Bio-Active Heterocyclic Compounds

As Potential Antimicrobial Agents

A Thesis Submitted
To
BHAVNAGAR UNIVERSITY

UNDER THE GUIDANCE OF

Dr. N. C. DESAI
(PROFESSOR IN CHEMISTRY)

UNIVERSITY DEPARTMENT OF CHEMISTRY

BHAVNAGAR UNIVERSITY
BHAVNAGAR- 364 002 (INDIA)

Date : 17/05/08
 Objective

The fundamental goal of this work

is to try to assist the chemist in his
search for what to make next.
 Contents
• Synopsis
• General introduction
• Studies on 4-oxo-quinazoline and 4-
oxo-thiazolidine derivatives
• Studies on 5-imidazolone
derivatives
• Studies on [1,2,4]-triazole
derivatives
• Biological evaluation and
Characterization of compounds
• Conclusion
 General Introduction
Medicinal chemistry is the design and synthesis
of novel drugs, based on an understanding of how
they work at the molecular level. A useful drug must
interact with a molecular target in the body and also
be capable of reaching that target.

Most of the activity in this discipline is directed

to new natural or synthetic organic compounds.

A logical approach to the study of drugs and

their activities is the recognition of the basic
principles behind the biochemical events leading to
drug actions.
 Perspectives of Drugs
In a broad sense Medicinal Chemistry is a science of
Drugs.

It deals with the exogenously administrated

chemical molecules with living systems.

The fundamental objective of drug research is to

learn the cellular language in order to devise
meaningful and specific message.

Pharmacotherapy is usually concerned with two

factors:
Drug and Organism
 Drugs Discovery
• Our society is faced with challenges such as new disease like AIDS,
drug resistance, and aggressive agriculture pest control processes,
which can have a chemical agreement.

• To design new drug with improved properties and diminished side-

effects, and to assess the safety of some chemicals.

• The assessment of the risk of chemicals released to the environment

and the evolvement of environmentally benign synthesis method is
strongly required.

• There is also a demand on scientific methods that replace or at least

refine and reduce the use of laboratory animals.

• It should be Cost effective.

• Many of the concepts and methodologies applied to the design of

medicinally interesting compounds
 Drugs Development
It takes on average about 12-15 years to develop a drug from
an idea through to a product that can be sold for use in patients.

A little more than half of this time is spent testing the drug in
the laboratory (known as 'pre-clinical' testing).

The remaining is spent testing the product in humans (known as

'clinical‘ testing) and having the data reviewed by the regulatory
authorities.
Ideal Characteristics of Chemotherapeutic
Agents
• Selective toxicity
• Antimicrobial spectrum
• No side effects
• No killing effect on normal flora
• No inactivation
• Solubility in body fluids
• Sufficient concentration of the drug in target tissues
• Low break-down rate of drug
• No development of drug-resistance
• Stable viability
• Easily availability at affordable cost/price
Different Possible Route For Synthesis of
4-Oxo-quinazoline Moiety
COOH COOH

RCOCl
NH2 NHCOR
(i) (ii)

RCONHR'
(CH3CO)2O

O O

COOH R'
N H2NR' O
SOCl2 / DMF
Formamide
NH2 (R, R' =H) R N R
N
(i) (iii)
R' (A)
N

Cl SnCl2 HCl / R-CH2OH

R
reflux,2 days
O
CO2CH3
R'
NH

NH2
NO2
(v)
(vi)

Anthranilic acid is heated with excess of formamide in presence of

thionyl chloride, water is expelled and a nearly quantitative conversion of
4-oxo-quinazoline (A) is achieved. (A) are the formal condensation products
of anthranilic acid and amides, and these can also be prepared in this fashion
through the Niementowski (1866-1925) quinazolinone synthesis.
 Pharmaceutical Importance of
4-Oxo-quinazolines
Zaroxolyn (metolazone) is a currently available drug in
market which belongs to quinazoline class having
diuretic/antihyper-tensive potency.H
Cl N CH3

N
H2NO2S

O
CH3

Shingo Hirai and coworkers synthesized Quinazolinone type

alkaloids, febrifugine and isofebrifugine isolated from Dichroa
febrifuga roots, show powerful antimalarial activity against
Plasmodium falciparum.
N OH N
O OH O

