1,3,4-Oxadiazole DHPM Derivatives As Potent

Antimicrobial Agent , A Review

A Graduation Research Submitted To

College Of Pharmacy / AlBayan University
As Partial Fulfillment Of The Requirement For The Degree
Of Bachelor In Pharmacy

By:
Sarah Maan AlDouri
Fatimah Nazar Hashem

Supervised By:
Dr.Ali Majeed Hantoush

2021 AD 1442 AH
 Acknowledgment

At the first it’s a lovely gift, that my dream came true so I’ll start by
thanking the God for this beautiful and amazing destiny. Reaching to this
stage requires toughness, effort, encourage and support, and for that I’d like
to thank my best two people in my life Mom & Dad
“Dr.Maan AlDouri & Dr.Ruqayah Aljuboury”
And my supervisor Dr. Ali Majeed, who had the greatest credit in this stage.
Also I can’t forget people who had a great influence in what I have become
“My aunt Lec. Nibrass AlDouri , Dr.Ameer Alwash ,my whole family and
all who taught me a thing or stood up with me .
Every success in my life will be thanks to you all, and I promise you that I’ll
not disappoint you .
In the end, pray for me to become Pharmaceutical Chemistry professor
Written by the student Sarah Maan AlDouri in 2021 March 30.

------------------------------------------------------------------

I would like to thank everyone who stood beside me, especially my mother
and father who stood beside me at every moment in all stages of my
scientific and practical life. It is difficult to reach this stage. It took effort,
persistence, patience, and I still persistence to become a pharmacist. of
course not forget to thank my doctors,
Dr . Ali Majeed and Dr Ameer Alwash theirs study look like a father to a
son .
I will do all my best
By Fatimah Nazar .

In the End, smile it’s only the Beginning.

I
 LIST OF CONTENTS

Contents Page
Acknowledgment I
List of Contents
II

List of Schemes II

Abstract IV

1. Introduction 1

2. Biginelli reaction 2
2.1.Mechanistic details 3
3.Result and Discussion 4
4.Antimycobacterial Activity 19
5.References 20

LIST OF SCHEMES
No. Title Page
1 Synthesis of DHPMs by Biginelli reaction. 2
2 The proposed mechanisms of Biginelli reaction 3
3 Synthesis of 1,3,4-oxadiazole derivatives. 4
4 Synthetic route for the preparation of 1,3,4-oxadiazole 6
derivatives.
5 Synthesis of targeted comound . 7
6 General synthesis of target compounds. 8

LIST OF SCHEMES
No. Title Page
7 Synthetic protocol for the title compounds. 10

II
 8 Synthetic pathway of novel compounds 11
9 Synthetic track for the preparation of title compounds 13
10 Synthesis of polysubstituted 2-amino-thiophenes 1,3,4- 15
oxadiazole-substituted thiophenes and 1,2,4-triazole
11 For synthesis of 1,3,4-oxadiazole derivatives. 16

12 Synthesis of compounds 3-arylaminomethyl-5-(3,4- 18

dimethoxyphenyl)-1,3,4-oxadiazole-2(3H)-thiones and 3-[(4-
substituted piperazin-1-yl)methyl]-5-(3,4-dimethoxyphenyl)-
1,3,4-oxadiazole-2(3H)-thiones .

Abstruct
Heterocyclic chemistry is the interesting area for the effective approach
and for the invention of biologically active 1,3,4-oxadiazole cores. 1,3,4
Oxadiazole is a five membered heterocyclic ring which plays key role in the

III
development of new medicinal species for the treatment of numerous
diseases .Nowadays researchers have developed the innovative methods for
the synthesis of 1,3,4-oxadiazole derivatives and their medicinal
applications.

