Synthesis and characterization of flurbiprofen hydrazide derivatives as potential anti-HCV, anticancer and antimicrobial agents

Article abstract of DOI:10.1007/s00044-013-0550-3

A novel series of new flurbiprofen hydrazide derivatives 2-(2-fluorobiphenyl-4-yl)-N′-[(substituted phenyl/5-nitro-2-furyl) methylene]propanehydrazide (3a-k), 2-(2-fluorobiphenyl-4-yl)-N-(2-substituted-4- oxo-1,3-thiazolidine-3-yl)propanamide (4a-b, 4d-k), 2-[2-(2-fluorobiphenyl-4-yl) propanoyl]-N-substituted hydrazinecarbothioamide (5a-h) and 2-(2-fluorobiphenyl-4-yl)-N′-[(3-methyl-4-oxo-1,3-thiazolidin-2-ylidene] propanehydrazide (6a-b, 6e and 6g) has been synthesized in this study. All synthesized compounds were screened for antimicrobial activity against various bacterial and fungal strains. Additionally, compounds were evaluated for the ability to inhibit Hepatitis C virus NS5B polymerase. The most active 4-thiazolidinone compound was 4k (SGK119) with 67.0 % and thiosemicarbazide compound was 5d (SGK123) with 69.50 % inhibition at 200 μM against hepatitis C virus NS5B RNA polymerase. Anticancer activity of the selected compounds (3i, 3j, 3h, 4d, 4i and 6b) was determined at a single dose towards the full panel of 60 human cancer cell lines by the National Cancer Institute (NCI). 2-(2-Fluoro-4-biphenylyl)-N-[2-[4-(trifluoromethyl)phenyl]-4-oxo-1, 3-thiazolidine-3-yl]propanamide 4d, containing thiazolidinone ring, demonstrated the most marked effect with 20.80 % growth percent on leukaemia cancer cell line SR at 10^-5 M. The results demonstrated that none of the compounds tested have anticandidal and antifungal activities, but two of them (4a and 4i) showed antibacterial inhibition against Micrococcus luteus, and Staphylococcus cohnii and Staphylococcus aureus, respectively.

Full text of DOI:10.1007/s00044-013-0550-3

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res
DOI 10.1007/s00044-013-0550-3
ORIGINAL RESEARCH
Synthesis and characterization of flurbiprofen hydrazide
derivatives as potential anti-HCV, anticancer
and antimicrobial agents
˙
•
•
•
•
Pelin C¸ ıkla Esra Tatar Ilkay Ku¨c¸u¨kgu¨zel Fikrettin S¸ahin
•
Dils¸ad Yurdakul Amartya Basu Ramalingam Krishnan
•
•
•
•
Daniel Brian Nichols Neerja Kaushik-Basu S¸. Gu¨niz Ku¨c¸u¨kgu¨zel
Received: 8 May 2012 / Accepted: 18 February 2013
Ó Springer Science+Business Media New York 2013
Abstract A novel series of new flurbiprofen hydrazide
derivatives 2-(2-fluorobiphenyl-4-yl)-N0-[(substituted phenyl/
5-nitro-2-furyl)methylene]propanehydrazide (3a–k), 2-(2-flu-
orobiphenyl-4-yl)-N-(2-substituted-4-oxo-1,3-thiazolidine-3-
yl)propanamide (4a–b, 4d–k), 2-[2-(2-fluorobiphenyl-4-yl)
propanoyl]-N-substituted hydrazinecarbothioamide (5a–h)
and 2-(2-fluorobiphenyl-4-yl)-N0-[(3-methyl-4-oxo-1,3-thi-
azolidin-2-ylidene]propanehydrazide (6a–b, 6e and 6g) has
been synthesized in this study. All synthesized compounds
were screened for antimicrobial activity against various
bacterial and fungal strains. Additionally, compounds were
evaluated for the ability to inhibit Hepatitis C virus NS5B
polymerase. The most active 4-thiazolidinone compound
was 4k (SGK119) with 67.0 % and thiosemicarbazide
compound was 5d (SGK123) with 69.50 % inhibition at
200 lM against hepatitis C virus NS5B RNA polymerase.