N N
NH NH
O O
 Chemistry and Mechanism of
4-Oxo-thiazolidines
Most of the thiazolidines have been prepared by the
condensation of the required aldehydes with hydrazides and
2-sulfanyl acetic acid. Here, electronegative sulphur acquires
negative charge, which in turn is responsible for the attraction
between sulphur and the electropositive imino carbon. To maintain
the stability carbon breaks the π-Bond with nitrogen atom, leading
to the development of negative charge on nitrogen. This reactive
nitrogen attacks on the carbonyl carbon, which has positive charge
thus eliminating hydroxy group and forming the Thiazolidinone
ring. The hydroxyl group and the hydrogen atom released
combines together to give a water molecule.

O OH Ar O Ar
N N
H
S R R
H S
H
 Pharmaceutical Importance of
4-Oxo-thiazolidines
N C Desai et al have synthesized several 4-
oxo-quinazoline and thiazolidine derivatives and
O tested them for their anti HIV, anticancer and
O S
antitubercular activities.
N
N

N C6H 5

Shanker and coworkers

synthesized several quinazolines
as potential anti-inflammatory
O
O S
agents. Panamkant and Saksena
R
N S N synthesized 2-phenyl-3-p-(2′ -
O ethyl-3′ -aryl-4′ -oxo- thiazolin-
O
N CH 3 Ar 2′ -yl)phenyl quinazolin-4-ones
and studied their antimicrobial
activity.
Reaction Scheme of 4-Oxo-quinazoline

COCl O NH 2

COOH
Pyridine O
+
+
0-4oC, 2 hrs.
NH 2 N

CH 3 COOC2H 5
CH 3
Compound A

Pyridine
8-9 hrs.
O

NH 2 COOC2H 5
O NH O

N NH 2NH 2H 2O N
Methanol, 6-7 hrs.
N N

CH 3 CH 3
Compound C Compound B
Reaction Scheme of 4-Oxo-thiazolidines
O

NH 2
O NH

CH 3
Compound C

(1) R-CHO (2) SHCH 2COOH

Methanol, 6-7 hrs. 1:4 Dioxane, 6-7 hrs.

O NH R
N
N
S
O
N

CH 3
Compound D
O
NaOMe, 8-9 hrs.
Ar H
O

O NH R

N
N
S
O
N

CH 3 Ar
Compound E
Where, Ar = Different aryl groups
 Studies on 5-Imidazolone Derivatives
Many of the azoles comprise the ring system of several
natural and synthetic compounds which are important for the
living systems and also as important drugs, dyes and
agricultural chemicals.

IMPORTANT ANTIBACTERIAL AND ANTIFUNGAL DRUGS

One of these {2-butyl-5-chloro-3-

[2’-(1H-tetrazol-5-yl)- biphenyl-4-yl
HO methyl] -3H-imidazol-4-yl}methanol
Cl [Losartan] is undergoing extensive
clinical evaluation. This pioneer work
N N
N NH initiated a flurry of activity in
N N pharmaceutical research, and many
n-Bu
Losartan other nonpeptide orally active AT1
angiotensin-II receptor antagonists have
been reported.
Different Possible Route For Synthesis of
5-Imidazolone Moiety
X
CO
NH S A Siddiqui et al introduced
CH 3COONa
some new imidazolone (D),
COOH
OH CHO
+ (CH 3CO)2O
synthesized by the condensation of
aromatic and substituted aromatic
CH 3O (A) (B)
amines with 5-oxazolone
X X
derivatives (C) in presence of (Y-H)
OH O Zeolite (Y-H) O
Zeolite. The 5-oxazolone derivatives
OH
Ar-NH 2 are prepared by the condensation of
CH 3O N N N O
hippuric acid with 5-bromo/iodo-4-
CH 3O
hydroxy-3-methoxy benzaldehyde in
presence of sodium acetate and
R (C)
(D)
acetic anhydride.
Where X = Br / I

R O O R1
R1-NH 2 R NH
N O O
NH
Ar Ar
Amides of acylamino acrylic acids
R O
R1-NH 2
K CO POCl3
Ethanol 2 3
N N
R1
Ar
4-arylidine-imidazolin-5-ones
Ar = -C6H 5, R = Different aryl groups, 1= R Alkyl or aryl groups
 Sequence of Transformation of
Imidazolone From Oxazolone
(a) Cyclisation of hippuric acid and aromatic aldehydes to form Oxazolones.
(b)Proton abstract by base results in the generation of carbanion.
(c) Carbanion attacks on carbonyl carbon of aldehyde, followed by
dehydration .
(d)Ring opening by attack of amine.
(e) Ring closure results in the formation of imidazolone ring.