In the other hand, molecules containing dihydropyrimidinone (DHPMs)

structures are very important in pharmaceutical and medicinal chemistry due
to their excellent biological activities application in synthesis of natural
products. Biginelli condensation reaction between an aldehyde, urea/
thiourea and a carbonyl compound is the most popular strategy for the
synthesis of dihydropyrimidinones by three mechanisms involving the first
mechanism, the so-called ‘iminium route’, the second mechanism involving
‘enamine route’ and the third mechanism involves a ‘Knoevenagel type
reaction mechanism’.

Antibacterial activity of the newly synthesized compounds was

evaluated by the well diffusion method against Staphylococcus aureus,
Bacillus subtilis (gram positive) and Escherichia coli, Klebsiella
pneumoniae (gram negative) strains of bacteria .They both possess diverse
biological activities such as antimicrobial, antiinflammatory, antitubercular,
antimalarial,etc.

IV
1.Introduction
The 1,3,4-oxadiazole derivatives are very well-known biologically
active N-containing heterocycles and represent a versatile lead molecule for
designing potential bioactive agents [1]. The oxadiazole derivatives have
been found to exhibit varied biological activities; viz, anti-microbial [1],
antifungal [2], anti-malarial [3], anti-inflammatory [4], and anti-tubercular [8].

However, the fact which cannot be overlooked is that, with the increase in
the number of a specific pattern of drugs, resistance towards them also
increases.Thus, owing to this increased drug resistance, new classes of
pharmacological agents with novel mechanisms are a crucial need so as to
combat with this multidrug resistivity. Thus , we have synthesized the 1,3,4-
oxadiazole derivatives incorporating the dihydropyrimidinone unit with a
view of developing new potentially bioactive compounds [6]. Needless to
mention, the dihydropyrimidinones have emerged as potential calcium
channel blockers, anticancer, antihypertensive, α-1a adrenergic antagonists,
and neuropeptide antagonists [7].

The dihydropyrimidinone skeleton thus has been studied extensively to

expand the existing structure to get new chemically and biologically
enhanced entities [6]. On the interesting site ,the combination of the two
works as antitubercular agents by design hybrid molecules by molecular
hybridization of different bioactive substances [8]. Dihydropyrimidines are
potential inhibitors of dihydrofolate reductase (DHFR), a promising drug
target for the development of anti-microbial agents. Although DHFR does
not represent a novel target, there is still enthusiasm for the development of
DHFR inhibitors, particularly with regard to mycobacteria [9]. Further, 1,3,4-
oxadiazole heterocycles are very good bioisosteres of amides and esters and
possess a phenomenal pharmacokinetic property, lipophilicity that
influences the drug’s ability to reach the target by transmembrane diffusion
and may demonstrate potent activity against resistant TB by inhibiting the
biosynthesis of lipids .Recognizing these facts and in continuation to our
endeavors toward the development of anti-infective agents , it was envisaged
that the design and synthesis of such novel compounds which include

1
advantage of dual pharmacophore of DHPMs and 1,3,4-oxadiazoles in single
molecular framework are worth the attempt [10].

2.Biginelli Reaction
Biginelli reaction is a multicomponent reactions (MCRs) involve a
combination of three or more reactants in a single vessel to produce a final
product with features of all the reactant components [11] , that involves acid-
catalyzed one-pot synthesis of 3,4-dihydropyrimidin-2(1H)-ones (DHPMs)
using easily-accessible starting materials, namely, aldehyde, active
methylene compound and (thio)urea ( Scheme 1) .DHPMs have stimulated
resurgence of interest in the past two decades due to their wide ranging
pharmacological activities and presence of diverse natural products.
Presently, green approaches to asymmetric Biginelli reaction are being
explored for bioactive chiral DHPMs. In materials chemistry, DHPMs are
increasingly finding applications in the development of materials such as
polymers, adhesives, fabric dyes, etc [12].

Scheme 1. Synthesis of DHPMs by Biginelli reaction.

2
2.1. Mechanistic Details:
As shown in (Scheme 2.), three mechanisms involving protonated
intermediates have been proposed [12].