Anticancer activity of the selected compounds (3i, 3j, 3h, 4d,
4i and 6b) was determined at a single dose towards the full
panel of 60 human cancer cell lines by the National Cancer
Institute (NCI). 2-(2-Fluoro-4-biphenylyl)-N-[2-[4-(trifluo-
romethyl)phenyl]-4-oxo-1,3-thiazolidine-3-yl]propanamide
4d, containing thiazolidinone ring, demonstrated the
most marked effect with 20.80 % growth percent on leu-
kaemia cancer cell line SR at 10-5 M. The results demon-
strated that none of the compounds tested have anticandidal
and antifungal activities, but two of them (4a and 4i)
showed antibacterial inhibition against Micrococcus luteus,
and Staphylococcus cohnii and Staphylococcus aureus,
respectively.
Keywords Anticancer activity ꢀ
Hepatitis C NS5B polymerase ꢀ 4-Thiazolidinone ꢀ
Hydrazide-hydrazone ꢀ Flurbiprofen
Introduction
Hepatitis C virus (HCV) infection is a global public health
problem with *200 million cases worldwide (Wasley and
Alter, 2000; Grakoui et al., 2001; Lauer and Walker, 2001).
Chronic HCV infection is often associated with hepatitis,
cirrhosis and hepatocellular carcinoma (Choo et al., 1989;
Saito et al., 1990; Ruiz et al., 1992; Hoofnagle and di
Bisceglie, 1997; Lauer and Walker, 2001; Pawlotsky,
2004). HCV activates multiple signalling pathways
including those related to inflammation such as cyclooxy-
genase-2 (COX-2) (Bae et al., 2001; Rahman et al., 2001;
Waris and Siddiqui, 2005). Activation and upregulation of
COX-2 has been implicated in a number of cancers,
including HCV-associated hepatocellular carcinoma (Sano
et al., 1995; Uefuji et al., 1998; Chan et al., 1999;
Yip-Schneider et al., 2000; Bae et al., 2001; Rahman et al.,
2001; Waris and Siddiqui, 2005).
This work was partly presented at the 5th International Symposium on
˙
Pharmaceutical Chemistry, Istanbul—Turkey, 5–7 September 2007
and the Third International Meeting on Pharmacy & Pharmaceutical
˙
Sciences, Istanbul, Turkey, 9–12 June 2010.
˙
P. C¸ ıkla ꢀ E. Tatar ꢀ I. Ku¨c¸u¨kgu¨zel ꢀ S¸. G. Ku¨c¸u¨kgu¨zel (&)
Faculty of Pharmacy, Department of Pharmaceutical Chemistry,
˙
Marmara University, 34668 Haydarpas¸a, Istanbul, Turkey
e-mail: gkucukguzel@marmara.edu.tr
F. S¸ahin ꢀ D. Yurdakul
Genetics and Bioengineering Department, Faculty of
Engineering and Architecture, Yeditepe University,
˙
˘
Kayıs¸dagı, Istanbul, Turkey
A. Basu ꢀ R. Krishnan ꢀ D. B. Nichols ꢀ N. Kaushik-Basu
Department of Biochemistry and Molecular Biology,
UMDNJ-New Jersey Medical School, 185 South
Orange Avenue, Newark, NJ 07103, USA
123

Med Chem Res
Immunosuppression in cancer and as a result of che-
motherapy often leads to infections caused by opportunistic
pathogens. The array of these pathogenic organisms is
diverse. Therefore, an ideal cancer therapy should inhibit
cellular proliferation and also antagonize opportunistic
infections.
