World Journal of Pharmaceutical Research

Sharma et al. SJIF Research

World Journal of Pharmaceutical Impact Factor 6.805

Volume 5, Issue 7, 2003-2016. Research Article ISSN 2277– 7105

SYNTHESIS, CHARACTERIZATION AND BIOLOGICAL

EVALUATION OF SOME NOVEL FLUOROQUINOLONE CLUBBED
THIADIAZOLE ANALOGS

*Prabodh Chander Sharmaa, Monika Chaudharyb, Mona Piplania, Archana Sharmaa,

Harish Rajakc

a
Institute of Pharmaceutical Sciences, Kurukshetra University, Kurukshetra-136119, India.
b
Hindu College of Pharmacy, Sonepat, India.
c
Institute of Pharmaceutical Sciences, Guru Ghasidas University, Bilaspur-495009, India.

ABSTRACT
Article Received on
22 May 2016, An efficient and facile procedure for synthesis of novel series of
Revised on 12 June 2016, fluoroquinolone analogs clubbed with thiadiazoles is described herein.
Accepted on 02 July 2016
Antibacterial screening of synthesized compounds was carried out
DOI: 10.20959/wjpr20167-6689
against a range of Gram-positive and Gram-negative bacterial strains.
Some of synthesized compounds 5i (MIC: 0.625 µg/mL) and 5k (MIC:
*Corresponding Author 0.3125 µg/mL) exhibited promising activities against the E. coli and S.
Dr. Prabodh Chander
aureus respectively. The analogs 5c, 5d, 5f and 5j displayed good
Sharma
Institute of
anthelmintic activity against Eisemia foetida when compared with
Pharmaceutical Sciences, standard drug.
Kurukshetra University,
Kurukshetra-136119, KEYWORDS: Fluoroquinolones; thiadiazole; infection; antibacterial;
India. anthelmintic.

INTRODUCTION
Heterocyclic moieties have been found in a large number of compounds exhibiting a great
biological potential.[1] Thiadiazole is a heterocyclic moiety commonly known to exhibit
diverse antiviral, antibacterial, antifungal and antitubercular potential.[2-5] In recent years,
scientific researchers across the world are taking considerable interest in this important
biological scaffold due to its wide spectrum of applications and accordingly have been
actively involved in the progression of thiadiazole chemistry.[6]

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Bacterial infections had been the chief cause of death across the world since the time
immemorial.[7,8] Antimicrobial agents provide an effective way for the management and
treatment of infectious diseases caused by bacteria as well as other microbes.[9]
Fluoroquinolones, a series of synthetic antibacterial agents are being utilized as a great
weapon against infection by the clinicians and has been proved one of the most regularly
prescribed categories of antibacterial drugs.[10] The mechanism of action by which they act,
slightly differs from other antibacterial drugs.[11,12] They act on two enzymes primarily
involved in DNA synthesis: DNA gyrase and topoisomerase IV and thus obstruct DNA
replication, transcription as well as cell repair, leading to cell death.[13] Some other
fluoroquinolones derivatives with anti-fungal, anti-tubercular and anti-viral activities are well
recognized.[14-22] The promising role of fluoroquinolones in carcinogenesis and mutagenesis,
an immense research endeavors are being carried out.[23,24] Despite of availability of various
accepted fluoroquinolones for curing a number of infections, continuous efforts have been
carried out for the invention of fluoroquinolones having desired pharmacokinetic profile and
therapeutic index for solving the problems of rising bacterial resistance.[25,26]

Microbial resistance is the serious concern in the medical community to antimicrobial agents.
This global crisis has caught the attention of medicinal chemists to overcome the pitfalls of
presently accessible antimicrobial therapy against microbial infections.[27] Therefore,
development of novel derivatives/analogs of antimicrobial agents having significant potency
and broad spectrum of activity is highly desired.[28]

Keeping in view, the biological significance of thiadizole moiety as well as fluoroquinolones,

it is the task of curiosity to keep these both moieties in a singular framework which would
results in compounds of interesting biological profile. As a part of our ongoing research in
developing new bioactive molecule, we tried to study the influence of fluoroquinolone and
thiadiazole scaffold combination on the antimicrobial activity. In this present study, we have
attempted to incorporate both these biologically active moieties together to give a confined
structure for evaluating their anti-bacterial and anthelmintic activities.

