1 ^ ' 5 4' 3I

FF2A (11)

„ Hz

Structure Elucidat ion of FF^C (12)

It was a white needle shaped crystalline solid, m.p 182 °C. The

structure of FFjC has been established by DCI-MS, ' H - N M R and '^C-NMR

spectra. The DCI-MS spectrum (Fig. 7a) showed a set of [M+1]"^ peaks as the

base peak at m/z 617/619/621 confirming its molecular weight 616/618/620

[ C 3 4 H 3 4 N 4 O S C I 2 ] which is equal to the molecular weight of (11)

(634/636/638) minus one molecule of water. This indicated that

cyclocondensat ion has occurred between the amino group of t r iazole moiety

and carbonyl group of benzopyrone moiety forming th iadiazepine ring. The

peak at m/z 492/494 was obtained by the cleavage on 9-decenyl side chain at

the y^-position to the 5-triazole ring. The peaks at m/z 380/382/384 [(M+1)-

C,2H2iN3S]^ and at m/z 240 [M-C25HigN4SCl2]^ are the diagnost ic f ragment

ion peaks arised by the cleavage of 3-4 and 1-7 bonds of thiadiazepine ring

as shown in Chart 7.

8 4

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8 5

[M+1 ]m/z 61 lie 19/621 (100.0/82.9/22.4)

1-7 and 3-4 cleavage m/z 492/494 (5.4/4.8)

iWz 240 (4.3)

m/z 380/382/384 (23 4/14 2/4 4)

CHART 7

All the signals in ' H - N M R and '^C-NMR spectra (Fig. 7b and 7c) have

been ass igned (Table 8) on the basis of 5 and J va lues and also by

compar ison with the spectra of ( l l ) ( T a b l e 7). The absence of signals

cor responding to amino protons at 5 5.22 and carbonyl g roup at 6 189.69 in

the spectra of FF2C and in place, the appearance of C-4 signal at higher

f ields, 5 156.24 (C-8b) in ' ^C-NMR spectrum is s trongly in agreement with

the fo rmat ion of a thiadiazepine ring by the cyclocondensation of NH2 group with

the carbonyl group. The chemical shifts of H- and C-atoms generally go to higher

fields in the formation of the ring. The same trend was observed in this case also.

The shifts of C-2 and C-3 protons to higher fields from 5 6.42 to 5 5.90 and from

6 4.37 to 5 4.14 in the 'H-NMR spectrum and also from 6 79.04 to 6 76.98 and 6

55.08 to 5 45.22 in '^C-NMR spectrum showed that these positions are

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8 9

involved in ring formation. Also the shift of carbonyl group f rom 5 189.69 to

6 156.24 confirmed that intramolecular cyclocondensat ion has occurred

between the NH2 group of triazole and > C = 0 group of pyrone forming

thiadiazepine ring. The other signals corresponding to tr iazole and

benzopyrone moieties appeared as in (11).

On the basis of above spectral evidence, FF2C was character ised as

2,3-bis(4-chlorophenyl)-l l-(9-decenyl)-2a,3-dihydro[l]benzopyrano[3,4-e]-

2, 2a-dihydro-l, 2,4-triazolo[3,4-b][ 1,3,4]thiadiazepine (12).

FF2C (12)

Mechanism of the formation of FF,A (11) and FF^C (12)

The reaction occurred via nucleophilic attack of thiol group to the

vinyl carbon of 3-benzylidene flavanone which resulted in the format ion of

Michael adduct (11). This was followed by int ramolecular dehydrat ive

cyclocondensat ion between the amino group and the carbonyl group forming

the cyclised product, thiadiazepine (12) (Scheme 4).

9 0

N N N N

H" transfer

N N

H^ transfer

H' FF2A (11)

N- N N

-CI

FF2C (12)

SCHEME 4

9 1

2,3-Bis(phenyl)-ll-(9-decenyl)-2a,3-dihydro[l]benzopyrano[3,4-e]-l,2,4-

triazolo-2,2a-dihydro[3,4-b][l,3,4]ihiadiazepine FTZXg (15) along with the

adduct, 2-(phenyl)-3-[4-amino-3-(9-decenyl)-5-(benzylthio)-l,2,4-triazolyl]-

2,3-dihydro-4-benzopyrone FFjA (14) [Reaction of the s - t r iazo ie (1) with

E-3-ben2yl idene f lavanone (13)]

A solut ion of 1,2,4-triazole (1) and ^ - S - b e n z y l i d e n e f l avanone (13)

(molar ratio, 1:1) in dry benzene was re f luxed for 35 h in presence of a

catalyt ic amount of / )-TsOH. The products on pur i f i ca t ion by column

chromatography (sil ica gel, pet. e ther-EtOAc, 8:2 v/v) a f f o r d e d F F , A (14)

(39 .10%) as pale yel low oily liquid. Further e lut ion of the c o l u m n (pet. ether-

EtOAc. 7:3 v/v) gave white solid mass which on recrys ta l l i sa t ion (benzene-

acetone) a f fo rded FTZX5 (15) as white crystal l ine needles in 46 .36% yield.

N N

'SH

NH2

F T Z (1) O (13)

(i) /7-TsOH/dry benzene Reflux 35 h

FF,A(14) FTZX5 (15)

9 2

Structure Elucidat ion of FFjiA (14)

It was a pale yellow oily liquid. The structure of F F i A has been

established by FTIR, EI-MS and 'H-NMR spectra. The FTIR spectrum

displayed characteristic signals at 3402.9 (NH), 2927.0 (CH), 1694.5 (C=0) ,

1630.8 (C=C), 1607.2 (C=N), 1578.0 (phenyl) and 1495.1 (C-S) cm"'.

EI-MS spectrum of F F i A showed a [M+l]"" peak at m/z 567 confirming its

molecular weight 566 [C35 H38N4O2S], which is equal to the sum of the

molecular weights of the 5-triazole (1) (254) and £-3-benzyl idene f lavanone

(13) (312). This indicated the Michael addition of thiol group to the

or,/?-unsaturated exocyclic system of £-3-benzyl idene f lavanone. The

fragmentat ion pattern was found to be similar to that of FF2A (11). The

characterist ic retro-Michael addition fragment ion peaks were observed at

m/z 313 [M-C25H2oN4S]^ and m/z 255 [M-CnHj iN jS ] " . It also showed the

retro-Diels Alder fragment ions at m/z 191 and m/z 121 in addition to the

common ion peaks as in (11) at m/z 255,143 and 130 as shown in Chart 8.

The 'H-NMR signals (Fig. 8) have been assigned (Table 9) to

individual hydrogen atoms by comparing with the spectra of a previously

described compound (11) (Table 7). The absence of signals corresponding to

thiol proton at 6 12.25 of the triazole and the vinyl proton of the £ -3 -

benzyl idene f lavanone at 5 8.09 and appearance of this proton as doublet at

higher f ield 5 5.52 confirmed the addition of thiol group to a,/?-unsaturated

exocyclic system of £'-3-benzylidene f lavanone. As in (11), the large coupling

constants of protons at C-2 and C-3 positions (12,3 11.40) revealed that the

hydrogen atoms are oriented at the axial disposition. In addit ion, the signals

9 3

fe S a " «_) a o Q 3

O T

Z X i u

3 M • S o . • 3 a

CTI ^ o

o

" a 1 r £88

O * i 5

J r J £ S £ g J g S — - ^ j ^ j a c o o o p o — o r u i n r

i c t o o O 4 j « - -

? g fe 1 lO o o> *

' . 11 i I I

S50E 0

LL<!<I 5

^ 6 6 5 0 —

g i 9 8 0

n e i 0

502Z 0

9 8 f l 0

i C L ^ o e o 0

0

901-9 0

0E99 0

^ e e E E 0 - - ^ ^ O t ' O p

PIPE _0

— ^ S f t - O 0 0

Bst-g 0

0E9e f 0

OPPO t

1201 p

- o

-OJ

-m

00 feC 'W*

(tjea-jui

-in

- I D

-co

a. D .

9 4

corresponding to i^-S-benzylidene f lavanone and tr iazole moiet ies also

appeared as shown in Table 9.

CH..

r ~

N N

1! 1 I

NH2 m/z 255 (100.0)

N N H

P-cieavage

I

NH2

m/z 143(10.6)

N N I 11 ^ N - ^ S H

NH2 m/z 130(12.5)

m/z567( 7.2)[ M+V


m/z 3 13 (18.4)


O

m/z 121 (13.3) m/z 193 (12.2)

CHART 8

9 5

Table 9: 'H-NMR spectral data of FF ,A (14)


Multi-plicity

J (Hz)

NH2 5.02 2H s

1"' 5.52 IH d J,... 3 1.80

2 6.38 IH d J2,3 11.40

3 4.38 IH dd h a 11-40,13.9 1-80

5 7.38 IH dd J5,6 7.80, J w 1-65

6 6.95 IH dd J6,7 7.80, J6 5 7.50

7 7.55 IH dd J7,e7.50J7 8 7.20

8 7.16 IH dd Jg,7 7.20, J8 6 1.09

1" 2.76 2H t J i " , 2 " 7 . 2 0

2" 1.73 2H br p J « 7 . 1 8

3"-7" 1.22 5x2H br s

8" 1.99 2H m

9" 5.79 IH tdd J9..8.. 6.68, J9",he 10.20. Jg-.Hz 17.06

He-10" 4.92 IH dd J h e , 9 " 10.20, Jhe. h z 2.29

Hz-10" 4.95 IH dd J h z , 9 " 17.06, J h z , . i e 2.29

Ar'+Ar" 7.17-7.29 2x5H m

Based on the above spectral evidence, F F , A was character ised as, 2-

(phenyl)-3-[4-amino-3-(9-decenyl)-5-(benzylthio)-l,2,4-triazolyl]-2J-dihydro

-4-benzopyrone (14).

9 6

Structure Elucidation of FTZX^dS)

It was a needle shaped white crystalline solid, m.p 167 °C. The

structure of FTZXg has been fully established by IR, DCI-MS, 'H-NMR,

'^C-NMR and further by COSY and HETCOR spectra. IR spectrum displayed

characterist ic absorption bands at 2900 (CH), 1630 (C=C), 1600 (phenyl),

1580 ( C - N ) and 1450 (S-C) cm"'. The DCI-MS spectrum displayed a [M+1]"

peak as the base peak at m/z 549 showing its molecular weight 548

[C34H36N4OS], which is equal to the molecular weight of (14) (566) minus

one molecule of water, indicating the formation of thiadiazepine ring by the

intramolecular cyclocondensat ion between the NH2 and > C = 0 groups of (14).

A less intense peak at m/z 424 [M-CgHp]"^' was obtained by the y9-cleavage

on the 9-decenyl side chain to the triazole ring. The peaks at m/z 310 [m/z

424-C2|H,6N0]^ and m/z 115 [m/z 424-C3H5N3S]^-, arised by the cleavage of

1-7 and 3-4-bonds are the diagnostic peaks of the thiadiazepine r ing obtained

by f ragment ing the ions at m/z 424. The retro-Diels Alder f ragment ion peaks

were observed at m/z 191 and m/z 121 as shown in Chart 9.

All the signals in 'H-NMR and '^C-NMR spectra (Fig. 9a and 9b) were

assigned to individual H and C atoms (Tables 10a and 10b) on the basis of

chemical shifts , multiplicity, relative integrations and by compar ison with the

spectra of a previously established compound FF2C (12) (Table 8). The

absence of the signals corresponding to NH2 protons at 5 5.02 and carbonyl

carbon at 5 189.69 in the 'H- and '^C-NMR spectra and the appearance of C-4

signal at higher field 5 156.09 indicated that the intramolecular

cyclocondensat ion between the two groups has occurred, forming the

thiadiazepine ring. The shifts of C-2 and C-3 protons of pyrone ring to higher

9 7

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T

(S ON

9 8

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w« lo to t! M « V C « N C 0 « >

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too

Table 10b: ^^C-NMR and HETCOR data of FTZXj (15)

C-nr 5(ppm) HETCOR

13a 164.43

11 157.47'

8b 156.09'

8a 119.03 H-8

8 125.88 H-7

7 122.40 H-6

6 136.00 H-5

5 118.59

4a 144.45 H-3

3 77.47

2a 45.36 H-2

2 60.35

r 25.45 H - r

2' 27.67 H-2'

y-7 29.20-30.40 H-3' - T

8' 34.46 H-8'

9' 139.93

10' 114.64

Ar'-l 136.46

Ar'-2,6 130.11 H-Ar'-2,6

Ar'-3,5 128.72 H-Ar'-3,5

Ar'-4 129.85 H-Ar'-4

Ar" 126.72-128.86 Ar"

' Assignment may be interchanged

i n i

f ields f rom 6 6.38 to 5 5.83 and f rom 5 4.38 to 6 4 .07 in the 'H -NMR

spect rum conf i rmed the format ion of the ring. The other s ignals appeared as

in the adduct (14). Also in '^C-NMR spectrum, the s ignals appeared were

ass igned as 5 157.47 (C-11), 156.09 (C-8b), 77.47 (C-3), 45 .36 (C-2a) , 60.35

(C-2) and 164.43 (C-13a) in addit ion to the signals of a romat ic carbons and

9-decenyl s ide chain as shown in Table 10b.

H \

N N N N

^ ^ ^ ^ P-cleavage^ N ^ .

[ M + l f m / z 549 (100.0) m/z 494 (8.7)

1-7 and 3-4 bond cleavage

H \

N N

H ^SH

m/z 115 (16.0) m/z 310 (11.8)


m/z 121 (5.5)

m/z 193 (6.0)

CHART 9

1 0 2

The ass ignments of ' H - N M R and '^C-NMR signals (Tables 10a and

10b) and the coupl ing of the protons were fur ther con f i rmed by a ' H - ' H

COSY and a H E T C O R spectra. In ' H - ' H C O S Y spec t rum (Fig. 9c), the

double doublet at 5 7.06 (J=8.16, 8.42, H-7) showed two cross peaks each

with a double doublet at 8 7.41 ( J - 8 . 4 2 , 8.24, H-6) and 5 8.16 (J=8.16, 1.65,

H-8) indicat ing the coupl ing of H-7 proton wi th both H-6 and H-8 protons.

Similar ly, the signal at 5 7.41 (H-6) showed two cross peak mult iple ts at

5 7.06 (H-7) and 5 6.54 (J=8.42, 1.10, H-5) , indicat ing the coupl ing of H-6

proton with both H-5 and H-7 protons . The double doublet at 5 4 .07 (J=5.86.

1.10. H-2a) showed a cross peak with the signals at 5 5.83. 5.87 (H-3. H-2),

indicat ing the coupling between them. The signals for the terminal olefenic

methylene protons appeared as two double double ts at 5 4 .90 (J= 10.26. 2.20,

HE-10') and 5 4.98 (J=17.22, 2.20, H^-10') which were found to be coupled

with each other and also with the ad jacen t meth ine p ro ton at 6 5.80. The

signals appeared as triplet double doublet at 5 5.80 (J=6.70 , 10.26, 17.22)

were due to the coupl ing of the methine proton (CH-, H-9 ' ) with the protons

at H-8' and HE-10', H^-IO'. In addit ion, COSY spect rum also showed the cross

peaks cor responding to the other s ignals of 9-decenyl side chain and aromatic

protons (Table 10a).

In H E T C O R spectrum (Fig . 9d , T a b l e 10b) the ' H - N M R signal at

5h 8.16 (H-8) is correlated to 6c 125.88 (C-8) , 5h 7.06 (H-7) to 8c 122.40

(C-7), 8h 7.41 (H-6) to 8c 136.0 (C-6), 8h 6 .54 (H-5) to 8c 118.59 (C-5), 8h

5.83 (H-3) to 8c 77.47 (C-3) and 8h 5.87 (H-2) to 8c 60.35 (C-2) . In addition,

it also showed the cross correlat ion peaks of 9-decenyl s ide chain and the

aromat ic rings.

1 0 3

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On the basis of the above discussion, F T Z X 5 was characterised as 2,3-

bis(phenyl)-l l-(9-decenyl)-2a, 3-dihydro[ 1 JbenzopyranofS, 4-e]-2,2a-dihydro

-1,2,4-triazolo[3,4-b][ 1,3,4]thiadiazepine (15)

F T Z X 5 (15)

1 0 8

EXPERIMENTAL

4-Amino-3-(9-decenyl)-5-mercapto-l,2,4-triazole (1) was prepared

following a literature procedure and spectral studies performed as described in

Chapter 1.

E-3-(4-Chlorobenzylidene)-4'-chloroflavanone AF2 (10).

It was prepared according to a published procedure^^ To a mixture of

2-hydroxyacetophenone (2.0 g, 0.0147 mol) and jf?-chlorobenzaldehyde (6.0 g,

0.0441 mol), a warm solution (45 °C) of KOH (15%) in aqueous-methanol

(1:1) was gradually added with stirring. A clear solution resulted in after the

addition of 23 ml of this solution. Turbidity was removed by adding 1 ml of

methanol on cooling to room temperature. The reaction mixture was then kept

s toppered for 4 days with occasional shaking. The solid mass appeared was

filtered and washed with excess of water fol lowed by 10 ml of alcohol. It was

then recrystal l ised from chloroform-acetone to give AF2 (10) as yellow

crystall ine needles, 3.40 g (61%), m.p. 224-226 °C, Rf 0.66 (pet. ether-

benzene, 8:2 v/v).

Spectral data of AF7 (10)

IR (KBr): v^ax 2850 (C-H), 1665 ( > C = 0 ) , 1605, 1550 (C=C, phenyl) , 1130,

( C - O - C ) , 1095, 1080, 1005, 950, 905, 875, 845, 805, 740 cm"'.

' H - N M R (400 MHz, CDCI3): 6h 6.52 ( IH, s, H-2), 8.03 ( I H , s, H-1'"), 7.85

( IH , dd, J = 7.93, 1.68, H-5), 6.98 ( IH, ddd, J= 7.93, 7.17, 168, H-6), 7.43

( IH , ddd, , J= 7.17, 8.39, 1.68, H-7), 6.95 ( IH , dd, J = 8.39, 1.68, H-8), 7.24-

7.48 (8H, br m, H-Ar '+H-Ar") .

1 0 9

'^C-NMR (CDCI3, 75 MHz,): 5c 76.86 (C-2), 133.28 (C-3), 182.08 (C-4),

138.28 (C-1'"), 122.65 (C-4a), 127.64 (C-Ar-5), 122.10 (C-Ar-6), 136.33 (C-

Ar-7), 118.58 (C-Ar-8), 121.92 (C-8a), 129.16 (C-Ar'-2,6), 131.05 (C-Ar'-

3,5), 128.93 (C-Ar"-2,6), 129.21 (C-Ar"-3,5), 132.37 (C-Ar '-4) , 132.25 (C-

Ar"-4), 136.30 (C-Ar ' - l ) , 134.86 (C-Ar"-1).

E-3-Benzylidene flavanone AFJ (13)

To a mixture of 2-hydroxyacetophenone (5.0 g, 0.0367 mol) and

benzaldehyde (11.70 g, 0.1103 mol), a warm solution (45 °C) of KOH (15%)

in aqueous-methanol (1:1) was gradually added with stirring. A clear solution

resulted in after the addition of 44 ml of this solution. On cooling to room

temperature the reaction mixture became turbid and 1.5 ml of methanol was

added to remove the turbidity. The reaction mixture was then kept at room

temperature. After Ca 3-4 h, an oily liquid began to deposit at the bottom and

the reaction mixture gradually turned into dark green. The reaction mixture

was then kept stoppered for 5 days with occasional shaking. The progress of

reaction was closely monitored by TLC (silica gel G, pet. ether-benzene, 8:2

v/v). The liquid that separated was changed into a semi solid mass, which

showed a mixture of three components (TLC). The reaction mixture was then

extracted with ethyl acetate, washed successively with water to make the

solution neutral and dried over anhydrous sodium sulphate overnight. The

solvent was then distilled off under reduced pressure. The product obtained

was separated and purified by column chromatography (silica gel, pet. ether-

benzene, 8:2 v/v) to give a greenish yellow mass which on crystallisation

(pet. ether-chloroform) yielded £'-3-benzylidene f lavanone (13), 8.0 g (67%),

m.p 104-106 °C, Rf 0.73 (pet. ether-benzene, 8:2 v/v).The other two

1 1 0

components, corresponding to chalcone and flavanone were also obtained in

very low yields on further elution of the column.

Spectral data of AFj (13)

IR (KBr): Vn,ax 2900 (C-H), 1710 (>C=0) , 1625, 1590 (C=C, phenyl), 1110

( C - O - C ) , 1005, 940, 860, 805, 720 cm-'.

'H-NMR (400 MHz, CDCI3,):5h 6.65 ( IH, s, H-2), 8.09 ( IH , s, H-1'"), 7.92

( IH. dd, J = 8.24, 1.83, H-Ar-5), 6.95 ( IH, ddd, J=8.24, 7.14, 1.10, H-Ar-6).

7.47 ( IH, ddd, J=7.14, 7.33, 1.83, H-Ar-7), 6.92 ( IH, dd, J=7.33, 1.10. H-Ar-

8), 7.02-7.43 (lOH, br m, H-Ar' + H-Ar").

'^C-NMR (100 MHz, CDCI3): 5c 77.69 (C-2), 133.28 (C-3). 182.65 (C-4).

l21.63(C-4a), 126.17 (C-Ar-5), 121.75 (C-Ar-6), 136.05 (C-Ar-7), 118.65 (C-

Ar-8). 158.97 (C-8a), 139.32 (C-1'"), 128.63 (C-Ar'-2,6) 129.94 (C-Ar'-3.5),

127.64 (C-Ar"-2,6), 128.85 (C-Ar"-3,5), 129.70 (C-Ar'-4), 128.63 (C-Ar"-4),

138.18 (C-Ar ' - l ) , 134.17 (C-Ar"-1).

2,3-Bis(4-chlorophenyl)-ll-(9-decenyl)-2a,3-dihydro[l]benzopyrano[3,4-e]-

2,2a-dihydro-l,2,4-triazolo[3,4-b][l,3,4]thiadiazepine FF2C (12) alongwith

the adduct, 2~(4-chlorophenyl)-3-[4-amino-3-(9-decenyl)-S-(4-chlorobenzyl-

thio)-l,2,4-triazolyl]-2,3-dihydro-4-benzopyrone FF2A (11). [Reaction of

the 5-triazole (1) with £'-3-(4-chlorobenzylidene)-4'-chloroflavanone (10))

A mixture of ^-triazole (1) (500 mg, 1.96 mmol) and £-3-(4-chloro-

benzylidene)-4'-chloroflavanone (10) (744 mg, 1.96 mmol) in dry benzene

(20 ml) containing catalytic amount of/7-TsOH (5 mg) was ref luxed for 38 h, %

monitoring the progress of the reaction by TLC (silica gel G, pet. ether-

EtOAc, 8:2 v/v). After completion, the reaction mixture on TLC examination

showed two spots, labelled as FF2A (11) (upper) and FF2C (12) (lower)

I l l

which on usual work up and column chromatography (si l ica gel, pet. ether-

EtOAc, 8:2 v/v) afforded F F j A (11) as glassy flakes, 504 mg. (40.5%), m.p.

93 °C, Rf 0.65 (pet. ether-EtOAc, 8:2 v/v). Further elut ion of the column

using pet . ether-EtOAc, (6:4 v/v) as eluent yielded a white solid mass which

on crystal l isat ion (benzene-EtOAc) afforded FF2C (12) as whi te crystalline

needles, 530 mg (44.72%), m.p. 182 °C, RfO.29 (pet. e ther -EtOAc, 6:4 v/v).

Spectral data of FF7A (11)

IR (KBr): 2900 (C-H), 1630, 1600 (C=C, phenyl) , 1580 (C=N), 1560,

1500, 1490, 1450 (S-C), 1420, 1340, 1270, 1200, 1 140, 1 1 10, 1050, 990, 900,

850, 750 cm"'

'H-NMR (400 MHz, acetone-Jg): 5h 5.22 (2H,s, NH2), 5.65 ( I H , d, J=2.75,

H-1'") , 4.37 ( IH , dd, J=10.98, 2.75, H-3), 6.42 ( IH , d, J=10.98, H-2), 7.22

( IH , dd, J=8.39, 1.07, H-8), 7.67 ( IH, ddd, J=8.85, 8.39, 1.68, H-7), 7.06

( IH, ddd, J=8.24, 8.85, 1.07, H-6), 7.51 ( I H , dd, J=8.24, 1.68, H-5), 7.28 -

7.36 (2x4H, m, Ar+Ar') , 2.84 (2H, t, J=7.48, H-1"), 1.83 (2H, br p, J~ 7.43,

H-2"), 1.26 (lOH, br s, H-3'-7'), 2.01 (2H, m, H-8"), 5.81 ( I H , tdd, J=6.71,

10.22, 17.09, H-9"), 4.91 ( IH, dd, J=10.22, 2.29, 10"-HE), 4.99 ( I H , dd,

J=17.09, 2.29, Hz-lO").

'^C-NMR (100 MHz, ncetone-ds): 5c 153.73 (C-3'), 168.72 (C-5') , 25.31

(C-1"), 26.54 (C-2"), 29.20-30.40 (C-3"-7"), 34.44 (C-8") , 139.85 (C-9"),

114.65 (C-10"), 60.01 (C-1"'), 55.08 (C-3), 79.04 (C-2) , 160.10_ (C-8a),

119.00 (C-8), 137.84 (C-7), 122.76 (C-6), 127.46 (C-5), 121.74 (C-4a),

189.69 (C-4), 137.39 (C-Ar"-1), 130.89 (C-Ar"-2,6) , 129.13 (C-Ar"-3,5),

134.66 (C-Ar"-4) , 136.05 (C-Ar ' - l ) , 129.57 (C-Ar'-2,6), 128.85 (C-Ar'-3,5),

134.45 (C-Ar '-4) .

1 1 2

DCI-MS (NHj) : m/z (%) 635 (5.4), 636 (2.5), 637 (4.1), 638 (2.4), 380 (8.4),

381 (22.6), 382 (10.3), 383 (14.7), 384 (4.3), 301 (4.5), 295 (13.4), 254 (5.0),

255 (100.0), 256 (17.6), 257 (26.6), 258 (5.1), 259 (14.3), 261 (6.8), 225

(6.0), 189 (5.0), 149 (4.7), 143 (6.0), 130 (9.4), 121 (12.4), 94 (4.8).

Spectral data of FF^C (12)

' H - N M R (400 MHz, acetone-«f^); 5h 5.90 ( IH, d, 1.07, H-3), 4.14 ( IH, dd,

J=6.41, 1.07, H-2a), 5.92 ( IH, d, J=6.41, H-2), 8.15 ( IH , dd, J=7.94, 1.68, H-

8), 7.10 ( IH, ddd, J=8.85, 7.94, 1.07, H-7), 6.62 ( IH, dd, J=8.39, 1.07, H-5),

5.92 ( IH, d, J=6.4L H-2), 7.17 (2H, d, J=8.69, H-Ar-2,6), 7.50 (H-Ar'-2,6),

7.28-7.29 (4H, m, H-Ar'-3,5, H-Ar'-3,5), 2.76 (2H, t, J=7.48, H-1'), 1.45 (2H,

br p, J~7.47,H-2'), 1.23 (lOH, br s, H-3'-7'), 2.00 (2H, m, H-8'), 5.79 ( IH,

tdd, J=6.71,10.22, 17.09 H-9'), 4.89 ( IH, dd, J=10.22, 2.29, HE-IO'), 4.97

( IH, J=17.09, 2.29, Hz-10').

' ^C-NMR (100 MHz, acetone-^/^): 5h 157.07 (C-11), 156.24 (C-8b), 118.94

(C-8a), 126.02 (C-8), 122.82 (C-7), 136.30 (C-6), 118.56 (C-5), 144.17

(C-4a), 76.98 (C-3), 45.22 (C-2a), 59.39 (C-2), 160.83 (C-13a), 138.63 (C-

Ar-1), 131. 77 (C-Ar-2,6), 128.75a (C-Ar-3,5), 135.42 (C-Ar-4), 134.41 (C-

Ar ' - l ) , 129.61 (C-Ar'-2,6), 128.59 (C-Ar'-3,5), 135.29 (C-Ar'-4), 25.48 (C-1'),

27.81 (C-2'), 29.20-30.40 (C-3'-7'), 34.46 (C-8'), 139.89 (C-9'), 114.63

(C-10').

D C I - M S (NHs): m/z (%) 616[M^ 5.4], 617 (100.0), 618 (50.8), 619 (82.5),

620 (38.2), 621 (22.4), 622 (7.6), 583 (2.6), 585 (2.8), 492 (5.4), 494 (3.8),

378 (7.1), 380 (23.4), 381 (8.1), 382 (14.2), 383 (4.3), 240 (4.3), 199 (3.8),

149 (3.8).

1 1 3

2,3-Bis(phenyl)-ll-(9-decenyl)-2a,3-dihydro[lJbenzopyrano[3,4-e]-2,2a-

dihydro-l,2,4-triazolo[3,4-b][l,3,4]thiadiazepine FTZXg (15) alongwith the

adduct, 2-(phenyl)-3-[3-(9-decenyl)-4-amino-S-(benzylthio)-l,2,4-triazolyl]-

2,3-dihydro-4-benzopyrone FF,A (14) [Reaction of the s-triazole (1) with

E-3-benzyIidene flavanone (13)]

A mixture of the 5-triazole (1) (500 mg, 1.96 mmol) and £ -3 -

benzylidene f lavanone (13) (610 mg, 1.96 mmol) in dry benzene (20 ml) was

ref luxed with stirring on an oil bath at 80 °C using a Dean Stark apparatus for

35 h in presence of a catalytic amount of jtP-TsOH (5 mg). TLC examination

(silica gel G, pet. e ther-EtOAc, 8:2 v/v) of the reaction mixture revealed the

presence of two spots, labelled as FFjA (14) (upper) and FTZX5 (15) (lower).

The solvent was distilled off in vacuum and the residue thus obtained was

dissolved in EtOAc, washed several times with water and the organic phase

dried over anhydrous Na2S04. The solvent was removed under reduced

pressure and the residue left was chromatographed over a silica gel column

using pet. e ther-EtOAc (8: 2 v/v) as eluent which a f forded FF ,A (14) as an

oily liquid, 434 mg (39.10%), Rf 0.66 (pet. ether-EtOAc, 8:2 v/v). Further

elution of the column (pet. ether-EtOAc, 7:3 v/v) yielded FTZXg (14) as a

white solid compound which on recrystallisation (EtOAc-benzene) as white

crystall ine needles, 498 mg (46.36%), m .p . l67 °C, R, 0.31 (pet. ether-EtOAc,

8:2 v/v)

Spectral data of FFj A (14)

FTIR (KBr): v^ax 3402.9, 3068.2 ( N H 2 ) , 3032.9, 2927.0, 2853.9(CH),

2362.6, 1694.5(C=0) , 1630.8 (C=C), 1607.2 (C=N), 1578.1(phenyl) , 1495.1,

1460.5, 1420.1, 1357.6, 1296.0, 1210.7 cm-'

1 1 4

' H - N M R (300 MHz, acetone-f/g): 5h 5.02 (2H, s, NH2), 5 .52 ( I H , d, J=1.80,

H-1'") , 6 .38 ( I H , d, J=11.40, H-2), 4.38 ( IH , dd, J=1 1.40, 1.80, H-3) , 7.16

( IH , dd, J=7.50, 1.09, H-8), 7.55 ( I H , ddd, J=7.50, 7.20, 1.65, H-7) , 6.95

( IH , dd, J=7.80, 7.20, H-6) , 7.38 ( l H , d d , 7.80,1.65, H-5) , 2 .76 (2H, t, J=7.20,

H-1 ' ) , 1.73 (2H, br p, J=7.18, H-2") , 1.22 ( lOH, brs, H-3" -7" ) , 1.99 (2H, m,

H-8") , 5 .79 ( I H , tdd, J=, 6.68, 10.20, 17.06 H-9") , 4 .92 ( I H , d, J=10.20, He-

10"), 4.95 ( I H , d, J=17.06, H^-IO"), 7.17- 7.29 ( lOH, m, 2xAr) .

E I - M S (NH3): m/z (%) 567 [(M+1), 7.2], 568 (3.1), 312 (6.7) , 313 (18.4),

314 (4.5), 254 (6.3), 255 (100.0), 256 (19.8), 191 (12.2) , 192 (5.8), 193

(12.2), 143 (10.6), 144 (2.9), 130 (12.5), 131 (4.1), 121 (13.3) , 122 (3.7)

Spectral data of F T Z X S (15)

' H - N M R (400 MHz, acetone-rf^): 5 h 5.83 ( I H , d, J= poor ly resolved, H-3) ,

4.07 ( I H , dd, J=5.86, 1.10, H-2a) , 5.87 ( IH , d, J=poor ly reso lved , H-2) , 8.16

( I H , dd, J=8.16 , 1.65, H-8) , 7.06 ( I H , dd, J=8.16, 8.42, 1.29, H-7) , 7.41 ( I H ,

dd, J=8 .42 , 8.24, 1.65, H-6) , 6.54 ( I H , dd, J=8.24, 1.10, H-5) , 7.48 (2H, d,

J=7.88, H-Ar ' -2 ,6) , 7.14 (2H, dd, J=7.88, 7.32, H-Ar ' -3 ,5 ) , 7.26 ( I H , d,

J=7.32, H-Ar ' -4) , 7.19-7.24 (5H, m, H-Ar") , 2.76 (2H, t, J=7 .47 , H-1 ' ) , 1.45

(2H, br p, J~7.37, H-2') , 1.24 ( lOH, brs, H-3 '-7 ' ) , 2.01 (2H, m, H-8 ' ) , 5.80

( I H , tdd, J=17.22, 10.25, 6.78, H-9') , 4.90 ( I H , dd, J=10 .25 , 2 .20, HE-10'),

4.98 ( I H , dd, J=17.22, 2.20, Hz-lO') .