performed at ambient temperature using a normal phase
Waters, lPorasil column (part number : 27477G). All
experiments were performed in isocratic mode. The mobile
phase was prepared by mixing M1; hexane:ethanol (95:5
v/v) for 3a–k, 4a–b and 4d–k and M2; hexane:etanol
(90:10 v/v) for 5a–h, 6a–b, 6e and 6g filtered through a
0.45-lm pore filter and subsequently degassed by ultra-
sonication, prior to use. Solvent delivery was employed at a
flow rate of 1 mL/min. Detection of the analytes was car-
ried out at 254 nm. FTIR spectra were recorded on Schi-
madzu FTIR-8400S FT-IR spectrometer. NMR spectra
were recorded on Bruker AVANCE-DPX 400 at 400 MHz
and Bruker 300 at 300 MHz for 1H-NMR and 75 MHz for
13C-NMR (Decoupled). The chemical shifts were expres-
sed in d (ppm) downfield from tetramethylsilane (TMS)
using DMSO-d6 as solvent. MALDI-TOF HR-MS spectra
using EI and FAB ionization techniques were performed
using a Jeol JMS-700 instrument. Elemental analysis
(CHNS) of the compounds was performed on VarioMI-
CRO V1.5.7. and CHNS-932 (LECO).
Flurbiprofen, hydrazide-hydrazones and thiazolidinones
represent promising categories of compounds with versatile
properties and biological activities. Flurbiprofen is a non-
steroid anti-inflammatory drug (NSAID) that blocks
inflammation by inhibiting COX-2. Flurbiprofen and its
R-enantiomer have also been attributed with anti-prolifer-
ative properties against several different cell lines (Wechter
¨
et al., 1997, 2000a, b; Grosch et al., 2003; Jin et al., 2010).
Hydrazide-hydrazones are well-characterized antimicrobial
agents and are known to inhibit a wide range of bacterial
species (Ku¨c¸u¨kgu¨zel et al., 2003; Rollas and Ku¨c¸u¨kgu¨zel,
˘
˘
2007; Koc¸yigit-Kaymakc¸ıoglu et al., 2009). Thiazolidi-
nones possess anti-proliferative and anti-inflammatory
properties as well as antimycobacterial, antibacterial,
antifungal and antiviral activities (Ku¨c¸u¨kgu¨zel et al., 2002,
2006; Tatar et al., 2010). Recently, we demonstrated that
thiazolidinones inhibit HCV NS5B RNA-dependent RNA
polymerase (RdRp), and suggested that this scaffold may
be optimized for generating new analogues with improved
anti-NS5B potency (Kaushik-Basu et al., 2008a).
Methyl 2-(2-fluorobiphenyl-4-yl)propanoate (1)
Flurbiprofen (0.01 mol) was refluxed in methanol con-
taining concentrated H2SO4 for 3 h. The contents of the
flask were subsequently cooled and neutralized by
NaHCO3 (5 %). The oily crude product was extracted with
diethyl ether twice and diethyl ether was evaporated to
obtain compound (1). This compound has been previously
studied by Amir and Kumar, (2005).
In search of compounds with anti-proliferative, antimi-
crobial and anti-HCV NS5B properties, we have synthe-
sized a series of novel flurbiprofen hydrazide dervivatives
with some compounds bearing the thiazolidinone ring.
Here we describe the chemical characterization of these
compounds. We have also explored the anti-HCV NS5B,
anticancer and antimicrobial properties of these flurbipro-
fen hydrazide derivatives as discussed below.
2-(2-Fluorobiphenyl-4-yl)propanehydrazide (2)
Compound 2 was prepared by refluxing methyl 2-(2-fluo-
robiphenyl-4-yl)propanoate (1) (0.01 mol) with hydrazine
hydrate (80 %) (0.05 mol) in ethanol for 2 h. The reaction
mixture was allowed to cool and the precipitate was
washed with ice-cold water and recrystallized from meth-
anol. This compound has been previously studied by Amir
and Kumar, (2005).
Materials and methods
Experimental
All chemicals were purchased from Sigma-Aldrich or
Fluka. Reactions were monitored by thin-layer chroma-
tography (TLC) on silica gel plates purchased from Merck.