The first mechanism, the so-called ‘iminium route’[13] involves

condensation between aldehyde and urea to give rise to an iminium
intermediate, which undergoes a nucleophilic addition with a β-keto ester
leading to DHPM [12].

The second mechanism involving ‘enamine route’[14] is based on

condensation between urea and β-keto ester leading to a protonated enamine
intermediate, which subsequently reacts with aldehyde to give rise to the
DHPM [12].

The third mechanism involves a ‘Knoevenagel type reaction

mechanism’[15] The reaction between aldehyde and β-keto ester results in
the formation of a carbenium ion intermediate, which reacts with urea to
afford the DHPM [12].

Scheme 2. The proposed mechanisms of Biginelli reaction .

3
3. Result and Discussion
In 2009 Kishor H. Chikhalia and his team tried to discover new
candidates with improved antimicrobial activities, here the synthesis and in
vitro biological evaluation of various series of 2-{(3,4,5-trimethoxyphenyl-
1,3,4-oxadiazolyl)-5-thio}-4-(morpholino)-6-(phenylureido)-striazine,and 2-
{(3,4,5-trimethoxy phenyl-1,3,4-oxadiazolyl)-5-thio}-4-(morpholino)-6-
(phenyl thioureido)-s-triazine. Antimicrobial properties of the title
compoundswere investigated against two Gram ( + ve) bacteria (S. aureus,B.
subtilis), two Gram (-ve) bacteria (P. aeruginosa, E. coli) and yeast-like
fungi (C. albicans) using the broth microdilution method. The commercially
available 3,4,5-trimethoxy benzoicacid (compound 1) was converted into
3,4,5-trimethoxy phenyl methyl ester (compound 2) using methanol and
sulphuric acid in acceptable yields (Scheme 3).

Scheme 3. Synthesis of 1,3,4-oxadiazole derivatives.

4
 Ester (compound 2) which on treatment with hydrazine hydrate yielded
3,4,5-trimethoxybenzoicacid hydrazide (compound 3) in good yield.
Antibacterial activity here is focused on the synthesis of novel heterocyclic
compounds as possible antibacterial agents. A series of novel compounds
were prepared and tested for their in vitro antibacterial activity against the
four strains of bacteria (gram + ve, gram –ve).

Five compounds of the obtained series showed high in vitro antimicrobial

activity. 2-{(3,4,5-trimethoxy phenyl1,3,4-oxadiazolyl)-5-thio}-4-
(morpholino)-6-(2-chloro phenyl ureido)-s-triazine showed excellent activity
against E. coli and P. aeruginosa, 2-{(3,4,5-trimethoxy phenyl-1,3,4-
oxadiazolyl)-5-thio}-4-(morpholino)-6-(4-chloro phenyl ureido)-s-triazine
showed excellent activity against P. aeruginosa indicated in vitro
antibacterial activity comparable to slightly lower than the ampicilline 2-
{(3,4,5-trimethoxy phenyl-1,3,4-oxadiazolyl)-5-thio}-4-(morpholino)-6-(4-
methyl phenyl thioureido)-s-triazine and 2-{(3,4,5-trimethoxy phenyl-1,3,4-
oxadiazolyl)-5-thio}-4-(morpholino)-6-(4-nitro phenyl thioureido)-s-triazine
showed excellent activity against C. albicans in vitro antifungal activity
comparable or slightly lower than that of Fluconazole.

The presence of electron-withdrawing group on the aromatic ring in

general increases the antimicrobial activities of the tested compounds
compared to compounds having electron donating groups. Based upon the
results it will also be necessary to optimize the led compound by substitution
in the C2 and C4 position in of phenyl ring by chloro and polar group
(phenolic or nitro moiety) seem to be very important for antibacterial effect,
as well as the presence and the positionof –NHCSNH- group in the
connecting linker between the aromatic ring seems to be very important for
antibacterial effect [16].