EXPERIMENTAL
General
Laboratory chemicals and solvents used were procured from Merck AG (Mumbai, India),
Qualigens (Navi Mumbai, India), Sigma Aldrich (Bangalore, India) and SD Fines (Mumbai,
India). Melting points (uncorrected) were recorded on a Labindia MR-VIS visual melting

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range apparatus (Mumbai, India). The infrared (IR) spectra were obtained on a Perkin Elmer
(Waltham, MA), IR spectrophotometer (KBr disk). 1H NMR spectra were run on Bruker 400
(Fallanden, Switzerland) spectrometer using tetramethylsilane as an internal standard in
DMSO.

Chemistry
Synthesis of 2-amino-5-aryl-1, 3, 4-thiadiazole (2a-e)
Thiosemicarbazide (9.1 g, 0.1 mol) and aryl carboxylic acid (0.1 mol) was taken in round
bottom flask and allowed to reflux for 4-5 h in presence of concentrated sulphuric acid (10
drops). The mixture was poured over crushed ice. The solid thus obtained was filtered and
given washing with water. The product was purified by recrystallization process from
ethanol.[3]

Synthesis of substituted N-(5-aryl-1, 3,4-thiadiazole-2-yl)-2-chloroacetamide (3a-e)

To a solution of 2-amino-5-aryl-1, 3, 4-thiadiazoles (2a-e) (0.1mol) in 25 mL of N,N dimethyl
formamide (DMF). Chloroacetyl chloride was added by drop wise manner (7.9 mL, 0.1 mol)
to avoid any vigorous reaction to take place. The reaction mixture was refluxed for 2 h and
poured into crushed ice. The precipitates separated out were filtered and washed with cold
water. It was purified by recrystallization from absolute alcoh.[4]

Synthesis of fluoroquinolone derivatives bearing acetamide linkage (5a-l)

Equimolar quantity of thiadiazole 3a-e (0.03 mmol), fluoroquinolone 4a-e (0.03 mmol) and
NaHCO3 (0.03 mmol) in DMF (10 mL), was taken in round bottom flask and heated at
temperature 85-90˚C. After completion of reaction as observed by TLC, 15 mL of water was
added in reaction mixture. The precipitates were isolated and washed with water. The product
was purified by passing through silica gel column (solvent: chloroform-ethanol; 19:1) and
recrystallized from DMF-H2O to yield compounds 5a-l.[5] The new analogs were synthesized
following the versatile synthetic method, given in scheme 1 (Figure 1).

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R= -H, -Cl, -NO2,- OCH3 Ciprofloxacin R3= cyclopropyl, R4= -H, R5= -H
R1= -OCH3, -NO2 Norfloxacin R3= -CH2CH3, R4= -H, R5= -H
R2= -NO2 Gatifloxacin R3= cyclopropyl, R4= -OCH3, R5= -CH3

Scheme 1
Molecular
Compd R R1 R2 R3 R4 R5
Formula
5a -Cl -H -H -H -H C27H24ClFN6O4S
5b -NO2 -H -H -H -H C27H24FN7O6S
5c -OCH3 -OCH3 -H -H -H C29H29FN6O6S
5d -OCH3 -H -H -H -H C28H27FN6O5S
5e -H -NO2 -NO2 -H -H C27H23FN8O8S
5f -NO2 -H -H -CH2CH3 -H -H C26H26FN7O5S
5g -Cl -H -H -CH2CH3 -H -H C26H24ClFN6O4S
5h -OCH3 -H -H -CH2CH3 -H -H C27H27FN6O5S
5i -H -NO2 -NO2 -CH2CH3 -H -H C26H23FN8O8S
5j -Cl -H -H -OCH3 -CH3 C29H28ClFN6O5S
5k -NO2 -H -H -OCH3 -CH3 C29H28FN7O7S
5l -H -NO2 -NO2 -H -H C29H27FN8O9S
Figure 1. Synthetic scheme for preparation of fluoroquinolone compounds 5a-l