Melting points of the synthesized compounds were deter-
General procedure for the synthesis 2-(2-fluorobiphenyl-4-
yl)-N0-[(substituted phenyl/5-nitro-2-
furyl)methylene]propane hydrazide 3a–k
¨
mined in Schmelzpunktgerat SMP II melting apparatus and
were uncorrected. The liquid chromatographic system
consists of an Agilent Technologies 1100 series instrument
equipped with a quaternary solvent delivery system and a
model Agilent series G1315 A photodiode array detector.
A Rheodyne syringe loading sample injector with a 50-lL
sample loop was used for the injection of the analytes.
Chromatographic data were collected and processed using
Agilent Chemstation Plus software. The separation was
2-(2-Fluorobiphenyl-4-yl)propanehydrazide (2) (0.01 mol)
was dissolved in boiling absolute ethanol. Equimolar
amounts of the appropriate aromatic aldehyde were added
and refluxed for 2 h. The flask content was allowed to cool,
and the filtered and dried precipitates were recrystallized
from ethanol.
123

Med Chem Res
2-(2-Fluorobiphenyl-4-yl)-N0-[(5-nitro-2-furyl)methylene]prop-
anehydrazide 3a m.p., 241 °C. HPLC tR (min.): 12.218
(M1) IR (vmaks, cm-1): 3215 (NH), 1682 (C=O), 1622
135.35, 138.64, 141.89, 144.07, 145.70, 157.71, 169. 96
(Ar–C), 126.18 (Ar–CF3), 160.97 (=CH), 175.13 (C=O).
Analysis for C23H18F4N2Oꢀ1/2 H2O (423.395). Calcd: C,
65.24; H, 4.23; N, 6.62 %. Found: C, 64.61; H, 5.07; N,
5.84 %.
1
(C=N). H NMR (400 MHz, DMSO-d6) d ppm: 1.44–1.47
(m, 3H, CH3); 3.72 and 4.53 (2q, 1H, CH); 7.22–7.80 (m,
10H, Ar–H); 7.90 and 8.19 (2s, 1H, CH=N); 11.81 and
11.98 (2s, 1H, CO–NH). 13C-NMR (MHz) (DMSO-d6/
TMS) d ppm: 18.72 (CH–CH3), 44.11 (CH–CH3), 114.24,
114.24 115.98, 124.67, 127.19, 128.28, 129.07, 131.48,
135.33, 143.58, 152.27, 157.72, 170.25 (Ar–C), 160.98
(N=CH), 175.21 (C=O). Analysis for C20H16FN3O4
(381.357). Calcd: C, 62.99; H, 4.23; N, 10.85 %. Found: C,
62.68; H, 4.055; N, 11.08 %.
2-(2-Fluorobiphenyl-4-yl)-N0-[(2-fluorophenyl)methylene]prop-
anehydrazide 3e m.p., 194–195 °C. HPLC tR (min.):
4.666(M1). IR (vmax, cm-1): 3227 (NH), 1647 (C=O), 1616
1
(C=N). H NMR (400 MHz, DMSO-d6) d ppm: 1.43–1.47
(m, 3H, CH3); 3.77 and 4.76 (2q, 1H, CH); 7.26–7.97 (m,
12H, Ar–H); 8.18 and 8.46 (2s, 1H, CH=N); 11.52 and
11.74 (2s, 1H, CO–NH). 13C-NMR (75 MHz) (DMSO-d6/
TMS) d ppm: 18.75 (CH–CH3), 44.12 (CH–CH3), 116.29,
12.24, 124.53, 124.53, 126.80, 128.19, 129.07, 131.15,
132.37, 136.40, 140.15, 144.12, 157.71, 159.53, 162.84,
169.73(Ar–C), 160.97 (N=CH), 174.97 (C=O). Analysis
for C22H18F2N2O (364.387). Calcd: C, 72.51; H, 4.98; N,
7.69 %. Found: C, 72.10; H, 5.07; N, 7.43 %.