Ar O
H Ar O Ar O

N O .
N N N
H
N N Ar

.
H Ar OH Ar
 Pharmaceutical Importance of
5-Imidazolone Derivatives
Imidazolone ring system is of biological and chemical interest since long. The
imidazolinones are associated with a wide range of therapeutic activities such as
anticonvulsant, sedative and hypnotic, potent CNS depressant, antihistamine,
antimalarial, bacteriocidal, fungicidal, anti-inflammatory, MAO inhibitory,
antiparkinsonian, antihypertensive and anthelmintic.

Hooshang et al synthesized 3-methyl-2-{[5-oxo-2-phenyl-4-(phenylmethylene)(2-

imidazolinyl)]amino}-3- hydroquinazolinones which is an important compound in
chemistry and pharmacology.

O
Ph
N CH 3 N

N
N NH
C6H 5
O
Reaction Scheme of 5-Imidazolone
CONHCH 2COOH

Ar H
+
N-benzoyl amino acetic acid

Anhydrous
(CH 3CO)2O
CH 3COONa
Ar O O
O

N O N
+H N NH N
2
R
Pyridine
150oC, 6-8 hrs.
O
Ar O O

N
N N NH N
R

Where, Ar = Different aryl groups,R = 3-Nitrophenol, 4-Nitrophenol

Studies on [1,2,4]-Triazole Derivatives

The presence of additional nitrogen atoms in the five-

membered ring has important effects on the properties of the
ring system.

Triazoles have been found to exhibit properties such as

anticonvulsant, analgesic, anti-inflammatory, herbicidal,
anthelmintic, CNS depressant, antitumor agent and
tranquilizing. Fluconazole, Ribavirin, Triazolam, Itraconazole,
Viramidine, Voriconazole and Terconazole are leading
N-substituted 1,2,4-triazole derivatives and these are used on

tropical antifungal infections.

Molecular Geometry of [1,2,4]-Triazole
Studies carried out at 1600C proves the molecular dimensions
as shown in following table for one unit of a pleated sheet linked
by hydrogen bridges. Slightly different values are obtained at
room temperature or in substituted aromatic triazoles.
5 4
NH

1N N 3

Angle (°) Bond Bond length (m)

5-1-4 110.2 1-2 135.9

1-2-3 102.1 2-3 132.3
2-3-4 114.6 3-4 135.9
3-4-5 103.0 4-5 132.4
4-5-1 110.1 5-1 133.1
N(4)-H 103.0
C(2)-H 93.0
C(5)-H 93.0

a
The numbering refers to annular centers in 1,2,4-triazole.
 Different Possible Route For Synthesis of
[1,2,4]-Triazole Moiety
Route (1) 1,3-dipolar cycloadditions
leading to a great variety of heterocyclic
systems are applicable to the synthesis of
triazoles and derivatives. Nitrilimines
formed by dehydro-halogenation of C-
halobenzylidenephenyl- hydrazones, which
react with phenylcyanide, phenyl
PhC NNHPh isocyanate and benzylidenephenylamine to
Cl afford triazoles derivatives.
C-halobenzylidenephenylhydrazones

Et3N

+ -
PhC N NPh
PhCN Nitrilimines PhCH NPh

Ph Ph
PhNCO H N
N
Ar N
N
Ph
N Ph N
Ph N Ph Ph
O N
N
Ph Ph
Route (2) Ring closure of acyl
derivatives of semicarbazides,
thiosemicarbazides or aminoguanidines
in alkaline solutions is a method widely
applied for the preparation of 1,2,4-
triazoles. Gehlen reported that 3-
hydroxy-5-alkyl–1,2,4-triazoles are
produced in 65-75% yield by this
method.