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1-Cyclopropyl-6-fluoro-7-(4-{2-(5-(4-chloro-phenyl-1,3,4-thiadiazol-2-yl-)amino)-acetyl}-
piperazin-1-yl)- 4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5a). Yield 62%, m.p. 298-
301°C, IR (KBr) (cm-1): 3374 (NH str), 3063 (C-H str), 1744 (C=O str), 1682 (CONH str),
1636 (C=O str), 1535 (C=C str), 1311 (C-O str ), 1111 (C-N str); 1H NMR (DMSO-d6) δ
ppm: 2.34 (m, 4H, -CH2CH2- cyclopropyl), 2.62-2.73 (m, 9H, piperazine-H and cyclopropyl-
H), 3.30 (s, 2H, -CH2 methylene bridge), 7.28-8.14 {m, 6H, aromatic (H5, H8- quinolone and
H2’, H3’, H5’ and H6’- phenyl thiadiazole)}, 8.06 (s, 1H, H2-quinolone).

1-Cyclopropyl-6-fluoro-7-(4-{2-(5-(4-nitro-phenyl-1,3,4-thiadiazoyl-)amino)-acetyl}-4-
oxoethyl) piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5b). Yield 52%,
m.p. 280-282°C. IR (KBr) (cm-1): 3342 (NH str), 1744 (C=O str), 1682 (CONH str), 1628
(C=O str), 1535 (C=C str), 1227 (C-O str ), 1196 (C-N str); 1H NMR (DMSO-d6) δ ppm:
1.45-1.60 (m, 4H, -CH2CH2- cyclopropyl), 2.62 (m, 9H, piperazine-H and cyclopropyl-H),
4.09 (s, 2H, -CH2 methylene bridge), 7.97-8.02 {m, 2H, aromatic (H5, H8- quinolone) 8.806
(s, 1H, H2-quinolone), 9.13-9.20{m, 4H, aromatic (and H2’, H3’, H5’ and H6’- phenyl
thiadiazole)}.

1-Cyclopropyl-6-fluoro-7-(4-{2-(5-(3,4-dimethoxy-phenyl-1,3,4-thiadiazol-2-yl)amino)-
acetyl}-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5c). Yield 57%, m.p.
272-274°C. IR (KBr) (cm-1): 3370 (NH str), 3145-2850 (C-H str), 1715 (C=O str), 1672
(CONH str), 1633 (C=O str), 1550 (C=C str), 1262 (C-O str ), 1160 (C-N str); 1H NMR
(DMSO-d6) δ ppm: 0.98 (m, 4H, -CH2CH2- cyclopropyl), 3.43-3.70 (m, 9H, piperazine-H
and cyclopropyl-H), 3.73 (s, 6H, -OCH3 of phenyl ring of thiadiazole), 3.95 (s, 2H, -CH2
methylene bridge), 6.92-7.78 {m, 5H, aromatic (H5, H8- quinolone and H2’, H5’ and H6’-
phenyl-thiadiazole)}, 7.96 (s, 1H, H2-quinolone).

1-Cyclopropyl-6-fluoro-7-(4-{2-(5-(4-methoxy-phenyl-1,3,4-thiadiazol-2-yl)amino)-acetyl}-
piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5d). Yield 65%, m.p. 287-
288°C. IR (KBr) (cm-1): 3380 (NH str), 3160-2850 (C-H str), 1750 (C=O str), 1665 (CONH
str), 1628 (C=O str), 1551 (C=C str), 1227 (C-O str ), 1183 (C-N str); 1H NMR (DMSO-d6)
δ ppm: 0.87 (m, 4H, -CH2CH2- cyclopropyl), 3.30-3.60 (m, 9H, piperazine-H and
cyclopropyl-H), 3.65 (s, 3H, -OCH3 of phenyl ring of thiadiazole), 4.11 (s, 2H, -CH2
methylene bridge), 7.11-7.78 {m, 6H, aromatic (H5, H8- quinolone and H2’,H3’,H5’ and H6’-
phenyl-thiadiazole)}, 8.26 (s, 1H, H2-quinolone).