2-(2-Fluorobiphenyl-4-yl)-N0-[(4-nitrophenyl)methylene]prop-
anehydrazide 3b m.p., 205 °C. HPLC tR (min.): 8.328
(M1) IR (vmax, cm-1): 3181 (NH), 1670 (C=O), 1613
1
(C=N). H NMR (400 MHz, DMSO-d6) d ppm: 1.45–1.48
(m, 3H, CH3); 3.80 and 4.80 (2q, 1H, CH); 7.29–7.98 (m,
12H, Ar–H); 8.06 and 8.33 (2s, 1H, CH=N); 11.41–12.26
(b, 1H, CO–NH). 13C-NMR (75 MHz) (DMSO-d6/TMS) d
ppm: 18.77 (CH–CH3), 44.04 (CH–CH3), 115.46, 124.48,
128.26, 129.07, 131.19, 135.32, 141.00, 141.23, 143.64,
144.10, 145.00, 148.09, 157.72, 170.10 (Ar–C), 160.98
(N=CH), 175.28 (C=O). Analysis for C22H18FN3O3
(391.395). Calcd: C, 67.51;H, 4.64; N, 10.74 %. Found: C,
66.83; H, 4.47; N, 10.56 %.
2-(2-Fluorobiphenyl-4-yl)-N0-[(3-fluorophenyl)methylene]prop-
anehydrazide 3f m.p., 171–173 °C. HPLC tR (min.): 5.461
(M1). IR (vmax, cm-1): 3171 (NH), 1663 (C=O), 1645
1
(C=N). H NMR (400 MHz, DMSO-d6) d ppm: 1.43–1.47
(m, 3H, CH3); 3.79 and 4.77 (2q, 1H, CH); 7.23–7.55 (m,
12H, Ar–H); 7.96 and 8.24 (2s, 1H, CH=N); 11.02 and
12.10 (2s, 1H, CO–NH). 13C-NMR (75 MHz) (DMSO-d6/
TMS) d ppm: 18.63 (CH–CH3), 43.99 (CH–CH3), 115.60,
117.13, 123.78, 124.46, 127.10, 128.25, 129.13, 131.21,
135.36, 137.32, 142.13, 144.27, 146.10, 157.71, 164.54,
169.84 (Ar–C), 160.97 (N=CH), 175.05 (C=O). Analysis
for C22H18F2N2O. 1/2 H20 (373.395). Calcd: C, 70.77;H,
5.12; N, 7.50 %. Found: C, 71.32; H, 4.76; N, 7.58 %.
2-(2-Fluorobiphenyl-4-yl)-N0-[(2,4,6-trimethyl phenyl)methy-
lene]propanehydrazide 3c m.p., 180 °C. HPLC tR (min.):
4.172 (M1). IR (vmax, cm-1): 3190 (NH), 1649 (C=O),
1614 (C=N). 1H NMR (400 MHz, DMSO-d6) d ppm:
1.42–1.47 (m, 3H, CH3); 2.23 (s, 3H, Ar–CH3); 2.35–2.38
(d, 6H, Ar–CH3); 3.76 and 4.62 (2q, 1H, CH); 6.90–7.53
(m, 10H, Ar–H); 8.36 and 8.50 (2s, 1H, CH=N); 11.82 and
11.95 (2s, 1H, CO–NH). 13C-NMR (75 MHz) (DMSO-d6/
TMS) d ppm:18.89 (CH–CH3), 19.18, 21.12, 21.58 (Ar–
CH3), 44.02 (CH–CH3), 115.68, 124.41, 127.04, 128.19,
129.13, 129.91, 131.12, 135.38, 137.57, 138.62, 144.17,
147.34, 157.71, 169.30 (Ar–C), 160.97 (N=CH), 174.53
(C=O). Analysis for C25H25FN2O (388.477). Calcd: C,
77.29; H, 6.49; N, 7.21 %. Found: C, 77.38; H, 6.76; N,
7.44 %.