5
 In the end of 2011 , Habibullah Khalilullah and partners works on a
series of 1,3,4-oxadizole derivatives containing 1,4-benzodioxane ring
system were synthesized starting from 2,3-dihydro-1,4-benzodioxane-2-
carbohydrazide. The synthesized compounds were characterized and
evaluated for antibacterial activity against Staphylococcus aureus,
Escherichia coli and Bacillus subtilis and antifungal activity against
Aspergillus niger, Aspergillus flavus and Candida albicans by two fold serial
dilution technique. The synthetic route used to prepare starting materials and
the title compounds is outlined in (Scheme 4). The starting material ethyl-
l,4-benzodioxane-2-carboxylate (compound 1) was prepared by reaction
between catechol and ethyl-2,3-dibromopropionate in dry acetone in the
presence of anhydrous potassium carbonate, which on treatment with
hydrazine hydrate afforded the corresponding hydrazide (compound 2). The
reaction of hydrazide (compound 2) with substituted aryl carboxylic acids
in phosphorus oxychloride (POCl3) gave the cyclized products 2-
(substituted-phenyl)-5-(2,3-dihydro-1,4-benzodioxane-2-yl)-1,3,4-
oxadiazoles [17].

Scheme 4. Synthetic route for the preparation of 1,3,4-oxadiazole derivatives.

6
 Then in 2015, Manoj N. Bhoi and his group of co-researchers, were
synthesized some novel N'-(7-chloroquinolin-4-yl)-6-methyl-2-oxo-4-
phenyl-1,2,3,4-tetrahydropyrimidine-5-carbohydrazide derivatives via three
step reactions by convectional method. It have been evaluated it inhibition
capacity for various gram positive and gram negative bacterial strain. All
compounds were found to be good to excellent active against all four
bacterial strains.

The title compounds, N'-(7-chloroquinolin-4-yl)-6-methyl-2-oxo-4-

phenyl-1,2,3,4-tetrahydro pyrimidine-5-carbohydrazide derivative were
synthesized via three step reactions from biginelli reaction reaction,
hydrazine hydrate and 4,7-dichloro Quinoline as outlined in (Scheme 5).
Biginelli reaction is a versatile method for the preparation of
Dihydropyrimidines derivative by reaction between aldehydes derivative,
ethyl acetoacetate (compound 2) and urea (compound 3) in ethanol solvent
in the presence of Alumino silica as prompt catalyst. Here we first time
reported Alumino silica catalysed synthesis of Dihydropyrimidines
derivative. All the compounds were screened for their antibacterial activity
out on Nutrient-agar plates by well–diffusion assay against test culture.
Cultures were stimulated in Nutrient broth. Isolates inhibits the above
mentioned organisms or not were studied [18].

Scheme 5. Synthesis of targeted comound .

7
 Then in the same year of 2015 Ambareen Shaikh and Jyotsna Meshram
synthesis of a series of some novel 5-(5-(aryl)-1,3,4-oxadiazol-2-yl)-3,4-
dihydro-6-methyl-4-styrylpyrimidin-2(1H)-one derivatives. The synthetic
protocol applied for the synthesis of target compounds is sketched in
(Scheme 6). Ethyl 1,2,3,4-tetrahydro-6-methyl-2-oxo-4-styrylpyrimidine-5-
carboxylate (compound 1) , the starting material, was prepared according to
the reported method using cinnamaldehyde, β-keto ester, and urea.
Refluxing this styrylpyrimidine carboxylate with hydrazine hydrate in
absolute ethanol furnished the corresponding carbohydrazide; viz, 2,3,4-
tetrahydro-6-methyl-2-oxo-4-styrylpyrimidine-5-carbohydrazide
(compound 3). The reaction of this carbohydrazide with varying substituted
aromatic aldehydes in the presence of ceric ammonium nitrate (CAN) in
dichloromethane solvent afforded 5-(5-(aryl)-1,3,4-oxadiazol-2-yl)-3,4-
dihydro-6- methyl-4-styrylpyrimidin-2(1H)-one. Antibacterial activity of
this newly synthesized compounds was evaluated by the well diffusion
method against Staphylococcus aureus, Bacillus subtilis (gram positive) and
Escherichia coli, Klebsiella pneumoniae (gram negative) strains of bacteria
[11].