'^C-NMR (100 MHz, acetone-^/^): 5c 157.47 (C-11) , 156.09 (C-8b) , 119.03

(C-8a) , 125.88 (C-8), 122.40 (C-7), 136.00 (C-6) , 118.59 (C-5) , 144.45

(C-4a) , 77.47 (C-3), 45.36 (C-2a), 60.35 (C-2) , 164.43 (C-13a) , 136.46

( C - A r ' - l ) , 130.11 (C-Ar ' -2 ,6) , 129.85 (C-Ar '-4) , 126.72-128.86 (C-Ar"-1 , C-

Ar"-2,6 , C-Ar"-3 ,5 , C-Ar"-4) .

1 1 5

D C I - M S (NH3): m/z (%) 549 [(M+1), 100.0], 550 (50.0), 551 (17.9), 424

(8.7), 425 (6.9), 310 (11.8), 312 (38.8), 313 (14.3), 314 (8.7), 208 (5.0), 191

(4.6), 193 (6.0), 121 (5.5), 115 (6.0), 91 (12.8).

b l . f'jL Via )

Hii .

Long alkyl chain substituted-s- Triazolo- Th iadiazoles

1 1 6

DISCUSSION

SUMMARY

The work described in this chapter includes the reaction of 4-amino-3-(9-

decenyl)-5-mercapto-l,2,4-triazole (1) with:

(i) p-Anisaldehyde which yielded 4-(N-4-methoxybenzylidene amino)-3-(9-

decenyl)-5-mercapto-J,2,4-triazole (16) and 2'(4-methoxyphenyl)-5-(9-

decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4]thiadiazole (17)

(ii) p-Chlorobenzaldehyde which afforded 4-(N-4-chlorobenzylidene amino)-

3-(9-decenyl)-5-mercapto-l,2,4-triazole (18) and 2-(4-chlorophenyl)-5-

(9-decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4]thiadiazole (19).

(Hi) Thiophen-2-aldehyde which furnished 4-(N-2-thienylidene amino)-3-{9-

decenyl)'5-mercapto-l,2,4-triazole (20) and 2-(2-thienyl)-5-(9-decenyl)-

2.3-dihydro-l,2,4-triazolo[3,4-b][l,3,4] thiadiazole (21).

(iv) Furfur aldehyde which gave 4-(N-2-furfurylidene amino)-3-(9-decenyl)-5-

mercapto-1,2,4'lriazole (22) as the sole product

INTRODUCTION

1,2,4-Triazole and its derivatives have been found to play vital role in

medicine, agriculture and industry^^"^^. The fused ring systems such as

i-- tr iazolo[3,4-b][l ,3,4]thiadiazoles have varied range of biological activities

as antibacterial^'•^^ antifungaP^'^®, a n t i i n f l a m m a t o r y ^ a n t i h y p e r t e n s i v e ^ ^ ' ^ ^

a n a l g e s i c " and anthelmintic^'"^^ agents. The 5-tr iazolo-thiadiazole system can

be viewed as a cyclic analogue of two very important components .

thiosemicarbazide^° and biguanide^' which often display diverse biological

activities probably due to the presence of -N=C-S group responsible for their

activities. Also, the Schiff bases and their complexes prepared by the

condensat ion of various aldehydes and ketones with substi tuted or

1 1 7

unsubsti tuted 1,2,4-triazoles have been found to be potential biologically

active agents^^'^^ as well as substrates for the syntheses of various

heterocycles of medicinal importance^^. These f indings have tempted us to

undertake this problem. It consists of the syntheses of three new compounds,

(i) 2-(4-methoxyphenyl)-5-(9-decenyI)-2,3-dihydro-l ,2 ,4- t r iazolo[3,4-b]

[ l ,3 ,4] thiadiazole (17) (ii) 2-(4-chlorophenyl)-5-(9-decenyl)-2,3-dihydro-

1.2,4-tr iazolol3,4-b][ l ,3 ,4]thiadiazole (19) and (iii) 2-(2-thienyl)-5-(9-

decenyl)-2 ,3-dihydro- l ,2 ,4- t r iazolo[3,4-b][ l ,3 ,4] thiadiazole (21) from (i)

p-anisaldehyde (ii) /?-chlorobenzaldehyde and (iii) th iophen-2-aldehyde

respectively using 4-amino-3-(9-decenyl)-5-mercapto-1.2,4-tr iazole (1) in

the presence of a catalytic amount of p -TsOH. In each reaction, the

corresponding Schiff bases (16), (18), and (20) have also been obtained. The

reaction of fur fura ldehyde with the triazole (1) yielded only the Schiff base

(22). no cyclised product could be isolated due to the decomposi t ion of the

products.

Structural assignment and biological assay are discussed. Screening

results of the compounds (16), (17), (20), (21) and (22) are summarised for

the anticancer activity against 3-cell lines of three types of human cancers:

lung, breast and CNS (details in chapter 6)

(1)

1 1 8

(16) R = 0CH3 ( 1 8 ) R = C1

( 1 7 ) R = 0 C H 3

( 1 9 ) R = C1

( 2 0 ) X - S

( 2 2 ) X = 0

( 2 1 ) X = S

1 1 9

RESULTS AND DISCUSSION

The synthesis of compounds (17), (19) and (21) was carried out by

reacting the triazole (1) (obtained by fusing 10-undecenoic acid with

thiocarbohydrazide) with /?-anisaldehyde, /7-chlorobenzaldehyde and

thiophen-2-aldehyde respectively in dry benzene using p - T s O H as catalyst. It

is believed that the corresponding Schiff base is first formed which undergoes

intramolecular cyclocondensation to give the desired product alongwith the

uncyclised Schiff base in each case (Scheme 4). In the reaction of triazole

with furfural , only the Schiff base could be isolated from the products as on

prolonging the reaction, the product underwent decomposi t ion.

N N

(16) R = 0CH3 (18) R = C1

+ N N

(20) X = S (22) X = 0

(17) R = 0CH3 (19) R = CI

(21) X = S

SCHEME 4

1 2 0

2-(4-Methoxyphenyl)-5-(9-decenyl)-2,3-(iihydro-l,2,4-triazolo[3,4-b][1,3,4]

thiadiazole FTZX' (17) along with the Schiff base, 4-(N-4-

methoxybenzylidene amino)-3-(9-decenyl)-5-mercapto-l,2,4-triazole FTZX

(16) [Reaction of the 5-triazoIe (1) with p-anisaldehyde in presence of

/j-TsOH]

A solution of the 1,2,4-triazole (1) and p-anisaldehyde (molar ratio.

1:1) in dry benzene was refluxed with stirring for 5 h with catalytic amount of

jD-T s OH. T L C examination of the reaction mixture revealed the presence of

two spots, labelled as FTZX (16) and FTZX' (17). The products on column

chromatography (pet. ether-EtOAc, 9:1 v/v) as eluent first afforded a pale

yellow crystalline solid, FTZX (18%) and then a colourless oily liquid.

FTZX' (71%) in yields. The outline of the reaction is as follows:

N N

II II

I

NH2

(1) N N

+ OHC-

FTZX (16)

i p-TsOW dry benzene Reflux 5 h

OCH3

N N

OCH3 \

HN-

FTZX' (17) Q '0CH3

structure Elucidation of FTZX (16)

It was a pale yellow needle shaped crystalline solid, m.p. 66 °C. The

structure of FTZX has been established by IR, DCI-MS, 'H-NMR and '^C-

NMR spectra. The IR spectrum (KBr) exhibited characteristic peaks at 2900

(CH), 2350 (SH), 1630 (C=C), 1600 (C=N), 1580 (phenyl) and 1480 (S-C) cm"'.

iw HflilC: 'lUi ' -VTi'r'.'"" " " r iTi111 " T T I "1K: Tu RI r wuMr . nr~ur u nciTr • i?i ^ ^ i -s7«

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1 ie!0-'i= 450560 ! I

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The DCI-MS spectrum of FTZX (Fig. 10 a) showed a [M+1]^ peak at m/z

373 confirming its molecular weight 372 [C20H28N4OS], which is equal to the

sum of the molecular weights of the ^-triazole (1) (254) and /p-anisaldehyde

(136) minus one molecule of water. This indicated the format ion of the Schiff

base by the condensation of/7-anisaldehyde with ^-triazole (1). The prominent

fragment ion peak at m/z 133 (base peak) [(M+l)-Ci2H2iN3S]"'' and a less

intense peak at m/z 240 [(M+l)-C8H8NO]'^ were arised by the exocyciic N-N

bond cleavage. The peak at m/z 206 [m/z 239-SHj'^ and another peak at m/z

115 [m/z 239-C9Hi6]'^' were obtained from the f ragment ion m/z 239 as

shown in Chart 10.

N N II

"SH

N = C H — / ( ^ N — O C H 3 m/z 373 (84.1)[M+1]

exocyciic N —N bond cleavage

N N 11

m/z 206

I 'SH

H

m/z 240 (5.0)

P-cleavage

N N

H

NEC- o -OCH,

m/z 133(100.0)

m/z 115 (26.0)

CHART 10

1 2 3

All the signals in ' H - N M R and '^C-NMR spect ra (Fig . lOb and 10c)

were ass igned to specif ic H- and C- atoms (Table 11a and l i b ) f rom their

typical chemical shif t values, mult ipl ici ty, relat ive in tegra t ions and also by

compar ing with the spectra of F T Z (1). The absence of NHa pro tons at 6 5.18

in ' H - N M R spectrum of F T Z X and appearance of new s ignals at 5 10.16 and

at 5 162.51 for benzyl idene proton and benzyl idene ca rbon conf i rmed the

format ion of the Schif f base. The other signals for p ro tons and carbons

appeared as in F T Z spectra as shown in Table 11a and l i b .

Table l ib : Comparison of the "C-NMR spectral data of FTZX (16) with that of FTZ (1)

FTZX (16) FTZ (1)

C-nr 8(ppm) C-nr 5(ppm)

3 152.85 3 153.34

5 164.22 5 168.26

6 162.51

1' 25 .62 1' 25 .40

2' 26 .73 2' 26 .68

y-T 2 9 . 2 3 - 3 0 . 3 8 y-T 29.40-30.20

8' 34 .43 8' 34 .40

9' 139.85 9' 139.82

10' 114.63 10' 114.61

Ar -1 126.36

A r - 2 , 6 131.24

Ar -3 ,5 115.45

A r - 4 163.73

OCH3 55 .98

L T

1 2 4

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Q . 1 o Q . I- "

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Based on the above evidence, the structure of FTZX was characterised

as 4-(N-4-methoxybenzylidene amino)-3-(9-deceny l)-5-mercapto-1,2,4-

triazole (16).

FTZX (16) OCHi

Structure Elucidation of FTZX' (17)

It was a colourless oily liquid. The consti tution of FTZX' has been

fully established by DCI-MS, 'H-NMR and '^C-NMR spectra. The DCI-MS

spectrum of FTZX' (Fig. 11) showed [M+1]^ peak at m/z 373 as the base

peak corresponding with the molecular formula [C20H28N4OS], indicating thai

the compound FTZX' is different from FTZX (16) in stability but same in

molecular weight. The prominent fragment ion peak at m/z 240 [(M+1)-

CgHyNO]^ and another peak at m/z 133 [(M+l)-C,2H2iN3S]^- were arised by

the cleavage of 3-4 and 1-5 bonds of the thiadiazole ring. The peak at m/z 151

[ M - C i 2 H 2 o N 4 ] ^ obtained by the cleavage of 1-2 and 4-5 bonds of the fused

ring [which was absent in the non-cyclised adduct (16)] is the diagnostic peak

of a ^•-triazolo-thiadiazole ring^^. In addit ion, it also showed the usual

/^-cleavage f ragment ion peak at m/z 115 (Chart 11).

1 2 8

lu 'JuHr : ' -L ' ' -'I"' - - Ti ' r f "vfi r ^ U f J " Z. T I f J J. * 1 J" ' 1- ^ r 'lU" t ' L- Li i

^• t , 0

I • 1

ISi .0

1 'i> O 1 — V - w « * « . . ' . 1 1 . V

I 1 1 HM-- = ^ 'rtxi 1 -- - -n -I ' \ I ' 1 1 ' '-_ 1 , J ,

I I v

- - H

! iGG.- 1 GG 1 ^ 1 >

I u I n L : . . .

-Tfi r""rV-rvf T'

" ^ r r

II <1 <TOhl C- zr! • ' '

^ilr f C'l = M1 .1 1 fiM= " ; c: cr;

r—1 ^ • ; T > — —— ; 1 ^ \ r—I 1 1 r--a 'tifl (fifi ' i n 1 "•'f. ( <5 I afi i < i;;4 < ' C.C> t MA /1 fi

1 1 1

i ' I I

ii, , I _, , , , ^ , 1 1 ^ , , , ^ , , , ^ ^ J , [—, , r—1—•

-.-.fi '••da ••'n - :<i-t ' Cit, -u-iPi ''•i 1 Ci 'idiH "^Cfl

{Mr ny

'I Ml T" ~r -!—'—r - T 1 r -

^-'fi jr-ip j^ia j ^ s a3r. Ada 451; 4£,ri 4og a-ag GLii SCAN 36 STGfiA=5 3 BACKGL'=C'Xi00 lGO%= 3325?52 -TTfr PTTY&TC;

32 n n i w> w

00 1 * A A A -

35 0 0 ~ t 0(1 - " - J A

1 53 n n fi, n r S.O

p T 34< n n 373 00

A rt r .

5 7 inn n

Fig. 11

1 2 9

m/z373 (100.0)[M+H]



N N

N = C OCH3 H

SH

m/z 133(85.2) m/z240 (42.1)

P-cleavag

H N + N N

i II H

m/z 115(11 .4)

C H A R T 1 1

S = C - O -0CH3

m/z 151 (35.2)

The 'H-NMR and '^C-NMR spectra showed signals as assigned (Table

12). The assignments of all signals to individual H- and C- a toms were made

on the basis of chemical shifts, multiplicity, relative integrations and by a

comparison with the spectral data of the uncyclised Schiff base FTZX (16)

(Tables 11a and l i b ) . The signal corresponding to thiol proton at 6 12.40 of

the Schiff base was found to be absent in the ' H - N M R spectrum of FTZX',

showing that thiol group is involved in the newly formed ring. Also, the

benzylidene proton and carbon signals shifted upfield f rom 6 10.16 to 6 8.60

and f rom 5 162.51 to 5 70.18, confirming the formation of thiadiazole ring. In

addition, both the spectra showed H- and C- signals of aromatic protons,

5-triazole ring and 9-decenyl side chain as shown in Table 12.

Table 12. ^H-NMR and ^^C-NMR spectral data of FTZX' (17)

1 3 0


Multip-licity J (Hz) C-nr 5(ppm)

6 8.60 IH S 3 151.71

NH - 6 70.18

R 2.83 2H t h\T 7.32 7a 164.33

2' 1.75 2H br p J « 7.36 R 25.54

y-T 1.30 5x2H br s 2' 26.65

8' 2.01 2H m y-T 29.22-30.37

9'

10'-HE

5.79

4.90

IH

IH

tdd

dd

J9.HZ 17.09, J9'.HB 10.23, J9',8' 6.72 JHE,9' 10.26, JHE, HZ 2.29

8'

9'

34.45

139.82 I

l O ' - H z 4.97 IH dd W 17.09, JHE, Hz 2.29 10' 114.49

Ar-2,6 7.43 2H d J A r - 2 , 6 , A r - 3 , 5 8 . 8 5 Ar-1 126.70

Ar-3,5 6.97 2H d J A r - 2 , 6 5 A r - 3 , 5 8.85 Ar-2,6 131.04

O C H 3 3.83 3H s Ar-3,5

Ar-4

O C H 3

114.65

161.19

55.67

1 3 1

Based on the above facts, the compound FTZX' was assigned the

structure as 2-(4-methoxyphenyl)-5-(9-decenyl)-2,3-dihydro-l ,2,4-triazolo

[3,4-b][ 1,2,4]thiadiazole (17).

FTZX' (17)

2-(4-Clilorophenyl)-5-(9-decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4]

thiadiazole FTZY' (19) along with the Schiff base, 4-(N-4-

chlorobenzylidene amino)-3-(9-decenyl)-5-mercapto-l,2,4-triazole FTZY

(18) [Reaction of the 5-triazole (1) with p-chlorobenzaldehyde in presence of

p-TsOH]

A solution of j-triazole (1) and /?-chlorobenzaldehyde (molar ratio,

1:1) in dry benzene was refluxed with stirring for 12 h in presence of

catalytic quantity of p-TsOH. TLC examination revealed the presence of two

spots, labelled as FTZY (18) and FTZY' (19). The products on column

chromatography (silica gel, pet. ether-EtOAc, 8:2 v/v) as eluent first

furnished a white solid, FTZY which was recrystall ized (benzene-acetone)

as white crystalline needles in 59% yield. Further elution of the column

yielded a colourless oily liquid, FTZY' in 27% yield. The react ion sequence

is as follows.

1 3 2

N- -N

+ OHC-N I

NH2

(1)

^SH o

N- -N

I N :

SH

:CH-

FTZY (18) o

p-TsOH/ dry benzene Reflux 12 h

-CI

-CI +

FTZY' (19)

Structure Eiucidation of FTZY (18)

It was a white needle shaped crystalline solid, m.p. 102 °C. The IR

spectrum (KBr) displayed characteristic absorption bands at 2850 (CH), 2400

(SH), 1640 (C=C), 1620 (C=N), 1570 (phenyl) and 1480 (S-C) cm' ' . The

DCI-MS spectrum showed a set of [M+1]"^ peaks at m/z 377/379 confirming

its molecular weight 376/378 [ C 1 9 H 2 5 N 4 S C I ] which is equal to the sum of the

molecular weights of the 5-triazole (1) (254) and p-chlorobenzaldehyde (140)

minus one molecule of water, suggesting the formation of the Schiff base. All

the signals in the 'H-NMR and '^C-NMR spectra have been assigned to

individual H- and C- atoms (Table 13) by comparing with the spectral data of

a previously established structure of compound (16) (Table 11a and l i b ) . In

'H-NMR spectrum, the absence of NH2 signals at 5 5.18 and appearance of

new signals at 5 10.49 and 5 160.28 for benzyl idene proton and carbon,

conf i rmed the formation of Schiff base. In addition, both the 'H- and '^C-

N M R spectra gave usual signals for the 9-decenyl side chain and A2B2 signals

of /7-chlorophenyl ring as shown in Table 13.

Table 12. ^ H - N M R and ^C-NMR spectral data of FTZX' (17)

1 3 3



SH 12.50 IH brs 3 153.08

6 10.49 IH s 5 163.78

1' 2.84 2H t h ' T 7.32 6 160.28

2' 1.78 2H br p J « 7.46 1' 25.58

y-T 1.31 5x2H br s 2' 26.74

8' 2.01 2H m 3'-7' 29.23-30.39

9' 5.80 IH tdd J9.,8-6.78, J9. he 10.25, J9',Hz 17.03

8' 34.43

j 10'-HE 4.90 IH dd JHE,9'10.25,JHE, Hz 2.38 9' 139.85

1

lO'-Hz 4.98 IH dd JHZ,9' 17.21, JHE, HZ 2.38 10' 114.64

Ar-2,6 7.96 2H d JAr-2,6>Ar-3,5 8-42 Ar-1 132.87

Ar-3,5 7.59 2H d JAr-2,6)Ar-3,5 8.42 Ar-2,6

Ar-3,5

Ar-4

130.82

130.21

138.60

1 3 4

Based on the above discussion, compound FTZY was character ised as 4-

(N-4-chlorobenzylidene amino)-3-(9-decenyl)-5-mercapto-l,2,4-triazole (18).

FTZY (18)

Structure Elucidation of FTZY' (19)

It was a pale yellow oily liquid. The DCI-MS spectrum of FTZY' showed a

set of [M+l]"^ peaks as the base peak at m/z 377/379 confirming its molecular weight

376/378 [C19H25N4SCI]. The fragment ion peaksat m/z 240 [(M+1)-C7H4NC1] ^ and at

m/z 137/139 [(M+O-CuHaiNBS]""' were obtained by the cleavage of 3-4 and 1-5

bonds of the thiadiazole ring. The set of peaks at m/z 155/157 [M-C|2H2oN4]^ was

obtained by the cleavage of 1-2 and 4-5 bonds of the thiadiazole ring as shown in

Chart 12.

N- -N

\ ^ 5 •

HN—

m/z 377/379 (100.0/47.3)

N-

N = C- C1 +

1 ,5 and 3,4 • • bond 1,2 and 4,5-- bond cleavage cleavage

1

- N

A

+

S = C -< -CI

m/z 137/139 (43.7/21.2)

SH H m/z 240 (28.3)

m/z 155/157 (16.4/6.8)

H

N-

P-cleavage

— N

H

A SH

m/z 115 (9.3)

CHART 12

1 3 5

All the signals in the and '^C-NMR spectra have been assigned

(Table 14). The assignments have been made on the basis of chemical shifts,

multiplicity, relative integrations and by comparison with the spectra of the

uncyclised Schiff base FTZY (18) (Table 13). The absence of signal

corresponding to the thiol proton at 5 12.50 and the shif t of benzylidene

proton (H-6) to higher field from 5 10.49 to S 8.84 suggested the formation of

thiadiazole ring. In '^C-NMR spectrum, the benzylidene carbon also shifted to

higher field f rom 5 160.28 to 5 69.23, confirming the format ion of the ring. In

addition, other signals for j:7-chlorophenyl ring and 9-decenyl side chain

appeared as usual as shown in Table 14.

Based on the above spectral evidence, FTZY' was characterised as 2-(4-

chlorophenyl)-5-(9-decenyl)-2,3-dihydro-l, 2,4-triazolo[3,4-b ] [1 ,S,4]thiad-

iazole (19)

FTZY' (19)


1 3 6



6 8.84 IH S 3 151.78

r 2.86 2H t J,. 2. 7.33 6 69.23

2' 1.76 2H br p J=«7.35 7a 163.69

y-T 1.31 5x2H br s r 25.56

8' 2.01 2H m 2' 26.62

9' 5.79 IH tdd J9.8.6.72, J9.,HE 10.23, y-T 29.22-30.37

J9.HZ 17.06 10'-HE 4.90 IH dd Jhe,9'10.23,JHE, HZ 2.28 8' 34.42

l O ' - H z 4.97 IH dd W 17.06, JHE, Hz 2.28 9' 139.83

Ar-2,6 7.54 2H d JAr-2,Ar-3,5 8.35 10' 114.51

Ar-3,5 7.37 2H d JAr-2,6?Ar-3,5 8.35 Ar-1 130.12

Ar-2,6 127.25

Ar-3,5 129.14

Ar-4 134.58

1 3 7

2-(2-Thienyl)-5-(9-decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4Jthm^^

azole FT' (21) along with the Schiff base, 4-(N-2-thienylidene amino)-3-(9-

decenyl)-5-mercapto-l,2,4-triazole FT (20) [Reaction of the s-triazole (1)

with thiophen-2-aldehyde]

A solution of the 5-triazole (1) and thiophen-2-aldehyde (molar ratio,

1:1) in dry benzene was ref luxed with stirring for 8 h with catalytic amount of

/?-TsOH. TLC examination of the reaction mixture revealed the formation of a

major and a minor components, labelled as FT (20) and FT' (21). The

products on column chromatography over silica gel using pet. e ther-EtOAc

(9:1 v/v) as eluent yielded a pink crystalline solid, FT (16%) and a colourless

oily liquid FT' (77%). The reaction sequence is as fol lows.

N N

11 II

I

NH2

(1) N N

+ OHC- W

+

/7-TSOH / dry benzene Reflux, 8 h

N N

FT' (21) I

Structure Elucidation of FT (20)

It was a globule shaped pink crystalline solid, m.p. 74 °C. The IR

spectrum (KBr) of the compound displayed characteristic bands at 2800 (CH),

2300 (SH), 1640 (C=N) and 1580 (C=C) cm"'. The DCI-MS spectrum showed

a [M+1]"^ peak as the base peak at m/z 349, showing its molecular weight 348

1 3 8

[C17H24N4S2] which is equal to the sum of the molecu la r weights of the

5-tr iazole (1) (254) and thiophen-2-aldehyde (112) minus one molecule of

water. This indicated the format ion of the Shif f base. The other f ragment ion

peaks were observed at m/z 240 [(M+1)-C5H4NS]^-, m/z 206 [m/z 239-SH] ^

and m/z 115 [m/z 239-C9Hi6]^'^in addition to the peak at m/z 109 [(M+1)-

C,2H2iN3S]^- as shown in Chart 13.

m/z349(100.0)[M+H]

N—N bond cleavage

N N

H

N = C-

SH

m/z 240 (4.3)

m/z 109(5.2)

m/z 206 (8.5) H

P-cleavage

N N

^ N - ^ S H H

m/z 115(11.6)

CHART 13

The ' H - N M R and '^C-NMR spectra (Fig. 12a and 12b) of FT showed

signals as ass igned (Table 15). The absence of the signal for amino proton at

5 5.18 and appearance of a sharp singlet for the benzy l idene proton at 5 10.60

indicated the format ion of Schiff base. In ' ^C-NMR spec t rum a signal for the

benzyl idene carbon at 5 155.88 due to the format ion of C=N bond appeared.

1 3 9

J I e

3

a 1—( ei)

1 4 0

E

a

r bS t L r t

— ^ o

< « 0 O w w . . X O C

•) M V W £ y o o o J t -<n C '^oi^tn >, uj

3 Q. < O

. c c

rf e — I

a — E u V <0 t- >

Q^tn O — O w o — 3 a :

> • 0 • - - - - — c o — e n f f J o t o — in o r v> E w w > eo V o 3 w •ooe»na vOwsOO e

** u u j ^ C « Q ( & N « > « - > X *> C > Q . t - — N i O . C M '-^tuoL-Z} « > • ' » -

— « r - a w > u i o > C - i < V 30 — fN-tOO 00<t Ct CL < > « j « 3 i u a t > > c

0 0 o

« • J (S

W)

K A (n cn I X • • • • o eo « csi •

(n •-« ^ V) < ' <0 <n —• 4

1 o i n c j «> «

. rs, o ^ tn

o o o o o « f » i D f l o « © i n u i < n r ) n w n n < v j c g c s ( w f o < o c n i • 0) m CD U> I

S tf> w m UJ - to TT

** ^ \t> ^ *o f^ <s n

<M CJ «0 « fw ^

m tn «o V 0 V us r -

P^ CM CM tn

n n

in 0 <0 r«. cn ffi

W ^ 0 CM n CM ^

4 n n t r u s r » . t n c M « r r > . « > r > . o e i

0 w w w fp .H f ^ CM en tr> 0 0 CM CM tn CM « W 0 V m in C M « A V « > < H r v r < . ( O e ) C 3 ( e O o o rr an pw m - p^ A ft ts. f CM r - in 0 e •> 00 0 «S| «S( W

V <n (O <n in

0 •> fs. «J cn f n w

10 f ^ « <n

0 a> CO tn CM CM •H

^ 0 0 0 en n f n

^ ^ v lu Wi* w 1 1 i ^^ ^^ wj '

CnCMCMCMCMCMCMCMCMCMCMCMCM

o e j e i *r I

1 4 1

Table 12. ^ H - N M R and ^^C-NMR spectral data of FTZX' (17)

H-nr S(ppm) Integ-ration


SH 12.50 IH brs 3 152.82

6 10.60 IH s 5 163.73

1' 1.19 2H t h 'T 7.33 6 155.88

2' 1.77 2H br p J « 7.45 1' 25.57

y-T 1.32 5x2H br s 2' 26.72

8' 2.02 2H m 3'-7' 29.22-30.38

9' 5.80 IH tdd J9.,8'6.59,, J9.,he 10.26, J9. Hz 17.03

8' 34.41

10'-He 4.90 IH dd JHE,9' 10.26, JHE, HZ 2.38 9' 139.83

lO'-Hz 4.98 IH dd JHZ,9' 17.03, JHE; HZ 2.38 10' 114.61

Ar-3 7.83 IH d J Ar-3) Ar-4 4.94 Ar-1 138.45

Ar-4 7.25 IH dd Jat-S,Ar-4 4 . 9 4 , JAr-4,Ar-5 3 . 6 6 Ar-3 132.53

Ar-5 7.73 IH d JAr-45Ar-5 3 . 6 6 Ar-4

Ar-5

129.12

135.28

1 4 2

support ing the formation of the Schiff base. In addition, both spectra showed

the signals corresponding to 9-decenyl side chain and thiophene ring as

shown in Table 15.

Based on the above facts, compound FT was characterised as 4-(N-2-

thienylidene amino)-3-(9-decenyl)-5-mercapto-l,2,4-triazole (20).

FT (20)

Structure Elucidation of FT' (21)

It was a colourless oily liquid. The DCI-MS spectrum of FT' showed a

[M+l]"" peak as the base peak at m/z 349 confi rming its molecular weight 348

[C17H24N4S2]. The fragment ion peaks at m/z 128 [M-Ci2H2oN4]'^' was arised

by the cleavage of 1-2 and 4-5 bonds of the thiadizole ring. The peaks at m/z

240 [ (M+O-CsHjNS]^ and m/z 109 [(M+1)-C,2H2,N3S]^- were obtained by the

cleavage of 1-5 and 3-4 bonds as shown in Chart 14.

All the signals in the 'H-NMR and '^C-NMR spectra have been

assigned to individual H- and C- atoms (Table 16) by compar ing the spectral

data with the previously established compounds FTZX' (16) and FTZY' (18)

(Tables 12 and 14). Both the spectra fully conf i rmed the formation of the

thiadiazole ring. In 'H-NMR spectrum, the absence of signal for thiol proton

at 5 12.50 and the shift of benzylidene proton signal to up f ield f rom 5 10.60

to S 8.93 indicated the formation of the thiadiazole ring. Also in '^C-NMR

1 4 3

spectrum, the shift of benzylidene carbon signal to higher f ield f rom 5 155.88

to 5 66.37, supported the formation of the ring. Both the spectra also showed

the signals for thiophene ring and 9-decenyl side chain as shown in Table 15.

N N

m/z 349(100.0)[M+1]

] -5 and 3-4 bond cleavage

] -2 and 4-5 bond cleavage

1 . N N

N + H ^SH

fee- II \

m/z 128/129 (8.7/5.5)

m/z 109(13.9)

m/z 206(11.7) H

m/z 240(12.7)

p-cleavage

"N N

^ N ^ S H H

m/z 115(11.1)

CHART 14

On the basis of above spectral evidence, FT' was characterised as

5-(9-decenyl)-2-(2-thienyl)-2,3-dihydro-l, 2,4-tnazolo[3,4-b][ 1,3,4Jthaidia-

zole (19).

FT'(21)

Table 12. ^ H - N M R and ^^C-NMR spectral data of FTZX' (17)

1 4 4


Multip-licity J (Hz) C-nr S(ppm)

6 8.93 IH s 3 151.93

r 2.81 2H t Jr,2' 7.32 6 66.37

2' 1.77 2H br p J « 7.36 7a 163.48

y-T 1.31 5x2H br s 1' 25.53

8' 2.01 2H m 2' 26.58 j

9' 5.80 IH tdd J9.8'6.71, J9.,HH 10.22, J9-,Hz 17.09,

y-T 29.22-30.37

10'-HE 4.90 I H dd W 10.22, Jhe, Hz 2.29 8' 34.46

lO'-Hz 4.97 IH dd JHZ,9' 17.09, JHE, HZ2.29 9' 139.85

Ar-3 7.60 I H d JAr-3»Ar-4 5.19 10' 114.66

Ar-4 7.07 I H dd JAR-3,AR-4 5.19 , JAR-4,AR-5 3-67 Ar-1 135.77

Ar-5 7.39 IH d JAr-45Ar-5 3.67 Ar-3

Ar-4

Ar-5

129.06

127.02

130.08

1 4 5

4-(N-2-furfuryUdene amino)-3-(9-decenyl)-5-mercapto-l,2,4-triazole F F

(22). [Reaction of the 5-triazoIe (1) with furfuraldehyde]*

*[This react ion failed to yield the desired product, 5-(9-decenyl)-2-(2-furyl)-

2,3-dihydro-l, 2,4-triazolo[3,4-b][ 1,3,4] thaidiazole. ]

A solution of the 5-triazole (1) and fur fura ldehyde (molar ratio, 1:1)

was ref luxed in dry benzene containing a catalytic amount of p -TsOH. The

progress of the reaction was monitored by TLC, which revealed the presence

of only one spot labelled as FF. The refluxing continued for fur ther 5 h. but

no progress in the reaction was observed and the solution gradually changed

into dark brown in colour. The products on purif icat ion by column

chromatography (pet. ether-EtOAc, 9:1 v/v) yielded a grey coloured

crystall ine solid, F F as the sole product in 57% yield.

N N + OHC-

'SH

(1) N H ,

N N

(

N = C H -

FF (22)

O

p-TsOH/dry benzene Reflux, 11 h

N N

1 4 6

Structure Elucidation of FF (22)

It was a globular shaped grey crystalline solid, m.p. 98 °C. The

structure of FF has been fully established by IR, DCI-MS, 'H-NMR and

'^C-NMR spectra. The IR spectrum (KBr) showed characteris t ic peaks at

2850 (SH), 1640 ( C - C ) and 1610 (C=N) cm''. The DCI -MS spectrum

displayed [M+1]"^ peak as the base peak at m/z 333 conf i rming its molecular

weight [ C 1 7 H 2 4 N 4 O S ] , which is equal to the sum of the molecular weights of

the 5-triazole (1) (254) and furfura ldehyde (96) minus one molecule of water.

This indicated the formation of the Schiff base. Other prominent f ragment ion

peaks were observed at m/z 240 [(M+1)-C5H4NS]\ m/z 206 [m/z 239-SH]"

and m/z 115 [m/z 239-C9H,6]^; in addition to the peak at m/z 93 [(M+1)-

CnH.iNsS] ' ' - as shown in Chart 15.

N N

'SH N = C H - i )

m/z333(100.0)[M+l] O Exocyclic N —N bond cleavage

m/z 206 (4.8) H^

m/z 240 (4.3)

P-cleavag

N N

m/z 115(14.7)

N ^SH

^EC-O

m/z 93(7.0)

CHART 15

1 4 7

All the signals in 'H- and '^C-NMR spectra have been assigned to

individual H-and C-atoms (Table 17) by comparing the spectra with a

previously established compound FT (20) (Table 15). The absence of NH2

proton signal at 5 5.18 in 'H-NMR spectrum and appearance of new signal at

6 10.35 for CH=N proton confirmed the formation of the Schiff base. This is

also supported by the appearance of carbon signal for C H - N at 5 150.11 in

'^C-NMR spectrum. In addition, both the spectra showed characterist ic peaks

of furan ring and 9-decenyl side chain as shown in Table 17.