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1-Cyclopropyl-6-fluoro-7-(4-{2-(5-(3,5-dinitro-phenyl-[1,3,4]-thiadiazol-2-yl)amino)-
acetyl}-piperazin-1-yl)-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5e). Yield 58%, m.p.
283-285°C. IR (KBr) (cm-1): 3360 (NH str), 3056-2875 (C-H str), 1720 (C=O str), 1674
(CONH str), 1630 (C=O str), 1535 (C=C str), 1227 (C-O str ), 1145 (C-N str); 1H NMR
(DMSO-d6) δ ppm: 0.9 (m, 4H, -CH2CH2- cyclopropyl), 3.45-3.87 (m, 9H, piperazine-H and
cyclopropyl-H), 3.95 (s, 2H, -CH2 methylene bridge), 7.56-8.56 {m, 5H, aromatic (H5, H8-
quinolone and H2’,H4’and H6’- phenyl-thiadiazole)}, 8.26 (s, 1H, H2-quinolone).

1-Ethyl-6-fluoro-7-(4-{2-(5-(4-nitro-phenyl-[1,3,4]-thiadiazol-2-yl)amino)-acetyl}-piperazin-
1-yl)- 4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5f). Yield 69%, m.p. 256-258°C. IR
(KBr) (cm-1): 3618 (NH str), 1744 (C=O str), 1682 (CONH str), 1628 (C=O str), 1535 (C=C
str), 1227 (C-O str ), 1196 (C-N str); 1H NMR (DMSO-d6) δ ppm: 1.06 (s, 3H, -CH3 ethyl),
1.04 (s, 2H, -CH2 ethyl), 2.50-2.90 (m, 9H, piperazine-H and cyclopropyl-H), 3.46 (s, 2H, -
CH2 methylene bridge), 8.048-8.067 {m, 6H, aromatic (H5, H8-quinolone and H2’,H3’,H5’ and
H6’- phenyl-thiadiazole)}, 8.508 (s, 1H, H2-quinolone).

1-Ethyl-6-fluoro-7-(4-{2-(5-(4-chloro-phenyl-[1,3,4]-thiadiazol-2-yl)amino)-acetyl}-
piperazin-1-yl)- 4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5g). Yield 59%, m.p. 269-
272°C. IR (KBr) (cm-1): 3350 (NH str), 3068-2876 (C-H str), 1713 (C=O str), 1668 (CONH
str), 1625 (C=O str), 1550 (C=C str), 1257 (C-O str ), 1157 (C-N str); 1H NMR (DMSO-d6)
δ ppm: 1.27 (s, 3H, -CH3 ethyl), 2.24 (s, 2H, -CH2 ethyl), 3.16-3.67 (m, 9H, piperazine-H
and cyclopropyl-H), 3.57 (s, 2H, -CH2 methylene bridge), 7.35-8.09 {m, 6H, aromatic (H5,
H8-quinolone and H2’,H3’,H5’ and H6’- phenyl-thiadiazole)}, 8.52 (s, 1H, H2-quinolone).

1-Ethyl-6-fluoro-7-(4-{2-(5-(4-methoxy-phenyl-[1,3,4]-thiadiazol-2-yl)amino)-acetyl}-
piperazin-1-yl)- 4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5h). Yield 61%, m.p. 263-
265°C. IR (KBr) (cm-1): 3340 (NH str), 3055-2876 (C-H str), 1721 (C=O str), 1678 (CONH
str), 1649 (C=O str), 1557 (C=C str), 1254 (C-O str ), 1165 (C-N str); 1H NMR (DMSO-d6)
δ ppm: 1.35 (s, 3H, -CH3 ethyl), 2.26 (s, 2H, -CH2 ethyl), 3.10-3.61 (m, 9H, piperazine-H
and cyclopropyl-H), 3.65 (s, 3H, -OCH3), 4.05 (s, 2H, -CH2 methylene bridge), 6.83-7.37 {m,
6H, aromatic (H5, H8-quinolone and H2’,H3’,H5’ and H6’- phenyl-thiadiazole)}, 7.96 (s, 1H,
H2-quinolone).

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1-Ethyl-6-fluoro-7-(4-{2-(5-(3,5-dinitro-phenyl-[1,3,4]-thiadiazol-2-yl)amino)-acetyl}-
piperazin-1-yl)- 4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5i). Yield 63%, m.p. 263-
265°C. IR (KBr) (cm-1): 3350 (NH str), 3085-2834 (C-H str), 1712 (C=O str), 1666 (CONH
str), 1628 (C=O str), 1535 (C=C str), 1255 (C-O str ), 1160 (C-N str); 1H NMR (DMSO-d6)
δ ppm: 1.31 (s, 3H, -CH3 ethyl), 2.20 (s, 2H, -CH2 ethyl), 3.31-3.76 (m, 9H, piperazine-H
and cyclopropyl-H), 4.08 (s, 2H, -CH2 methylene bridge), 7.35-8.21 {m, 6H, aromatic (H5,
H8-quinolone and H2’,H4’ and H6’- phenyl-thiadiazole)}, 8.36 (s, 1H, H2-quinolone).