Scheme 6. General synthesis of target compounds.

8
 Later of 2015, Desai.N. C. and his co-workers synthesis a series of 5-(4-
acetyl-5-(aryl)-4,5-dihydro-1,3,4-oxadiazol-2-yl)-4-(2-fluorophenyl)-6-
methyl-3,4-dihydropyrimidin-2(1H)-ones . were synthesized in very good
yields. Biginelli adduct (compound 1) synthesized in the first step was
reacted with hydrazine hydrate to furnish carbohydrazide intermediate
(compound 2) which on further reaction with different aryl aldehydes
yielded 4-(2-fluorophenyl)-6-methyl-N'-(aryl)-2-oxo-1,2,3,4-
tetrahydropyrimidine-5-carbohydrazides. These intermediates were cyclized
with the help of acetic anhydride to give the titled compounds . All the
newly synthesized compounds were screened for their in vitro
antitubercular activity against Mycobacterium tuberculosis. The reaction
sequence for the preparation of 5-(4-acetyl-5-(aryl)-4,5-dihydro-1,3,4-
oxadiazol-2-yl)-4-(2fluorophenyl)-6-methyl-3,4-dihydropyrimidin-2(1H)-
ones is outlined in Scheme 3. Initially, ethyl 4-(2-fluorophenyl)-6-methyl-2-
oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (compound 1) was
synthesized .

Biginelli reaction of urea, ethylacetoacetate and 2-fluorobenzaldehyde in

methanol with catalytic amount of HCl. (Compound 1) was further reacted
with hydrazine hydrate in the presence of catalytic amount of con. H2SO4 to
furnish 4-(2-fluorophenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-
carbohydrazide (compound 2). In the next step, intermediate 2 was reacted
with different aryl aldehydes in the presence of catalytic amount of glacial
acetic acid to yield N'-arylidene-4-(2-fluorophenyl)-6-methyl-2-oxo-1,2,3,4-
tetrahydropyrimidine-5 carbohydrazide intermediates. Finally, these
intermediates were condensed with acetic anhydride to produce the targeted
compounds [19].

9
 Scheme 7.Synthetic protocol for the title compounds.

At the beginning of 2018, Desai.N. C.and his co-workers back with

another newly synthesized compounds were characterized using infrared,
proton nuclear magnetic resonance, carbon-13 nuclear magnetic resonance
and mass spectral techniques. From structure activity relationship studies, it
could be concluded that electron withdrawing groups played a crucial role in
enhancing antimicrobial and cytotoxic effects of title compounds. They have
synthesized new analogues in which pyrazole scaffold was linked to the
DHPMs systems. The synthetic pathway for final compounds is illustrated in
(Scheme 8).

(Compound 1) on reaction with hydrazine hydrate furnished 4-(2-

fluorophenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydro-pyrimidine-5-
carbohydrazide (compound 2). Cyclic condensation of (compound 2) with
ethyl acetoacetate resulted the compound 4-(2-fluorophenyl)-6-methyl5-(3-
methyl-5-oxo-4,5-dihydro-1H-pyrazole-1-carbonyl)-3,4-dihydropyrimidin-
2(1H)-one 3. This was further converted into 5-(4-(arylmethylene)-3-methyl-
5-oxo-4,5-dihydro-1H-pyrazole-1-carbonyl)-4-(2-fluorophenyl)-6-methyl-
3,4-dihydropyrimidin2(1H)-ones by Knoevenagel condensation with
different aryl aldehydes in the presence of piperidine catalyst [20].