Based on the above facts, compound FF was characterised as 4-(N-2-

fwfurylidene amino)-S-(9-decenyl)-5-mercapto-l,2,4-triazole (22).

FF (22)


1 4 8



SH 12.52 IH brs 3 153.03

6 10.35 IH s 5 163.62

V 2.79 2H t J,. 2' 7.32 6 150.11

2' 1.77 2H br p J « 7.46 r 25.52

y-T 1.32 5x2H br s 2' 26.61

8' 2.02 2H m y-T 29.22-30.38

9' 5.80 IH tdd J9'8'6.59, J9',HE 10.25, Hz 17.21,

8' 34.42

lO'-H,: 4.90 IH dd JHE ,9' 10.25, JHE, HZ 2.38 9' 139.85

l O ' - H z 4.98 IH dd Jh2,9' 17.21, JHC, HZ 2.38 10' 114.62

Ar-3 7.90 IH d JAr-3>Ar-4 1-83 Ar-1 149.40

Ar-4 6.73 IH dd JAr-3!Ar-4 3.48 , Jam,Ar-5 1-83 Ar-3 148.03

Ar-5 7.24 IH d JAr-45Ar-5 3.48 Ar-4

Ar-5

113.50

118.84

1 4 9

EXPERIMENTAL

4-Amino-3-(9-decenyl)-5-mercapto-l,2,4-triazole FTZ (1):

It was prepared according to the literature method^^ as described in Chapter 1.

2-(4-Methoxyphenyl)r5-(9-decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4]

thiadiazole FTZX' (17) along with the Schiff base FTZX (16) [Reaction of

the 5-triazoie (1) with p-anisaldehyde in presence of /7-TsOH]

A solution of i - t r iazole (1) (SOOmg, 1.96 mmol) and jO-anisaldehyde

(266 mg, 1.96 mmol) in dry benzene (20 ml) containing p - T s O H (5mg) as

catalyst was refluxed with stirring on an oil bath at 80 °C for 5 h using an

azeotropic water collector. The progress of the reaction was monitored by

TLC (silica gel G, pet. ether-EtOAc, 8:2 v/v) which revealed the presence of

two spots, one minor (upper) and another major (lower), labelled as FTZX

and FTZX'. The reaction mixture was then concentrated under reduced

pressure, extracted with diethyl ether and washed several t imes with water

until the solution was neutral. The ethereal solution was then kept over

anhydrous Na2S04. The solvent was evaporated under reduced pressure and

the residue left was chromatographed over a silica gel column using

(pet .ether-EtOAc, 9.5:0.5v/v as eluent). Elution of the column first afforded a

yellow solid which on crystallisation from pet .e ther-benzene yielded FTZX

as pale yellow crystalline globules, 130 mg (18.2%), m.p. 66 °C, Rf 0.76 (pet.

e ther-EtOAc, 9:1 v/v). Further elution of the column yielded a colourless oily

liquid which was further purif ied by repeated column chromatography using

pet. ether-diethyl ether (8:2 v/v) as eluent to yield FTZX', 518 mg (71.0%),

Rf 0.42 (pet. ether-EtOAc, 9:1 v/v).

1 5 0

Spectral data ofFTZX (16)

IR (KBr): 2900 (CH str), 2350 (SH), 1630 (C=C) 1600 (C-N) , 1580, 1480

(phenyl), 1500, 1460, 1410, 1290, 1250, 1150, 1000, 950, 820 cm-'.

'H-NMR (400 MHz, acetone-rf^): 6h 12.40 ( IH, br s, SH), 10.16 ( IH, s,

H - 6 ) , 2 . 8 1 ( 2 H , t , J = 7 . 3 3 , H - 1 ' ) , 1 . 7 7 ( 2 H , b r p , J « 7 . 3 1 , H - 2 ' ) , 1 . 3 2 (LOH, b r s,

H - 3 ' - 7 ' ) , 2 . 0 1 ( 2 H , m , H - 8 ' ) , 5 . 8 0 ( I H , t d d , J = 6 . 6 0 , 1 0 . 2 5 , 1 7 . 2 1 , H - 9 ' ) , 4 . 9 0

( I H , d d , J = 1 0 . 2 6 , 2 . 3 9 , H E - 1 0 ' ) , 4 . 9 8 ( I H , d d , J = 1 7 . 2 I ; 2 . 3 9 , H , - 1 0 ' ) , 7 . 8 9

( 2 H , d , J = 8 . 9 8 , H - A r - 2 , 6 ) , 7 . 1 0 ( 2 H , d , J = 8 . 9 8 , H - A r - 3 , 5 ) , 3 . 9 1 ( 3 H , s,

O C H 3 ) .

'^C-NMR (100 MHz, acetone-rf^): 5c 152.85 (C-3), 164.22 (C-5), 162.51

(C-6), 25.62 ( C - r ) , 26.73 (C-2'), 29.23-30.38 (C-3'-7'), 34.43 (C-8'), 139.85

(C-9'), 114.63 (C-10'), 126.36 (C-Ar-1), 131.24 (C-Ar-2,6), 115.45 (C-Ar-

3,5), 163.73 (C-Ar-4).

DCI-MS (NH3): m/z (%) 373 (84.), 374 (28.8), 375 (9.8), 239 (4.9), 240

(5.0), 21 1 (1 1.7), 206 (6.9), 133 (100.0), 13 (12.7), 135(7.6).

Spectral data o f F T Z X ' (17)

^H-NMR (400 MHz, acetone-rfg): 6 h 8 . 6 0 ( I H , s, H - 2 ) , 2 . 8 3 ( 2 H , t, J = 7 . 3 2 ,

H - 1 ' ) , 1 . 7 5 ( 2 H , b r p , J = 7 . 3 6 , H - 2 ' ) , 1 . 3 0 (LOH, b r s, H - 3 ' - 7 ' ) , 2 . 0 1 ( 2 H , m , H -

8 ' ) , 5 . 7 9 ( I H , t d d , J = 6 . 7 2 , 1 7 . 0 9 , 1 0 . 2 3 , H - 9 ' ) , 4 . 9 0 ( I H , d d , J = 1 0 . 2 3 , 2 . 2 9 ,

H E - 1 0 ' ) , 4 . 9 7 ( I H , d d , J = 1 7 . 0 9 , 2 . 2 9 , H , - I O ' ) , 7 . 4 3 ( 2 H , d , J = 8 . 8 5 , H - A r - 2 , 6 ) ,

6 . 9 7 ( 2 H , d, J = 8 . 8 5 , H - A r - 3 , 5 ) , 3 . 8 3 ( 3 H , s, O C H 3 ) .

'^C-NMR (100 MHz, acetone-rf^): 5c 151.71 (C-5), 70.18 (C-2), 164.33

(C-7a), 25.54 (C-1'), 26.65 (C-2'), 29.22-30.37 (C-3'-7'), 34.45 (C-8'), 139.82

(C-9'), 114.49 (C-10'), 126.70 (C-Ar-1), 131.04 (C-Ar-2,6), 114.65 (C-Ar-

3,5), 161.19 (C-Ar-4,) 55.67 ( O C H 3 ) .

1 5 1

DCI-MS (NH3): m/z (%) 372 (100.0), 373 (29.5), 374 (9.7), 341 (5.7), 239

(7.6), 240 (42.1), 241 (6.8), 206 (8.3), 151 (35.2), 152 (5.4), 153 (8.0), 133

(85.2), 134 (11.6), 135 (8.6), 136 (50.2), 137 (5.5), 128 (9.1), 115 (11.4), 80

(8.1).

2-(4-Chlorophenyl)-5-(9-decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4]

thiadiazole FTZY'(19) along with the Schiff base FTZY (18) [Reaction of

the 5-triazoIe (1) with p-chlorobenzaldehyde in presence of /7-TsOH]

A solution of the 1,2,4-triazole (1) (500 mg, 1.96 mmol) and /?-chloro-

benzaldehyde (274 mg. 1.96 mmol) in dry benzene (20 mi) containing

catalytic amount o f / ' - T s O H (5mg) was refluxed with stirring on an oil bath at

80°C for 12 h using an azeotropic water collector. The products on usual

work up gave a light yellow coloured oily residue. On examinat ion by TLC

(silica gel G, pet. ether-EtOAc, 8:2 v/v), it was found to be a mixture of two

compounds labelled as FTZY (18) (minor, upper) and FTZY' (19) (major,

lower). The oily residue was then chromatographed over a silica gel column

using pet. ether - EtOAc, 8:2 v/v as eluent. Elution of the column first

furnished a white solid, FTZY' which was crystallized f rom benzene-acetone

as white crystall ine needles, 440 mg (59.1%), m.p. 102 °C, Rf 0.68 (pet.

e ther-EtOAc, 8:2 v/v). Further elution of the column yielded a colourless oily

liquid, FTZY', 203 mg (27.3%), Rf 0.37 (pet. e ther-EtOAc, 8:2 v/v).

Spectral data of FTZY (18)

IR (KBr): 2850 (CH str.), 2400 (SH), 1640 (C=C), 1620 (C=N), 1570, 1480

(phenyl), 1450, 1410, 1320, 1240, 1160, 980, 910, 820, 750 cm"'.

'H-NMR (400 MHz, acetone-rfg): 1250 ( IH, s, SH), 10.49 ( I H , s, H-6), 2.84

(2H, t, J=7.32, H - r ) , 2.84 (2H, br p, J«7.46, H-2'), 1.31 ( lOH, br s, H-3'-7') ,

1 5 2

2 . 0 1 ( 2 H , m , H - 8 ' ) , 5 . 8 0 ( I H , t d d , J = 6 . 7 8 , 1 0 . 2 5 , 1 7 . 0 3 , H - 9 ' ) , 4 . 9 0 ( I H , d d ,

J = 1 0 . 2 5 , 2 . 3 8 , H E - 1 0 ' ) , 4 . 9 8 ( I H , d d , J - 1 7 . 2 1 , 2 . 3 8 , H , - 1 0 ' ) , 7 . 9 6 ( 2 H , d ,

J = 8 . 4 2 , H - A r - 2 , 6 ) , 7 . 5 9 ( 2 H , d , J = 8 . 4 2 , H - A r - 3 , 5 ) .

'^C-NMR (100 MHz, acetone-rf,): 6c 153.08 (C-3), 163.78 (C-5), 160.28 (C-

6), 25.58 ( C - r ) , 26.74 (C-2'), 29.23-30.39 (C-3'-7'), 34.43 (C-8'), 139.85 (C-

9'), 114.64 (C-10'), 132.87 (C-Ar-1), 130.82 (C-Ar-2,6), 130.21 (C-Ar-3,5),

138.60 (C-Ar-4).

DCI-MS (NH3): m/z (%): 3 7 7 ( 1 0 0 . 0 ) , 3 7 8 ( 2 9 . 2 ) , 3 7 9 ( 4 1 . 9 ) , 3 8 0 (1 1 .1) ,

2 3 9 ( 4 . 3 ) , 2 4 0 ( 4 . 1 ) , 2 0 6 ( 1 1 . 6 ) , 1 1 5 ( 3 . 6 ) .

Spectral data of FTZY'(19)

'H-NMR (400 MHz, acetone-^/^): 5h 8.84 (IH, s, H-2), 2.86 (2H, t, J=7.33, H-l'),

1.756 (2H, br p, J= 7.35, H-2'), 1.31 (lOH, br s, H-3'-7'), 2.01 (2H, m, H-8'), 5.80

(IH, tdd, J= 6.72, 10.23,17.06, H-9'), 4.97 (IH, dd, J=17.06, 2.29, H,-10'), 7.54 (2H,

d, J=8.35, H-Ar-2,6), 7.37 (2H, d, J-8.35, H-Ar-3,5).

'^C-NMR (100 MHz, acetone-^): 5c 151.78 (C-5), 69.23 (C-2), 163.69 (C-7a),

25.56 (C-r), 26.62 (C-2'), 29.22-30.37 (C-3'-7'), 34.42 (C-8'), 139.83 (C-9'), 114.51

(C-IC), 130.12 (C-Ar-1), 127.25 (C-Ar-2,6), 129.14 (C-Ar-3,5), 134.58 (C-Ar-4,)

DCI-MS (NH3): m/z (%) 377 (100.0), 378 (29.7), 379 (51.5), 240 (28.3), 241 (5.6),

155 (16.4), 157 (6.8), 137 (43.7), 138 (40.8), 139 (21.2).

1 5 3

2-(2-Thienyl)-5-(9-decenyl)-2,3-dihydro-l,2,4-triazolo[3,4-b][l,3,4]thiadi-

azole FT' (21) along with the Schiff base FT (20) [Reaction of the

s-triazole (1) with thiophen-2-aldehyde]

A mixture of 1,2,4-triazole (1) (500 mg, 1.96 mmol) and thiophen-2-

aldehyde (247 mg, 1.96 mmol) in dry benzene (20 ml) containing catalytic

amount of p -TsOH (5mg) was refluxed with stirring on an oil bath at SO' C for

8 h using an azeotropic water collector. The progress of the reaction was

monitored by TLC (pet. ether-EtOAc, 8:2 v/v), which revealed the formation

of one minor (upper) and another major (lower) spot labelled as FT (20) and

FT ' (21). The products on usual work up and column chromatography (silica

gel, pet. ether-EtOAc, 9:1, v/v) first afforded a pink coloured solid, FT

which was recrystallized from pet.ether-benzene as pink crystalline globules,

115 mg (16.3%), m.p. 74°C, Rf 0.82 (pet. ether-EtOAc, 8:2 v/v). Further

elution of the column using (pet. ether-EtOAc, 8:2 v/v), yielded FT' as a

colourless oily liquid, 548 mg (77.2%), Rf 0.44 (pet. ether-EtOAc, 8:2 v/v).

Spectral data of FT (20)

IR (KBr) :2800 (CH str), 2300 (SH), 1640 (C=C), 1690 (C=N), 1580, 1490

(phenyl), 1420, 1400, 1350, 1270, 1200, 1150,1 100, 1040, 980, 890, 830,

800, 750, 700 cm"'.

'H-NMR (400 MHz, acetone-RF^): 5h 12.50 ( IH, s, SH), 10.60 ( I H , s, H-6),

2 . 7 9 ( 2 H , t , J = 7 . 3 3 , H - 1 ' ) , 1 . 7 7 ( 2 H , b r p , J « 7 . 4 5 , H - 2 ' ) , 1 . 3 2 ( L O H , b r s , H - 3 ' -

7 ' ) , 2 . 0 2 ( 2 H , m , H - 8 ' ) , 5 . 8 0 ( I H , t d d , J = 6 . 5 9 , 1 0 . 2 6 , 1 7 . 0 3 , H - 9 ' ) , 4 . 9 0 ( I H ,

d d , J - 1 0 . 2 6 , 2 . 3 8 , H E - 1 0 ' ) , 4 . 9 8 ( I H , d d , J = 1 7 . 0 3 , 2 . 3 8 , H , - 1 0 ' ) , 7 . 8 3 ( I H , d ,

J = 4 . 9 4 , H - A r - 3 ) , 7 . 2 5 ( I H , d d , J = 4 . 9 4 , 3 . 6 6 , H - A r - 4 ) , 7 . 7 3 ( I H , d , J = 3 . 6 6 , H -

Ar-5).

1 5 4

'^C-NMR (100 MHz, acetone-cffi): 5c 152.82 (C-3), 163.73 (C-5), 155.88

(C-6), 25.57 ( C - r ) , 26.72 (C-2'), 29.22-30.38 (C-3'-7'), 34.41 (C-8'), 139.83

(C-9'), 114.61 (C-IC) , 138.45 (C-Ar-1), 132.53 (C-Ar-3), 129.12 (C-Ar-4),

135.28 (C-Ar-5).

D C I - M S (NHj) : m/z (%) 349 (100.0), 350 (26.2), 351 (13.2), 240 (4.3), 206

(8.5), 128 (6.7), 115 (11.6).

Spectral data of FT'(21)

• H - N M R (400 MHz, acetone-rf^): 5h 8.93 ( IH, s, H-2), 2.81 (2H, t, J=7.32,

H-1'), 1.77 (2H, br p, J= 7.36, H-2'), 1.31 (lOH, br s, H-3'-7') , 2.01 (2H, m,

H-8'), 5.80 ( IH, tdd, J=6.71,10.22, 17.09 H-9'), 4.90(1H, dd, J= 10.22, 2.29,

HE-10'), 4.97 ( IH, dd, J=17.09, 2.29, H,-10'), 7.60 ( IH, d, J=5.19, H-Ar-3),

7.07 ( IH, dd, 1=5.19,3.67, H-Ar-4) 7.39 ( IH, d, J=5.19, H-Ar-5).

' ' C - N M R (100 MHz, acetone-^/,): 6c 151.93 (C-5), 66.37 (C-2), 163.48 (C-

7a). 25.53 (C-1'), 26.58 (C-2'), 29.22-30.37 (C-3'-7'), 34.46 (C-8'), 139.85 (C-

9'), 114.66 (C-10'), 135.77 (C-Ar-1), 129.06 (C-Ar-3), 127.02 (C-Ar-4),

130.08 (C-Ar-5)

DCI -MS (NH3): m/z (%) 349 (100.0), 350 (25.4), 351(12.5), 334 (7.8), 239

(4.4), 240 (12.7), 206 (11.7), 129 (5.5), 128 (8.7), 1 15 (11.1), 1 12 (9.1), 109

(13.9).

4-(2-N-furfurylidene amino)-3-(9-decenyl)-5-mercapto-l,2,4-triazole, FF

(22). [Reaction of the 5-triazole (1) with furfuraldehyde]

A solution of 1,2,4-triazoIe (1) (500 mg, 1.96 mmol) and

furfuraldehyde (188 mg. 1.96 mmol) in dry benzene (20 ml) containing

catalytic amount of /?-TsOH (5mg) was refluxed with stirring on an oil bath at

80 °C for 6 h using an azeotropic water collector as described above. The

1 5 5

progress of the reaction was monitored by TLC (pet. ether-EtOAc, 8:2 v/v) at

every 15 min. TLC revealed the formation of only one spot, labelled as FF.

So, the refluxing continued for further 5 h to see the development of another

spot but no progress in the reaction was observed and the colour of the

solution gradually changed into dark brown and a black precipitate started

depositing on the sides of the round bottom flask. This may be due to the

decomposition of the furfural moiety in presence of strong acid (p-TsOH).

The reaction was stopped and the colour of the solution was removed by

passing through the activated charcoal. It was worked up as usual and the

brown oily residue thus obtained was chromatographed over a silica gel

column using pet. ether-EtOAc, 9:1, v/v as eluent which furnished a grey

coloured crystalline solid, FF (22). it was recrystallized from pet. ether-

benzene as grey crystalline globules, 371 mg (57.0%), m.p. 98°C, Rf 0.69

(pet. ether-EtOAc, 8:2 v/v).

Spectral data of FF (22)

IR (KBr): 2850 (CH str.), 2250 (SH), 1640 (C=C), 1610 (C=N), 1580, 1490

(phenyl), 1460, 1400, 1320, 1260, 1220, 1150, 1100, 1070, 1020, 980. 820,

750 cm"'.

'H-NMR (400 MHz, acetone-^/^): 5h 12.52 ( IH, s, SH), 10.35 ( IH, s, H-6).

2 . 7 9 ( 2 H , t , J = 7 . 3 2 , H - T ) , 1 . 7 7 ( 2 H , b r p , J « 7 . 4 6 , H - 2 ' ) , 1 . 3 2 (LOH, b r s , H - 3 ' -

T ) , 2 . 0 2 ( 2 H , m , H - 8 ' ) , 5 . 8 0 ( I H , t d d , 1 = 6 . 5 9 , 1 0 . 2 5 , 1 7 . 2 1 , H - 9 ' ) , 4 . 9 0 ( I H ,

d d , J = 1 7 . 2 . 1 , 2 . 3 8 , H E - 1 0 ' ) , 4 . 9 8 ( I H , d d , J = 1 0 . 2 5 , 2 . 3 8 , H ^ - I O ' ) , 7 . 9 0 ( I H , d ,

J = 1 . 8 3 , H - A r - 3 ) , 6 . 7 3 ( I H , d d , J = 3 . 4 8 , 1 . 8 3 , H - A r - 4 ) , 7 . 2 4 ( I H , d , J = 3 . 4 8 , H -

Ar-5).

1 5 6

' ^C-NMR (100 MHz, acetone-rfg): 5c 153.03 (C-3), 163.62 (C-5), 150.11

(C-6), 25.52 ( C - r ) , 26.61 (C-2'), 29.22-30.38 (C-3'-7'), 34.42 (C-8'), 139.85

(C-9'), 114.62 (C-10'), 149.40 (C-Ar-1), 148.03 (C-Ar-3), 113.50 (C-Ar-4),

118.84 (C-Ar-5).

DCI-MS (NHa): m/z (%) 333 (100.0), 334 (23.9), 335(8.3), 240 (3.2), 206

(4.8), 128 (8.4), 115 (14.7), 96 (12.5), 93 (7.0).

1 5 7

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1 6 4

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0 A T ^ l A l i

2,3-DISUBSTITUTED THIAZOLIDIN-4-ONES/THIAZAN-4-ONES

OAni 11 J r ,

1 6 5

THEORETICAL

4-Thiazolidinones and tetrahydro-l,3-thiazin-4-ones (called as 4-

Thiazanones) are two important classes of biologically active five and six

membered sulphur and nitrogen containing heterocyclic compounds with a

carbonyl group in the 4-position (I) and (II).

3 4 0 H N — f

3 4 0 3 HN ^ HN

I III V

0 0 0 3 IJ4 3

h n ^ N hn-2L Ja

1 J 6

1

II IV VI

0 , (2,4-thiazolidine-dione / th iazan-2,4-dione) V B / V I B , X = S, (2-thiono-4-thiazolidinone or rhodanine /2- th iono-4- th iazanone) V C / V I C , X = NR, (2- immo-4-thiazol idinone or pseudoth iohydantom / 2- imino-4- th iazanone) R V D / V I D , X = N - N = C ^ (2-hydrazmo-4- th iazol id inone/2-hydraz ino-4- th iazanone)

Substituents in the 2, 3 and 5-positions may be varied, but the greatest

difference in structures and properties is exerted by the group attached to the

carbon atom in the 2-position (Rj and R2 in formula II I , IV and X in formula

V, VI). Variations in the substituents attached to the nitrogen atom and the

methylene carbon atom are possible for the structures represented by

formulas III , IV, V and VI.

4-Thiazolidinones have been the subject of extensive study in the

recent past. Numerous reports in the form of chemical reviews''^ and review

articles^'"''^ have appeared in the literature which highlight their chemistry and

1 6 6

use, especially syntheses, reactions and biological activity. 4-Thiazolidinone

derivatives have been found to be associated with diverse biological activities

such as bactericidal^"'", fungicidal^"'", insecticidal"" '^, pesticidal''*,

herbicidal '^ ' '^ antitubercular'^"'^, antiinflammatory^""^^ antitumor^'',

antipsychotic^^"^^, antiviral^^"^", anti-convulsant^'"^'*, anthelmintic^^'^^

respiratory activity^^'^^ antidiarrhoeal^^ and antiarthritic^^. A number of

thiazolidinone derivatives has also been found to exhibit highly potent anti-

anti-histaminic'^^"'''', antidiabetic'^^''"', oxygenase inhibitory^^, calcium

antagonist'^^ potassium inhibitory"^', PPAR agonist '", cardioprotect ive ' ' ,

antiischemic activity" ' '^ and as promising agent for treating Alzheimer

disease'^^, cancer^'* and viral diseases like AIDS^^'^"'^"' and hepatitis-B^'.

Although 4-thiazolidinones have established chemistry, there is only

sporadic reports on the anaologous 4-thiazanones. They are also associated

with wide range of biological activities such as treatment of cardiac

arrhythmia''^, tranquilizer^^ anticoagulant^^, treating peptic u lcer '^

antimicrobial^^"^', antiarthritic®^, anti-rheumatic^^, pesticidal activity'"''

immunoregulatory agent^^ and for treating occular inflammation^®.

Several methods for the preparation of 4-thiazolidinones and 4-

thiazanones have been reported in the literature which are based mainly on

the reaction of:

1. a/p-mercaptoalkanoic acids with

(a) Schiff bases

(b) isothiocynates

(c) isocynates

(d) cyanamideS

1 6 7

2. a/p-halo- or a/p-hydroxy alkanoic acids, a , p-unsaturated acids or their

derivatives with

(a) dithiocarbamates

(b) thiocarbamates

(c) thioureas

(d) thiosemicarbazones

(e) alkali thiocynates

a/p-Mercaptoalkanoic acids have been extensively used for the

syntheses of 4-thiazolidinones/4-thiazanones. The substituted and

unsubstituted a/p-mercaptoalkanoic acids react conveniently with Schiff

bases of aromatic or heterocyclic aldehydes or ketones and aliphatic or

aromatic amines in different solvents to give a variety of 2-substituted-4-

thiazolidinones

Rj HS-C-COOH

I R.

Ry C = N - R

R2-

HS—CH-CH2-COOH

H R - N : ^ >

0

R3

S" R4

VII

-H20

0 R A

N ' RK

R /

VIII

R = alkyl, aryl or heterocyclic Rj = H, alkyl, aryl or heterocyclic R2 = alkyl, aryl or heterocyclic R3 = H, alkyl

R4 = H, alkyl

Hydrazones of aldehydes or ketones react with substituted or

unsubstituted a/p-mercaptoalkanoic acids to form 2-substituted or 2,2-

disubstituted 4-thiazolidinones (IX) ' "'®' or 4-thiazanones (X)®'.

1 6 8

R - N H - N = C R,

•Ri

HS-CH-COOH

- H 2 O

R

H S - C H - C H 2 - C O O H

-H2O

R-NH.

R,

R2-

N R,J R2

,0 'N-

S' IX

•R3

o

"s"

X

~R3

R = alkyl, aryl or heterocyclic R2 = alkyl, aryl or heterocyclic

Ri = H, alkyl, aryl or heterocyclic R3 = H, alkyl R4 = H, alkyl

a-Mercaptoacetic acid or p-mercaptopropionic acid reacts with the

aldehyde or ketone in the presence of ammonium salts such as ammonium

carbonate or acetate in inert solvent. The reaction seems to proceed by the

formation of an aldimine or a ketimine intermediate followed by the

cyclisation with a-mercaptoacetic/p-mercaptopropionic acid to form 2,2

disubstituted-4-thiazolidinones (XI)^''^^-^^/4-thiazanones (XII)

HS CHj COOH/NH3 HN-

O

-H20

HS CH2 CH2COOH/NH3

-H2O

Ri ^ S - ^

XI

o

HN R

R, ^S-^ R3

XII

R = Her alkyl Rj-a lkyl or aryl

a-Mercaptoacetamide reacts with the carbonyl function of aldehydes in

the presence of catalytic amounts of p-toluenesulphonic acid or

1 6 9

borontrifluoride ethereate in inert solvent to give 2-substituted-4-

thiazolidinones (XIII) '^ '^^

O H I HS.CH2.CONH2

R - C = 0

H N -

-H2O R-

S"

XIII

R = alkyl, aryl

a-Mercaptothiocarboxylic acids can also be used for the synthesis of 4-

thiazolidinones (XIV) by the reaction with different Schiff bases^°.

.0 HN-

R, R HS.CH^.COSH^

-"2S Ri "S' XIV

R = alkyl or aryl Ri = H or alkyl R2 = alkyl or aryl

A number of spiro-4-thiazolidinones (XV, XVII ) and 4-thiazanones

(XVI, XVII I ) have been prepared by the reaction of cyclic ketones with

a-mercaptoacet ic acid or P-mercaptopropionic acid in presence of ammonium

salts or substituted amines^'"^'*.

HS.CH2.COOH

-(CH2)

o

(NH4)2C03/ R-NH2

HS.(CH2)2.C00H

(NH4)2C03/ R-NH2

XV, XVI: R = H

XVII, XVIII: R = alkyl or aryl

1 7 0

Substituted and unsubstituted a/p-mercaptoalkanoic acids, their esters

react smoothly with the compounds containing activated nitrile groups in the

presence of an equivalent amount of alcoholate to give 4-thiazolidinones

(XIX)^^ or 4-thiazanones (XX)^^

R 4 R 3 C H - C E N -

HS-C-COOR I R2

.0 X I

®0R R4R3CH

Ri

R2

0

H S - C H - C H ; - COOR

®0R

N

RfRjHC^S'

.0 H N -

R4

RI

S" R2

XIX

0

H N ' "

XX

R = Alky],

R3 = H or aryl ,

Ri = H or alkyl, R2 = H or alky! or aryl

R4 = COOEt, CONH2, CN, Pyridyl, Indoyl or Ph

Monforte et al.^^ have synthesized 4-thiazolidinones by reacting

carbodiimides with a-mercaptopropanoic acid. The acid reacts with one of the

two carbodiimidic C=N groups to give 3-substi tuted-2-imino-5-methyl-4-

thiazolidinones (XXI).

c„, H S — C H - C O O H

R - N = C = N - R — -H2O R _ N ' ^s"

R = alkyl or aryl XXI

Substituted 2-imino-4-thiazolidinones (XXII) or 2-imino-4-thiazanones

(XXIII) are obtained in good yields by the reaction of symmetrical and

unsymmetrical thioureas with various substituted and unsubsti tuted a /p -

1 7 1

haloalkanoic acids, their esters, acid chlorides, amides and carbamates. The

reaction proceeds via the intermediate isothiourea which cyclise in the

presence of sodium acetate or pyridine^^''°^

R - N H - C - N H R ,

SH I

R ^ N H - C = N R ,

X - C - C O Y I R3

- H X

O N H

R , N ^s-' 'R2

XXII

X - C H - C H 2 - C O Y

o

- H X , - H Y R i N ^ S '

XXIII

R = alkyl, aryl or heterocycHc R2 = H or alkyl

R] = alkyi, aryl R3= H or alkyl

A number of fused thiazolidin-4-ones (XXIV) or fused thiazan-4-ones

(XXV) have been prepared by the reaction of cyclic thioureas with various

substituted a//?-haloalkalnoic acids and their derivatives^^"^' '®'*"'®'.

X-CH2-C0Y

HN NH

R = H or alky! X = Halo-Y = CI or OR

- H X , - H Y

ACmrf.

N N

X.CH2.CH2.COY

-HX, -HY

X X I V

x(CH2)n>

N N V

X X V

.0

2-Imino-4-oxo-5-thiazolidinoneacetic acid (XXVI) can be conveniently

prepared from dicarboxylic acids'"^. a-Bromosuccinic acid on treatment with

thioureas in the presence of sodium acetate in methanol gives 2-imino-4-oxo-

S-thiazoUdinoneacetic acid.

1 7 2

R N H - C - N H 2 R N = C - N H 2 + H 0 0 C - C - C H , C O O H -I ^

0 Br

- HBr

HN ^ "

XXVI

R = H, alkyl or aryl

Recently, Madhukar S. Chande et have synthesized 3-amino-2

alkyl/arylimino-5-carbethoxymethyl thiazolidin-4-one (XXVII) by the

reaction of diethylbromomalonate (DBM) with 4-arylsubstituted

thiosemicarbazide in ethanol in the presence of pyridine.

H2N

RNH-C-NHNH2+H5C200C"^~CH2COOC2H5 I \

XXVII

R = H or aryl

Thioureas add to the unsaturated carbon-carbon linkage of maleic,

fumaric, citraconic acids, their esters, imides and anhydrides probably by a

Michael type reaction and cyclisation to give a 2-imino-4-oxo-5-

thiazolidineacetic acid (XXVII I ) """"^

n HN

s R N H - C - N H 2 + H O O C - C H = C H ^ O O H ~ = Y 2 0

RN/ X g X ^CHjCOOH

R = H, alkyl

1 7 3

Dimethyl acetylenedicarboxylate (DMAD) reacts readily with

substituted thioureas, thiosemicarbazides and thiosemicarbazones to give 4-

thiazolidinones (XXIX)""

R N H - C - N H R + H3COOC - C = C -COOCH3

R=H, C H 3 , C g H j C H j or C ^ H j

C H C O O C H ,

Dihydro-l,3-thiazin-4-one derivatives (XXX) and (XXXII) are formed

when alkyl propiolates were added to thioureas and dithiocarbamic acids

respect ively"^ In the latter case, it is necessary to cyclize the initial products

(XXXI) with acetic anhydride.

o

R i - C = C — C O O E t -

R2NRCS.NHR2 - E t O H

NHt.CS2H

Ri - alkyl

NR2

Ri ^ S ' ^ N R z

XXX

[ - E t O H

Rr - S ^ S

XXXI

o N H

XXXII

When the thiosemicarbazone of aldehyde or ketone is allowed to react

with substituted or unsubstituted a-haloalkanoic acids, their esters, amides in

the presence of sodium ethoxide or sodium acetate, the product formed is

aldehyde or ketone derivative of 2-hydrazino-4-thiazoiidinone (XXXIII)"^'"^

or 4-thiazanone (XXXIV)"'".

R.NH,CS.NH.N=C .Ri

R2

SH I

R - N H - C = : N - N R2

R = H or aryl R2 = alkyl or aryl

X-CH-COY Ri

- H X , - H Y R2 \

X-CH,-CH,-COY

-HX, - H Y RI

R] = H, alkyl or aryl R3 = H, alkyl or aryl

1 7 4

R. .0 •N-

: N - N

XXXIII

O

HN '

R3

> XXXIV

A number of exonuclear thiazolidin-4-ones (XXXV) have been

prepared by the reaction of thiosemicarbazones of cyclic ketones with

substituted and unsubstituted a-haloalkanoic acids in the presence of sodium

acetate or sodiumethoxide"^"'^".