O
NH N
NH NH 2
N KHCO 3 OH
R' O N

Where R'=H, 6CH 5

 Pharmaceutical Importance of
[1,2,4]-Triazole Derivatives

Svensson et al synthesized Y X W
NH
2,4-dihydro-[1,2,4] triazole-3-thione. S
N
These compounds are inhibitors of the NH
enzyme myeloperoxidase (MPO) and Where W = H, CH
3, F, OH, CH2OH, Ph
are thereby particularly useful in the X = O, CH 3
2, NR . R3 = H/Cl/6 alkyl
treatment of prophylaxis of Y = phenyl, naphthyl
neuroinflammatory disorders.

R.R.Kamble et al have developed

O S
R a 3’-substituted-2-aryl-5-methyl- 5’-
N NH thioxo- [4,4’-bi-4H-1,2,4-triazol]-
N N
N N 3(1’H,2H)-ones and found intresing
properties, such as antinociceptive,
CH 3 R'
anticancer and plant growth regulative
Where R=Aryl groups, R'=Alkyl groups activities.
Commercially Available [1,2,4]-Triazoles As
Antimicrobial Agents
O
N Cl
NH 2
F N
N
N N
HO CH 3
F OH N
O N
N
N N N
Cl N
N HO OH
1. Fluconazole 2. Ribavirin 3. Triazolam

CH 3 H
O HO O N
O N
H 3C N
N N
N N N O O NH
N N
N
HO OH NH 2

4. Itraconazole Cl Cl 5. Viramidine

N
N
N
N N O O
OH CH N
3
F
F O
Cl N
N Cl
N N CH 3
F
6. Voriconazole 7. Terconazole CH 3
Reaction Scheme of [1,2,4]-Triazole
O NH
NH 2

CS2
KOH
C2H 5OH

O NH
NH S- K +

R
Compound A

C2H 5OH
Ar-CONHNH 2

SH O

N Ar

N N NH

R
Where, Ar = Different aryl groups
R =Naphthalene , 4-Chloro-1-methoxybenzene,
Methoxynaphthalene , 1-Methyl-3-nitrobenzene
 Biological Evaluation
ANTIBACTERIAL AND ANTIFUNGAL ACTIVITIES OF THE
COMPOUNDS
SYNTHESISED IN PART - I , II & III.

Escherichia coli (Gram negative) MTCC-443

Pseudomonas aeruginosa (Gram negative) MTCC-1688
Staphylococcus aureus (Gram positive) MTCC-96
Streptococcus pyogenes (Gram positive) MTCC-442
Candida albicans MTCC-227
Aspergillus niger MTCC-282
Aspergillus clavatus MTCC-1323
Important Antibacterial & Antifungal Drugs
H 2N OH
OH OH
H 3C O OH
NH O HO O
HO O OH OH OH OH O O
O CH 3
O NH 2 NH OH
H 3C

H 2N O O CH 3

Gentamycin K. Nystatin
HO OH

NH 2

Experimentation For Antibacterial Screening

 Characterization

We have determined the structure of the

newly synthesized compounds by their Infrared
spectroscopy (IR), Gas Chromatography Mass
Spectroscopy (GCMS) and Proton Magnetic
Resonance Spectroscopy (PMR).
 Conclusion
•Discussion of antibacterial activity

Part-I deals with the synthesis and antibacterial activities of

quinazoline which bears the 4-oxo-thiazolidine. On the basis of the
results of antibacterial activity, it has been observed that compounds
PI-07, PI-12, PII-05, PII-06, PII-07, PII-10, PII-14, PIII-05, PIII-11, PIV-
01, PIV-04, PIV-06, PIV-07, PIV-09, PIV-11 and PIV-13 were found to be
moderately active. PI-07, PI-13, PII-07, PII-08, PII-11, PIII-11 and PIV-
02 showed good activity against E. coli, while compound PII-01, PII-
10, PII-14, PIII-02, PIII-06, PIII-07 and PIII-13 showed excellent activity
against E. coli and P. aeruginosa.