10
 Scheme 8. Synthetic pathway of novel compounds

They have used the pyrazole bearing dihydropyrimidinone motif for the
development of potential antimicrobial agents. Both the pharmacophores are
therapeutically very important as dihydropyrimidines are potential inhibitors
of dihydrofolate reductase, which is a promising drug target for treatment of
mycobacterial infections. Similarly pyrazole moiety is also very promising
as it possesses varieties of biological activities .

Electron donating groups on aromatic ring, such as methyl, methoxy and

hydroxy, and electron withdrawing groups from aromatic ring, such as
fluoro, chloro and nitro, were chosen as substituents in the molecular
diversities of the targeted compounds. Results of antimicrobial activity of
final compounds showed that the presence of hydrophobic substituent at
ortho and para position of phenyl ring provided a positive impact on
antimicrobial activity. The amplified activity was due to the hydrophobic
nature of fluorine, which was responsible for the influence of substituent
group’s physicochemical properties.

11
 The presence of electron withdrawing functional groups, specifically
-F, -Cl and -NO2 exhibited excellent activity against all type of bacterial
strains, which is better than the electron donating groups like -OH, -CH3 and
-OCH3 . It was noted that the formation of pyrazole ring (compound 3)
from hydrazide precursor slightly improved the activity than its precursor
but the drastic change was observed in antimicrobial activity of final
Knoevenagel adduct . The newly targeted compounds presented here clearly
vary in their corresponding antimicrobial activity depending on the type of
substituents [20].

Newly 2019 Dinesh R. Godhani, make a series of dihydropyrimidine

substituted 1,3,4-oxadiazole derivatives was synthesized by cyclization of
carbohydrazide by phosphoryl chloride and benzoic acid in acidic condition.
The structures were perceived on the establishment of spectral tools and
their purity by elemental analysis. Every compound was primary assessed
for their in vitro antimicrobial activities against 5 bacterial strains viz
[Staphylococcus aureus , Klebsiella pneumoniae ,Escherichia coli ,
Acinetobacter baumannii, Pseudomonas aeruginosa ] and 2 fungi Strains viz.
[Candida albicans , Cryptococcus neoformans var. grubii ].New
dihydropyrimidine substituted 1,3,4-oxadiazole derivatives are set up
according to standard conventions with minor alterations as displayed in
(Scheme 9).

Biginelli reaction was transferred by solvent-free condition in presence of

Ionic Liquid ([BBI][BF4]). Hydrazinolysis of the ester in presence of
hydrazine hydrate in 1,4-dioxane at 110 °C for 6 hours gave 4-(2-
chloroquinolin-3-yl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-
carbohydrazide.

Cyclization of carbohydrazide by benzoic acid derivatives in phosphoryl

chloride afford 4-(2-chloroquinolin-3-yl)-6-methyl-5-(5-(substituted
phenyl)-1,3,4-oxadiazol-2-yl)-3,4-dihydropyrimidin-2(1H)-one, derivatives
[21].

12
 Scheme 9. Synthetic track for the preparation of title compounds

The biological advancement of synthesized compounds was evaluated

against varied bacterial and fungal strains by a standard broth-dilution
technique. The active compounds were additionally screened for cytotoxicity
against human embryonic kidney cell line. The compounds were likewise
screened for haemolysis of human blood cells. Fluconazole was utilized as a
positive fungal inhibitor standard for fungi. Colistin was utilized as positive
bacterial inhibitor guidelines for Gram-negative and Vancomycin for Gram-
positive bacteria. Melittin and tamoxifen were utilized as a positive
heamolytic and cytotoxicity standard, separately. All of the compounds that
were tested confirmed to no mentionable cytotoxicity against the human
embryonic kidney cell line, HK293 and no haemolytic activity observed
against human whole blood cells [21].

13
 Recently in 2020, with the increasing level of antimicrobial resistance in
pathogenic bacteria, together with the lack of new potential drug scaffolds in
the pipeline, make the problem of infectious diseases a major public health
concern.