S R2 ^ II r - H X A A

N - N H - C — N H ~ R I + X - C H - C O Y — ^ ( C H J ) , , J =

Rlx O

N - N Rj

Ri=H,Aiyl, alkyl R2 = H, alkyl, aiyl XXXV

When a,p-unsaturated acyl isothiocynates were reacted with alkyl

amines or aromatic amnes, it gave the corresponding thioureas (XXXVI)

which on intramolecular cyclization under different conditions (BF3, NaOEt

etc.) yielded the l,3-thiazin-4-one^^'- '" (XXXVII)

o

NCS + ^NH nhcs.n;

XXXVI

.R2 -R3

N R7

R3 XXXVII

R = Alkyl, aryl, heterocyclic Ri = H, alkyl, aryl R2 = Alkyl, aryl R3 = H, alkyl, aryl

1 7 5

A.F.S. Ahmad'^'^ et al. have reported that the reaction of cinnamoyl

isothiocyanate with 4-pyridine carbohydrazide yielded N-cinnamoyl triazole

(XXXIX) presumably via the non-isolable thiosemicarbazide (XXXVIII ) .

Base induced cyclization of (XXXIX) resulted in the intramolecular

cycloaddition of thiolate to the cinnamoyl moiety affording triazolo-thiazin-

4-one (XL).

o o

^NCS N

'NH.NHj

HN - N H

P h -

NHo""

O N

(XXXVIII)

N N T N N

HS^^N'

o ,

(XL) (XXXIX)

A number of 2-pyridyl/aryl substituted 4-thiazolidinones (XLI) have

been synthesized by Tanabe et al" ' in 1995 following the method of surrey^^

by condensing a-mercaptoalkanoic acids with aldehydes and primary amines

or with corresponding Schif f s bases in refluxing benzene or toluene with

continuous removal of water being formed and found to exhibit highly potent

and selective anti-Platelet Activating Factor activity both in vitro and in vivo.

Me H Ri I

R — C H = N - M e HS.CH.COOH

0 Me XLI

R = 3-pyridyl, 4-CI.C6H4, 4-N02 C6H4, 4-MeO.C6H4

1 7 6

S.M. Osman and coworkers have prepared some long alkyl chain

substituted thiazoIidin-4-ones and thiazan-4-ones from modified fatty acids as

part of their programme of preparing fatty acid derivatives^^"^^ They^^ have

reported the synthesis of long alkyl chain substituted thiazolidin-4-ones

(XLII) and thiazan-4-ones (XLIII) by reacting 10-oxoundecanoic acid and

12-oxooctadecanoic acid with mercaptoacetic acid and 3-mercaptopropionic

acid in presence of ammonium carbonate and dry benzene.

HS.CH2.COOH

R V o

RI

(NH4)2C03 /benzene

R Ri X "

H N ^ ^ S

O

HS .(CH2)2 .C00H

XLII

R ^ R , H N ^ ^ S

(NH4)2C03 /benzene O'

XLIII R =CH3, CH3(CH2)8,CH3 (CH2),6

Ri = (CH2)g.COOCH3, (CH2)7.C00CH3

Similar reaction of 9,10-dioxooctadecanoic acid with mercaptoacetic

acid or 3-mercaptopropionic acid and ammonium carbonate gave two

isomeric thiazolidin-4-ones (XLIVa) and (XLIVb)^^'^"' or thiazan-4-ones

(XLVa) and (XLVb). 0

R i

HS .CH2 .COOH

O

Ri

O

(NH4)2C03 /benzsne

R - X H N ^ S

W o

XLIVa O

H S . ( C H 2 ) 2 . C 0 0 H

RI

(NH4)2C03 /benzene 0 ' XLVa XLVb

R = C H 3 ( C H 2 ) 7

R i = ( C H 2 ) 7 . C 0 0 C H 3

1 7 7

J. Mustafa and S.M. Osman^^ have reported the preparation of a mixture

of isomeric thiazan-4-ones (XLVIa) and (XLVIb) by reacting a,P-

unsaturated fatty acid, (£')-methyl-4-oxooctadec-2-enoate with 3-

mercaptopropionic acid and ammonium carbonate.

O

R

H

A r , H

R=CH3(CH2),3 RI = COOCH3

HS.(CH2)2.C00H

(NH4)2C03 ^enzene

XLVIb

They have also prepared a mixture of thiazolidinedione (XLVII) and a

thiazole (XLVIII) by heating a solution of (£)-methyl-4-oxooctadec-2-enoate

with thiourea, sodium acetate, and dilute hydrochloric acid under reflux

condition 85

o H

R

H

H2N.CS.NH2

AcONa, EtOH' dil. HCI, reflux

R=CH3(CH2)I3

RI = COOCH3

R.

O

O 7 /

R

+ S NH

O

XLVII

o 7 /

o

S NH

O

XLVIII

W.H. Ansari and co-workers^®"^^ have synthesised two novel

compounds 2-[3-carboxyethylthio-2-(phenyl)ethyl]-2-(4-chlorophenyl)tetra-

hydro-l,3-thiazin-4-one^^ (XLIX) and 2-[2-carboxymethylthio-2-(4-chloro-

1 7 8

phenyl)ethyl]-2-(4-chlorophenyl)thiazolidin-4-one^^''' ' ' (L) as two novel

compounds from 4,4'-dichlorochalcone using 3-mercaptopropionic acid/

mercaptoacetic acid in presence of ammonium carbonate. In 2002, they have

synthesised a new compound, 2-[2,2-bis(4-chlorophenyl)ethyl]-2-(4-Q n

chlorophenyl)thiazolidin-4-one (LI) from 4,4'-dichlorochalcone via 1,3,3-

tris(4-chlorophenyl)propan-l-one using mercaptoacetic acid in presence of

ammonium carbonate in dry benzene. Compounds (L) and (LI) were studied C 7K B B

for cyctotoxic activity ' against 60 cell lines of nine types of human

cancers. Noteworthy results were obtained in the case of melanoma, colon

and renal cancers where in (LI) the reduction in growth was 52, 80 and 91

respectively.

bll — /m

. i ^ J I .

2-Aryl'3-N-Long alkyl chain substituted-Thiazolidin-4-ones/Thiazan'4'One

179

DISCUSSION

S U M M A R Y

The work described in this chapter includes the reaction of

m-nitrobenzaldehyde-10-undecenohydrazone (24) with:

(i) Mercaptoacetic acid in dry benzene which furnished an adduct, m-nitro-

benzaldehyde-1 l-(carboxymethylthio)undecanohydrazone (25) and a

novel compound, 2-(3-nitrophenyl)-3-[ll-(carboxymethylthio)undecana-

mido]thiazolidin-4-one (26).

(ii) 2-Mercaptopropiomc acid in dry benzene which yielded two novel

compounds, 2-(3-nitrophenyl)-5-methyl-2-(10-undecenamido)thiazolidin-4-

one (27) and 2-(3-nitrophenyl)-5-(methyl)-3-[l l~{(l-carboxy)ethylthio}

undecanamido]thiazolidin-4-one (28).

(iii) 3-Mercaptopropionic acid in dry benzene which afforded an adduct,

m-nitrobenzaldehyde-11 -(2-carboxyethylthio)undecanohydrazone (29) and

a novel compound, 2-(3-nitrophenyl)-3-[ll-(2-carboxyethyl-thio)undecan-

amido]thiazan-4-one (30).

The hydrazone (24) as a substrate was prepared from 10-undecenoicacid

by treatment with hydrazine hydrate in the presence of MeOH and acid to give

10-undecenoacid hydrazide (23) which on condensation with m-nitro-

benzaldehyde yielded the hydrazone (24) as yellow globules in 87% yield. It was

found to exist in two stereoisomeric forms as synperiplanar (major) and

antiperiplanar (minor) as are evident from its stereochemical studies.

1 8 0

INTRODUCTION

A survey of literature revealed that in recent years attention has been

focussed on the synthesis of analogs of naturally occurring long alkyl chain

substituted heterocycles'^^""® and the interest in substituted thiazolidin-4-ones

and thiazan-4-ones in particular for medical application is increasing

strongly. The compound, (-) 2-(5-carboxypentyl)-4-thiazolidinone known as

actithiazic acid is an antibiotic which was isolated'^^®'*' from the culture broth

of a strain of streptomyces and found to show highly specific in vitro activity

against Mycobacterium tuberculosis. Another compound, 2-(4-chlorophenyl)-

3-methyl-4-thiazanone-l,l-dioxide, known by the generic name

chlormezanone, is an important pharmaceutical compound and is widely used

as a tranquilizer^^ which is effective in the treatment of anxiety and tension

states. It has also been claimed to be of beneficial in muscle spasm"' and

effective as anticoagulant^^.

CI

Q o,s ' N - M e

O

Actithiazic acid Chlormezanone

During the past few years, several 2-sbustituted thiazolidin-4-ones and

thiazan-4-ones have been synthesized and found to possess anticonvulsant^'*,

antidiarrhoeal^^, antihistaminic'*^''*'', antimicrobial^"'®'^^ oxygenase

inhibitory''^, calcium antagonist'*^ and K" channel inhibitory activity'*^

Recently, a number of 2-pyridyl substituted 4-thiazolidinones has been

synthesized'*' and found to exhibit highly potent and selective anti-Platelet

1 8 1

Activating Factor activity both in vitro and in vivo. Although a large number

of reports''^ are available for the aromatic and heterocyclic substituted

thiazolidin-4-ones and thiazan-4-ones, there is only sporadic reports on long

alky] chain substituted thiazolidin-4-ones and thiazan-4-ones^^'^^ Also a

number of long chain alkyl thioethers have been reported for their

b a c t e r i c i d a l f u n g i c i d a l ' ^ ^ \ tranquilizer'^^, anti-cholestermic'^^

analgesic'^® and antidepressant'^® activities.

As part of our programme in search of biologically active compounds

with sulphur and nitrogen containing heterocycles, we have synthesised 2-[2-

carboxymethylthio-2-(4-chlorophenyl)ethyl]thiazolidin-4-one as a novel

compound^^. Because 4-thiazolidinones substituted in the 2-position were

proven to be biologically very potent and selective"" and in anticipation that

by incorporating a long chain alkylthioether moiety to thiazolidin-4-one ring,

the biological activity may be enhanced due to lipophilization of the resulting

molecule, we have undertaken this problem. It consists of the synthesis of the

following four novel compounds, (i) 2-(3-nitrophenyl)-3-[l l-(carboxy-

methylthio)undecanamido]thiazolidin-4-one (26), (ii) 2-(3-nitrophenyl)-5-

(methyl)-3-(10-undecenamido)thiazolidin-4-one (27) and 2-(3-nitrophenyl)-5-

(methyl)-3-[l l-{(l-carboxy)ethylthio}undecanamido]thiazolidin-4-one (28),

(iii) 2-(3-nitrophenyl)-3-[l l-(2-carboxyethylthio)undecanamido]thiazan-4-

one (30) from m-nitrbenzaldehyde-lO-undecenohydrazone (24) using

mercaptoacetic acid in (i), 2-mercaptopropionic acid in (ii) and

3-mercaptopropionic acid in (iii) in dry benzene. An adduct,

m-nitrobenzaldehyde-ll-(carboxymethylthio)undecanohydrazone (25) in the

former and w-nitrobenzaldehyde-11-(2-carboxyethylthio)undecanohydrazone

(29) in the latter was also obtained in good yields.

1 8 2

The hydrazone (24) as a substrate has been prepared by the reaction of

10-undecenoacid hydrazide (23) with w-nitrobenzaldehyde and its

stereostructure discussed. Structural assignment, stereochemistry and

biological assay are discussed. Screening results of the compounds (24), (26),

(27), (28) and (30) are summarised for cytotoxic activity against 3-cell lines

of three types of human cancers: lung, breast, CNS and then 60 cell lines of

nine types of human cancers: leukaemia, lung, colon, CNS, melanoma,

ovarian, renal, prostate and breast. Screening results of the above compounds

are also summarised for antimicrobial activity against four bacteria,

Escherichia coli. Bacillus subtilis, Staphylococcus aureus, Pseudomonas

aeruginosa and one fungi Candida albicans (details in Chapter 6)

(24)

1 8 3

N02

H CH3

1 8 4


The syntheses of compounds, (25)-(30) have been performed in two

steps (Scheme 1). The hydrazone FB (24) was first prepared following a

published procedure^ by refluxing a solution of 10-undecenoacid hydrazide

(23) in dry benzene with w-nitrobenzaldehyde (molar ratio, 1:1), as yellow

crystalline needles in 87% yield. The hydrazone FB (24) was then reacted

with (i) mercaptoacetic acid, (ii) 2-mercaptopropionic acid and (iii)

3-mercaptopropionic acid (molar ratio, 1:3) in dry benzene. The products on

column chromatography over silica gel using pet. ether-diethyl ether as eluent

yielded compounds FBX,B (25) and FBX,C (26) in (i); F B X 3 A (27) and

F B X 3 B (28) in (ii), and FBX^B (29) and FBX2C (30) in (iii). In the reaction

of (24) with 2-mercaptopropionic acid, it was observed (TLC examination)

that condensation competes addition, yielding (27) as the first product and

then (28). The increased nucleophilicity of the thio group in 2-mercapto-

propionic acid probably facilitates condensation.

1 8 5

( N CN -C

S I X' X . CN 3 1) <U ^ S u S JJ c ^ y

, rn -O

2 d « i o E

I U

I u O O X u u

u u lyi cA ly^ XXX

o t i Ci ^

^ ^ w w

c

u X u C/2

m-Nitrobenzaldehyde-lO-undecenohydrazone FB (24)

The outl ine of its synthesis is as fol lows

1 8 6

O O MeOH/H""

'OH NH2.NH2.H2O ^NH-NH2 (23)

dry benzene Reflux, 5 h

O

N07

s tructura l Elucidation of FB (24)

It was a yellow globule shaped crystalline solid, m.p. 128 °C. The

structure of FB has been fully confirmed by IR, DCI-MS. 'H-NMR, NOE,

'^C-NMR and HETCOR spectra. IR spectrum exhibited characteristic bands at

3100 (N-H), 2950, 2850 (C-H), 1660, 1640 (C=0) , 1580 (phenyl), 1550

(C=N), 1510 (NO2, asym. str) and 1330 (NO2, sym. str) cm"'. The DCI-MS

spectrum showed a [M+l]'^ signal as the base peak at m/z 332 conforming its

molecular weight 331 which corresponded with the molecular formula

[C18H25N3O3]. It is equal to the sum of the molecular weights of (24) (349)

and m-nitrobenzaldehyde (151) minus one molecule of water. This suggested

that the formation of a Schiff base has occurred by the condensation of

10-undecenoacid hydrazide (23) with m-nitrobenzaldehyde.

The 'H-NMR and '^C-NMR spectra (Fig. l a and l b ) dissolved in

acetone-<i^ showed signals as assigned (Table la) . The assignments of all

signals to specific H- and C-atoms have been performed on the basis of

typical 5-values, multiplicity, relative integrations, a HETCOR spectra and

also by comparing the spectra with reference spectra (SDBS) '^ ' of

1 8 7

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1 O ru

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m o ~ ro

in • (N i Os <N

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R- <N X X

(N (N

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S Q Cl vi

u •M X) •o •a T3 "O -o -o •a •o •o

T 3

I s bJO S 01 e « (N X (N

a <N m

E E a: K X X

s c. a. w to

l i 00 „ N? f- ON .-H in

m c- ^

Tt ^ r o ON • in r m NO ^ • 1 (N 00 00 ND Os (N — rM

O oo On OJ ' — '

<N in 00 00 00

c I

K (N oo

I

ON

o b CN w N V-. i-

K X < < m u-<

N O I k.

<

1 9 0

10-undecenoic acid, 10-undecenamide and w-nitrobenzaldehyde. The 'H-

NMR and '^C-NMR spectra dissolved in acetone-iig showed several double

signals, especially of NH, H-7 and H-2' in the 'H-NMR spectrum and of C-7,

C - r , C-2', C-3', C-2 and C-6 in the '^C-NMR spectrum which indicated that

this molecule is present in two different forms. The two forms were found to

be in the ratio ( -3 :1) as calculated from the integration values of the N-H and

H-2' signals in the 'H-NMR spectrum. This can be thought of about the

geometrical {E/Z) isomers of the C=N bond or stabile conformers around

single bond such as in the CO-NH group. Irradiation of the NH protons (Fig.

Ic) resulted in a NOE-enhancement of H-7 (both signals), showing that the

N-H proton and H-7 (in both forms) are very near to each other. This can only

be the case in ^-isomer. Further, in case of H-2' signals, a NOE enhancement

was observed only for the largest H-2' signal ( -75%) and absolutely no NOE

enhancement was seen for the small H-2' signal ( -25%) (Table la). This

showed that only in the major form, the H-2' protons are near to the NH-

proton. It was therefore, concluded that both forms of m-nitrobenzaldehyde-

10-undecenohydrazone (24) are due to a major synperiplanar CO-NH

conformation (~75%>) and a minor antiperiplanar CO-NH conformation

( -25%) which are in equilibrium with each other as presented in (Fig.l) . An

additional evidence for this conclusion is the fact that in the 'H-NMR

spectrum of this compound dissolved in DMSO-t/g (Table lb) , again two

forms are observed but in a (~ 4:6) ratio. This observation again is an

indication that the two forms are stabile conformers which are in equilibrium

with each other, an equilibrium which is dependent on the polarity of the

solvent.

1 9 1

en

g; •

C O - ; f e' . o «

m 5 I

ig f X u ^ ^ ^ ^ Z V

I s p> o

y , 8 £

U « ti

ti

•> "O u c « c o o

CD « V -M S - " S g s s ' C w U P'

• « I . C • • > — » — t

-i ^ a

UJ • «/ c

> i« « t K

1gi5 8 .5 JS I z 6 «/i 5

iil Q ^ tJ O Q c - s

L '

^ tl t «. o

1 9 2

Table l b : 'H-NMR spectral data of FB (24) in DMSO-t/g

H - n r 5 ( p p m ) Integ-ration

Mult i -plicity

J (Hz)

7 8.09, 8.50 ( -60%, 40%)

IH s

NH 11.41, 11.54 (60%,40%)

IH s

2' 2 . 22 , 2 .64 ( 4 0 % , 6 0 % )

2H t J r j ' 7.30, 7.41

3' 1.60 2H br p J~7.35

4'-8' 1.28 5x2H br s

9' 2.00 2H m

10' 5.76 IH tdd J,o..9' 6.77, J,O',hz10.26, Jio'.Hz 17.22,

HE 4.93 IH dd Jhe,io' 10.26, JHE, HZ 2.01

Hz 5.00 IH dd 1

J H , I O ' 17.22, Jhe, Hz 2.01

Ar-2 8.47 IH dd J - 2.03, 2.02

Ar-4 8.26 IH dd J= 8.23, 2.02

Ar-5 7.72 IH dd J= 8.23, 7.51

Ar-6 8.09 IH d J= 7.51

The assignments of the protonated carbons in (Table l a ) was further

confirmed by HETCOR spectra (Fig. Id). It was found that 5h signals are

correlated with 5c signals at 6h 8.17, 8.46 ( -75%, 25%; H-7) to 5c 140.79,

144.53 ( -75%, 25%; C-7); 6h2.74, 2.33 ( -75%, 25%; H-2') to 6c 33.09, 35.55

( -75%, 25%; C-2'); 6h 1.69 (H-3'), to 5c 25.41, 26.07 ( - 7 5 % , 25%; C-3'); 5h

1.31 (H-4'-8') to 6c 29.41-30.38 (C-4'-8'); 6h2.02 (H-9') to 6c 34.42 (C-9'); 6h

5.78 (H-10') to 5c 139.81 (C-10'), two signals at 5h 4.88 (He-1 1') and 5h 4.96

( H z - l l ' ) to 5 c 114.59 (C-IV). In aromatic region, the signals at 5 h 8.49

1 9 3

O T3

O ru

o U5

O CD

O o

o nj

E o a C\1 u _

o l O

s

y - nn

1 9 4

(H-Ar-2) correlated to 5c 121.80 (C-Ar-2); 6h 8.22 (H-Ar-4) to 5c 124.56

(C-Ar-4); 6h 7.71 (H-Ar-5) to 6c 130.98 (C-Ar-5) and 6h 8.12 (H-Ar-6) to

5c 133.30 (C-Ar-6) as shown in Table la.

On the basis of these spectral evidence, the structure of compound FB

formulated as m-nitrobenzaldehyde-lO-undecenohydrazone (24) was deduced

to be in its two forms, synperiplanar (major) and antiperiplanar (minor) as

represented in Fig. 1

H H 1 I 1 1

i' N

T NO2

1 9 5

m-Nitrobenzaldehyde-ll-(carboxymethylthio)undecanohydrazone FBX,B

(25) and 2-(3-nitrophenyl)-3-[ll-(carboxymethylthio)undecanamido]thia-

zolidin-4-one FBXjC (26) [Reaction of the hydrazone (24) with

mercaptoacetic acid]

A solution of the hydrazone (24) in dry benzene was refluxed with

mercaptoacetic acid (molar ratio, 1:3) for 16 h collecting the generated water

in an azeotropic water collector. The solvent was distilled off under reduced

pressure and the orange oily residue left was chromatographed over a silica

gel column using (pet.ether-diethylether, 1:1 v/v) as eluent which yielded the

two products, FBX,B (25) as crystalline yellow globules (EtOH) in 31.2%

yield and a yellow oily liquid FBXiC (26) in 54% yield respectively.

o N-

FB (24)

HS.CH2.COOH

"NO2 dry benzene, reflux, 16 h

HO'

0 O S N

FBXiB (25)

HO

+

O O N02 0 NH —N S

FBX,C(26)

Structure Elucidat ion of FBX^B (25)

It was a yellow globule shaped crystalline solid, m.p. 120 The

structure of FBXjB has been established by IR, DIC-MS, 'H-NMR and

'^C-NMR spectra. IR spectrum (KBr) showed characteristic bands at 3250

(NH), 3075 (OH), 2900, 2850 (C-H), 1725 (COOH), 1645 (CO), 1610

(phenyl), 1555 (C=N), 1520 (NO2, asym str), 1350 (NO2, sym str) and 1465

( S - C H 2 ) cm' ' . The DIC-MS spectrum showed a [M+1]^ peak at m/z 424

confirming its molecular weight 423 [C20H29N3O5S], which is equal to the

1 9 6

sum of the molecular weights of (24) (331) and mercaptoacetic acid (92)

suggesting that (25) is an adduct of (24) and mercaptoacetic acid. The peak

appeared at m/z 164 [M-C13H23O2S]'' was due to the cleavage of CO-NH

bond. The base peak at m/z 148 [M-Ci3H25N03S]"^ arised by the cleavage of

N-NH bond and other peaks at m/z 134 [M-Ci5H2oN303]^ and at m/z 118 [M-

C i 6 H 2 3 N 3 0 3 ] ^ formed by the cleavage of 8'-9' bond and 9'-10' bond of the long

alkyl chain respectively as shown in Chart 1.

O O

HO' N.

NH " CH m/z 424 [M+I]

NO2

0

HO"

0

W ^ N H 2

m/z 276 (5.2)

O

m/z 134 (58.8)

-CH3

- H

N. NH" ^ C H ' ^ ^N02

m/z 164(11.8)

m/z 118(13.0) "NO2

m/z 148 (100.0)

CHART 1

The signals in 'H-NMR and '^C-NMR spectra dissolved in DMSO-c?^

were assigned to individual H and C atoms (Table 2a) by comparing with the

spectra of (24) (Table la) . The 'H-NMR spectrum dissolved in acetone-ii?^

showed clearly that this compound was formed by the addition of

mercaptoacetic acid to terminal C=C double bond of compound (24) as the

signals for the terminal olefmic protons at 5 4.88 (Hg-l 1') and 5 4.96 (H^-l 1')

1 9 7

B a a

CO

(N f ^ ^ ^

~ o ro ^ Tf o t-- o • w ^ Tt ^ vo

, m

ON

od ON m »N

N? #N oo oo o o o

ON On VO o o 1—1 LY-) f—t r- CO NO

ON o od O o

NO ( N NO O od CO d ( N NJD T—< od I—1 M <r) FS CN CO RO

CNl (N M M

u C I

U

Csl (N

<N m m vo ^ VH ll kH < < < < < <

f 6 CT! S Q c

s ><

(m 0 «

C! -a

u u D.

0! ^

Z 1

u f) TS c c:

Pi

z

C3

a> !q « H

N (N

ro r-ro

»

(N CN

n

r o CO ON 00 oo NO

m —• CN Cv »N

ON m m ON o q o

r - oo od

l—5 II II H->

II

CN 00

ON l>

-3

S a X)

on a T 3 -o

T 3 •a

-a -o

•o -o

(ii) o

C «« I — I

(N (N (N X

(N p: (N (N

s a a

to (N ^ od sp

00 ^

m in ^ O O ^ ^ ^

„ S? CN sP ON ^ - ^

^ (N ^ Tt m 00 in r — ( o C--CS vn m (N o

CN CO od od od

c 1 X ON

I

(N

(N in NO 1 u 1 1 1 r—1 < < < <

1 9 8

in (24) have disappeared and a new pealc corresponding lo the methylene

protons at C- I" of the terminal thioether moiety appeared as a singlet at

5 3.22. The signals for H-10' appeared as a broad pentet nt 5 1.69 (J~7.28)

and for H-11' appeared as a triplet at 5 2.64 (J=7.32). This was further

confirmed by '^C-NMR spectrum which also showed the disappearance of the

signal corresponding to the olefmic carbons at 5114.59 (C-1 1') and 6 139.81

(C-10') in (24) and the appearance of the new signal corresponding to the

methylene carbon of the terminal thioether moiety at 5 33.20. It also showed

a signal for the carboxylic carbon (C-2") at 5 171.47. The 'H-NMR and

'^C-NMR spectra dissolved in DMSO-^/^/acetone-c/^ showed several double

signals indicating that this molecule is also present in two Ibrms. The signals

corresponding to C-1, C- l ' , C-2' in '^C-NMR spectrum and that of NH, H-7

and H-2' in the 'H-NMR spectrum dissolved in in DMSO-f^, the two forms

were found to be in the ratio (--6:4). From the NH, H-7 and 11-2' signals in the

'H-NMR spectrum in acetone-t/^, the two forms were found in the ratio of

( -3:1) as in (24) (Table 2b). This observation again is an indication that the

two forms are stabile conformcrs which are in equilibrium with each other as

shown in Fig. 2, an equilibrium which is dependent on Ihc polarity of the

solvent.

On the basis of the above discussion, the compound FBXiB was

characterized as m-nitrobenzaldehyde-1 l-(carboxymethylthio)undecano-

hydrazone (25) and its two forms deduced are shown in Fig. 2

1 9 9

H H

HO 2

9 anti-periplanar ^ NO2

Fig. 2

Table 2b: ^H-NMR spectral data of FBX,B (25) in acetone-j/e

H-nr 5(ppin) Integ-ration

Multi-plicity

J (Hz)

7 8.18, 8.48 ( -75%, 25%)

IH s

NH 10.29, 10.66 ( -75%, 25%)

IH s

T 2.31, 2.74 ( -25%, 75%)

2H t J2.,3' 7.30, 7.41

y 1.59 2H brp U1.2S

1.31 6x2H brs

10' 1.69 2H brp J=^7.28

i r 2.64 2H t J,,MO'7.32

1" 3.22 2H s

Ar-2 8.51 IH dd J= 1.98, 1.83

Ar-4 8.24 IH dd J=7 .93 , 1.98

Ar-5 7.73 IH dd J= 7.94, 7.93

Ar-6 8.14 IH dd J= 7.94, 1.98

2 0 0

Structure Elucidation of FBXi^C (26)

It was a yellow oily liquid. The structure of FBXjC has been

confirmed by IR, DCI-MS, 'H-NMR, '^C-NMR and COSY spectra. IR

spectrum gave characteristic peaks at 3250 (NH), 3050 (OH), 2900, 2850 (C-

H), 1725 (COOH), 1685 (CON) 1660 (CONH), 1610 (Phenyl), 1525 (NO2

asym. str) 1355 (NO2 sym. str) and 1430 ( S - C H 2 ) cm-'. The DCI-MS

spectrum (Fig. 3a) showed a [M+1]"^ peak at m/z 498 corresponding with the

molecular formula [C22H31N3O6S2] which is equal to the sum of the molecular

weights of compound (25) (423) plus mercaptoacetic acid (92) minus one

molecule of water. This suggested that the formation of thiazolidinone ring

has occurred by the cyclocondensation of (25) with mercaptoacetic acid. The

mode of fragmentation is presented in Chart 2. The peak at m/z 452 was due

to [M-COOH]"'. The other structure revealing fragment ion peak was

observed at m/z 424 [(M+1)-C0CH2S]^ arised by the cleavage of 1-2 and 3-4

bonds is the diagnostic peak of the thiazolidinone ring^^. Two a-cleavage

fragments to the thiazolidinone ring on the long alkyl chain were observed at

m/z 223 [M-C,3H24N03S]^ and m/z 276 [(M+1)-C9H7N203S]^ along with two

y9-cleavage fragments at m/z 240 [(M+l)-Ci3H22 03S]^ and m/z 258 [M-

C9H9N303S]^ The base peak was observed at m/z 210 [(M+l)-Ci3H24N203S]

arised by the cleavage of 2-3 and 3-4 bonds of the ring. The peaks at m/z 231

[M-C8H9N302S]^ and m/z 215 [M-Ci4H26N03S]^ were formed by the cleavage

of V-2' and 2'-3' bonds of the long alkyl chain. It also showed some common

fragment ion peaks to that of (25) at m/z 166, 148, 134 and m/z 118.

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N

;r 1 .1 .1 z : r F 6 X , c

"'J '-fUU -i'. O V^C Ii I.. ^ :: Z JJ,) - nil : - '^iiii.'ii

.1 1 Ij r n . 1 F i g . 3 a ^

! : I .

.. I,

-. r .

HO

, H a

o o

HO- NH m/z 276 (72 2)

0 "NH—N S

m/z 498 [M+1]

C 1 1 1 N

Q m/z 210 (100 0)

m/z 148(15 0)

CHART 2

N O z

„ o NHj-N S

NO2

m/z 240 (91 8)

2 0 2

m/z 1 18 (32 2)

c I I I s

Q

m/z 223 (27 2)

NO2

m/z 166 (40 4)

The signals in 'H-NMR and '^C-NMR spectra dissolved in acetone-^^^

(Fig. 3b and 3c) were assigned to individual H-and C-atoms (Table 3). The

assignments were made from their typical chemical shift values, coupling

constants, relative integrations and also by comparing with the spectra of

FBXiB (25) (Table 2a and 2b). The 'H-NMR spectra are strongly in agreement

with the formation of a thiazolidinone ring as the H-7 signal of (25) has shifted

from 5 8.18, 8.48 ( -75%, 25%) to higher field 6 6.04 (H-2 in FBXiC) due to

the disappearance of C=N double bond and the appearance of two

diastereotopic hydrogen atoms of thiazolidinone ring in the form of double

doublets by coupling with H-2 at 5 3.84 (Jjtrans.a 15.87, 1.68, H-5trans) and S

3.75 ( Jsc.s,215.87, 0.92, H-5cis). Also, the '^C-NMR spectrum (acetone-J^)

indicated the formation of a thiazolidinone ring as the C-7 signal of FBX ,A

has shifted from 5 139.99, 143.24 (-60%, 40%) to 6 62.23 (C-2 in FBX,B).

The assignment of signals as depicted in Table 3 and the coupling of the

2 0 3

G a C i

••KSf

1 J 5

XXXVII

-JS

2 0 4

e. Q • a

C C «

r fe

o w

o

o ID

> o • ^

Ka

'J ff I O ^ <

— ^ c c «J r i » o t " • i >. - o 5 w " U U IJ

c n V <3 a

cc O 3 w bi _ IS- Ui < o . — ^

o a «a c Al O O C

- « •J 3 a : I ^ ® «) O r-i - •

0> O- O ( ' r t c .. ' i;

» t< (O ( . L

- i o a a I S A . . a 3 3 rs< 4J a t D C O O l - - - * « < «

o . U U _J CI C

— If) O ro 01 u j r - a> o« o* n r j i n 8

^ ^ u ) <0 r o

O ffi

o o

O fO

WD

r u o CO t o ^ 01 ^

( W O l t O Cj r^ fXi O 1*1— J

O CVi

r - e o o ' o — J X i f o - ^ i o u s r - m o ' o — c v m « 5 » n 4 0 f -< v « 4 n i n i n i n i A m i n t n i n « n t o ( 0 U ] ( 0 ( O i o i o i o

o ( D

—I ft»

«-• 01 o ) «« C9 lO r^

a i 40 n t o rsi ^ o j

i I t ,

g U. (O

U) H W • I ® jl» * — <»» i O • K «-« O) O <J

Ol <0 ^ <n ID "T o cn t o

i

: V r M • T i

J o m

o o fu

o u j

R I

s a a

CO (N ON

(N rsi O r-CN ON

r--00 m cn

O m o m

00 1 cc m (N r--00 ON 00 ,—1 in CNj

<N ON TT ON od O ON ON d >0 m CN m

<N CN r-) ' — '

C I U U

(N (N l O (N ^ (N

^ (N CO Ti-I I I I

< < < <

1 l U

S o <L) u 03

X CO

o R «

" r t

u 00 iri

(N ON

XI >

"o <U d

»

ro m

CN 2 « ::

00 m ON (N ^ CM O N

ON ON

ON O ON — — oo 11 II

O tT ^ ON oo t^

§ "E X i -o xl T3 -D -T3 -o -o -a

CO "O T3 T3 T3

W) o 01 -4- t

a c3 (N

rN (N vo (N (N

E a a

to q

m • r-; 00 cn rn

-a <u KJ ID CI, a

O O O ON ON

m (N (N cn

Tt <N

(N ( (N 00 00

IT)

o- r-

a I

K (N

ON I o ^

(N

(N lO ^ I I I I l - l t H i H < < < <

2 0 6

protons were further confirmed by COSY spectra (Fig. 3d). The broad signal at

5 6.04 (H-2) showed a weak cross peak with two double doublets at 5 3.75

(J=15.87, 0.92, H-5c,s) and at 5 3.84 (J-15.87, 1.68, H-5trans) indicating the

coupling of H-2 proton with both the ring methylene protons at C-5. In contrast

to FB (24) and FBXjB (25), only one form of FBXjC (26) appeared in the

spectra.