1-Cyclopropyl-6-fluoro-7-(4-{2-(5-(4-chloro-phenyl-[1,3,4]-thiadiazol-2-yl)amino)-acetyl}-
3-methyl-piperazin-1-yl)-8-methoxy-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5j).
Yield 49%, m.p. 164-166°C. IR (KBr) (cm-1): 3350 (NH str), 3049-2864 (C-H str), 1713
(C=O str), 1669 (CONH str), 1627 (C=O str), 1549 (C=C str), 1255 (C-O str ), 1160 (C-N
str); 1H NMR (DMSO-d6) δ ppm: 1.27 (m, 4H, -CH2CH2- cyclopropyl), 2.34 (m, 3H, -CH3),
3.21-367 (m, 9H, piperazine-H and cyclopropyl-H), 3.77 (s, 3H, -OCH3), 4.14 (s, 2H, -CH2
methylene bridge), 6.96-7.85 {m, 6H, aromatic (H5, H8-quinolone and H2’,H3’ H5’ and H6’-
phenyl-thiadiazole)}, 8.12 (s, 1H, H2-quinolone).

1-Cyclopropyl-6-fluoro-7-(4-{2-(5-(4-nitro-phenyl-[1,3,4]-thiadiazol-2-yl)amino)-acetyl}-3-
methyl-piperazin-1-yl)-8-methoxy-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5k). Yield
43%, m.p. 175-179°C. IR (KBr) (cm-1): 3350 (NH str), 3020-2765(C-H str), 1711(C=O str),
1665(CONH str), 1630 (C=O str), 1545(C=C str), 1245(C-O str ), 1165 (C-N str); 1H NMR
(DMSO-d6) δ ppm: 1.08 (m, 4H,-CH2CH2- cyclopropyl), 2.28 (m, 3H, -CH3), 3.45-3.82 (m,
9H, piperazine-H and cyclopropyl-H), 3.75 (s, 3H,-OCH3), 4.12 (s, 2H,-CH2 methylene
bridge), 7.96-8.85 {m, 6H, aromatic (H5, H8-quinolone and H2’,H3’H5’ and H6’- phenyl-
thiadiazole)},8.12 (s, 1H, H2-quinolone).

1-Cyclopropyl-6-fluoro-7-(4-{2-(5-(4-nitro-phenyl-[1,3,4]-thiadiazol-2-yl)amino)-acetyl}-3-
methyl-piperazin-1-yl)-8-methoxy-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (5l). Yield
56%, m.p. 171-173°C. IR (KBr) (cm-1): 3320 (NH str), 3056-2841 (C-H str), 1718 (C=O str),
1665 (CONH str), 1635 (C=O str), 1555(C=C str), 1215(C-O str ),1160 (C-N str); 1H NMR
(DMSO-d6) δ ppm: 1.0 (m, 4H, -CH2CH2- cyclopropyl), 2.28 (m, 3H, -CH3), 3.34-3.65 (m,
9H, piperazine-H and cyclopropyl-H), 3.89 (s, 3H, -OCH3), 4.18 (s, 2H, -CH2 methylene
bridge), 7.10-7.96 {m, 5H, aromatic (H5, H8-quinolone and H2’,H4’ and H6’- phenyl-
thiadiazole)}, 8.22 (s, 1H, H2-quinolone).

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Biological evaluation
Anti-bacterial activity
Antibacterial screening for all the synthesized compounds 5a-5l, against two Gram-negative
bacteria i.e. E. coli (MTCC 119), P. aeruginosa (MTCC 7453) and two Gram-positive strains
i.e. B. pumilus (MTCC 7411), S. aureus (MTCC 96) was carried out. The antibacterial
evaluation of synthesized compounds was carried out by minimum inhibitory concentration
(MIC) method. Microorganisms were inoculated and incubated according to procedure
provided by NDRI, Karnal. The bacterial cultures were grown on nutrient agar plate and kept
at 4°C. The experimental MIC values of the target compounds against selected bacterial
strains have been shown in table 1.