Thus, Nishu Singla and her colleagues were synthesized.a novel series
of 1,3,4-oxadiazole-substituted thiophenes and 1,2,4-triazole substituted
thiophene derivatives . The synthesis of novel polysubstituted thiophenes
comprising 1,3,4-oxadiazole and 1,2,4-triazole heterocycles is outlined in
(scheme 10). The 2-aminothiophene derivatives which were further
converted to the respective hydrazides using excess of hydrazide hydrate in
ethanol. The cyclization of resultant hydrazides in the presence of potassium
hydroxide and carbon disulphide afforded the desired 2-thienyl-1,3,4-
oxadiazole-5-thiones . Whereas, the reaction of hydrazides with allyl-
isothiocyanate furnished the N-ethynyl hydrazine carbothioamides , which
underwent cyclization using NaOH to afford the desired 4-ethynyl-5-
thienyl-1,2,4-triazole-3-thiones. This influence of flouro- substitution on
enhancement of antibacterial activity is reminiscent of significance of fluoro
substitution in antibacterial activity of fluoroquinolone class of compounds
such as norfloxacin, ofloxacin, ciprofloxacin etc., wherein fluoro substituent
is shown to improve the DNA gyrase inhibition up to 18-fold over its 6-
hydrogen analog.[22].

14
Scheme 10. . Synthesis of polysubstituted 2-amino-thiophenes 1,3,4-oxadiazole-substituted
thiophenes and 1,2,4-triazole

In the end of 2020, Katiyar Pratima and Manjul P. Singh synthesis a new
series of benzyl moiety possessing 1,3,4-oxadiazole derivatives have been
synthesized by the reaction of 4-(benzylamino) benzohydrazide with
substituted aromatic acids in the presence of cyclizing agent phosphorus
oxychloride. Antibacterial activity of synthetic compounds were assayed
against two gram-positive strains Bacillus subtilis, Escherichia coli, and two
gram-negative strains Staphylococcus aureus, Pseudomonas aeruginosa.
Antifungal activity of synthetic compounds was screened against Candida
albicans and Aspergillus spp. Synthetic approach for a series of 2,5-
disubstituted 1,3,4-oxadiazole analogs holding benzyl moiety is depicted in
(Scheme 11).

15
 The para aminobenzoic acid was used as a starting material and masking
the carboxylic group by esterification process (Fischer method) before
benzylation [23] .Benzylated product was converted into respective hydrazide
by using hydrazine hydrate through simple reactions. Further, respective
hydrazide was made to react with various substituted aromatic acids in the
presence of cyclo-dehydrating agent POCl3 under specified conditions and
the desired products were obtained. The completion of reactions and purity
of synthesized compounds were examined by thin-layer chromatography
(TLC) method. The synthetic strategy for obtaining benzyl moiety bearing
2,5-disubstituted 1,3,4-oxadiazole derivatives is depicted in (Scheme 11)[24].

Scheme 11. For synthesis of 1,3,4-oxadiazole derivatives.

Finally in 2021, Lamya H. Al-Wahaibi and her team works on the

reaction of 5-(3,4-dimethoxyphenyl)-1,3,4-oxadiazole-2(3H)-thione 3 with
16
formaldehyde solution and primary aromatic amines or 1-substituted
piperazines, in ethanol at room temperature yielded the corresponding N-
Mannich bases 3-arylaminomethyl-5-(3,4-dimethoxyphenyl)-1,3,4-
oxadiazole-2(3H)-thiones or 3-[(4-substituted piperazin-1-yl)methyl]-5-(3,4-
dimethoxyphenyl)- 1,3,4-oxadiazole-2(3H)-thiones . The in vitro inhibitory
activity of compounds and was assessed against pathogenic Gram-positive,
Gram-negative bacteria [25]. The 3,4-Dimethoxybenzohydrazide (compound
2) was prepared from the commercially-available methyl 3,4-
dimethoxybenzoate (compound 1) via treatment with hydrazine in ethanol
[26].