Based on the above discussion, compound F B X , C was characterized as

2-(3-Nitrophenyl)-3-[l l-(carboxymethylthio)undecanamido]thiazolidin-4-one.

(26) and is represented as

H 2- . 1 , H , ...

F B X i C ( 2 6 ) ° H H

2-(3-Nitrophenyl)-5-(methyl)-3-(10-undecenamido)thiazolidin-4-one FBX3A

(27) and 2-(3-nitrophenyl)-5-(methyl)-3-[ll-{(l-carboxy)ethylthio}undecan-

amido]thiazolidin-4-one FBX3B (28) [Reaction of the hydrazone (24) with

2-mercaptopropionic acid]

A solution of FB (24) in dry benzene was refluxed with

2-mercaptopropionic acid (molar ratio, 1:3) in dry benzene for 22 h after

which TLC examination revealed the presence of two spots labelled as

FBX3A (27) (upper) and FBX3B (28) (lower). The products on usual work up

and column chromatography yielded two liquid compounds, FBX3A (42.3%)

as colourless oily liquid and FBX3B (34.5%) as a pale yel low oily liquid. The

reaction sequence is as follows:

0 7

V

• •

-

1

B

U 1

—1 I 1 T -—1 I 1 T - 1 1 I I 1 1 1 1 1 ^ t — i — 1 — 1 —

6 Q

iT) a

•a (T)

e a ru a a. ~

V m r (XI 01 o

a S :

« Q <

i a .

T

M t n Pt N (

t o c O t J ^ CNJ O

. If a — a

, » ^ O c «• o o «

" o in a ^ ^ " - - " - .

; 5 S 21 S X i I

^ I u 2 UJ <t «l o . .

as tt

(iJ ID ^ -•

a — rg Q. - U

£«» c ^ « « •

Q C

[ V) u . i n u . c o

o N

NH'' '^CH'

FB (24)

CH3 HS.CH.COOH

"NO2 ''O' benzene, reflux, 22 h

O H \ O x

NH—N' "S

M FBXjA (27) CH3

Structure Elucidation of FBX^A (27)

It was a colourless oily liquid. The structure of FBX3A has been fully

established by DCI-MS, 'H-NMR, and '^C-NMR spectra. The DCI-MS

spectrum (NH3 as a reagent gas) (Fig. 4a) showed [M+1]'^ peak at m/z 420

and a less intense [M+18]"^ peak at m/z 437, confirming its molecular weight

419 [C21H29N304S], which is equal to the sum of the molecular weights of

compound (24) (331) and 2-mercaptopropionic acid (106) minus one

molecule of water. This indicated that the thiazolidinone ring has been

formed by the cyclocondensation of (24) with 2-mercaptopropionic acid but

no addition of 2-mercaptopropionic acid to the terminal C=C double bond of

the 10-undecenoyl side chain has occurred. The characteristic structure

revealing fragment ion peak was observed at m/z 332 [(M+1)-C0CH(CH3)-

S]" arised by the cleavage of 1-2 and 3-4 bonds of the thiazolidinone ring.

The a-cleavage fragment ion peaks to the ring were observed at m/z 237

[(M+l)-CnH2oNO]^ and at m/z 184 [M^-C,oH9N303S]^ the /^-cleavage

fragment ion peaks were observed at m/z 254 [(M+l)-CiiHi90]"^ and at m/z

167 [ ( M + 1 ) - C i o H i ] N 3 0 3 S ] " ^ along with some lower mass ion peaks as shown

in Chart 3,

iij -jDMr. ,:ii"ii:) jrniir^ r Tiriu r o f r t/cor . r.rrL.fu nuir . rwf i c,-,-> Vw.A

209

, i v i f i - t - 51

' ; '-.ON - n iooe .. J — . - - •• . - • • 1 ^ .0 i ' , • j

1 luO%- ir20?2 A i I t s / \ 1 / \ 1 : ioo^-1 t

-j r- I -1-i i 4 C 0 0

1 ' ' ' ' - ' 1 1 ' ' • H i

-, - ' jTV . 0

' . ' s j 1 i 0 Oi "i -1 j

1 Ci'3 1 00 I ' 1 I' 1 ' ' ! • ! ' Hi

• / ^^

irCc.-iCi Ml - J \ J . ) i

—,... , , • ,• T—T—r—t-i—T -1 1 1 1 1 ' 1 1 1 1 I ' 1 1 1 I • I 1 1 1 1 I I 1 J J -i

1 1 - ' • ' \ •

(

1 * VI V- 'JJ L ^ -- ' ^ 1 ' ^ X 1

^ •• • • .

JLHH 'LU -'''. 2CRH 21 '"liLf iT'sxas'

iiori/r.n^ct ' t OiCi 0-5-1'j.'.

11 : II- I I . Ill I I.ilil.lii ilili il.l I III I iiii'ii'

•I il II ililli II 1.1 I.,

II II. .Ill r 1 '• --^i' • - •"V"1 1 1 1 1 ; ; i .i- •••••• . M KM -jy lyy 1 1 M 1 rjM 1 4y i .y 1 1/-w 1 KM 1 wy

'tft d'

lilL k /

1 1 '1 !•••••••••! r"' !"••'••'!••• 'i 1 1"''"; I r" •• f" " •• I r" "" i cvy cvy :'yy -I'ly f.-y .•" y ••y'' '•'y " ly ' r y •'' y

GUYiQ SCAN 2i SIGttf'i-? RT=Q'3C r(/iri<'r-ri=nY^ nn onTi-^i,

M no 1 0 c; ,1 n 1 1 nn Oil

Fig. 4a

210

NOj

NH — N S

m/z420 (M+lf

NHj m/z 183 (3 6)

NHj ^ I-

„ d NO2 NH2-N^S

m/z 184(11 6) m/z 167(60 5) N H '

m/z 332 (8 0)

NH2-N "S

m/z 254 (82 1)

K •CH3

m/z 130 (16 5)

mJz 149 (26 9) +H

Q -co

m/z 209 (3 3)-<-

I I I N m/z 148 (4 1)

NOT O r

CHj

m/z 237 (100 0)

^ ^ NO,

m/z 166 (8 7)

'+H-

m/z 167 (60 5)

CHART 3

All the signals in the 'H-and '^C-NMR spectra dissolved in acetone-fig

(Fig. 4b and 4c) have been assigned to specific H-and C-atoms (Table 4).

The 'H-NMR spectrum is in strong agreement with the formation of a

thaizolidin-4-one ring as the H-7 signal of (24) has shifted from 6 8.17, 8.46

( -75%, 25%) to 5 6.01-6.04 (position 2 in F B X 3 A ) due to the disappearance

of C=N double bond and formation of ring. Also the NH signal has shifted

from 5 10.41, 10.76 ( -75%, 25%) to 5 9.28 due to the ring formation. The

signal for the thaizolidin-4-one ring protons at C-5 appeared at 5 4.04 (IH,

qd, J -6 .86 , 0.91, H-5cis) and 5 4.11 ( IH, qd, J=7.17, 1.53, H-5uans)- Also, in

'^C-NMR spectrum, the C-7 signal of (24) has shifted f rom 8 140.79, 144.53

( -75%, 25%) to 5 61.16. Contrary to FBXjC (26), the FBX3A (27) 'H-NMR

spectrum showed signals for two terminal olefmic protons (=CH2) at 5 4.90

( IH, dd, J=10.23, 2.13 for HE-11') and 5 4.98 ( IH , dd, J=I7.09, 2.13 for

211

J e a.

1 " 3

fci

5

212

6 a

3 3

u

a c a — a S

« i - " ! : o o ^ c «

*-> W t » w

5 t» Cr-OWMO 5 r I« f ? i? r » a 6 o «fi o oo a

H « — t • «<«

_ • • • • • • • • « . . , , , , , . . .

^ i i i y i i i i i i l i i i i i i y i i i i i i i i i i i H

Q V .

213

I o c o ^ a u c:

fS < X M b

0

T5

u u C, W3 PS

z 1 U

-a c c: a S Z

I

S « H

c I U

E ON cn

Ci. nj d o. nj « m X) XI m xi x> CJ O m •o m •o 00 m ON m (N m to NO 00 (N NO O (N O 00 4 oo NO in oo in 00 o in to r—< oi in 00 Tf VO oi ON (N ON <N ON ro ^ in r-H ON ON ON d in ON m H «N (N m m t—1 rn m r—1 (N m (N <N •—1 •—1 1 •—1 '—1 >—1 '—1 1

C/D O

C c3 l-H

on "o

on C Ki

N

0 O 1 I (N m u-1 m lo c l ro

00 J . b ^ ON --

in a m c3 0 i: ' <N r<-i in 1 I I 1 I I I V-l kH l-H VM VH < < < < < < <

ON O

CO

ON o

m

vo 00 r--

00

OS D X u

X u (N

3 .Si S a

' B OX) o —i m m rn <N (N

N (N N d X w X

lil OJ w X X 1—) X 1—) b

>—i ri (N oC o OO ON ON m ON

00 (N CNT d L >

—< R- ' (N R - ^ ON" ON" ON m m o NO CN

b b ON O 00

o 00

ON

I > 11 b w X "N X I I I I I I II i—> > 1—5 1—1 1—> 1 — >

-a O CO

X) ^ ^ Id -d t3 T3 -a T3 -o -a -a

FFI K FFI K FFI ^ ^ ^ ^ rn ffi ffi ffi ffi X ffi ffi rl r-c (N (N VO CN

--H Tt Tt O O O —I ON ^ IT) vo 00 ^ O (N —' <N M CM 00

VO NO ^ ON (N

O 00 NO m m ON ON rn r- ON 4 00 00

00 "o

C/2 d cfl C/5 C VI a

M rt on rt ffi E O O i! u o ^ _ ^ „ „ „ ( N C N i n u r i i r i i n Z r ^ r n ^ l - ^ E K <1

ON (N M NO W N vL >-i

E K < < < <

T3 U 00 a X o

<u

<u

cd a o on <D a on (U a cn (U ^

c <u S c

't/3 C/3 <

214

Hz-11') and the signal for olefinic methine proton (=CH-) at C-10' appeared

at 5 5.81 ( IH, tdd, J = 6.72, 10.23, 17.09 H-10') indicating the presence of a

C=C terminal double bond. This strongly suggested that the addition of

2-mercaptopropionic acid to the terminal double bond has not occurred in this

case. Also, in '^C-NMR spectrum, the signals of the o lefmic carbons appeared

at 5c 139.88 (C-10') and 114.63 (C-11') further confirmed that addition has

not occured. It is probably due to the increased nucleophilicity of the thio

group in 2-mercaptopropionic acid which facilitated condensation rather than

addition. Furher in 'H- and '^C-NMR spectra (Fig. 4a and 4b, Table 4), the

signals corresponding to thiazolidinone ring protons/carbons at C-2, C-5, C-

5-CH3 indicated that this compound contained a mixture of isomers with cis

and trans standing methyl and 3-nitrophenyl groups on the thaizolidin-4-one

ring. However, it is not clear which isomer is the major and which one, the

minor.

Based on the above spectral evidence, compound F B X 3 A was

characterized as 2-(3-Nitrophenyl)-5-(methyl)-3-(10-undecenamido)thiazo-

lidin-4-one and is represented as (27)

0 FBX3A (27)

215

Structure Elucidation of FBXjB (28)

It was a pale yellow oily liquid. The structure of F B X 3 B has been fully

established by DCI-MS, 'H-NMR, and '^C-NMR spectra. The DCI-MS

spectrum (Fig. 5a) showed a set of intense [M+1]"^ and a less intense [M+18]"^

peaks at m/z 526 and m/z 543 confirming the molecular weight 525

[C24H35N3O6S2] which is equal to the sum of the molecular weights of

compound F B X 3 A (419) and 2-mercaptopropionic acid (106). This suggested

that compound F B X 3 B was formed by the addition of 2-mercaptopropionic

acid to the terminal C=C double bond of F B X 3 A . The mode of fragmentation

is presented in Chart 4. The peak at m/z 480 [M-COOH]"^ confirmed the

presence of terminal thioether moiety in the compound. The peak at m/z 419

was due to the loss of [M-HSCH(CH3)C00H]^. The peaks observed at m/z

290 [(M+O-CioHgNjOsSJ^and at m/z 237 (base peak) [(M+1)-C,4H28N03S]^

were due to a-cleavage to the ring. The peaks at m/z 271 [M-CioHnN303S]^

and at m/z 254 [(M+l)-Ci4H25 03S]"' due to the y?-cleavage to the ring. The

characteristic peak for thiazolidinone ring was obsereved at m/z 436

[M-C0CH(CH3)S]^ as shown in Chart 4.

All the signals in the 'H-NMR and '^C-NMR spectra dissolved in

acetone-fig (Fig. 5b and 5c) could be easily assigned to specific H- and C-

atoms (Table 5) by comparison with the spectra of F B X 3 A (27) (Table 4).

All the signals in both the spectra were found compatible with that of F B X 3 A

(27) except H2C=CH- signals which didn' t appear here. In 'H-NMR spectrum,

the signals at 6 4.90 (IH, dd, J=10.23, 2.13, HE - IT) and 6 4.98 ( IH, dd,

J=17.09, 2.13, H z - l l ' ) of F B X 3 A disappeared and in place a new peak

corresponding to H-1" of the terminal thioether moiety appeared as a quartet

at 5 3.42 (J=7.02). This strongly supported that compound F B X 3 B has been

216

ii.. • • " [ • • • • I ' " r - r ' - I " ' . , . . . . . , , . , . . , , • 1 " • • ! r " ' ' ••] r — r

• 4(3 25C i'60 270 260 300 3!0 320 330 3'?0 ?50 360 3?P 390

... .. . i., 1. . . Irll. . 1. I -t—I r-~| ,—, r -i-T 1 p-H 1 1 (—I r-^ ; r—1 1 , , ,—t—, 1 r"-1 1 1— 400 410 420 430 440 453 460 4?0 4S? 4?0 500 510 525 530 540 550

'"BKSB SCRH 35 S!6!1P.= 18 RT^l:? SP.CKGD=S :i?''| ionv= t;cijo;-o rTjLF ."f3x35" - • • - . . - . - w ^ . . .

-r-r ; 1 . . , i ! . ; ,1.(11. ,i..ii. ii,i iiii.i, ii.i>i.i(.i.i.ii.i,iiiii.ii<(i(iiiiU

l " " l • I I . . . .1 • j . . . . [ • I iiiiiiil...... <(4((iiii.. I. i..t.tiiiiui. (.(., . III., tiui ,((li....(..i t. l-'T' -l"-;' h . , . . - I . , . . , , , . . , . . , . . . . , . . . . 1 . . • . • . . . . , . . . . , . . . . . . . . 1 ' - | — I " -1

80 ?0 100 110 120 130 140 150 160 170 150 190 200 210 220 230 rr-i

! 1 ; H I . ! : I. I II 1 ,1)!, )!i. •I""!'"''-I •"•"• ,1., t

T"-'—I" iiiii. I,

'-"I" • p i J . j 1 t • i ' V ' . 1 1 1 240 250 260 270 260 2?0 30-5 310 320 33 ^ 349 350 360

- r T I.. I. li, Mil "T ' : ^ I i ' r""""—I——'—;—'—i——•—r 4(30 4ig <135 iidQ 45g d^a <igg jqo cng Rig c^g q^g pao

5'E'X3l! SCAN C'TC-S .7 r(Ari<'r;n=nY< nn <nny= TITLE: FBXIB"

75 QO 16, c; ISO 00 ^ s * w . A . 245 0 0 24 1 524 n n V V — w/ . A g7 ,00 17 2 184 00 17 253 0 0 17 t . 525 n n

W V 22 ij i l l ; 0 0 17 5 1 ? ? 00 20 0 : 254 0 0 32 A 526 u n w w

A 130. nn 20 k 7>ri7 nn V w 37 2 : 0 C' 33 527 0 0 31 u 138. 00 2B ? H s 00 15 ' g . 30B fin i ? 543 CO 31 7 147, .00 22 7 .•t^-xi 00 100 0 41?, 0 0 i 7 2

CO

A ¥ rw

37APT t: i Fwt) = 45 TnTAi_ " 620Oi

76 0 1022764 237.0 . . . . . . . . . ' "^iii?!-!) 254.0 '"5806 2?Q.O 432?I-524 0 inc;fi.q.q

Fig. 5a

?17

1 ,

I/) bJD

218

6 CL CL

i

n r j o

O tri

c N com v o o : o — o i

u o e u j >-i> • - - I C --O — o CO « A « JCfv. e> w^^r-o •«

UlCU u i « Ol t . - f s » a .

•f a : : 3 > — <t — w t s w w o > c « i u j V- u Z 3 U — -r 3 OL < o « 0. Ol. < 31 ajiua. o 3 c-

« a . o o v » > o o c a S

E c a e

— « e c —

o

ei)

1

H o r«. e» . > « n M « ro

« ) n -M rs. n «

- • ^^ f^ a0 n ^ t AM ^^ ^ ^^ « . rf , fc ® ^^ ^^ ^ i o l e

< fs. fx.

I (M

O r* r ) V I •aimtowtciavvitor-^mwto

219

formed by the addition of 2-mercaptopropionic acid to the terminal C=C

double bond of F B X 3 A .

HO Y >- 0 NH2 CH3

m/z 289 (9 2)

+H

m/z 290 (33 1)

m/z 147 (22 7)

m/z 148 (10 3) m/z 237 (100 0)

o K . CH3

m/z 130(20 5)

s NO,

m/z 166(15 5)

CHART 4

This was further confirmed by '^C-NMR spectrum which also showed the

disappearance of the signals corresponding to the terminal C=C double bond

at 6 114.63 ( C - i r ) and 6 139.88 (C-10') of F B X 3 A and the appearance of the

new signals corresponding to the terminal thioether moiety at 6 41.35 (C-1"),

174.33 (COOH) and 5 17.78 ( r ' - C H j ) indicating that the compound F B X 3 B

contains both thiazolidinone and terminal thioether moieties. The signals of

H-2, H-5, 5-CH3 in 'H-NMR spectrum and C-2, C-5, 5-CH3 in '^C-NMR

spectrum (Table 5) again indicated that this compound also contains mixture

of isomers with cis and trans standing methyl and 3-nitrophenyl group on the

thiazolidinone ring as in F B X 3 A .

220

VC I

Oi c o 41 O «

00 rs CO X ca <*-

o a cs TS n « c.

z u

13 s cs

Z I K

Qi

rt H

S o.

CO

c I

U

N X

.JL = .ii § a

§ G cs I—I u

a m XI O O OO M CO ^ '—I ^ ^ m 00

1—I r- ON ON o ^ CO m < fS

(N fN) OO o

o o cn

(N Tf to ON

CO •a T3 o lo <N NO 1—< OO m CO m OO m OO O to ON CO (N CO ^ to

r - r - Tf- fS CM CO CO CO 1—H

C/2 O

C/3 'o

C/5 c

00 C 00 cd C C<1

X 00

"o cd

K b u r - H CN) U 1—N 1 ^ 1 1 - V-c 1 1 m ^ CN ^ 1—1 ( ^ CN < < OO 00

CO to NO NO

ON m I u ly-i 1 w-i 1

I lO

o u-1 I I u

lO

"O Crt o o VI c ra u crt c. CO X I O (N tN

O lAt u o (N 11 1—) >—1 1—1 1—> 1—1

<N O (N O -o (U > 1 1 o s X c/) «

rn K a: « O u 1—1

CO ON fO ON OO ON ON <N

o i rs rv

00 On ON CO o . O O CJV

II od II

00 II

t--II II

1—1 II

>—1 II

"-1 11

1—1

Td -a T3 -o o cr T3 T3 00

Ui 1- T3 cr T3

K ffi ffi K K ffi ffi r^ r^ ^

X

CN (N vo cs cs m K ffi ^ m —I

-a •a -a

•o (U M c C3 CJ VM a>

(U c o

o IZI VH (U a c/l (U s Kl M (U >

0 c 01 £ £ Ofl M tn

s a. a

CO

o o NO NO

ON 00 O CN NO <N 00 NO CO TO LO NO (N >—1 >O (N M NO CO CO CN R-- ON

F—( ON CN 1—1 CNI CO 00 0C5

k. c I

"o

to c fS

I -o ^

t/3 c u u

(N (N lO >/-) u-i X z

ON u 1 CN 1 1 NO t . . J o ^ : CN CO —' ^ ^ < S-H < < <

2 21

On the basis of the above discussion, the deduced structure of F B X 3 B

as 2-(3-nitrophenyl)-5(methyl)-3-[l l-{(l-carboxy)ethylthio}undecanamido]

thiazolidin-4-one (28) is represented as:

H CH3

FBX3B (28)

m-Nitrobenzaldehyde-ll-(2-carboxyethylthio)undecanohydrazone FBXjB

(29) and 2-(3-nitrophenyl)-3-lll-(2-carboxyethylthio)undecanamidoj-

thiazan-4-one FBX2C (30) [Reaction of the hydrazone (24) with


A solution of the hydrazone FB (24) in dry benzene was refluxed with

3-mercaptopropionic acid (molar ratio, 1:3) for 26 h, monitoring the reaction

by TLC which showed it to be a mixture of two components, labelled as

FBX2B (29) (major, upper) and FBXjC (30) (minor, lower). The products on

purification By column chromatography (silica gel, pet. ether-diethyl ether.

1:1 v/v) first yielded a yellow solid which on crystallisation (benzene-

acetone) furnished FBXjB (58.0%) as white crystalline globules and then a

yellow oily liquid of FBX2C in (26.0%) yield.

HS . (CH2)2 .C00H 0 0

NO2 dry benzene, ^ H i ^ n H ' reflux, 26 h

F B X j B (29)

NO2

0 O

H O H ^ - W ^ N H — N

0 FBX2C (30)

N O ,

222

Structure Elucidation of FBX7B (29)

It was a white globule shaped crystalline solid, m.p. 98 °C. The

structure of F B X j B has been fully established by IR, DCI-MS, 'H-NMR and

'^C-NMR spectra. IR spectrum (KBr) showed characteristic peaks at 3175

(N-H), 3040 (OH), 2900, 2800 (C-H), 1730 (COOH), 1645 (CONH), 1605

(Phenyl), 1555 (C=N), 1520 (NO2, asym. str.), 1480 (S-CH2), and 1340

(NO2, sym. str.). The DCI-MS spectrum strongly supported the structure

which showed a [M+1]"^ peak at m/z 438 confirming its molecular weight 437

[C21H31N3O5S]. This is equal to the sum of the molecular weights of (24)

(331) and 3-mercaptopropionic acid (106). This indicated that FBX2B is an

adduct of (24) and 3-mercaptopropionic acid. The base peak at m/z 134

[(M+1)-Ci6H23N303]^ and another fragment ion peak at m/z 304

[M-C5H902S]^ were formed by the cleavage of 9'-10' bond of the long alkyl

chain. The peaks at m/z 218 [M-Ci2Hi302S]^ and at m/z 186 [M-C,2H,2N303]"

were obtained by the cleavage of 3'-4' and 5'-6' bonds. It also showed some

fragment ion peaks common to (25) at m/z 164, 166, 148, 118 and 119 as

shown in Chart 5.

o o

m/z 438 [M+

O O

HO'

m/z 134 (23.6) -CH3

m/z 118(37.7)

Q

m/z 218 (76.1)

NH ^ C H C" ^ NO2 I I I N m/z 148

m/z 164(11.8)

CHART 5

2 21

All the signals in the 'H and '^C-NMR spectra have been assigned to

individual H-and C-atoms (Tables 6a and 6b) by comparison with the NMR

spectra of a previously established compound FB (24) (Tables l a and lb ) and

FBX,B (25) (Tables 2a and 2b). The 'H-NMR spectrum dissolved in acetone-

di clearly showed that this compound was formed by the addition of 3-

mercaptopropionic acid to terminal C=C double bond of compound (24) as

the olefmic proton signals of HE- l l ' - and Hz-11' of (24) at 6 4.88 ( IH, dd,

J=10.22, 2.14, HE-11') and 5 4.96 ( IH, dd, J=17.09, 2.14, Hz-11') have

disappeared and in place two additional triplets at 5 2.74 (J=6.87, H-1 ) and 5

2.57 (J=6.87, H-2 ) corresponding to two methylene protons of the thioether

moiety appeared. The triplet at 5 2.55 (J=7.33) was attributed to the

methylene proton at C-11'. Also, in '^C-NMR spectrum (Table 6a), C-11'

signal of (24) has shifted from 5 114.59 to 6 31.00 and an additional peak was

observed at 5 178.50 for the carboxylic carbon at C-3". Here again, the 'H-

NMR and '^C-NMR spectra showed several double signals, the signals

especially of C-7, C-l", C-2' in '^C-NMR spectrum and of NH, H-7 and H-2' in

'H-NMR spectrum. This indicated that FBX2B is also present in two different

forms as synperiplaner and antiperiplanar conformers of CONH bond in a

55/45 ratio in DMS0-g?6 (Table 6a) and 85/15 ratio in acetone- d^ (Table 6b).

Thus, the two forms are the stabile conformers in equilibrium with each other

as in (24) and (25), and is dependent on the polarity of the solvent.

224

s a c. to

CN li (N ^ WO ^ r- " ^ ^ o

UO !>• UO .IT) —' in Tj-c ^ - . rn _

o 00 06

ON (T)

CN I. N 1 0 0 00 ON 00 as m in

0 0 m m m r — 1 <n — I M3 01 >0 06 r- m in 0 00 d 00 cn d (N vo 06 . — 1 m (N Tt- CM m m

(N (N ro (N m H

1 o

Q c

oT fS ea X oa ta 0 « C! -a 13 u u 0 I I I I I I Li Vh i-i < < < < < <

N K

o

m m ro O »

. — I 00 . — 1

00 0 00 0 rsi (N

r-- r\ ON m i n 00 b ON 0 0 00 — r — 1 06 00

11 rNi I I I I I I I I <—> > — 5 )

•S .t; "a S d

D, CO Qi Li Li J2 Li X) TJ TJ T3 -O -O Td

M) 0 -w <<_> C iss

CN (N X (N p: <N E (N <N Dh K K K

s & c.

CO

sO 10 ^ ^

<N ^ cn ^

00 „ X© (N s p

^ m >n pr, in 00 ' I 7 7 fN ^ 06 w ^ w (N w

ON >n ON m (N 00 <N m NO (N 0

(N (N CN 00 00 00

C I K

(N a\ I o (N

(N tn ^ I I I I iH Vi Li < < < <

225

Table 6b: 'H-NMR spectral data of FBXjB (29) in acetone-rf<5

H - n r 5 (ppm) Integ-ration

Multi-plicity

J (Hz)

7 8 .18 , 8 .48 I H s ( - 8 5 % , 15%)

NH 10.30,10.66 I H s ( - 8 5 % , 15%)

2' 2 .31,2.73 2H t J2.3. 7.31, 7.48 ( - 8 5 % , 15%)

3' 1.56 2H br p J«7 .28

4'-9' 1.31 6x2H br s

10' 1.69 2H br p J^7 .25

i r 2.55 2H t JnMO'7.33

1" 2.74 2H t Jr . 2" 6 .87

2" 2.57 2H t J2M" 6.87

C O O H 12.20 IH br s

Ar-2 8.51 IH dd J= 1.99, 1.82

Ar-4 8.24 IH dd J= 8.08, 1.98

Ar-5 7.73 IH dd J= 8.08, 7.94

Ar-6 8.14 IH dd J= 7.94, 1.86

226

On the basis of the above spectral evidence, compound FBX2B which

was characterized as m-nitrobenzaldehyde-1 l-(2-carboxyethylthio)undecano

hydrazone (29) is represented as in Fig. 4

H H

synperiplanar

H H

Structure Elucidation of FBX^C (30)

It was a pale yellow oily liquid. The structure of FBX2C has been fully

established by IR, DCI-MS, 'H-NMR and '^C-NMR spectra. IR spectrum

(Neat) displayed characteristic absorption bands at 3200 (NH), 3050 (OH),

2910, 2840 (CH), 1710 (COOH), 1680 (CON), 1650 (CONH), 1610 (Phenyl),

1525 (NO2, asym. str), 1430 (S-CH2) and 1375 (NO2, sym. str) cm"'. The

DCI-MS spectrum (Fig. 7a) showed a [M+1]^ peak at m/z 526 confirming its

molecular weight 525 [C24H35N3O6S2] which is equal to the sum of the

227

1 ~ J " < n r*~T—r~>"""rrT i~i I''"I

1."ill's - J I

) M M ' . J ' r rt ( J

II 1 7 1 i i i 1 ! i TTTTTTT C 4 , «

- n — t - i — 1 ; 1 I I I - I r T r - ' " ' ' " 'jzz:

1 MM = 1 ^ ^ u < \ ! - _ """ - 1

. 1 i GIJ ^ f i . M u = I j ' u. r kj. 'i = I ^ i ^ V '' • ' S1 w w

..! i.! -H. • • nil! . . . J . . . . . . ^ - J , . ,

,1, , 111 1 1 . I ' , . 1 1 i I ' (

. . i n I 1/1 1 J l ' i t I ' t t \ 1 - 3 1 1 . - . / l i t J • t/i ^ '^Ln ' ^

nil 1 •II

J A ' T y - t "p lA ' ^ 1,'t n i ' i J 0 < JWW 376 ^ ^

: ; ; i i j i i = : T i i i ~ aia aj ij .laA 45|j ujfi} ilyy 51313 51^ 52)3 553 Jjjy 55^ I T 1 I i i IM

rlH T < jii" If r.Ti= IIJ Ci ii

-0 "

1 ~< •J I i r "* I i t 1 "y 0 ley M h V <->

It 1 ' t M 1 1 . -I j ^ 1 < Ll a 1 w I

J j 1 1 l l 'A 1 fi 1 0 0 A J j w 1 a .

_ _ - J J r< ( X 1 / It i J _ V W W J \J w A.

I j II t i V K i 11 - < 1 M i 1 i i H n 11 I j II w 1 v-*

T 111, - ~ 1 It f — t 1 1 S. A ' X H h i A n .t. . k

It J . - 4_ W / w

Fig. 7a

228

molecular weights of FBX2B (437) and 3-mercaptopropionic acid (106) minus

one molecule of water. This indicated that the cyclocondensation has

occurred between FBX2B and 3-mercaptopropionic acid forming thiazanone

ring. The base peak at m/z 341 [M-CgHgNOaS]"^ was resulted by the cleavage

of 2-3 and 5-6 bonds of the thiazanone ring. The fragment ion peak at m/z

438 [(M+1)-C0CH2CH2S]^ arised by the cleavage of the 1-2 and 3-4 bonds, is

the diagnostic peak of a thiazanone ring®" . The fragment ion peaks at m/z 290

[(M+O-CioHgNjOsS]^ and m/z 237 [M-CnHjeNOsS]^ were obtained by the a -

cleavage to the ring. The fragment ion peaks at m/z 273 [M-CioHjoNaOsS]^

and m/z 254 [M-Ci4H2503S]"^ were due to the /^-cleavage to the ring. These

peaks strongly supported the formation of thiazanone ring. In addition to

these, some peaks common to FBX2B appeared at m/z 220, 165, 121 as shown

in Chart 6.

m/z 438 (41 2)

0

m/z 220 (76 1)

m h 148

+2H ^ m/z 150 (15 7) g

C m/z 166 (23 1)

N

CHART 6

m/z 231 a \ 4)

2 29

All the signals in the 'H-NMR and '^C-NMR spectra dissolved in

acetone-<i6 (Fig. 7b and 7c) have been assigned to individual H-and C-atoms

(Table 7) by comparison with the NMR spectra of FBXzB (Tables 6a and 6b).

The 'H-NMR spectrum is in agreement with the formation of a thiazanone ring

as the H-7 signal of FBXzB has shifted to higher field from 5 8.18, 8.48 (85%,

15%) to 5 6.14 (position 2 in FBX2C) due to the disappearance of C=N double

bond signal and also the NH signal has shifted to higher field from 5 10.30,

10.66 (75%, 25%) to 5 9.12, due to the ring formation. It also showed two

additional triplets for two ring methylene protons (-CH2-CH2-S-) at 5 2.94

(J=6.71, H-5) and 5 3.10 (J=6.71, H-6). Also, in '^C-NMR spectrum, the C-7

signal of FBXjB has shifted to higher field from 5 139.97, 143.22 ( -55%.

45%) to 5 66.02 (position C-2 in F B X 2 C ) . It also showed an additional peak at

6 173.14 for the carbonyl carbon at C-4. Here again, in both the NMR spectra,

only one form of FBX2C appeared

Based on the above spectral evidence, compound FBX2C was

characterized as 2-(3-mtrophenyl)-3-[ll-(2-carboxyethylthio)undecanamido]

thiazan-4-one (30).

NO2

FBX2C (30)

In conclusion, it is stated that the course of addition of mercaptoacetic

acid, 2-mercaptopropionic acid and 3-mercaptopropionic acid to terminal

olefinic bond followed anti-Markovnikov's rule which is attributed to the free

radical mechanism^^. The increased nucieophilicity of thio group in

2-mercaptopropionic acid facilitates condensation rather than addition thereby

yielding FBX3A (27) as the first product and then F B X 3 B (28).

230

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231

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233

EXPERIMENTAL

lO-Undecenoicacid hydrazide (23)

It was prepared from 10-undecenoic acid by a well known common

procedure. A mixture of 10-undecenoic acid (10.Og, 0.0542 mol), methanol

(40 ml) and sulphuric acid (1 ml) was refluxed on a water bath for 6 h. To

this was added freshly distilled hydrazine hydrate (4.08g, 0.0813 mol) and

refluxed for further 8 h. The reaction mixture was cooled, concentrated in

vacuum and the solid residue left was extracted with ethylacetate, waslicd

several times with water and the organic phase dried over anhydrous Na^SOj-

The solvent was distilled off under reduced pressure to a white solid residue

which on crystallization from ethanol afforded (23) as white globules, 9.02 g

(84%). m.p.88 °C, Rf 0.76 (benzene-diethyl ether, 6:4 v/v)

IR (KBr): v,,, , 3300 (N-H), 2950, 2850 (C-H), 1630 (C=0) , 1600 (C-C) .