Table 1: Minimum inhibitory concentration (MIC) of target compounds 5a-l in µg/mL

against B. pumilus, S. aureus, E. coli and P. aeruginosa bacterial strains.
Compounds Gram (+) Gram (-)
B. pumilus S. aureus E. coli P. aeruginosa
5a 0.3125 0.3125 0.625 1.25
5b 0.625 0.625 0.625 2.50
5c 0.3125 0.3125 0.3125 0.3125
5d 0.625 0.3125 0.625 0.625
5e 0.625 0.3125 0.625 1.25
5f 0.625 0.3125 1.25 1.25
5g 2.50 2.50 2.50 2.50
5h 10 10 10 10
5i 0.625 0.3125 0.625 0.3125
5j 0.625 0.625 0.3125 0.3125
5k 0.3125 0.3125 0.3125 0.3125
5l 10 2.50 2.50 10
Ciprofloxacin <0.3125 <0.3125 0.3125 0.3125
Norfloxacin 0.625 <0.3125 1.25 0.625
Gatifloxacin 0.625 0.625 <0.3125 0.3125

Anthelmintic evaluation
The anthelmintic activity is a test to determine the capability of a chemical agent to eliminate
worms. The newly synthesized compounds were screened for anthelmintic activity in vitro
using adult Indian earth worm E. foetida due to its analogous anatomical and physiological
features with the intestinal roundworm, parasites of human beings.[29,30] Earthworms (E.
foetida) were purchased from Department of Agriculture, Gurukul, Kurukshetra. The
suspensions of the synthesized compounds were prepared by using 0.5% tween 80 as a
suspending agent and distilled water. The mechanical stirrer was utilized for stirring the

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resulting suspension for half an hour. Anthelmintic activities of these suspensions were
determined. Piperazine citrate in the suspension form was also used as reference standard and
taken in the equal concentration as the vehicle control.

Petri dishes having 50 mL suspension of the standard drug (piperazine citrate) for
comparison, 50 mL suspension of distilled water and 0.5% tween 80 as control and 50 mL
suspension of each test samples were taken at room temperature. Five earthworms having
comparable sizes and varieties were taken in each dish and examined for paralysis and death.
The death time was recorded after ascertaining that earthworm didn’t move on dropping the
paralysed worm in water having temperature up to 50°C.

The mean paralysing time and mean death time was calculated. All the outcomes of
anthelmintic activity have been summarized in Table 2.

Table 2. Anthelmintic activity of synthesized derivatives 5a-l.

Concentration of Mean paralysing Mean death time
Compounds
compounds (mg/100mL) time (min)+S.E. (min)+S.E.
5a 200 26.00±2.0* 38.40±0.9**
5b 200 32.60±1.1 43.20±1.2
5c 200 22.80±1.9** 38.20±0.6**
5d 200 23.20±1.3** 44.40±1.2
5e 200 27.80±0.7 44.40±1.3
5f 200 23.60±0.7** 39.20±1.2**
5g 200 26.80±1.5* 43.20±2.1
5h 200 27.00±1.4 40.40±1.0*
5i 200 28.40±1.2 47.80±1.5
5j 200 23.60±1.8** 42.00±1.0
5k 200 24.60±1.6 43.60±1.9
5l 200 29.00±2.7 44.80±1.4
Control - - -
Standard 200 24.40±1.0 34.80±1.0

Statistical analysis: The results were calculated as mean ± (SEM) of five worms in every
group. Dunnett’s ‘t’ test have been performed after one way ANOVA method for statistical
analysis.

RESULTS AND DISCUSSION

Chemistry
Detailed synthetic pathways of various fluoroquinolone derivatives 5a-l are depicted in
scheme 1. Compounds 2a-e were synthesized by refluxing substituted aryl carboxylic acid

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1a-g and thiosemicarbazide in acidic environment for 4-5 hrs. The intermediate compounds
2a-e after reaction with chloroacetyl chloride in DMF gave substituted N-(5-aryl-1, 3,4-
thiadiazole-2-yl)-2-chloroacetamide 3a-e. Finally the compounds 3a-e were allowed to reflux
with fluoroquinolones 4a-e in DMF as solvent and basic environment provided by sodium
bicarbonate at 85-90°C. Product obtained was collected, dried and recrystallized from DMF
and water to give 5a-l.