2-(2-Fluorobiphenyl-4-yl)-N0-[(4-fluorophenyl)methylene]prop-
anehydrazide 3g m.p., 173–175 °C. HPLC tR (min.): 5.765
(M1). IR (vmax, cm-1): 3171 (NH), 1660 (C=O), 1601
1
(C=N). H NMR (400 MHz, DMSO-d6) d ppm: 1.43–1.47
(m, 3H, CH3); 3.77 and 4.76 (2q, 1H, CH); 7.25–7.78 (m,
12H, Ar–H); 7.96 and 8.22 (2s, 1H, CH=N); 11.40 and
11.61 (2s, 1H, CO–NH). 13C-NMR (75 MHz) (DMSO-d6/
TMS) d ppm: 18.77 (CH–CH3), 43.98 (CH–CH3), 166.21,
127.57, 127.07, 128.25, 139.16, 131.29, 135.38, 142.41,
144.25, 146.33, 157.71, 160.97, 165.20, 169.68 (Ar–C),
161.91 (N=CH), 174.88 (C=O). Analysis for C22H18F2N2O
(364.387). Calcd: C, 72.51;H, 4.98; N, 7.69 %. Found: C,
72.38; H, 4.85; N, 7.64 %.
2-(2-Fluorobiphenyl-4-yl)-N0-{[4-(trifluoromethyl)phenyl]meth-
ylidene}propanehydrazide 3d m.p., 164–166 °C. HPLC tR
(min.): 5.582 (M1). IR (vmx, cm-1): 3184 (NH), 1666
1
(C=O), 1605 (C=N). H NMR (400 MHz, DMSO-d6) d
ppm: 1.44–1.47 (m, 3H, CH3); 3.81 and 4.80 (2q, 1H, CH);
7.27–7.93 (m, 12H, Ar–H); 8.03 and 8.29 (2s, 1H, CH=N);
11.35 and 12.11 (2b, 1H, CO–NH). 13C-NMR (75 MHz)
(DMSO-d6/TMS) d ppm: 18.74 (CH–CH3), 44.01 (CH–
CH3), 122.75, 124.63, 127.12, 128.19, 129.13, 131.17,
2-(2-Fluorobiphenyl-4-yl)-N0-[(2,6-difluorophenyl)methy-
lene]propanehydrazide 3h m.p., 193–195 °C. HPLC tR
(min.): 4.614 (M1) IR (vmax, cm-1): 3175 (NH), 1651
1
(C=O), 1624 (C=N). H NMR (400 MHz, DMSO-d6) d
123

Med Chem Res
2-(2-Fluoro-4-biphenylyl)-N-[2-(5-nitrofuran-2-yl)-4-oxo-
1,3-thiazolidine-3-yl]propanamide
ppm: 1. 43–1.48 (m, 3H, CH3); 3.77 and 4.69 (2q, 1H, CH);
7.17–7.52 (m, 11H, Ar–H); 8.14 and 8.38 (2s, 1H, CH=N);
11.55 and 11.75 (2s, 1H, CO–NH). 13C-NMR (75 MHz)
(DMSO-d6/TMS) d ppm: 18.63 (CH–CH3), 44.11 (CH–
CH3), 11.83, 113.01, 115.90, 124.34, 128.25, 129.06,
131.19, 133.84, 135.36, 137.83, 157.71, 159.10, 162.47,
169.67 (Ar–C), 160.92 (N=CH), 174.99 (C=O). Analysis
for C22H17F3N2O (382.378). Calcd: C, 69.10;H, 4.48; N,
7.33 %. Found: C, 68.78; H, 4.21; N, 7.38 %.
4a m.p.,
HPLC tR (min.): 10.729, 15.752 (M1) IR (vmax, cm-1
99 °C.