1575, 1530, 1460, 1440, 1380, 1 190, 1160, 910 cm-1.

m-Nitrobenzaldehyde-lO-undecenohydrazone, FB (24)

It was prepared following the literature method®. A solution of 10-

undecenoacid hydrazide (23) (4.0g, 0.0202 mol) and m-nitrobenzaldehyde

(3.052g, 0.0202 mol) in dry benzene (50 ml) was refluxed with stirring on an

oil bath at 80 °C for 5 h, collecting the generated water in a Dean Stark

apparatus. Progress of the reaction was monitored by TLC (silica gel G, pet.

ether-diethyl ether, 6:4 v/v) at every 15 min. TLC examination showed the

disappearance of the spot corresponding to the substrate and the appearance

of a new spot corresponding to the product, labelled as FB (24). The solution

was then cooled, washed several times with water, and the benzene layer was

dried over Na2S04. The solvent was removed under reduced pressure and the

234 yellow solid residue was crystallized from ethanol to yield FB (24) as yellow

crystalline globules, 5.81g (87%), m.p. 128 °C, Rf 0.70 (benzene-diethylether,

6:4 v/v)

IR ( K B r ) : v„ax 3100 (N-H), 2950, 2850 (C-H), 1660, 1640 (C=0) , 1580

(phenyl, C=C), 1550 (C=N), 1510 (NO2, asym str), 1330 (NO2, sym. str),

1420, 1255, 1225, 1150, 1100, 1055, 970, 935, 885 810,790, 715 cm"'

' H - N M R (, 400 MHz, acetone-^/^): 5h 8.17, 8.46 ( - 7 5 % , 25%); IH, s, H-7),

10.41, 10.76 ( -75%, 25%; IH, s, NH), 2.33, 2.74 ( - 2 5 % , 75%; 2H, t, J= 7.32,

7.48, H-2'), 1.69 (2H, br p, J= 7.32, H-3'), 1.31 (5H, br s, H - 4 ' - 8 ' ) , 2.02 (2H,

m, H-9) , 5.78 ( IH, tdd, J=| 6.72,10.22, 17.09 H-IO'), 4.88 ( I H , dd, J=10.22,

2.14. H r - l l ), 4.96 ( IH, dd, J=17.09, 2.14, l l ' -Hz) , 8.49 ( IH , dd, J=1.99, 1.83

Ar-2), 8.22 ( IH, dd, J=7.94, 1.83, Ar-4), 7.71 ( IH, dd, J=7.78, 7.94, Ar-5)

8.12 ( IH , dd, J=7.78, 1.99, Ar-6).

' H - N M R (400 MHz, DMS0-rf<5): 5h 8.09, 8.50 ( - 6 0 % . 40%; IH. s. H - 7 ) .

1 1.41, 11.54 ( -60%, 40%; IH, s, NH), 2.22, 2.64 ( - 4 0 % , 60%; 2H, t, J= 7.30,

7.41, H-2'), 1.60 (2H, br p, J=7.14, H-3'), 1.28 (lOH, br s, H-4'-8'), 2.00 (2H.

td, J=7.14, 6.60, H-9') 5.76 ( IH, tdd, 1=6.77,10.26, 17.22, H-10'), 4.93 ( ]H,

dd, J=10.26, 2.01, HE), 5.00 ( IH, dd, J=17.22, 2.01, Hz), 8.47 ( IH , dd,

J=2.02. 2.03, Ar-2), 8.26 (IH, dd, J=8.23, 2.02, Ar-4), 7.72 ( I H , dd, J=7.51.

8.23, Ar-5), 8.09 ( IH, d, J -7 .51 , Ar-6)

'^C-NMR (100 MHz, acetone-rf^,): 5c 140.79, 144.53 ( - 7 5 % , 25%; C-7),

170.07, 172.12 ( -75%, 25%; C-l ' ) , 33.09, 35.55 ( - 7 5 % , 25%; C-2'), 25.41,

26.07 ( -75%, 25%; C-3'), 29.22-30.38 (C-4'-8'), 34.42 (C-9'), 139.81 (C-10) ,

114.59 (C- l l ' ) , 137.68 (Ar-C-1), 121.80 (Ar-C-2), 149.72 (Ar-C-3), 124.56

(Ar-C-4), 130.98 (Ar-C-5), 133.30 (Ar-C-6).

235

DCI-MS (NH3): m/z 332 [(M+1), 100.0], 333 (48.0), 334 (19.0), 335 (7.0).

316 (6.4), 317 (3.6), 165 (5.0), 149 (4.4), 118 (4.0), 59 (3.8)

m-Nitrobenzaldehyde-ll-(carboxymethylthio)undecanoltydrazone FBXiB

(25) and 2-(3-nitrophenyl)-3-[ll-(carboxymethylthio)undecanamido]thia-

zolidin-4-one FBX,C (26) [Reaction of the hydrazone (24) with

mercaptoacetic acid]

To a solution of the hydrazone (24) (1.20g, 0.0030 mol) in dry benzene

(20 ml) was added mercaptoacetic acid (0.832g, 0.0090 mol) and refluxed

with stirring on an oil bth at 80 °C for 16 h, collecting the generated water

using a Dean Stark apparatus. The products on TLC examination (silicagel G,

benzene-diethyl ether, 6:4 v/v) showed a major (lower) and a minor (upper)

spot labelled as FBXiB (25) and FBXjC (26). The solution was cooled,

washed several times with water to make the solution neutral and the organic

phase dried over anhydrous Na2S04. The solvent was distilled off under

reduced pressure and the orange oily residue left was chromatographed over a

silica gel column using (pet.ether-diethylether, 1:1 v/v). Elution of the

column first furnished a pale yellow solid FBXiB which was recrystallised

from ethanol as crystalline yellow globules, 217 mg (31.2%), m.p 120 °C, Rf

0.43 (benzene-diethyl ether, 6:4 v/v). Further elution of the column yielded

FBXiC as a yellow oily liquid which was further purified by repeated column

chromatography using the same solvent, 810 mg (54%), Rf 0.38 (benzene-

diethyl ether, 6:4 v/v).

236

Spectral data of FBX,B (25)

IR (KBr): v^^a, 3250 (N-H), 3075 (OH), 2900,2850 (C-H), 1725 (COOH),

1645 (CO), 1610 (Phenyl), 1555 (C=N), 1520 (NO2, asym str), 1350 (NO2,

sym str) 1465 (S-CH2), 1400, 1380,1190 cm"'.

^H-NMR (400 MHz, acetone-^/g,): 5h 8.18, 8.48 ( - 7 5 % , 25%; IH, s, H-7),

10.29,10.66 ( -75%, 25%; IH, s, NH), 2.31, 2.74 ( - 2 5 % , 75%; 2H, t, J=7.30,

7.47, H-2 ') , 1.59 (2H, br p, J «7.25, H-3') , 1.31 (12H, br s, H-4 ' -9 ' ) , 1.69 (2H,

br p, J«7.28, H-10'), 2.64 (2H, t, J=7.32, H - l l ' ) , 3.22 (2H, s, H-1") , 8.51

( IH, dd, 1.98, 1.83, Ar-2), 8.24 ( IH, dd, 7.93,1.98, Ar-4), 7.73 ( IH, dd,

J=7.94, 7.93, Ar-5), 8.14 ( IH, dd, J=7.94, 1.98, Ar-6).

' H - N M R (400 MHz, DMSO-^/fi,): 5H 8.07, 8.27 ( - 6 0 % , 40%; IH, s, H-7),

11.39, 11.53 ( -60%, 40%; IH, s, NH), 2.21, 2.62 ( - 4 0 % , 60%; 2H, t, J=7.33,

7.42, H-2') , 1.48 (2H, br p, J=7.37, H-3'), 1.58 (2H, br p J«7.22, H-10') , 1.25

(12H, br s, H-4 ' -9 ' ) , 2.54 (2H, t, J -7 .34 , H - l l ' ) , 3.17 (2H, s, H-1") , 8.45

( IH, dd, J=1.99, 1.83, Ar-2), 8.21 ( IH, dd, J= 8.05, 1.83, Ar-4), 7.70 ( IH, dd,

J=8.05, 7.69, Ar-5), 8.07 ( IH, dd, J -7 .69 , 1.82, Ar-6).

^^C-NMR (lOOMHz, DMSO-rf^,): 5c 139.99, 143.24 ( - 6 0 % , 40%; C-7),

168.95, 174.57 ( -40%, 60%; C - l ' ) , 31.76, 34.12 ( - 6 0 % , 40%; C-2 ') , 24.17

(C-3'), 28.04-28.79 (C-4'-10') 31.67 (C-11'), 171.47 (C-2") , 33.20 ( C - l " ) ,

136.18 (Ar-C-1), 120.51 (Ar-C-2), 148.18 (Ar-C-3), 123.70 (Ar-C-4), 130.30

(Ar-C-5), 132.60 (Ar-C-6)

DCI-MS (NH3): m/z (%): 424 [ ( M + l ) ^ 50.9] 425 (28.6), 426 (10.2), 336

(10.6), 337 (4.2), 275 (3.8), 276 (52), 218 (5.8), 215 (4.3), 163 [ ( M > C , 3

H23O2S, 11.8], 149 (36.7), 150 (10.2), 165 (9.4), 166 (10.0), 167 (4.8), 147

237

(11.0). 148 [ ( M > C , 3 H25 NO3S, 10.0], 149 (36.7), 150 (10.2), 136 (16.6),

137 (5.3), 133 (9.8), 134 [(M+l)-C,5 H20 N3O3, 58.8], ,121 (9.5), 118 [M^-

C,6H23N303, 13.0], 119(6.3) .

Spectral data of FBX^C (26)

IR (Neat): v^ax 3250 (N-H), 3050(OH), 2900,2850 (C-H), 1725 (COOH),

1685 (CON), 1660 (CONH), 1610 (phenyl), 1525 (NO2 asym str), 1355 (NO.,

sym str), 1430 (S-CH2), 1405, 1260, 1165 cm"'

'H-NMR (400 MHz, acetone-c?^,): 5h 6.04 ( IH, s, H-2), 3.84 (IH, dd,

J=15.87, 1.68, H-5 trans), 3.75 ( IH, dd, J=15.87, 0.92, H-5cis) , 2.10 (2H, t,

J -7 .32 , H-2 ' ) , 1.50 (2H, br p, J=7.14, H-3 ' ) , 1.19 (12H, br s, H-4 ' -9 ' ) , 1.59

(2H. brp, J=7.26, H-10 ' ) 2.64 (2H, t, J=7.33, H-11 ' ) , 3.22 ( IH , s. H-1"), 8.34

( IH, dd, J=1.99, 1.98, Ar-2), 8.24 ( IH, dd, J=8.10, 2.23, Ar-2), 7.71 ( IH,

J=8.10, 7.94, Ar-5), 7.95 ( IH, dd, J=7.94, 1.99, Ar-6).

^^C-NMR (100 MHz, acetone-^/^,): 5c 62.23 (C-2), 172.17 (C-4), 34.09(C--5),

171.71 ( C - r ) , 33.09 (C-2') , 25.97 (C-3') , 29.50-30.00 ( C - 4 ' - l l ' ) , 33.80 (C-

1"), 169.67(C-2"), 142.18 (Ar-C-1), 124.76 (Ar-C-2), 149.54 (Ar-C-3),

123.63 (Ar-C-4), 131.01(Ar-C-5), 135.3l(Ar-C-6).

D C I - M S (NH3): m/z (%) 498 [(M+1)^ 14.5], 499 (10.1), 452 [(M^)-C02H,

10.2], 424 [(M+1)^-SCH2C0, 10.4], 425(4.8), 332(9.5), 273(10.4), 274 (7.6),

275(13.6), 276[(M+1)^-C9H7N203S, 72.2], 277 (31.3), 278 (14.0), 258

[(M+1)^-C9H9N303S, 10.0], 239 (7.2), 240 [(M+O^'-CoHssOsS, 91.8], 241

(35.9), 242 (11.8), 230 (10.9), 231 [(M^)-C8H9N302S, 25.4], 232 (16.3), 223

[(M^)-C,3H24N09S, 27.2], 224 (12.2), 210 [(M+1)^-C,3H24N203S, 39.5], 212

(24.0), 213 (13.1) 214 (6.4), 215 [(M>CI3H24N03S, 47.7], 216 (22.2), 217

(8.1), 218 (14.5), 193 (25.0), 194 (12.2), 195 (29.5), 196 (11.3), 176 (11.8),

2 38

178 (12.7), 162 (8.1), 163 (8.2), 164 (8.1), 165 (7.9), 166 (40.4), 148 (15.0),

149 (10.0), 150 (9.0), 151 (15.4), 152 (16.3), 153 (7.5), 134 (42.2), 135

(20.0), 136 (32.2), 137 (19.0), 138 (30.0), 139 (18.1), 1 18 (32.2), 1 19 (9.5),

1 10 (8.2), 81 (13.1), 82 (9.0), 83 (14.5), 87(1 1.8), 89 (10.4), 91 (12.7), 93

(13.6), 94 (29.0), 95 (17.7), 96 (8.4), 97 (8.6), 99 (20.0), 100 (9.5).

2-(3-Nitrophenyl)-5-(meihyl)-3-(10-undecenaniido)thiazolidin-4-one F B X 3 A

(27) and 2-(3-nitrophenyl)-5-(methyl)-3-[ll-{(l-carboxy)ethylthio}undecan-

amidoJthiazolidin-4-one F B X 3 B (28) [Reaction of the hydrazone (24) with


To a solution of the hydrazone (24) ( l .Olg , 0.0030 mol) in dry benzene

(20ml) was added 2-mercaptopropionic acid (0 .96Ig, 0.0090 mol) and

refluxed for 22 h as above, monitoring the progress of reaction by TLC (silica

gel G, pet. ether-diethylether, 3:7 v/v) which revealed the presence of two

spots labelled as F B X 3 A (27) (upper) and F B X 3 B (28) (lower) and then

worked up as usual. The product on purification by column chromatography

(silica gel, pet. ether-diethyl ether, 3:7 v/v) as eluent yielded F B X 3 A as a

colourless oily liquid, 536 mg (42.3%), Rf 0.46 (benzene-diethyl ether, 6:4

v/v). Further elution of the column using pet. ether-diethyl ether, 6:4 v/v

afforded F B X 3 B as a pale yellow oily liquid, 528 mg (34.5%), Rf 0.32

(benzene-diethyl ether, 6:4 v/v).

Spectral data of FBX,A (27)

'H-NMR (400 MHz, acetone-ds,): 5H 6.01 ( IH, d, J=0.91, H-2cis), 6.04 ( IH,

d, J=1.53, H-2trans), 4.04 ( IH, qd, J=6.86, 0.91, H-5cis), 4.11 ( IH, qd,

J=7.17, 1.53, H-5trans), 1.59 (3H, d, J=6.86, S-CHacis), 1.61 (3H, d, J=7.17,

5-CH3trans), 9.28 ( IH, s, NH), 2.11 (2H, t, J=7.25, H-2'), 1.50 (2H, br p.

239

J = 7 . 0 5 , H - 3 ' ) , 1 .22 ( 1 2 H , b r s, H - 4 ' - 9 ' ) 5 . 8 1 ( I H , t d d , J = 6 . 7 2 , 1 0 . 2 3 , 1 7 . 0 9 ,

H - 1 0 ' ) , 4 . 9 0 ( J = 1 0 . 2 3 , 2 . 1 3 , H E - 1 1 ' ) , 4 . 9 8 ( I H , d d . J = 1 7 . 0 9 , 2 . 1 3 ) , 8 . 3 4 ( I H ,

d d , J = 1 . 9 9 , 1 .83 , A r - 2 ) , 8 . 2 6 ( I H , d d , J = 8 . 0 9 ; 1 . 9 9 , A r - 4 ) , 7 . 7 3 ( I H , d d ,

J = 8 . 0 9 , 7 . 9 3 , A r - 5 ) , 7 . 9 5 ( I H , d d , J = 7 . 9 3 , 2 . 2 8 , A r - 6 ) .

'^C-NMR (ICQ MHz, acetone-</<j,): 5c 61.16 (Cc-2), 172.83 (C-4), 39.53 (C-

5cis), 39.82 (C-5tran), 19.46 (C-S-CHjcis), 20.60 (C-S-CHstran), 172.23 (C-

r ) , 34.30 {C-T), 25.98 (C-3'), 29.24-30.39 (C-4'-8'), 34.44 (C-9^), 139.88

(C-10'), 114.63 ( C - i r ) , 141.59 (Ar-C-lcis) , 142.41 (Ar-C-l t rans) , 124.84

(Ar-C-2), 149.55 (Ar-C-3), 123.82 (Ar-C-4), 131.05 (Ar-C-5).

DCI-MS (NH3): m/z (%) 420 [ ( M + l ) \ C21H29N3O4S, 82.1], 421 (36.0), 422

(26.9), 437 [(M+18)^ 6.6], 438 (3.7), 387 (9.1), 332 [(M+18)^-C3H40S, 8.7],

313 (9.1), 266 (4.9), 254 [(M+1)^-C,,H ,90, 82.1], 255 (13.6), 256 (4.9), 237

[(M+1)"-ChH2ONO, 100.0], 238 (20.7). 239 (7.8), 218 (4.1), 209 (3.3), 207

(4.5), 184 (11.6), 185 (6.6), 186 (3.3), 165 (7.0), 166 (8.7), 167(60.5), 168

(9.1), 169 (3.3), 147 (21.9), 148 (4.1), 149 (26.9), 150 (5.3), 151 (4.1), 137

(4.5), 130 (16.5), 131 (3.7), 121 (6.2), 116 (4.5), 117 (3.3), 118 (7.8), ] 10

(5.3), 105 (3.3), 107 (4.5), 95 (4.9), 94 (3.3), 93 (6.2), 91 (3.3), 89 (7.4), 87

(7.4), 85 (24.0), 83 (6.2), 82 (3.3), 81 (14.9), 80 (3.7), 76 (31.1), 74 (21.1),

72 (10.3), 71 (4.1).

Spectral data of FBX^B (28)

'H-NMR (400 MHz, acetone-dg): 5h 6.01 ( IH , d, J=0.91, H-2cis), 6.04 (IH,

d, J=1.53, H-2trans), 4.05 ( IH, qd, J=6.86, 0.91, H-5cis), 4.11 ( IH, qd,

J=7.17, 1.53, H-5trans), 1.59 (3H, d, J=6.87, 5-CH3cis), 1.61 (3H, d, J=7.]7 ,

5-CH3trans), 9.28 ( IH, s, NH), 2.11 (2H, t, J=7.25, H-2'), 1.50 (2H, brp,

J~7.09, H-3'), 1.22 (12H, brs, H-4'-9'), 1.59 (2H, brp, J~poorly resolved, H-

240

10'), 2.67 (2H, m, H-11 ' ) , 3.42 ( IH, q, J=7.02, H-1"), 1.60 (3H, d, J=7.02, 1"-

CH3), 8.34 ( IH, dd, J=1.98, 1.83, Ar-2), 8.26 ( IH, dd, J=8.09, 1.99, Ar-4),

7.73 ( IH , dd, J=8.09, 7.93, Ar-5), 7.95 (IH, dd, J -7 .93 , 2.29, Ar-6).

'^C-NMR (100 MHz, DMSO-rf^): 6c 61.16 (C-2), 172.83 (C-4), 39.53 (C-

5cis), 39.83 (C-5trans), 19.46 (C-S-CHscis), 20.61 (C-S-CHjuans), 172.22 (C-

r ) , 34.08 (C-2') , 25.97 (C-3'), 29.24-30.40 (C -4 ' -10 ' ) , 31.91 (C- IT) , 17.78

(C-1"-CH3), 41.35 (C- l") , 174.33 (C-COOH), 141.57 (Ar -C - lc is) , 142.40

(Ar-C-1 trans), 124.85 (Ar-C-2), 149.55 (Ar-C-3), 123.82 (Ar-C-4), 131.06

(Ar-C-5).

DCI-MS (NH3): m/z (%) 525 [ (M+l )^ C24H35N3O6S2, 22.0], 526 (98.6), 527

(31.0), 524 (58.6), 543 [(M+18)", 31.7], 544 (8.9), 541 (8.9), 480 [ ( M >

COOH, 15.0], 471 (9.8), 454 (9.6), 436 (8.2), 419 [ (M+1)^-SCH(CH3)C00H,

17.2], 405 (7.2), 308 [(M^)-(C|2H2302S. 17.2], 290 [(M+l)^-C,oH8N203S,

33.1], 289 (9.2), 288 (10.4), 280 [(M^)-Ci3H2502S, 10.0], 273 [ (Vf)-

C,OHioN303S, 9.6], 271 (9.4), 237 [ ( M ^ C u H j e N O j S , 100.0], 238 (15.8), 239

(10.3), 229 (14.6), 218 (15.8), 207 (37.2), 199 (20.0), 184 (17.9), 180 (18,6),

164 (15.2), 165 (15.0), 166 (15.5), 151 (14.9), 147 (22.7), 138 (28.9), 130

(20.6), 121 (17.9), 108 (15.2), 87 (17.2), 75 (16.5).

m-Nitrobenzaldehyde-ll-(2-carboxyethylthio)undecanohydrazone FBX2B

(29) and 2-(3-nitrophenyl)-3-[ll-(2-carboxyethylthio)undecanamido]-

thiazan-4-one FBXjC (30) [Reaction of the hydrazone (24) with 3-

mercapto propionic acid]

To a solution of the hydrazone FB (24) (1.02g, 0.0030 mol) in dry

benzene (20 ml) was added 3-mercaptopropionic acid (0.961g, 0.009 mol) and

refluxed for 26 h as described above. TLC examination (silica gel, pet.ether-

241

diethyl ether, 6:4 v/v) of the products showed it to be a mixture of two

compounds labelled as FBX2B (29) (major, upper) and FBX2C (30) (minor,

lower). The products on purification by column chromatography (silica gel,

pet. ether-diethyl ether, 1:1 v/v) first yielded a yellow solid which on

crystallisation from (benzene-acetone) furnished FBX2B as white crystalline

globules, 765 mg (58.0%), m.p 98 °C, RfO.46 (benzene-diethyl ether 6:4 v/v).

Further elution of the column using (pet.ether-diethyl ether, 3:7 v/v) yielded

FBX2C as a yellow oily liquid, 412 mg (26.0%),Rf 0.24 (benzene-d ie thy l

ether, 6:4 v/v).

Spectral data of FBX^B (29)

IR (KBr): 3175 (N-H), 3040 (OH), 2900, 2800 (C-H), 1730 ( ^ O H ) ,

1645 ( m N H ) , 1605 (Phenyl), 1555 (C=N), 1520 (NO2 asym str), 1480 (S-

CH2), 1340 (NO2 sym str), 1415, 1380, 1215, 1 1 10, 800 cm"' .

'H-NMR (400 MHz, acetone-t/^): 5h 8.18, 8.48 (~ 85%, 15%; IH, s, H-7),

10.30, 10.66 ( -85%, 15%; IH, s, NH), 2.31, 2.73 ( - 1 5 % , 85%; 2H, t, J=7.48,

7.31, H-2') , 1.56 (2H, brp, J«7.28, H-3') , 1.31 (12H, br s, H-4 '-9 ' ) , 1.69 (2H,

br p, 7.25,H-10'), 2.55 (2H, t, J=7.33, H-11') , 2.74 (2H, t, J=6.87, H-1") ,

2.57 (2H, t, J=6.87, H-2") , 12.20 ( IH, brs, COOH), 8.51 ( IH , dd, J=1.99,

1.82, Ar-2), 8.24 ( IH, dd, J -8 .08 , 1.98, Ar-4), 7.73 ( IH, dd, J=7.94, 7.63, Ar-

5), 8.14 ( IH, dd, J=7.94, 1.98, Ar-6).

'H-NMR (400 MHz, DMSO-rf^.): 5H 8.11, 8.29 ( - 5 5 % , 45%; IH, s, H-7),

1 1.41, 11.55 ( -55%, 45%; IH, s, NH), 2.23, 2.63 ( - 4 5 % , 55%; 2H, t, J=7.34,

H-2') , 1.48 (2H, brp, J^7.33, H-3') , 1.26 (12H, br s, H-4 ' -9 ' ) , 1.59 (2H, br p,

y^7.14, H-10') , 2.47 (2H, t, J=7.14, H-11') , 2.65 (2H, t, J= 6.77, H - l " ) , 2.49

(2H, t, J=6.77, H-2") , 8.47 ( IH, dd, J=1.99, 1.83, Ar-2), 8.23 ( IH , dd, J=

242

8.05, 2.01, Ar-4) 7.72 ( IH, dd, J=7.88, 8.05, Ar-5), 8.08 ( IH, dd, J=7.88,

2.01, Ar-6);

'^C-NMR (100 MHz, DMSO-rfe,): 5c 139.97, 143.22 ( - 5 5 % , 45%; C-7),

168.92, 174.55 ( -45%, 55%; C-1'), 31.75, 34.11 ( -55%, 45%; C-2'), 24.16

(C-3'), 28.40-28.80 (C-4'-10') , 31.00 (C-11'), 34.53 (C-2") , 26.35 (C-1"),

178.50 (C-3") , 136.17 (Ar-C-1), 120.50 (Ar-C-2), 148.18 (Ar-C-3), 123.69

(Ar-C-4), 130.28 (Ar-C-5), 132.61 (Ar-C-6).

D C I - M S (NH3): m/z (%) 438 [ ( M + l ) \ C21H31N3O5S, 63.5], 439 (41.7), 440

(16.5), 366 (8.2), 336 (24.6), 337 (13.0), 304 [ (M>C5H902S , 14.2], 305

(8.0), 218 [ (M>C,2Hi302S, 25.9], 219 (10.0), 220 (7.4), 201 (8.4), 186

[(M^)-C,2Hi2N303, 18.8), 187 (9.0), 163 (8.2), 164 (9.0), 165 (7.8), 166

(10.6). 148 [(M^)-C,5H2oN303, 24.8], 149 (16.4), 150 (15.2), 151 (15.2), 152

(7.8). 133 (23.6), 134 [(M+1)^-C,6H23N303, 100.0], 135 (67.0), 136 (38.7),

137 (16.8), 118 (37.7), 1 19 (18.2), 120 (26.3), 121 (88.3), 122 (35.1), 123

(15.4), 106 (11.4), 107 (1 1.0), 108 (13.6), 109 (7.8), 94 (9.2), 72 (21.0), 73

(9.0).

Spectral data of FBX^C (30)

IR (Neat): v^^, 3200 (NH), 3050 (OH), 2910, 2840 (C-H), 1710 (COOH),

1680 (CON), 1650 (CONH), 1610 (phenyl) ,1525 (NO2 asym. str), 1430 (S-

CH2), 1375,1350 (NO2 sym. str)1230, 1170,1070, 800, 720 cm"'.

'H-NMR (400 MHz, acetone-^/e): 5h 6.14 ( IH, s, H-2), 2.94 (2H, t, J=6.71,

H-5) 3.10 (2H, t, J=6.71, H-6), 9.12 ( IH, s, NH), 2.09 ( IH , t, J=unresolved,

H-2') . 1.58 (2H, br p, J=7.17, H-3 ' ) 1.29 (12H, m, H-4 ' -9 ' ) , 1.60 (2H, br p,

J= unresolved, H-10') , 2.56 (2H, t, J=7.38, H-11') , 2.75 (2H, t, J=7.17, H-1"),

2.58 (2H, t, J=7.17, H-2"), 8.28 ( IH, dd, J=2.14, 1.83, Ar-2), 8.23 ( IH, dd, J=

243

8.09, 2.29, Ar-4), 7.72 (IH, dd, J=8.09, 7.93, Ar-5), 7.90 ( IH, dd, J=7.93,

1.07, Ar-6).

'^C-NMR (100 MHz, acetone-^/g,): 5c 66.02 (C-2), 173.14 (C-4), 37.24 (C-

5), 27.59 (C-6), 172.35 (C-1'), 34.13 (C-2'), 25.97 (C-3'), 29.50-30.20 (C-4'-

11'), 24.53^ (C-1"), 35.25' (C-2"), 169.15 (C-3"), 141.89 (Ar-C-1), 122.94

(Ar-C-2), 149.36 (Ar-C-3), 124.17 (Ar-C-4), 130.76 (Ar-C-5), 134.65 (Ar-C-

6).

DCI-MS (NHj): m/z (%) 526 [ (M+l )^ C24H35N3O6S2, 18.8], 527 (9.3), 438

[(M+1)^-C0(CH2)2S, 41.2], 439 (15.6), 440 (6.0), 420 (10.5), 406 (4.6), 341

[(M^)-C8H7N02S, 100.0], 342 (31.7), 343 (10.2), 290 (13.8), 254 [(M+1)^-

C,4H2503S, 27.0], 255 (9.0), 237 [(M^)-C,4H26N03S, 71.4], 238 (15.1), 239

(9.5), 220 (76.1), 221 (16.5), 166 (25.3), 167 (17.5), 150 (15.7), 138 (1 1.4),

120 (1 1.4), 121 (33.1), 123 (23.8), 118 (9.1), 93 (12.1), 87 (17.2), 89 (14.0),

81 (12.5), 77 (9.3), 72 (15.5).

-w, fmrv^ WA xm

. ml 2-Aryl-2-Methyl-3-N'Long alkyl chain

substituted'Thiazolidin-4-one

244

DISCUSSION

SUMMARY

The work described in this chapter includes the reaction of p-chloroacetophenone-

10-undecenohydrazone (31) with:

(i) Mercaptoacetic acid in dry benzene which furnished an adduct, p-chloro-

acetophenone-1 l-(carboxymethylthio)undecanohydrazone (32) and a new

compound, 2-(4-chlorophenyl)-2-methyl-3-[l ]-(carboxymethylthio)undec-

anamido]thiazolidin-4-one (33) and a byproduct, bis (carboxymethylthio)-4-

chloroacetophenotal (34).

(ii) 3-Mercaptopropionic acid in dry benzene which afforded an adduct, p-chloro-

acetophenone-1 l-(2-carboxyethylthio)undecanohydrazone (35) as the sole product.

The hydrazone (31) was also found to exist in two stereoisomeric forms as syn-periplanar

(major) and anti-periplanar (minor) [as in (24)] as evident from its stereochemical studies.


As part of our programme in searcii of biologically active compounds with

sulphur and nitrogen containing heterocycles, we have synthesised four novel

compounds, 2-(3-ni t rophenyl)3-[n-(carboxymethy/ethyl thio)undecanamido

thiazolidin-4-ones/thiazan-4-ones (26), (27), (28) and (30) f rom w-nitrobenz-

aldehyde-10-undeceno-hydrazone (described in Chapter 4). In continuation of

the above mentioned work, we have undertaken this problem on the synthesis of

2 - (4-ch lorophenyl -2-methyl -3- [ l l - ( c a r b o x y m e t h y l / 2 - c a r b o x y e t h y l t h i o ) u n d -

e c a n a m i d o ] t h i a z o l i d i n - 4 o n e s / t h i a z a n - 4 - o n e f rom yc-ch lo roace tophenone-

1 0 - u n d e c e n o h y d r a z o n e (31) u s ing m e r c a p t o a c e t i c a c i d / 3 - m e r c a p t o -

p rop ion i c ac id .

245

Structural assignment, stereochemistry and biological assay are

discussed. Screening results of the compounds (31) and (32) are summarised for

cytotoxic activity against 3-cell lines of three types of human cancers: lung,

breast, CNS. Screening results of the above compounds are also summarised for

antimicrobial activity against four bacteria, Escherichia coli, Bacillus subtilis,

Staphylococcus aureus, Pseudomonas aeruginosa and one fungi Candida

albicans (details in Chapter 6)

(31)

246

R E S U L T S A N D D I S C U S S I O N

The synthesis of compounds (32)-(35) has been performed in two steps (Scheme

2). The hydrazone FD (31) was first prepared following a published procedure^ by

refluxing a solution of 10-undecenoicacid hydrazide (23) in dry benzene with j!?-chloro-

acetophenone (molar ratio, 1:1) as white crystalline needles in 83% yield. The hydrazone

(31) was then reacted with (i) mercaptoacetic acid and (ii) 3-mercaptopropiomc acid

(molar ratio, 1:3) in dry benzene. The products on column chromatography over a silica

gel column using pet.ether-diethyl ether as eluent, yielded compounds FDXjA (32),

FDX,B (33) and FDXjC (34) in (i) and FDXjA (35) a sole product in (ii). The failure of

the reaction to yield the cyclised product in (ii) may be attributed to the increased

nucleophilicity of the carbon at C-7 due to the presence of the electron donating methyl

group which retarded the nucleophilic attack by mercapto group at this carbon.

247

X z

m X M e

\ s

+

0 >=0

X o b.

. 00 (N g

(U u N C

IT) m

fS fi

o

'3!

c C/3

O I

"o a, K O O u

0 1 "o

K O O u

I u X u CO ^ E I

tn r^

a S w X u '73

248

p-Chloroacetophenone-lO-undecenohydrazone FD(31)

A solution of 10-undecenohydrazide (23) in dry benzene was refluxed with

;?-chloroacetophenone for 6 h as described in (24). The products on usual work up

followed by crystallization from ethanol furnished FD (31) as white crystalline needles

in 83% yield. The reaction sequence is as follows:

O O MeOH/H

NH2.NH2.H2O ^ N H - N H ,

(23)

dry benzene Reflux, 6 h

Structure Elucidation of FD (31)

It was a needle shaped white crystalline solid, m.p 64 °C. The stereostructure of

FD has been fully confirmed by IR, DCI-MS, 'H-NMR, '^C-NMR, APT and HETCOR

spectra. IR spectrum (KBr) exhibited characteristic absorption bands at 3175 (NH).

2900, 2850 (C-H), 1675, 1650 (CO), 1595 (phenyl) and 1540 (C=N) cm"'. The DCI-MS

spectrum of the compound showed an intense [M+1]"^ signal at m/z 335/337 which is in

consistent with the molecular formula [C19H27N2OCI]. The signals in 'H-NMR and '^C-

NMR spectra (Fig. 8a and 8b), dissolved in DMSO-c/^ could be easily assigned (Tables

8a and 8b) by comparing the spectra with the previously established hydrazone FB (24).