Part-II describes the preparations and antibacterial activities

of 5-imidazolones. From the results of antibacterial activity, it has
been observed that compounds PV-11, PV-12, PVI-01, PVI-03, PVI-04,
PVI-11, PVI-12, PVI-13, PVI-14, PVII-03, PVII-04, PVII-06, PVII-10,
PVII-11, PVII-12, PVII-13, PVII-14, PVIII-02, PVIII-04, PVIII-10, PVIII-11,
PVIII-12 and PVIII-13 are moderately active.
 Conclusion
•Discussion of antibacterial activity

Compound PIV-02, PVI-04, PVI-07, PVII-03, PVII-06, PVII-10,

PVII-12, PVII-14, PVIII-05, PVIII-06, PVIII-07, PVIII-12 and PVIII-14
exhibit good activity. Compounds PVI-05, PVI-07, PVI-09, PVI-10 and
PVIII-02 showed excellent activity against E. coli.

Part-III encompasses the preparations and antibacterial

activities of 1,2,4-triazoles. Results of antibacterial activity indicate
that compounds PIX-02, PIX-03, PIX-08, PIX-10, PIX-11, PIX-12, PX-04,
PX-08, PX-09, PX-12, PXI-03, PXI-04, PXI-08, PXI-09, PXI-12, PXII-01,
PXII-02, PXII-04, PXII-07 and PXII-11 are moderately active.
Compounds PIX-01, PIX-06, PX-01, PX-04, PXI-06, PXI-08, PXI-12, PXII-
01, PXII-04 and PXII-11 showed good activity, whereas compounds
PX-08, PXI-10, PXII-09 showed excellent activity against E. coli.
 Conclusion
•Discussion of antifungal activity

The results of the compounds synthesized in part 1 to 3 clearly

reveals that PIII-02, PIII-09, PIV-10, PV-12, PVI-06, PVI-13, PVIII-02, PVIII-
04 and PIX-11 were found to be comparable with standard drug
against C. albicans, A. niger and A. clavatus. The compounds PVIII-05,
PIX-06, PIX-07, PIX-10 were found to be quiet active compared to
standard drug against C. albicans while compound PVIII-05, PIX-07
and PIX-10 were found to be quiet active compared to standard drug
against A. niger. Moreover compound PVIII-05, PIX-07 and PIX-10 were
found to be quiet active compared to standard drug against A.
clavatus.

For antibacterial activity, in present protocol, 100 µg/ml is

considered as moderate activity and 50 µg/ml is considered as good
activity. 25 and 12.5 µg/ml are considered as excellent as compared to
the standard drug Gentamycin. For antifungal activity, 100 µg/ml is
considered as compared to standard drug. 50 µg/ml is reported as
active compared to standard drug K. Nystatin.
 In the present work, we found some of the interesting antifungal
agents and we may send these compounds for the in vivo screening. The
structures of the compounds are as under with their FBC value.
Ar O O
PVIII -05 Where Ar = -2-OH-C10 H6
NH
OC2H 5
C. albicans (FBC) = 50 µg/ml
N N NH
S
P
OC2H 5
A. niger (FBC) = 50 µg/ml
A. clavatus (FBC) = 50 µg/ml

SECTION : 8

SH O

N Ar
PIX -06 Where Ar = -2-Cl-C6H4
N N NH
C. albicans (FBC) = 50 µg/ml

PIX -07 Where Ar = -3-Cl-C6H4

C. albicans (FBC) = 50 µg/ml
A. niger (FBC) = 50 µg/ml
SECTION : 9 A. clavatus (FBC) = 50 µg/ml

PIX -10 Where Ar = -4-NO2-C6H4

C. albicans (FBC) = 50 µg/ml
A. niger (FBC) = 50 µg/ml
A. clavatus (FBC) = 50 µg/ml
The road ahead in Medicinal Chemistry
The 2nd half of the 20th century showed a remarkable
advance in the therapy of bacterial infections.
Unfortunately due to the excess use of some currently
available antibacterial agents, the bacteria begin to fight
back-developing mechanism for resisting the antimicrobial
agents. And due to this there is a urgent need to find out
some new antibacterial agents. Keeping this in mind, we
have focused on the synthesis of some bioactive molecules
like imidazolone with a quinazolone moiety, and triazole
derivatives. On the basis of the results of antimicrobial
activity, we conclude that still there is a wide scope for
these derivatives in Medicinal Chemistry and by the help of
these compounds we may generate lead molecules.