)
3140 (NH), 1720 (thiazolidinone, C=O), 1667 (amide,
C=O).1H NMR (400 MHz, DMSO-d6) d ppm: 1.35–1.38
(m, 3H, CH3); 3.77, 3.79 (2d and each 1H, S–CH2,
J = 16.0 Hz); 3.89–3.94 (m, 1H, CH); 5.96, 6.00 (2s, 1H,
S–CH–N); 6.80, 6.97 (2d, 1H, furan-H, J = 3.78 Hz);
7.14–7.21 (m, 2H, Ar–H); 7.38–7.55 (m, 6H, Ar–H); 7.61,
7.68 (2d, 1H, furan-H, J = 3.77 Hz); 10.52 and 10.59 (2s,
1H, CONH). 13C-NMR (75 MHz) (DMSO-d6/TMS) d
ppm: 19.01 (CH–CH3), 29.13 (S–CH2–), 42.81 (CH–CH3),
54.75 (S–CH–N), 114.07, 114.44, 115.58, 124.35, 127.25,
128.27, 129.07, 130.98, 135.33, 142.95, 152.01, 154.93,
157.64, 160.90 (Ar–C), 168.42 (NH–C=O), 172.35 (thia-
zolidinone C=O).
2-(2-Fluorobiphenyl-4-yl)-N0-[(4-chlorophenyl)methylene]propa-
nehydrazide 3i m.p., 179–181 °C. HPLC tR (min.): 5.518
(M1). IR (vmax, cm-1): 3171 (NH), 1659 (C=O), 1612
(C=N). Analysis for C22H18ClFN2O (380.842). Calcd: C,
69.38;H, 4.76; N, 7.36 %. Found: C, 69.53; H, 4.32; N,
7.57 %. This compound has been previously studied by
El-Sadek et al., (1996).
Analysis for C22H18FN3O5S (455.459). Calcd: C, 58.02;
H, 3.98; N, 9.23; S, 7.04 %. Found: C, 58.39; H, 4.91; N,
8.73; S, 7.09 %.
2-(2-Fluorobiphenyl-4-yl)-N0-[(4-methoxyphenyl)methylene]
propanehydrazide 3j m.p., 181–183 °C. HPLC tR (min.):
6.514 (M1). IR (vmax, cm-1): 3185 (NH), 1651 (C=O),
1605 (C=N). Analysis for C23H21FN2O2. C2H5OH
(422.423). Calcd: C, 71.07; H, 6.44; N, 6.63 %. Found: C,
71.34; H, 7.36; N, 6.92 %. This compound has been pre-
viously studied by El-Sadek et al., (1996).
2-(2-Fluoro-4-biphenylyl)-N-[2-(4-nitrophenyl)-4-oxo-1,3-
thiazolidine-3-yl]propanamide 4b m.p., 118–123 °C.
HPLC tR (min.): 8.895, 12.067 (M1). IR (vmax, cm-1):
3140 (NH), 1707 (thiazolidinone, C=O), 1678 (amide,
C=O).1H NMR (400 MHz, DMSO-d6) d ppm: 1.22–1.29
(m, 3H, CH3); 3.73, 3.75 (2d and each 1H, S–CH2,
J = 15.9 Hz) 3.89–3.93 (m, 1H, CH); 5.86, 5.90 (2s, 1H,
S–CH–N); 6.93–7.06 (m, 2H, Ar–H); 7.28–7.38 (m,
2H, Ar–H); 7.41–7.47 (m, 4H, Ar–H); 7.51 (d, 1H, Ar–H,
J = 8.79 Hz); 7.67 (d, 1H, Ar–H, J = 8.82 Hz); 8.06
(d, 1H, Ar–H, J = 8.78 Hz); 8.15 (d, 1H, Ar–H,
J = 8.79 Hz); 10.41 (s, 1H, CONH). 13C-NMR (75 MHz)
(DMSO-d6/TMS) d ppm: 18.86 (CH–CH3), 29.65 (S–CH2–),
42.61 (CH–CH3), 61.08 (S–CH–N), 115.35, 124.02,
127.11, 128.27, 129.11, 129.70, 130.89, 135.32, 143.11,
146.26, 148.16, 157.47, 160.80, 169.03 (Ar–C), 169.21
(NH–C=O), 172.12 (thiazolidinone C=O). Analysis for
C24H20FN3O4S (474.504). Calcd: C, 60.75; H, 4.46; N,
8.85; S, 6.76 %. Found: C, 60.33; H, 4.59; N, 8.60; S,
6.88 %.