(Table la and lb).

249

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251

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d ri ri w-i Tt-" i> K

-

c X 00 1

— r - M fsT fV CO

u K 00 1 d

r — (

w N Z CN ON

d r — ( X < <

2S2

Table 8b: " C - N M R , A P T and H E T C O R spectral data of FD (31) im D M S O - 4

C-nr 8 (ppm) APT H E T C O R correlat ion vt'ith

7-CH3 13.19,13.76 ( -60%, 40%) CH3 H-CH3

7 145.43, 149.35 ( -60%, 40%) C H-7 r 175.13, 169.16 ( -60%, 40%) C 2' 32.25, 33.92 ( -60%, 40%) CH2 H-2' 3' 24.20, 25.07 ( -60%, 40%) CH2 H-3' 4'-8' 28.18-28.87 CH2 H-4'-8' 9' 33.07 CH2 H-9' 10' 138.73 CH H-10' Ar-1 133.66 C Ar-4 137.11 C Ar-2,6 127.51 CH H-Ar-2,6 Ar-3,5 128.25 CH H-Ar-3,5

Table 8c: ' H - N M R spectral data of FD (31) in DMSO-^/^


Multi-plicity

J(Hz)

7-CH3 2.34,2.76(~75%, 25%) 3H s NH 9.37,9.68 ( -75%,25%) IH s 2' 2.28,2.73 ( -25%,75%) 2H t J2'.3' 6.71,7.62 3' 1.66 2H br p J ~ 7.48 4 ' -8 ' 1.31 5x2H br s

9' 2.04 2H m 10' 5.79 IH tdd J,O'H,17.09, J,O'he10.22,

Jjo'.g' 6.72 HE-11' 4.88 IH dd JHJ:, 10' 10.22, JH^, H,2.29

H z - i r 4.95 IH dd JHZ,10'17.09, JH , HE 2.29

Ar-2,6 7.84 2H d JAr-2, 6, Ar-3,5 8.55 Ar-3,5 7.42 2H d JAr-2,6 ,Ar-3,5 8.55

253

All the signals were found comparable with that of (24) except that it showed no signal

corresponding to the benzylidene proton at C-7 and in place a singlet at 5 2.21, 2.24 (~

60%, 40%) for the methyl group protons appeared. In addition, it also showed the

characteristic A2B2 signals for the /?-chlorophenyl ring. As for (24), here again both the

spectra showed several double signals for 7-CH3, C-7, C-l ' , C-2', and C-3' in the '^C-

NMR spectrum and for H-7-CH3, NH and H-2' in the 'H-NMR spectrum, indicating that

this molecule is also present in two different forms. Irradiation of NH-proton (Fig. 8c)

resulted in a NOE-enhancement of both H-8 signals corresponding with the methyl

group in both forms, suggesting that N-H proton and the methyl group in both forms are

very near to each other. This can only be the case in ^'-isomer. Further, only a very weak

enhancement of the H-2' signal is seen, so it could be impossible to make a definitive

conclusion about the conformation of the CO-NH bond. However, it could be assumed

that in analogy to (24), the major form is the synperiplanar conformer and the minor one

is the antiperiplanar conformer. As in (24), the 'H-NMR spectrum of this compound

dissolved in acetone-fig, again two forms were observed in -3 :1 ratio (Table 8c)

indicating that the equilibrium between the stabile conformers in this case also depends

on the polarity of the solvent.

The structure of the compound was further confirmed by HETCOR spectra

(Fig. 8d and 8d') . It showed cross correlation peaks between CH3-7 (60%, 40%)/CH3-7

(60%, 40%), H-2' (60%, 40%)/C-2' (60%, 40%), H-3' (60%,40%)/C-3' (-60%,40%), H-

4'-8'/ C-4'-8', H - 9 ' / C - 9 ' , H - I O ' / C - I O , HE-11', HZ-117C-11', H-Ar-2,6/C-Ar-2,6, and H -

Ar-3,5/C-Ar-3,5 as shown in Table 8b . An APT spectrum (Fig. 8e) further confirmed

the assignments of '^C-NMR signals (Table 8b).

254

6 Q

- in

00 bh

s

mi K

Ivi

(1) a

o o

f Isje ^ 5 P» W *» t? tri P £ cx N o >• o o

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o oi o • • fiosifegis X o • • » • ^

s t F ? 2 -

1

i s , i f 11 ! M f j i | 5 t i n M t i s i i l i s s i i j l i i M i O c i i

" SSSassgRRaRfcRsa^BHRRii i i i lRRRRpisR i I

L

s

I I P L I L L I Y Y I L I P ^ I I S S I S I N I I I I I Y I ^

i t ^ _ .. in rv ut ot o —

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o i£)

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o o

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R IR

258

Based on the above spectral evidence, the compound (31) was characterised as

p-chloroacetophenone-lO-undecenohydrozone and deduced to be its two forms as

synperipianar (major) and antiperiplanar (minor) (Fig. 8)

, H H

NOE NOE

H CH3

Q XI

synperipianar

NOE

(Fig. 8)

259

p-Chloroacetophenone-ll-(carboxymethylthio)undecanohydrazone FDXiA (32), 2-(4-

ch loroph enyl)-2-methyl-3-[ll-(carboxymethylth io) undecan amidojth iazolidm-4-one

FDXjB (33) and bis(carboxymethylthio)-4-chloroacetophenotal FDX,C (34)

[Reaction of the hydrazone (31) with mercaptoacetic acid]

A mixture of the hydrazone (31) and mercaptoacetic acid (molar ratio, 1:3)

dissolved in dry benzene was refluxed with stirring for 24 h and worked up as usual. The

products on column chromatography first afforded a yellow solid which on

crystallisation (EtOAc) gave FDXiA (32) (41.2%) as white amorphous solid . Further

elution of the column yielded a colourless oily liquid FDX,B (33) (33.3%) and then a

pale yellow solid, FDXjC (34) which on crystallization (EtOAc) afforded white

crystalline needles (13.2%).

HS.CH2.C00H dry benzene, HO reflux, 24 h

FD(31) FDX,A (32)

0 0

OH + HO

CHr

0 N H — N S

F D X , B (33)

Structure Elucidation of FDX^A (32)

It was a white amorphous solid, m.p. 68 °C. The structure of FDXjA has been

fully established by IR, DCI-MS, 'H-NMR and '^C-NMR spectra. IR spectrum exhibited

characteristic peaks at 3150 (N-H), 3050 (OH), 2900, 2850 (C-H), 1640 (CONH), 1700 (

( m O H ) , 1590 (phenyl), 1530 (C=N) and 825 cm"'. The DCI-MS spectrum showed a

260

set of [M+1]"^ peaks as the base peak at m/z 427/429 corresponding with the molecular

formula [C21H31N2O3SCI], which is equal to the sum of the molecular weights of (31)

(334/336) and mercaptoacetic acid (92). This suggested that FDXjA is an adduct of (31)

and HS.CH2.COOH. The two prominent fragment ion peaks at m/z 168/170 [(M)-

CnHssOsS]^ and m/z 154/156 [(M+O-CnHjsNOsS]^ were formed by the cleavage of

CO-NH and N-NH bonds respectively .

The 'H-NMR and '^C-NMR signals have been assigned to individual H-and C-

atoms (Table 9a) by comparing with the spectra of a previously established compound

FBXjB (25) (Table 2a). All the signals were found comparable with (25) except that it

showed no signal at 5 8.07, 8.27 (60%, 40%) corresponding to the benzylidene proton at

C-7 in (25) and in place a singlet at 5 2.22, 2.24 (~ 60%, 40%) for the methyl group at

C-7 appeared in (32). In addition, it also showed the characteristic A2B2 signals for the

jD-chlorophenyl ring. As for (25), here again both the spectra showed several double

signals for 7-CH3, C-7, C-1', C-2', and C-3' in the '^C-NMR spectrum and for H-7-CH3,

NH, H-2' in the 'H-NMR spectrum, indicating that this molecule is also present in two

different forms. The two forms were found to be in the ratio -6 :4 in DMS0-<i(5 (Table 9a

) and -3:1 in acetone-c/^ (Table 9b).

Based on the above spectral evidence, the structure of compounnd FDXiA was

characterised as 4-chloroacetophenone-l l-(carboxymethylthio)undecano hydrazone

(32)which is represented in its two forms as in Fig. 9

261

6 Q

r ?

Q to 0 03 cs -o

I. u O) a. iZl P!i §

u

-n c: «

S Z 1 a • • « OS a 3 cs H

S a

to

S3 I U

N K

s .H

^ £ bX) S « .2 -t-t a es

s a, & to'

u CJ

o r-

O

m" OS tri

O

o VO

O r .

O T

VO f^ cn —;

>n as

O

O >0

<N ON r-) in (N CN m

o oo 00 (N 1 t— IT) r—< o in

OS 00 (N 1 OS Tt in ^ (N q VO ^ f - H rn od 00 r - H en m (N m CN

(N <N " — ' ' — '

U I r- —1 <N on

o VO (N

t--l> oo

in m

as VO n

o 00 vd n

o

00 00

rr^ ri 4 ^ VO (H r n

CX t/: Vh u

a

K ffi ffi a X r<-i ^ CN CN VO ffi ffi ffi ffi ffi (N (N <N (N

O

0 1 (N oi (N CN oi

o

VO

O OS o

o s o

VO

O T —' VO

^ CN OS <N O o 00 m OS r-m in (N in in

(N 1— >—1 H (N rn

U (N r<-)

lO cn (S

I Si

in <

262

Table 9b: ^H-NMR spectral data of FDXiA (32) in acetone-rfg

H-nr 5(ppm) Integr-ation

Multip -licity

J (Hz)

7-CH3 2.30,2.34 3H s ( -25%, 75%)

NH 9.37, 9.68 IH s ( -75%, 25%)

2' 2.28,2.71 2H t J2.3'7.33, 7.32

( -25%, 75%) 3' 1.59 2H brp J«7.33

4'-9' 1.30 6x2H brs

10' 1.67 2H brp J»7.38

i r 2.64 2H t JIIMO'7.48

1" 3.22 2H s

Ar-2,6 7.84 2H d JAr-2,6 Ar-3,5 8.55

Ar-3,5 7.42 2H d JAr-3,5 Ar-2,6 8.55

2 38

H CH3

synperiplanar

H CH3

Structure Elucidation of FDX^B (33)

It was a colourless oily liquid. The structure of FDXjB has been fully

confirmed by IR, DCI-MS, "H-NMR, and '^C-NMR spectra. IR spectrum (Neat) gave

characteristic peaks at 3240 (NH), 3050 (OH), 2900, 2850 (C-H), 1700 (COOH), 1685

(CON) 1660 (CONH), 1600 (phenyl), 1420 (S-CH2), 825 and 780 cm"'. The DCI-MS

spectrum showed a set of [M+1]"^ peaks at m/z 501/503, confirming its molecular

weight 500/502 which corresponds with the molecular formula [C23H33N2O4S2CI].

This is equal to the sum of the molecular weights of compound (32) (426) plus

mercaptoacetic acid (92) minus one molecule of water. This suggested that the

formation of thiazolidinone ring has occurred by the cyclocondensation of (32) with

mercaptoacetic acid. The mode of fragmentations is shown in Chart 7, The set of

264

peaks at m/z 456/458 was due to [ (M+1) -C00H] . The diagnostic peak of the

thiazoiidinon ring was observed at m/z 427/429 [ (M+O-COCHjS] arised by the

cleavage of 1-2 and 3-4 bonds. The set of peaks at m/z 212/214 was due to

<2-cleavage (base peak) and an another set of peaks at m/z 229/231 was due to

/^-cleavage on the long alkyl chain to the thiazolidinone ring. It also showed

some common fragment ion peaks to that of (32) at m/z 276, 155, 134 and m/z

118.

nVz 456/548

O O s

HO" " f ^ T ^ N H m/z 275 (12.5)

O

HO"

O

HO'

0

^^-^fo^NH —N S

m/z 501/503 [M+1]

O

H ^ n h

m/z 427/429 (7.2/3.3)

S O m/z 155/157(31.2/13.1) m/z 197/199(13.4/4.1)

cleavage

- N "S w

o ^ m/z 229/231 (12.6/4.9)

- cleavage

m/z 212/214 (100.7/72.3)

CHART?

265

The signals in 'H-NMR and '^C-NMR spectra dissolved in acetone-c/^

have been assigned to individual H-and C-atoms (Table 10). The assignments

were made from their typical chemical shift values, multiplicity, relative

integrations and also by comparing with the spectra of the previously established

compound FBXjC (Tables 2a and 2b). All the signals in both the spectra were

found comparable except the signal for C-2 proton which didn ' t appear here at

5 6.04 and in place a new singlet at 5 1.98 for the methyl proton was observed. It

also showed a sharp singlet at 5 9.02 for the NH proton. Also in '^C-NMR

spectrum a new signal appeared at 6 20.21 for the methyl carbon at C-2. In

addition, both the spectra gave characteristic signals for the aromatic

protons/carbons of the /?-chlorophenyl ring.

Based on the above discussion, compound FDXjB was characterised as 2-

(4-chlorophenyl)-2-(methyl)-3-[l l-(carboxyrnethylthio)undecanamido]thiazo-

lidin-4-one (33).

.CI

FDX,B (33) H H

266

o c o <

Q

O - i - t re -a

u 0 u a IZl

csj s Z 1 u f)

PS s I

£ a H

S a a CO

<N ea 00 1 CO >n r-) t—1 ON O 00 ON 00 MD ,—1 00 (N <N 00 (N ON ON oo m l> MD a\ ro VO (N CM (N m CM (N ' — ' ' — ' ' — '

s U

N X

o m d m

I U CN (N

3 .Si S c .

W) S u .M C e« l-H

s c. a

CO 00 ON

U I (N

CN ^ (N (N rn ^ I I I I

>_ u< Uc < < < <

\D m m lO (N m

MD (N <N b LO r-1

r-' OO OO rN n n I I I I

>—> > 1—1 1—) I—)

a t/2 D. -D X) -a -o -a -a

(N X X ^ X X X X X < N ( N \ 0 ( N r M r M ( N ( N

(N o ON ON 00 M OL o in lO O q in (N RO ON (N - - - - (N

o. c 3 -o in 2 (N ro C\ J b ^ ^ ^

vo m fN r<-r I ( t l fcH

- < <

(U <u (D

JO

£ i C <u s d bJ3 t/2 [/3 <

67

Structure Elucidat ion of FDXj^C (34)

It was a white needle shaped crystalline solid. The structure of the compound was

established by IR, DCI-MS, 'H-NMR and '^C-NMR spectra. IR spectrum (KBr)

showed characteristic peaks at 3100 (OH, br), 1720 (COOH) and 1610 (phenyl) cm"'. In

DCI-MS spectrum the molecular ion peak was absent, but it showed a [(M)-

SCH2COOH] peak as the base peak at m/z 229/231 which corresponds with the stabile

fragment ion: HOOC-CH2-S-C^(CH3)-C6H4Cl. 'H-NMR spectrum dissolved in acetone-

de showed a singlet at 5 2.04 for the methyl proton. It also showed two doublets for two

methylene protons each at 5 3.36 (J=15.26, Hup) and 5 3.46 (J=15.26, Hdn). It gave

characteristic A2B2 signals for the p-chlorophenyl ring as doublets at 5 7.75 (J=8.85, Ar-

2,6), and 6 7.40 (J=8.85, Ar-3, 5). '^C-NMR spectrum dissolved in acetone-ii^ showed

the signals at 6c 30.25 (CH3), 61.77 (S-C-S), 34.04 (CHj), 170.80 (COOH), 142.46 (Ar-

C-1), 129.79 (Ar-C-2, 6), 129.28 (Ar-C-3, 5) and 134.03 (Ar-C-4).

Based on the above spectral evidence, compound FDX|C was characterised as

bis (carboxymethylthio)-4-chloroacetophenotal (34).

CI

H H C H j ^ ^ ^ H H HO. J X . . J X OH

O

F D X I C ( 3 4 )

268

p-Chloroacetophenone-ll-(2-carhoxyethylthio)Jundecanohydrazone FDX2A (35)

[Reaction of the hydrazone (31) with 3-mercaptopropionic acid]*

*[The target compound, 2-(4-chlorophenyl)-2-methyI-3-[l l-(2-carboxyethylthio)

undecanamido]thiazan-4-one could not be obtained probably because of increased

nucleophilicity of N=C-CH3 group carbon]

A solution of the hydrazone (31) in dry benzene was refluxed with 3-

mercaptopropionic acid (molar ratio, 1: 3) for 28 h. The products on purification by

column chromatography afforded a pale yellow solid, FDX2A (35) as the sole product

which was recrystallised (benzene-acetone) as white crystalline globules (54.2%).

Structure Elucidation of FDX^A (35)

It was a white globule shaped crystalline solid, m.p 72 °C. The structure of

FDXjA was fully established by IR, DCI-MS, 'H-NMR and '^C-NMR spectra. The IR

spectrum (KBr) gave characteristic peaks at 3175 (NH), 2900, 2850 (CH),

1680 (COOH), 1650 (CONH), 1595 (phenyl), 1540 (C=N) and 1485 (S-CH2) cm''.

The DCI-MS spectrum showed a set of [M+1]"^ peaks as the base peak at m/z 441/443,

confirming its molecular weight 440/442 which corresponds with the molecular formula

[C22H33N2O3SCI]. This is equal to the sum of the molecular weights of (31) (334/336)

and 3-mercaptopropionic acid (106). This indicated that FDX2A is an adduct of (31)

with HS.CH2.CH2.COOH. Other two prominent fragment ion peaks were observed at

m/z 169/170 [(M+1)-C,3H2303S] due to the cleavage of CO-NH bond and m/z 153/155

[(M+l)-Ci3H25N03S,] arised by the cleavage of N-NH bond.

The 'H-NMR and '^C-NMR signals could be easily assigned to individual H-and

C-atoms (Table 11) by comparing the spectra with that of previously established

269

I o

S Q a

(n <

X Q fe O a •M C5 -o

u u O) a

u

•o B «

s « H

e a.

60

L. C I

U

N ffi

W g C «

^ u

a a w to

o

o

VO r-i

O

O

o

CO ON

o Cp vO . o

o VO I

T

lO

On VO

(N O) rn m

r\

(N o4 m

00 r-00 (N 1 o 00 m

OO o 00 >—1 in o CS »—< p CO tn 00 rn 00

•tt 00 1-H rt r- m (N m CN (N m (N T—(

o r i

(N r^ rj- (N < <

in r-T < <

tn u-i I/O tn oo oo

rn (N p r-" »

(N »

r-r-' o

a\ a\ vd

tN <N ~ — (N

X) ^ t

( y - l ^

u — ^ X X X ^ ffi ffi E E E n: CN CS (N <N <N (N

O

o

(N <N

(N CN

O

o

o Tj-

o

o

rn o

(N m (N

-H Tt m (N ON 00 O 00 C--in IT) t-; Tt;

(N (S (N

S I U X

z tr-0 I

cs

(N ro I I

270

compounds FBX2B.(29)^ and FDXiA.(32) (Tables 6a, 6b and 9a, 9b). Both the spectra

revealed the double signals corresponding to synperiplanar and antiperiplanar

conformers around CO-NH bond as in (29) and (32).

Based on the above spectral evidence, the structure of compound FDX2A was

characterised as p-chloroacetophenone-1 l-(2-carboxyethylthio)undecano hydrazon (35)

which is represented as in Fig. 10

synpe r ip l ana r

H CH3

71

EXPERIMENTAL

p-Chloroacetophenone-10-undecenohydrazone (31)

A mixture of 10-undecenoicacid hydrazide (23) (4.00g, 0.0202 mol) and p-

chloroacetophenone (3.122g, 0.0202 mol) dissolved in dry benzene (50 ml) was refluxed

for 6 h as described in (24) and then worked up as usual. The products on crystallization

from ethanol furnished FD (32) as white crystalline needles, 5.624g (83%), m.p. 64 °C,

Rf 0.80 (benzene- diethylether, 6 : 4 v/v)

IR (KBr): 3175 (NH), 2900, 2850 (C-H), 1675, 1650 (CO), 1595 (phenyl,

C=C), 1540 (C=N), 1485, 1420, 1395, 1 180, 1110, 1090, 980, 900, 820 cm"'.

' H - N M R (400 MHz, acetone-^/^,): 8H 2.34, 2.76 ( -75%, 25%; 3H, s, CH;,-?).

9.37, 9.68 ( -75%, 25%; IH, s, NH), 2.28, 2.73 ( - 2 5 % . 75%; 2H, t, J=6.71, 7.62.

H-2'), 1.66 (2H, br p, J^7.48, H-3'), 1.31 (lOH, brs, H-4 '-8 ' ) , 2.04 (2H, m, H-9'),

5.79 ( IH, tdd, J -6 .72 , 17.09, 10.22, H-10') , 4.88 ( IH, dd, J=10.22, 2.29, Hg),

4.95 ( IH, dd, J= 17.09, 2.29, Hz), 7.84 (2H, d, J= 8.55,Ar-2,6), 7.42 (2H, d, J=

8.55, Ar-3,5).

'H-NMR (400 MHz, DMS0-</<5,): 5 H . 2 . 2 1 , 2 . 2 4 ( - 6 0 % , 4 0 % ; 3 H , s, 7-CH3), 10 .29,

10 .39 ( 4 0 % , 6 0 % ; I H , s, N H ) , 2 .31 , 2 . 6 4 ( 4 0 % , 6 0 % ; 2 H , t, J = 7 . 5 1 , 7 . 3 2 , H - 2 ' ) , 1.57

( 2 H , b rp , J - 6 . 9 6 , H - 3 ' ) , 1-28 (LOH, brs , H - 4 ' - 8 ' ) , 2 . 0 0 ( 2 H , td, J = 7 . 1 4 , 6 5 9 , H - 9 ' ) , 5 .76

( I H , ddt , J = 1 7 . 0 3 , 10.25, 6 .59 , H - 1 0 ) , 4 . 9 2 ( I H , dd , J = 1 0 . 2 5 , 2 . 1 4 , H E - 1 1 ' ) , 4 . 9 8 ( I H ,

dd , J - 1 7 . 0 3 , 2 .15 , H z - 1 1 ' ) , 7 .77 ( 2 H , d, J = 8 . 6 0 , A r - 2 , 6 ) , 7 .45 ( 2 H , d, J = 8 . 6 0 , A r - 3 , 5 ) .

'^C-NMR (lOOMHz, DMSO-rfg): 5c 13.19, 13.76 (-60%, 40%; C-7-CH3), 145.43,

149.35 (-60%, 40%; C-7), 175.13, 169.16 (-60%, 40%; C-1'), 32.25, 33.92 (-60%,

272

40%; C-2'), 24.20, 25.07 (-60%, 40%; C-3'), 28.18-28.87(C-4'-8'), 33.07 (C-9'),

138.73 (C-10'), 114.49 (C- l l ' ) , 133.66 (Ar-C-1), 127.51 (Ar-C-2,6), 128.25 (Ar-C-3,5),

137.11 (Ar-C-4).

DCI-MS (NH3): m/z 335/337 [(M+1) 100.0/48.4; CigHjyClNzO], 336 (49.6), 338 (21.8),

339 (7.7), 210 (7.8), 168/170 [(M+l)-CnH,90, 47.7/18.9], 169 (21.3), 171 (6.5), 167

(6.7), 152/154 [M-CnH2oNO, 10.6/7.5] and 153 (6.3).

p-Chloroacetophenone-ll-(carboxymethylthio)undecanohydrazone FDXjA (32), 2-(4-

cltloroplienyl)-2-methyl-3-[ll-(carboxymethylthio)undecanamido]thiazolidin-4-one

FDX,B (33) and bis(carboxymethylthio)-4-chloroacetophenotal FDX,C (34).

[Reaction of the hydrazone (31) with mercaptoacetic acid]

A mixture of the hydrazone (31), (0.782 mg, 0.0085 mol) dissolved in dry

benezene (20 ml) was refluxed with stirring for 24 h and worked up as described in (24).

The products on purification by column chromatography (silica gel, light pet.ether-

diethyl ether, 1:1 v/v) as eluent first afforded a yellow solid, labelled as FDX,A (32)

which on crystallization from EtOAc afforded crystalline globules, 520 mg (41.2%), m.p

68 °C, Rf 0.49 (benezene-diethylether, 6:4 v/v). Further elution of the column using light

pet.ether-diethylether, 3:7 v/v) furnished a colourless oily liquid, labelled as FDXjB (33)

490 mg (33.3%), Rf 0.39 (benzene-diethylether, 6:4 v/v) and then a pale yellow solid

labelled as FDXiB (34) which on crystallization from EtOAc gave white crystalline

needles, 120mg (13.2%), m.p.l04 °C, RfO.33 (benzene-diethylether, 6:4v/v).

273

Spectral data of FDX^A(32)

IR (KBr): v„ax 3150 (NH), 3050 (OH) 2900, 2850 (C-H), 1700 (COOH), 1640

(CONH), 1590 (phenyl), 1530 (C=N), 1480 (S-CH2) 1420, 1395, 1360, 1250,

1180, 1085, 825 cm"'

' H - N M R (400 MHz, acetone-fiTg): 5H 2.30, 2.34 ( - 2 5 % , 75%; 3H, s, CH,-7),

9.37, 9.68 ( - 7 5 % , 25%; IH, s, NH), 2.28, 2.71 ( - 2 5 % , 75%; 2H, t, J - 7.32.

7.33), 1.59 (2H, br p, J«7.33, H-3'), 1.30 (12H, br s, H-4 ' -9 ' ) , 1.67 (2H,br p.

J«7.48,H-10') , 2.64 (2H, t, J= 7.48, H-11') , 3.22 (2H, s, H-1" ) . 7.84 (2H, d, J=

8.55, Ar-2,6), 7.42 (2H, d, J=8.55, Ar-3,5).

' H - N M R (400 MHz, DMSO -RFG): SH 2.22, 2.24 ( - 6 0 % , 40%; 3H, s, CH,-7).

10.29, 10.39 ( - 4 0 % , 60%; IH, s, NH), 2.31, 2.64 ( -40%, 60%, 2H, t, J= 7.17, H-

2'), 1.50 (2H, br p, J «7.63, H-3'), 1.24 (12H, br s, H-4 ' -9 ' ) , 1.58 (2H, br p, J

-6 .87 , H-10 ' ) , 2.55 (2H, t, J =7.17, H - l l ' ) , 3.19 (2H, s, H-1" ) , 7.77 (2H, d, J =

8.55, Ar-2,6), 7.45 (2H, d, J = 8.55, Ar-3,5).

'^C-NMR (100 MHz, DMSO-rf^,): 5c 13.19, 13.76 ( - 6 0 % , 40%, CH3-7), 145.43,

149.36 ( -60%, 40%; C-7), 169.16, 175.13 ( -40%, 60%; C-1') , 32.25, 33.92

( -60%, 40%; C-2 ') 24.19 (C-3'), 28.0-28.80 (C-4'-10') . 31.64 (C-11') , 171.47

(C-2") , 33.19 (C-1") , 133.45 (Ar-C-1), 127.51 (Ar-C-2,6), 137.10 (Ar-C-4),

128.25 (Ar-C-3,5).

DCI-MS (NH3): m/z 427/429 [(M+1); 100.0/57.7 C21H3J CIN2O3S], 428 (72.5), 430

(31.6), 431 (11.3), 276 (11.1), 210 (9.3), 168/170 [(M)-Ci3H2303S, 40.4/18.9], 169

274

(24.0), 171 (10.1), 154/156 [M+1)-C,3H25N03S, 36.7/12.9], 155 (16.5), 152 (15.0), 153

(8.8), 73 (6.9).

Spectral data of FDXiB (33)

IR (Neat): v^^, 3240 (N-H), 3050(OH), 2900, 2850 (C-H), 1700 (COOH), 1660

(CON), 1640 (CONH), 1600 (phenyl), 1430 (S-CH2), 1240, 1160, 870, 780 cm"'

'H-NMR (400 MHz, acetone-cT^,): 5h 1-98 (3H, s, CH3-2), 3.45 ( I H , d, J=15.56,

H-5up), 3.53 ( IH , d, J=15.56, H-5dn), 9.02 ( lH,s , NH), 2.07 (2H, t, J=7.62, H-

2') , 1.49 (2H, br p, J=7.13, H-3') , 1.19 (12H, br s, H-4 ' -9 ' ) , 1.58 (2H, brp,

J=7.24, H-10 ' ) 2.63 (2H, t, J=7.33, H-11 ' ) , 3.22 ( IH , s, H-1"), 7.30 (IH, d.

J=8.53, Ar-2,6), 7.25 ( IH, d, J=8.53, Ar-3,5).

'^C-NMR (100 MHz, acetone-</<5,): 5c 25.21 (2-CH3), 67.19 (C-2), 172.12 (C-

4), 33.87(C-5), 172.48 (C-T) , 34.18 (C-2') , 25.97 (C-3 ' ) , 29.48-30.30 ( C - 4 ' -

11'), 33.77 (C-1"), 169.65(C-2"), 138.13 (Ar-C-1), 128.84 (Ar-C-2,6), 129.36

(Ar-C-3,5), 133.24(Ar-C-4).

D C I - M S (NH3): m/z (%) 501/503[(M+1)\ 12.3/5.7], 456/458[(M+l) -C02H,

11.7/4.8], 427/429[(M+1)-SCH2C0, 7.3/3.3], 273(10.8), 274 (10.2), 275(12.5),

276( 53.5), 277 (21.8), 278 (8.3), 258 ( 6.7), 259 (11.3), 239 (7.2), 240 (31.8),

241 (13.9), 242 (11.8), 230 (10.9), 231 ( 25.4), 232 (16.3), 223 (27.2), 224

(12.2), 210 ( 34.5), 212 (100.0), 213 (47.4) 214 (72.3), 215 ( 33.5), 216 (11.8),

217 (5.7), 218 (14.5), 197(13.4), 199 (4.1), 176 (11.8), 178 (12.7), 162 (8.1),

163 (8.2), 164 (8.1), 165 (7.9), 166 (40.4), 148 (15.0), 149 (10.0), 150 (9.0), 151

(15.4), 152 (16.3), 153 (7.5), 134 (42.2), 135 (20.0), 136 (32.2), 137 (19.0), 138

(12.0), 168 (18.4), 169 (13.0), 171 (8.1), 154 (22.9), 155 (13.5), 152 (17.0).

2 75

S p e c t r a l da ta of FDXi^C (34)

IR (KBr): v^^x 3100 (OH, br), 1720 (COOH), 1610 (phenyl), 1510, 1400, 1300, 1270,

1170, 1140, 1030, 840, 760 cm"'.

'H-NMR (acetone-rffi): 5h 2.04 (3H, s, CH3), 3.36 (2H, d, J = 15.26, H up), 3.46 (2H, d,

J= 15.26, H dn), 7.75 (2H, d, J=8.85, Ar-2,6), 7.40 (2H, d, J=8.85, Ar-3,5).

'^C-NMR (acetone-dg): 5c 30.25 (CH3) 61.77 (S-C-S), 34.04 (CH2), 170.80

(COOH), 142.46 (Ar-C-1), 129.79 (Ar-C-2,6), 129.28 (Ar-C-3,5), 134.03 (Ar-

C-4)

DCI-MS (NH3): m/z 229/231 [M-(SCH2.C00H), 100.0/231.0], 230 (39.8), 232 (14.8),

171 (23.7), 172 (9.2), 173 (9.5), 139 (14.9), 140 (6.0), 141 (6.0), 85 (5.0)

p-Chloroacetophenone-ll-[(2-carboxyethylthio)]undecanohydrazone F D X 2 A ( 3 5 )

[Reaction of the hydrazone (31) with 3-mercaptopropionic acid]

To a solution of the hydrazone (31) (1.0 g, 0.0028 mol) in dry benzene (20 ml)

was added 3-mercaptopropionic acid (0.903 g, 0.0085 mol) and refluxed for 28 h. The

products on usual work up and column chromatography (silica gel, pet. ether-diethyl

ether, 1:1 v/v) followed by crystallization from (benzene-acetone) afforded the sole

product F D X 2 A ( 3 5 ) as white crystalline globules, 705 mg (54.2%), m.p 72 °C , RF 0.48

(benzene-diethylether, 6:4 v/v).

Spectral data of FDX^A (35)

IR (KBr) : v„,ax 3175 (NH), 2900, 2850 (CH), 1680 ( m O H ) , 1650 (CONH), 1595

(Phenyl), 1540 (C=N), 1485 (S-CH2), 1420, 1460, 1400, 1250, 1190, 1110, 1090, and

825 cm-'.

276

'H-NMR (400 MHz, DMSO-^/tf ) : 6h 2.24, 2.27. ( -60%, 40%; 3H, s, 7-CH3),

10.31, 10.40 ( -40%, 60%; IH, s, NH), 2.32, 2.66 ( -40%, 60%; 2H, t, J =7.42, H-

2'), 1.51 (2H, brp, J;^7.12, H-3'), 1.24 (6x2H, br s, H-4'-9 ' ) , 1.59 (2H, br p, J

^7.02, H-10') , 2.48 (2H, t, J=6.96, H-11') , 2.65 (2H, t, J = 6.79, H - l " ) , 2.50 (2H,

t, J=6.79, H-2") , 7.78(2H, d, J=8.55, H-Ar-2, 6), 7.47 (2H, d, J= 8.55, H-2") ,

7.78 (2H, d, J=8.55, H-Ar-2, 6), 7.47 (2H, d, J -8 .55 , H-Ar-3,5);

'^C-NMR (100 MHz, DMSO-^/^,): 5c 13.20, 13.76 ( -60%, 40%; 7-CH3), 145.42,

149.34 ( - 6 0 % , 40%; C-7), 169.14, 175.11 ( -40%, 60%; C-1 ' ) 32.24, 33.92

( -60%, 40%; C-2') , 24.18(C-3'), 28.10-28.78 (C- 4 '-10') . 31.08 ( C - I T ) , 26.34

(C-1"), 34.51(C-2") , 178.47 (C-3") , 133.44 (C-Ar-1). 127.50 (C-Ar-2, 6),

137.08 (C-Ar-3, 5), 128.25 (C-Ar-4).