The structures, molecular formula of newly synthesized compounds have been supported and
confirmed by spectral analysis (IR and 1H NMR). Detailed study of IR spectra of the target
compounds 5a-l revealed the characteristic absorption band at 1720-1705 cm-1 and 1674-
1666 cm-1, corresponds to C=O and CONH stretching vibration respectively. The 1H NMR
spectra displayed proton of methylene bridge at about 3.35-4.12 (singlet 2H) ppm, evidently
demonstrating the occurrence of acetamide linkage in fluoroquinolone derivatives 5a-l.

Pharmacological screening
Antibacterial and anthelmintic screening of synthesized compounds was performed to find
out their pharmacological prospective.

Anti-bacterial activity
The synthesized compounds 5a-l were investigated for anti-bacterial activity by minimum
inhibitory concentration method. The activity was carried out against selected Gram-negative
bacteria i.e. E. coli (MTCC 119), P. aeruginosa (MTCC 7453) and Gram-positive strains i.e.
B. pumilus (MTCC 7411), S. aureus (MTCC 96). These novel derivatives demonstrated
varying antibacterial activities against different strains. Compounds 5i and 5k (MIC= 0.3125-
0.625 µg/mL) having nitro functional group at aromatic ring attached to thiadiazole moiety
showed significantly potent antibacterial activities against selected bacterial strains using
norfloxacin and gatifloxacin (MIC= 0.625-1.25 µg/mL) as standard antibiotics. These
compounds with nitro substituted derivatives were observed to be more potent against
bacterial strains compared to methoxy, chloro and dimethoxy substituted derivatives.
Ciprofloxacin derivative 5c (MIC= 0.3125 µg/mL) was found to exhibit equivalent MIC on
comparing with its standard drug (MIC= 0.3125 µg/ml) against Gram-negative bacteria like
E. coli and P. aeruginosa. Similarly when nofloxacin derivative 5f (MIC= 0.625-1.25 µg/mL)
against E. coli along with B. pumilus and gatifloxacin derivative 5j (MIC= 0.3125-0.625
µg/mL) against P. aeruginosa, B. pumilus and S. aureus were tested, they were found to
exhibit equivalent MIC against antibiotics norfloxacin and gatifloxacin used as standards.

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Sharma et al. World Journal of Pharmaceutical Research

The results of antibacterial activity reveal that norfloxacin and gatifloxacin analogs were
proved to be more efficacious as compared to ciprofloxacin derivatives.

Anthelmintic activity
The synthesized compounds were evaluated for anthelmintic activity against E. foetida. All
the compounds were found to exhibit significant anthelmintic activity. Compound 5c, 5d, 5f
and 5j possessed good activity having mean paralysis time within the range 22.80±1.9** to
23.60±1.8* min whereas majority of the compounds expressed death time varies from 38.20
±0.9** min to 39.20±1.2** min. Piperazine citrate used as standard drug at concentration of
200 mg/mL exhibited mean paralysis time 24.40± 1.0 min and mean death time 34.80 ± 1.0
min. Anthelmintic activity of synthesized analogs revealed that fluoroquinolone derivatives
bearing dimethoxy, methoxy, nitro and chloro group showed good activity compared to other
substitution at thiazdiazole ring. It has been proved from the results that activity of the
compounds can be improved by substitution of more electro negativity group.

CONCLUSION
In current investigation, a facile novel synthesis of fluoroquinolone clubbed thiadiazole
analogs with acetamide linkage is reported. The in vitro screening of newly synthesized
compounds showed improved therapeutic efficiency as compared to parent drugs. Some of
the synthesized compounds demonstrated more potent or equipotent antibacterial activities
against the selected bacterial strains. A few analogs have also possesses promising
antihelmintic activity. The significant findings of the present research work may be utilized
by the researchers for development of better antibacterial and antihelmintic agents in future.

Conflict of interest
The authors have declared no conflict of interest.

ACKNOWLEDGEMENT
We express sincere thanks to Prof. A. C. Rana, Director, Institute of Pharmaceutical
Sciences, Kurukshetra University, Kurukshetra, for providing necessary facilities.

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