1,3,4-Oxadiazole-2(3H)-thione (compound 3) was obtained via reaction of

the carbohydrazide (compound 2) with carbon disulfide in ethanolic
potassium hydroxide [27]. Furthermore, 1,3,4-Oxadiazole-2(3H)-thiones
were reported to undergo aminomethylation through reaction with primary
aromatic amines and formaldehyde to yield the corresponding N-Mannich
bases [28]. Consequently, treatment of (compound 3) with formaldehyde
solution and various primary aromatic amines or 1-substituted piperazines,
in ethanol at room temperature yielded their corresponding 3-
arylaminomethyl-5-(3,4-dimethoxyphenyl)-1,3,4-oxadiazole-2(3H)-thiones
or 3-[(4-substituted piperazin-1-yl)methyl]-5-(3,4-dimethoxyphenyl)-1,3,4-
oxadiazole-2(3H)-thiones N-Mannich bases, respectively, in good yields
(Scheme 12) .

17
 Scheme 12. Synthesis of compounds 3-arylaminomethyl-5-(3,4-dimethoxyphenyl)-1,3,4-
oxadiazole-2(3H)-thiones and 3-[(4-substituted piperazin-1-yl)methyl]-5-(3,4-
dimethoxyphenyl)-1,3,4-oxadiazole-2(3H)-thiones .

For the antibacterial activity, it could be concluded that the activity of the
piperazinomethyl N-Mannich bases is superior to their arylaminomethyl
analogues . The replacement of the arylaminomethyl moiety with a
piperazinomethyl moiety greatly enhanced the antibacterial activity and the
piperazinomethyl derivatives exhibited higher potency and broader
antibacterial spectrum compared to their arylaminomethyl analogues. In
addition, the antibacterial activity of the piperazinomethyl derivatives seems
correlated to their lipophilicity as the optimum antibacterial activity against
the tested Gram-negative bacteria was displayed by compounds [25].

4.Antimycobacterial Activity
18
 Making antimicrobial compounds is still a challenge, because resistance is
a very common drawback and infections that were previously easily treated
are now difficult to solve. The increasing prevalence of this resistance,
together with a low number of new drugs coming on to the market, causes an
important public health problem [29].

The presence of pyrimidine base in thymine, cytosine, and uracil, which

are the essential building blocks of nucleic acids DNA and RNA, is one
possible reason for their widespread therapeutic applications .Antibacterial
Agents, drugs which are included in this category are antifolates possessing
antagonistic activity against folic acid and sulfa drugs which are sulphur
containing pyrimidine derivative drugs [30]. A large number of 2,4-
diaminopyrimidines have been synthesized as antifolates and it was
eventually proved that these pyrimidines are inhibitors dihydrofolate
reductase (DHFR) [31]. Also pyrimidine containing sulfa drugs are
monosubstituted and disubstituted sulfa drugs include sulfadiazine ,
sulfamerazine , sulfadimidine (sulfamethazine) , sulfameythoxydiazine , and
methyldiazine [30].

Antibacterial activity of the newly synthesized compounds was evaluated

by the well diffusion method against Staphylococcus aureus, Bacillus
subtilis (gram positive) and Escherichia coli, Klebsiella pneumoniae (gram
negative) strains of bacteria [11].

In nutshell, we can conclude that the incorporation of electron donating

groups such as hydroxy, methyl, and methoxy diminished the antibacterial
property. Effect of halogen group’s as substitution was clearly visible in
activity enhancement [20].

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 ‫‪1,3,4-Oxadiazole DHPM Derivatives As Potent‬‬
‫‪Antimicrobial Agent , A Review‬‬

‫مشروع دراسة مقدمة الى‬

‫كلية الصيدلة \ جامعة البيان‬
‫كجزء من متطلبات الحصول على درجة‬
‫البكلوريوس في الصيدلة‬

‫‪ :‬إعداد‬
‫ساره معن الدوري‬
‫فاطمة نزارهاشم‬

‫‪ :‬بإشراف‬
‫م‪.‬م علي مجيد حنتوش‬

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