DCI-MS: m/z (%) 441 (100.0), 443949.7), 169 (18.3), 170 (5.9), 171 (8.9), 153

(16.7), 155 (6.8).

277

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J i J J J. \\ H H — I

SCREENING FOR BIOLOGICAL ACTIVITY

Hii

Study for Anticancer and Antimicrobial Activities

287

Biological Screening: Study for Anticancer and Antimicrobial

Activities

(A) A n t i c a n c e r activity against mal ignant human t u m o u r cells:


The compounds are evaluated as one dose primary anticancer assay in 3-cell

line panel consisting of three types of human cancers: breast (MCF7), Lung (NCI-

H460) and CNS (SF-268). In the screening protocol, each cell line is inoculated and

preincubated for 24-28 h on a microtiter plate. Test agents are then added at a single

concentration and the culture incubated for further 48 h. End-point determinations are

made with alamar blue'. Results for each test agent are reported as the percent of

growth of the treated cells when compared to the untreated control cells. Compounds

which reduce the growth of anyone of the cell lines to 32% or less (negative number

indicates cell kill) are then passed on for evaluation in the full panel of 60- cell lines

over a 5 -log dose range^.

In the fiill panel of 60- cell lines, 9 types of human cancers ; leukaemia, lung

cancer, colon cancer, central nervous system, melanoma, ovarian, renal, prostate and

breast are used. These test agents are added in five 10- fold dilutions and a 48 hours

continuous drug exposure protocol is used. Sulforhodamine B (SRB), a protein-

binding dye assay, is used to estimate cell viability or growth. Results are expressed

as log 10 GI50 which is the drug concentration (M) causing a 50% reduction in the net

protein increase in control cells during the drug incubation.

Data Analysis: The calculated Measurement of Effect: Percentage Growth (PG)

The measured effect of the compound on a cell line is currently calculated according

to one or the other of the following two expressions:

288

If (Mean ODtest - Mean ODtzero) ^ 0. then

PG = 100 X (Mean OD,est - Mean ODtzero)/ Mean OD, ero

Where:

Mean ODtzero^ The average optical density measurements of SRB-derived colour

just before exposure of cell to the test compound.

Mean OD,est= The average optical density measurements of SRB-derived colour after

48 hours exposure of cell to the test compound.

Mean ODcri = The average optical density measurements of SRB-derived colour after

48 hours with no exposure of cell to the test compound.

The results as obtained from experimental data are represented in three ways.

(i) The Data Sheet:

The first two columns of the Table describe the subpanei and cell lines

involved. The next two columns list the Mean ODtzero and Mean OCdri: the next five

columns list the Mean ODtest for each five different concentrations. Each concen-

tration is expressed as the logio (molar or ^g/ml). The next five columns list the

calculated PGs for each concentration. The response parameters GI50, TGI, and

LC50 are interpolated values represenfing the concentrations at which the PG is

+50.0. and -50. respectively.

(ii) Dose-Response Curves:

The dose-response curve page of the data package is created by plotting the

PGs against the logio of the corresponding concentration for every cell line. The cell

line curves are grouped by subpanei: Horizontal lines are provided at the PG values

of + 50.0.and -50. The concentrations corresponding to points where the curves cross

these lines are the GI50, TGI, and LC50 respectively,

(iii) The Mean Graphs:

The mean graphs show mean graphs at each of the principal response

parameters: GI50, TGI, and LC50. Bars extending to the right representing sensitivity

of cell lines to the test agent in excess of the average sensitivity of all tested cell lines.

289

Since the bar scale is logarithmic a bar 2 units to the right implies the compound

achieved the response parameters (eg. GI50) for the cell line at the concentration one-

hundredth the mean concentration required over all cell lines, and thus the cell line is

usually sensitive to that compound. Bar extending to the left correspondingly imply

sensitivity less than the mean.


The compounds [FTZ (1), FBCB (4), FTZX4B (7), FTCA (9), FFjC (12),

FZX (16), FTZX' (17), FT (20), FT' (21), FF (22), FB (24), FBX,C (26), FBX3A

(27), FBXjC (30), FD (31), FDXiA (32)] were evaluated as one dose primary

anticancer assay in a three cell line panel and the results are depicted in Table 1. Out

of the sixteen compounds screened, only one compound, F B X 3 A (27) was found to

be active. This compound was then screened for cytotoxic activity^ at five different

concentrations against 60 cell lines of nine types of human cancers: leukaemia, lung,

colon, CNS, melanoma, ovarian, renal, prostate and breast. A 48 h continuous drug

exposure protocol was used and a Suphorhodamine B protein assay was used to

estimate cell viability or growth. Results (Table 3) are expressed as logjoGI50' which

is the drug concentration (M) causing 50% reduction in the net protein increase in

control cells during the drug incubation. This compound showed activity only at

higher concentration. Noteworthy results was obtained for the compound F B X 3 A in

the case of non-small lung cancer, melanoma and renal where the reduction in growth

is 75, 74 and 70 and 77% respectively (Table 3).

290

Table 1. Results of anticancer activity of the compounds against 3-ceiI lines of human cancers

Test Compounds Concentration

Percent growth Test

Compounds Concentration (Breast) MCF7

(Lung) NCI-H460

(CNS) SF-268

Activity

FTZ (1) IxlQ-'M 69 93 95 Inactive

FBCB (4) IxlG'^M 72 98 1 12 Inactive

FTZX4B (7) 1x10"'M 79 99 106 Inactive

FTCA (9) IxlQ-'M 63 99 90 Inactive

FFjC (12) I x l C ' M 52 67 71 Inactive

FTZX(I6) IxlO'^M 49 82 113 Inactive

FTZX' (17) I x l C ' M 84 98 107 Inactive

FT (20) IxlQ-'M 72 96 109 Inactive

FT' (21) IxlG'^M 94 98 114 Inactive

FF (22) IxlQ-'M 82 96 112 Inactive

FB (24) I x l C ' M 59 95 107 Inactive

FBX,C (26) 1x10"'M 53 49 65 Inactive

FBX3A (27) lxlO"'M 14 1 1 Active

FBXjC (30) IxlQ-'M 48 53 75 Inactive

FD (31) 1X10"'M 61 98 114 Inactive

FDX,A (32) 1x10"'M 47 65 98 Inactive

The dose response curves of the compound F B X 3 A (27) treated with

malignant human tumor cell lines has been deciphered in Fig. l a (Table 2).

In F B X 3 A , the highest percentage of retardation of growth was observed in

NCI-H226 of non-small cell lung cancer (75%). Other significant activities

were observed in UACC-62 (74%), SKMEL-28 (70%) of melanoma cancer,

SN12C (72%) and U031 (70%) of renal cancer.

The overall anticancer effect of the compound (27) on all the malignant

human tumour cell lines is illustrated in Fig. lb .

Mean graphs of the compound (27) which facilitate visual scanning of

data for potential patterns of selectivity for particular cell lines with respect

to a selected response parameters are represented in Fig. Ic.

291

Table Z National Cancer Institute Developmental Therapeut ics P r o g r a m In-Vitro Testing Results

NSC: D- 722671 / I Exper iment ID: 0201NS46-28 Test T y p e : 0 8 Units: Molar

Report Date: February 4 , 2 0 0 2 Test Date: January 14, 2 0 0 2 Q N S : M C :

C O M ! : FBX 3A Stain Reagent: SRB Dual-Pass S S P L : OGVG

L o g l O C o n c e n t r a t i o n T i m e Mean O p t i c a l D e n s i t i e s

P a n e l / C e l l L i n e Z e r o C t r l - 8 0 - 7 0 - 6 0 - 5 0 L e u k e m i a

CCRF-CEM 0 2 1 0 0 7 8 4 0 7 7 6 0 . 7 5 7 0 7 8 9 0 6 1 8 H L - 6 0 ( T B ) 0 4 2 4 1 3 5 3 1 3 6 1 1 3 4 1 1 2 5 8 1 6 8 5 K - 5 6 2 0 1 9 6 1 0 7 2 1 0 6 2 1 0 7 9 1 0 9 8 0 7 3 6 MOLT-4 0 2 3 3 1 1 3 6 1 1 1 9 1 1 2 0 1 0 6 3 0 8 3 0 R P M I - 8 2 2 6 0 3 2 8 0 9 3 4 0 9 1 9 0 9 4 3 0 907 0 6 6 3 SR 0 3 4 0 0 7 0 2 0 7 3 2 0 6 6 7 0 6 3 0 0 4 3 0

N o n - S m a l l C e l l Lung C a n c e r A54 9/ATCC 0 1 6 0 0 7 2 5 0 7 7 5 0 7 8 2 0 737 0 6 3 3 EKVX 0 4 8 0 1 374 1 4 2 6 1 4 3 9 1 467 1 3 5 6 HOP-62 0 2 8 5 1 0 3 2 1 164 1 1 8 9 1 2 0 9 1 1 4 3 HOP-92 0 3 6 2 0 9 5 5 0 9 1 5 0 8 9 1 0 892 0 8 1 6 N C I - H 2 2 6 1 1 0 3 1 7 6 9 1 8 1 5 1 874 1 8 2 0 1 7 6 4 N C I - H 2 3 0 5 2 7 1 414 1 5 0 6 1 5 7 0 1 437 1 3 5 0 NCI-H322M 0 7 5 3 1 2 8 6 1 384 1 3 5 8 1 346 1 2 7 3 N C I - H 4 6 0 0 4 2 4 1 5 4 5 1 5 7 9 1 6 6 8 1 567 1 4 2 7 N C I - H 5 2 2 0 6 7 5 1 3 9 6 1 4 8 3 1 4 5 5 1 5 2 5 1 2 2 4

C o l o n C a n c e r COLO 20 5 0 1 3 5 1 2 4 8 1 2 6 6 1 3 2 2 1 192 1 1 6 0 HCC-2998 0 4 5 7 0 7 5 0 0 8 8 3 0 9 1 0 0 8 5 8 0 8 8 1 HCT-116 0 1 1 3 0 6 3 4 0 7 4 7 0 7 8 8 0 7 5 3 0 7 3 4 HCT-15 0 2 0 7 1 0 5 0 1 0 9 2 1 1 0 4 1 0 9 6 0 9 9 0 HT29 0 4 57 1 2 1 5 1 2 4 1 1 244 1 2 2 1 1 1 0 8 K«12 0 4 2 2 1 3 6 0 1 4 3 3 1 382 1 392 1 2 1 1 SW-620 0 2 5 9 1 1 4 3 1 2 8 5 1 3 3 1 1 3 6 8 1 3 3 9

CNS C a n c e r S F - 2 6 8 0 5 8 6 1 1 7 5 1 1 8 3 1 2 9 2 1 2 4 1 1 1 0 8 S F - 2 9 5 0 4 2 1 1 4 1 7 1 4 5 9 1 4 9 8 1 4 6 8 1 2 7 6 S F - 5 3 9 0 5 2 6 1 4 5 1 1 4 6 7 1 524 1 5 0 1 1 4 2 5 SNB-19 0 5 3 0 0 9 6 8 0 9 9 1 1 074 1 1 1 3 1 0 1 8 SNB-75 0 6 7 8 0 8 2 9 0 9 2 3 0 9 5 1 0 9 4 8 0 907 U2SI 0 2 5 3 1 0 9 0 1 144 1 1 1 0 1 0 2 9 1 0 0 2

Me lanoma LOX IMVI 0 3 3 3 1 0 1 0 1 0 7 3 0 914 0 914 0 8 4 0 KALME-3M 0 7 9 2 1 3 9 6 1 4 5 7 1 4 5 4 1 4 0 7 1 359 M14 0 3 7 6 0 9 4 4 1 0 2 9 1 1 1 3 1 0 5 8 0 9 4 5 SK-MEL-2 0 4 8 5 0 9 9 4 1 0 6 1 1 1 4 3 1 115 1 0 4 9 SK-MEL-28 0 7 7 1 1 4 3 3 1 4 1 8 1 4 7 6 1 474 1 4 5 1 SK-MEL-5 0 5 2 6 1 3 4 2 1 4 5 5 1 4 8 1 1 4 7 8 1 2 6 7 UACC-257 0 7 6 1 1 5 2 0 1 5 8 9 1 6 0 3 1 572 1 334 UACC-62 0 7 5 0 2 1 3 5 2 1 8 5 2 2 3 9 2 1 6 5 1 6 0 3

O v a r i a n C a n c e r IGROVl 0 329 0 9 3 1 1 0 1 8 1 0 1 2 0 8 8 8 0 9 4 9 OVCAR-3 0 2 9 3 0 9 4 8 0 9 9 2 0 9 9 5 0 9 6 9 0 8 3 6 OVCAR-4 0 8 0 1 1 5 8 8 1 5 8 5 1 6 8 1 1 5 9 3 1 3 8 4 OVCAR-5 0 3 1 3 0 9 9 8 1 0 6 9 1 0 9 5 1 036 1 0 1 8 OVCAR-B 0 3 7 9 1 0 5 0 1 1 2 0 1 1 4 2 1 1 4 0 1 0 7 8 SK-OV-3 0 1 4 7 0 9 9 4 1 0 2 7 1 0 9 0 1 0 5 5 1 102

R e n a l C a n c e r 7 8 6 - 0 0 2 4 2 1 1 5 9 1 1 9 9 1 2 6 2 1 237 1 1 4 3 A498 1 0 6 8 1 5 8 8 1 6 6 2 1 6 8 9 1 6 5 3 1 547 ACHN 0 3 8 8 1 4 5 0 1 4 6 7 1 4 8 3 1 4 8 8 1 2 8 3 C A K I - 1 0 4 8 0 1 5 6 7 1 6 1 9 1 6 6 8 1 598 1 3 3 0 SN12C 0 4 8 6 1 5 5 2 1 5 9 6 1 624 1 569 1 4 5 1 T K - 1 0 0 7 6 3 1 6 0 8 1 5 9 6 1 6 5 2 1 6 6 3 1 5 2 6 U C - 3 1 0 4 6 7 1 2 6 1 1 3 5 S 1 3 6 6 1 3 2 7 1 0 7 0

P r o s t a t e C a n c e r P C - 3 0 317 1 034 1 0 4 8 1 0 1 4 0 9 6 1 0 8 3 0 D U - 1 4 5 0 274 0 9 7 0 1 0 0 8 1 0 5 6 1 0 0 5 0 964

B r e a s t C a n c e r MCF7 0 5 6 7 1 1 0 7 1 0 8 3 1 2 0 9 1 1 6 1 1 0 6 1 NCI /ADR-RES 0 5 5 3 1 4 7 5 1 5 5 4 1 6 0 7 1 574 1 364 MDA-MP 231/ATCC 0 3 4 9 0 9 9 9 1 0 1 7 0 9 9 7 1 0 6 2 0 884 HS 5 7 s r 0 6 8 0 0 9 3 0 1 0 2 3 1 0 6 2 0 987 0 8 5 3 MDA-MB-435 0 5 1 8 1 8 1 1 1 9 9 7 2 0 3 2 2 0 0 9 1 6 6 1 MDA-N 0 544 1 7 2 1 1 7 8 7 1 S92 1 906 1 6 7 7 B T - 5 4 J 0 8 2 0 1 1 3 8 1 1 9 7 1 1 9 6 1 1 5 1 1 0 7 2 T - 4 7 D 0 2 5 8 0 8 0 0 0 8 6 1 0 8 1 5 0 8 1 5 0 7 0 9

-4 0

0 519 0 2 2 5 0 216 0 3 4 8 0 208

P e r c e n t G r o w t h

194 6 1 3 2 6 3 222 2 7 9 4 0 3 344 3 7 5

1 7 5 3 4 2 2 6 9 2 0 6 196 2 4 8 3 1 6

187 2 5 2 2 3 0 226 1 3 1

0 3 0 6 0 3 4 5 0 367 0 2 3 3 0 210 0 2 7 8

0 202 0 1 7 8 0 3 6 6 0 2 8 3 0 3 7 0 0 364

0 176 0 201 0 3 6 7 0 137 0 2 6 7 0 1 4 1

0 2 6 5 0 1 7 3

0 4 0 3 0 1 5 3 C 314 0 169 0 3 5 2 0 336

- 8 0 - 7 0 - 6 0 - 5 0 - 4 0 G I 5 0 T C I LC50

99 9 5 1 0 1 7 1 12 2 2 7 E - 0 5 > 1 OOE - 0 4 > 1 OOE - 0 4 1 0 1 99 90 136 10 4 8 2 E - 0 5 > 1 OOE - 0 4 >1 OOE - 0 4

99 1 0 1 1 0 3 62 3 1 e O E - C S >1 OOE - 0 4 >1 OOE - 0 4 ° 3 9 8 92 66 - 7 1 6 6 E - 0 5 7 95E - 0 5 >1 OOE - 0 4 98 1 0 1 96 55 3 1 2 6 E - 0 5 >1 OOE 04 >1 OOE - 0 4

lOS 90 80 2b 39 3 4 9 L 06 2 44L 05 >1 OOC 04

109 1 1 0 102 84 6 2 7 2 E - 0 5 >1 OOL - 0 4 >1 OOE 04 106 1 1 3 1 1 0 98 15 3 7 8 E - 0 5 >1 OOE - 0 4 >1 OOE 04 116 1 2 1 124 115 8 3 3 8 E - 0 5 8 62E - 0 5 >1 OOE 04

93 89 89 77 - 3 9 1 7 0 E - 0 5 4 61E - 0 5 >1 OOE 04 107 1 1 6 1 0 8 99 75 1 9 2 E 05 3 7 2 E 0 5 7 21E 05 1 ! 0 1 1 8 1 0 3 93 24 2 3 3 E - 0 5 b 27E 0 5 >1 OOE - 0 4 l i s 1 1 3 111 98 54 2 0 6 E 0 5 4 39E 0 5 9 37E 05 103 1 1 1 102 89 - 1 2 2 4 5 E - 0 5 7 66E - 0 5 >1 OOE - 0 4 112 1 0 8 l i e 76 - 5 4 1 5 8 E 0 5 3 B3E - 0 5 9 2 7 E - 0 5

102 1 0 7 95 92 4 2 9 9 E - 0 5 >1 OOE--04 >1 OOE 04 1 4 5 1 5 5 137 1 4 5 25 3 6 1 E - 0 5 7 l l E - •05 >1 OOE' -04 122 1 2 9 123 119 30 5 9 4 E 0 5 >1 OOE--04 >1 OOE' -04 1 0 5 1 0 6 1 0 5 93 2 8 8 E 0 5 9 8SE-- 0 5 >1 OOE--04 103 104 1 0 1 86 - 5 7 1 7 8 E 0 5 3 98E-- 0 5 3 90E^ - 0 5 108 1 0 2 1 0 3 84 - 4 1 1 8 7 E - 0 5 4 69E 0 5 >1 OOE 04 116 1 2 1 125 122 6 4 2 0 E 0 5 >1 OOE--04 >1 OOL •04

1 0 1 1 2 0 1 1 1 89 50 1 9 0 E 0 5 4 37E 05 >1 OOE 04 104 1 0 8 1 0 5 86 56 1 7 9 E - 0 5 4 04E 05 9 H E 05 102 1 0 8 1 0 5 97 52 2 0 7 E - 0 5 4 48E-•05 9 68E-•05 105 124 1 3 3 111 57 2 32E 0 5 4 60E 05 9 12E 05 163 1 8 1 179 152 67 2 9 3 E - 0 5 4 96E-- 0 5 8 39E-•05 106 1 0 2 93 89 - 4 8 1 9 3 E - 0 5 4 46E-•05 >1 OOE-•04

109 86 86 7 5 46 1 6 1 E - 0 5 4 18E 0 5 >1 OOE-•04 110 1 1 0 102 94 - 6 1 1 9 2 E - 0 5 4 0 2 E - •05 8 44E-•05 115 1 3 0 1 2 0 100 8 2 9 0 E - 0 5 8 37E-•05 >1 OOE-•04 113 1 2 9 124 1 1 1 - 2 4 2 8 2 E - 0 5 6 6 1 E - 0 5 >1 OOE-•04

98 1 0 6 106 1 0 3 - 7 0 2 0 2 E - 0 5 3 9 4 E - 0 5 7 67E-•05 114 1 1 7 117 9 1 - 6 0 1 8 6 E - 0 5 3 9 9 E - 0 5 8 S6E-•05 109 1 1 1 107 76 -D3 1 5 3 E - 0 5 3 4 9 E - 05 8 OOE--05 104 107 102 62 - 7 4 1 2 2 E - 0 5 2 B4E- 0 5 6 62E 05

114 1 1 3 93 1 0 3 - 3 9 2 3 6 E - 0 5 5 3 3 E - 0 5 > 1 OOE--04 107 1 0 7 1 0 3 83 - 3 9 1 8 6 E - 0 5 4 7 7 E - 0 5 >1 OOE- 04 1 0 0 1 1 2 1 0 1 74 - 5 4 1 5 4 E - 0 5 3 7 8 E - 05 9 26E 05 110 114 1 0 5 103 10 2 9 5 E - 0 5 8 1 9 E - 0 5 >1 OOE- 04 110 1 1 4 1 1 3 104 3 3 22E 0 5 9 4 7 t - 0 5 >1 OOL- 04 104 1 1 1 107 1 1 3 26 5 2 5 E - 0 5 >1 OOE- 04 >1 OOE- 04

104 1 1 1 109 9 8 11 2 7 7 E 0 5 1 9 7 E - OS >1 OOE--04 114 1 2 0 1 1 3 92 - 8 4 1 7 4 E - 0 5 3 3 5 E - 0 5 6 4 4 E - 05 102 1 0 3 104 84 - 4 8 1 8 1 E - 0 5 4 3 3 E - 0 5 ' 1 OOE- 04 105 1 0 9 103 78 24 1 8 9 E - 0 5 5 8 6 E - 0 5 >1 OOE- 04 104 1 0 7 102 90 72 1 7 e E - 0 5 3 6 1 E - 05 7 33L--05

99 1 0 5 106 90 - 6 5 1 8 2 E - 0 5 3 8 1 E - 0 5 8 OOE- 05 112 1 1 3 1 0 8 76 - 7 0 1 5 1 E - 0 5 3 3 2 E 0 5 7 3 1 E - 05

102 97 9 0 12 16 1 7 6 E 0 5 6 5 1 E 05 >1 OOL 04 105 1 1 2 1 0 5 99 - 3 7 3 0 F - 0 5 5 36E 0 5 >1 OOt- 04

96 1 1 9 1 1 0 91 - ? 9 2 2 1 E - 0 5 5 nc 0 5 >1 OOE 04 109 1 1 4 1 1 1 88 - 2 7 2 1 4 E - 0 5 5 80E 0 5 >1 oot- 04 103 1 0 0 110 82 - 5 6 1 7 1 E 0 5 3 9 2 E - 0 5 9 O l E - 05 137 1 5 3 1 2 3 69 - 5 4 1 4 3 E - 0 5 3 6 5 E 0 5 9 31E 05 114 1 1 7 1 1 5 88 - 6 7 1 7 6 E - 0 5 3 6 9 E - 0 5 7 72E 05 106 115 116 96 35 ? 2 5 E - 0 5 5 3 9 F 0 5 >1 OOr 04 118 118 104 79 59 1 6 3 E 0 5 3 7 4 L 05 8 6 0 E - 05 111 1 0 3 1 0 3 83 13 -1 9 9 E - 0 5 >1 OOE 04 >1 OOE- 04

29?

Table 3. Anticancer activity of F B X 3 A (27) against 60 cell line at the concentration of

1.0 X 10"^

Type of cancer Cell line Concen-tration

LogioGISO Retardation of Growth (%)

L e u k a e m i a CCRF-CEM 1 0 X 10^ -4 64 _ HL-60 (TB) l O x l O - ' -4 32 -

K-562 -4 80 -

MOLT-4 l O x l Q - ^ -4 78 7 RPMI-8226 1 0x10"^ -4 90 -

SR I Ox 10-* -5 64 39 Non- sma l l cell A549/ATCC - -4 57 -

L u n g cancer EKVX - -4 42 -

HOP-62 1 0 x 1 0 - " -4 47 8 HOP-92 1 Ox ID"' -4 77 39 NC1-H226 1 Ox 10"' -4 72 75 NCI-H23 - -4 63 24 NCI-H322M - -4 69 54 NC1-H460 1 Ox 10^ -4 61 12 NC1-H522 1 Ox 10"^ -4 80 54

Colon cancer COLO 205 -4 52 -

HCC-2998 1 Ox lO"" -4 44 25 HCT-116 I Ox 10"^ -4 23 HCT-15 1 Ox lO-" -4 54 HT29 1 Ox lO"* -4 75 57 KM 12 1 Ox 10"^ -4 73 41 SW-620 - -4 38 -

C N S cance r SF-268 1 Ox 10-* -4 72 50 SF-295 l O x l O - * -4 75 56 SF-539 1 Ox lO-* -4 68 51 SNB-I9 l O x l O - * -4 63 57 SNB-75 I Ox 10-' -4 53 67 U25I 1 Ox lO"' -4 71 48

M e l a n o m a LOX IMVI l O x l O - ' -4 79 46 MALME-3M -4 72 61 M14 - -4 54 8 SK-MEL-2 1 Ox lO-" -4 55 24 SK-MEL-28 1 Ox lO-' -4 69 97 SK-MEL-5 1 Ox lO-* -4 73 60 UACC-257 1 Ox lO-' -4 82 64 UACC-62 1 Ox 10-' -4 91 74

Ovar ian cancer IGROVI 1 Ox 10-* -4 63 39 OVCAR-3 1 Ox 10"' -4 73 39 OVCAR-4 1 Ox 10-* -481 54 OVCAR-5 - -4 53 10 OVCAR-8 1 Ox 10-* -4 49 3 SK-OV-3 1 Ox lO-' -4 28 -

Renal c ance r 786-0 ! 0 x 1 0 - ' -4 56 11 A498 1 0 x 1 0 - ' -4 76 84 ACHN 1 Ox 10-* -4 74 48 CAKI-1 - -4 72 24 SN12C 1 0 x 1 0 - ' -4 75 72 TK-10 l O x l O - " 4 74 65 UO-31 1 Ox lO-' -4 82 70

Pros ta te cance r PC-3 1 Ox lO-* -4 75 16 DU-I45 . -4 64 37

Breas t cance r MCF7 1 Ox 10"' -4 66 29 NCI/ADR-RES - -4 67 27 MDA-MB-231/ATCC -4 77 56 HS 578T 1 Ox 10-' -4 84 54 MDA-MB-435 - -4 75 67 MDA-N -4 65 35 BT-549 1 Ox 10-' -4 79 59 T-47D 1 Ox 10"' -4 52 13

293

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(B) A N T I M I C R O B I A L A C T I V I T Y

INTRODUCTION

The test organism used in the study included Escherichia coli UP2566

(Central Drug Research Institute, Lucknow, India), Bacillus subtilis MTCC-121

(Institute of Microbial Technology, Chandigarh, India) and clinical isolates of

Staphylococcus aureus (IOA-106), Pseudomoms aeruginosa (IOA-110) and

Candida albicans (IOA-109) (Department of Microbiology, J.N. Medical College,

AMU, Aligarh, India).

Culture Media and Inoculum

Sabour and Dextrose (SD), Nutrient broth and agar were obtained from

Hi-Media Pvt. Ltd. (Bombay, India) and were used for Canadida albicans and test

bacteria respectively. Freshly grown microbial cultures at 37° were appropriately

diluted in sterile normal saline solution to obtain a cell suspension of lO^CFU/ml.

Antimicrobial assay

Each synthetic compound was dissolved in DMSO to obtain a stock solution of

different concentrations ranging from 2000 - 5000 ng/ml. The agar well diffusion

method (Perez et. al^) as adopted by Ahmad and Arina'^ was used for our assay.

0.1 ml of the diluted inoculum (10^ CFU/ml) of test organism was spread on

NA/SDA (Nutrient Agar/Sabour and Dextrose Agar) plates. Wells of 8 mm diameter

were punctured into the agar medium and filled with 100 jil of compound, solvent

blank and an antibiotic (chloramphenicol, 100 |ig/ml) to which the test bacteria were

sensitive. Fluconazole at the concentration of 100 i^g/ml was used as the control

against Candida albicans. The plates were incubated for 18 h at 37 °C. Antimicrobial

activity was evaluated by measuring the zone of inhibition against the test organism.

297


A total of 10 compounds were tested against two Gram +ve bacteria

{Staphylococcus aureus, Bacillus subtilis) and two Gram -ve bacteria

{Estcherichia coli, Pseudomonas aeruginosa) and a yeast, Candida albicans. The

varying level of anti-microbial activity was detected against one or more test

organisms.

Antimicrobial activity of test compounds

The results for the antimicrobial study of the tested compounds detected

against the test organisms Escherichia coli, Bacillus subtilis. Staphylococcus aureus,

Pseudomonas aeruginosa and Candida albicans are depicted in Table 4. Solvent

control (DMSO) showed non-significant inhibition to test microorganisms.

Antimicrobial activity against Gram -ve bacteria was deduced in all compounds.

However such activity could be detected only in four compounds against Gram +ve

bacteria. The compounds (25), (26), (30) and (31) demonstrated activity against both

Gram +ve and Gram -ve bacteria.

The compounds exhibited antimicrobial activity in the concentration range

200 \ig to 500 ^g/100 fil. Two compounds, (30) and (31) also demonstrated

antifungal (anticandidal) activity. It was interesting to note that the compound (31)

which is structurally similar to (24) but only differs in substitution, resulted in the

enhanced activity against fungi. However, the compound (24) is devoid of antifungal

activity. Overall broad spectrum antimicrobial activity i.e. active against Gram +ve

and Gram -ve was deduced in (25), (26), (30) and (31). These compounds may be

directly explored in the preparation of topical antiinfective agents. However, further

exploration requires detailed study on exact MIC values and least toxicity to host cell

and system.

298

Table 4. Antimicrobial activity of some compounds by Agar well diffusion

method

Test Compounds

Effective concentration

Hg/well

Antimicrobial activity in terms of zone of inhibition in mm Test

Compounds

Effective concentration

Hg/well SA BS EC PA CA

FB (24) 500 - - 19 - -

FBX,B (25) 500 12 12 10 14 -

FBX.C (26) 250 - 21 13 12 -

FBXjC (30) 200 12 14 10 16 15

FBXjA (27) 500 - - 18 22 -

FD (31) 500 22 12 28 18 18

FDX,A (32) 500 - - 17 21 1 FTZX4B (7) 500 - - 10 12 -

FBX3B (28) 250 - - 14 16 -

FBCB (4) 300 - - 15 18 -

Chloram- 100 |ig/well 25 20 24 30 -

phenicol Fluconozole 100 ^ig/well - - - - 25

SA, Staphylococcus aureus; BS, Bacillus subtilis; EC, Escherichia coli; PA, Pseudomonas

aeruginosa; CA, Candida albicans.

299

REFERENCES

1. G.D. Gray and E. Wickstrom, Biotechniques, 1996, 21, 780.

2. A. Monks, D. Scudiero, P. Skehan, R. Shoemaker, K. Paull, D. Vistica, C. Hose,

J. Langley, P. Cronise and A. Vaigro-wolff, J. Natl, Cancer Inst., 1991, 38, 757.

3. C. Perez, M. Pauli and P. Bazerque, Acta Biologiae et Medicinae Experimentalis,

1990, 15 , 113.

4. Arina Z. Beg and I. Ahmad, World J. Microbiol, and Biotech., 2000, 16, 841.

300

CONCLUSION

The compounds synthesised in total of seventeen reactions and screened for anticancer/antimicrobial activities are as fol lows:

Chapters Compounds Synthesised

Total Chapters Simple Compounds

New Compounds

Total

I 4 2 6

II 2 2 4

III 4 3 7

IV 3 4 7

V 4 1 5

Grand Total 17 12 29

V I - Screening for Anticancer and Antimicrobial Activities:

(A) Anticancer Activity IG

(B) Antimicrobial Activity l 6

The techniques used for evaluation of structure are 'H-NMR, '^C-NMR,

APT, NOE, 2-DNMR (COSY, HETCOR) including DC-MS, IR and FTIR.

301

List of Publications and Presentations

1. Bifunctional derivatives of70,/>'-dichlorochalcone Part II.

Synthesis of a novel compound 2-[2-(carboxymethylthio-2-(4-chloro-

phenyl)ethyl]-2-(4-chlorophenyl)-4-thiazolidinone.

molecules, 4, 232 (1999).

2. Synthesis, stereochemical study and biological activity of some novel

thiazolidinones, 2-aryl substituted 3-[ l l - (carboxymethylthio)undecan-

amido]thiazolidin-4-ones alongwith l l - (carboxymethyl /e thyl th io undec-

anohydrazones from 10-undecenohydrazide.

Tetrahedron (2002) (communicated)

3. Synthesis and biological activity of novel long alkyl chain substituted-5'-

triazolo-thiadiazepines.

Eur. J. Med. Chem. (2002) (communicated)

4. Synthesis and biological activity of novel long alkyl chain substituted-

benzopyrano-i '-triazolo-thiadiazepines.

Tetrahedron Lett. (2002) (communicated)

5. Synthesis and biological activity of novel long alkyl chain substituted-

i--triazolo-thiadiazoles, Indian J. Chem. (2002) (communicated)

6. Synthesis, stereochemical investigation and biological activity of novel

cyclohexanols from chalcones and their stereoselective syn elimination

products, Eur. J. Med. Chem. (2002) (communicated)

7. Flavonol glycosides from Mallotus philippensis (Euphorbiaceae).

{33'''' Annual convention of chemists, Coimbatore, December 26-29, 1996).

8. Synthesis of 2-p-chlorophenyl -2- ( r -phenyl - r - th iapropionic acid ethyl)

tetrahydro-l ,3-thiazin-4-one from chalcone.

{33''' Annual convention of chemists, Coimbatore, December 26-29, 1996).

9. Synthesis and stereochemical study of E-3-benzylidene flavanones and

their thio-adducts.

{39'^ Annual convention of chemists, Nagarjunanagar , December 22-26,

2002).

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