2-(2-Fluorobiphenyl-4-yl)-N0-[(2-chlorophenyl))methylene]
propanehydrazide 3k m.p., 199 °C. HPLC tR (min.):
4.415 (M1). IR (vmax, cm-1): 3185 (NH), 1651 (C=O),
1624 (C=N). 1H NMR (400 MHz, DMSO-d6) d ppm:
1.43–1.47 (m, 3H, CH3); 3.78 and 4.77 (2q, 1H, CH);
7.27–8.03 (m, 12H, Ar–H); 8.36 and 8.61 (2s, 1H, CH=N);
11.59 and 11.84 (2s, 1H, CO–NH). 13C-NMR (75 MHz)
(DMSO-d6/TMS) d ppm: 18.81 (CH–CH3), 44.14 (CH–
CH3), 115.62, 124.54, 127.13, 128.18, 129.06, 130.38,
131.16, 131.66, 131.89, 133.57, 135.36, 139.58, 157.72,
144.23, 143.22, 169.82 (Ar–C), 160.98 (N=CH), 175.02
(C=O). Analysis for C22H18ClFN2O (380.842). Calcd: C,
69.38; H, 4.76; N, 7.36 %. Found: C, 69.15; H, 4.76; N,
7.29 %.
2-(2-Fluoro-4-biphenylyl)-N-[2-[4-(trifluoromethyl)phenyl]-4-
oxo-1,3-thiazolidine-3-yl]propanamide 4d m.p., 114–119 °C.
HPLC tR (min.): 6.41, 7.384 (M1). IR (vmax, cm-1): 3254
General procedure for the synthesis 2-(2-fluoro-4-
biphenylyl)-N-(2-substituted-4-oxo-1,3-thiazolidine-3-
yl)propanamide (4a–b and 4d–k)
1
(NH), 1709 (thiazolidinone, C=O), 1678 (amide, C=O). H
2-(2-Fluorobiphenyl-4-yl)-N0-[(substitutedphenyl/5-nitro-
2-furyl)methylene]propanehydrazide (3a–b and 3d–k)
(0.01 mol) in dried toluene was refluxed with thioglycolic
acid (mercaptoacetic acid) (0.1 mol) under a Dean-Stark
water trap for 10 h. Toluene was evaporated under vacuo and
the flask content was neutralized by addition of NaHCO3
(5 %) until CO2 release was completed. The precipitate was
washed with water and recrystallized from ethanol (50 %).
NMR (400 MHz, DMSO-d6) d ppm: 1.33 (m, 3H, CH3);
3.64, 3.79 (2d and each 1H, S–CH2, J = 16.0 Hz) 3.92–3.97
(m, 1H, CH); 5.85 (s, 1H, S–CH–N); 6.99–7.64 (m, 12H,
Ar–H); 10.41 (s, 1H, CONH). 13C-NMR (75 MHz) (DMSO-
d6/TMS) d ppm: 17.99 (CH–CH3), 29.61 (S–CH2–), 42.56
(CH–CH3), 60.81 (S–CH–N), 115.45, 122.63, 124.28,
125.83, 126.92, 128.27, 129.15, 129.56, 129.85, 130.91,
135.35, 143.38, 157.50, 160.77 (Ar–C), 124.46 (Ar–CF3),
123

Med Chem Res
169.26 (NH–C=O), 172.03 (thiazolidinone C=O). Analysis
for C25H20F4N2O2SꢀH2O (506.497). Calcd: C, 59.28; H,
4.38; N, 5.53; S, 6.33 %. Found: C, 59.55; H, 4.33; N, 5.34;
S, 6.27 %.
C, 63.14; H, 4.86; N, 6.14; S, 7.02 %. Found: C, 63.14; H,
4.86; N, 6.14; S, 7.02 %.
2-(2-Fluoro-4-biphenylyl)-N-[2-(2,6-difluorophenyl)-4-oxo-1,3-
thiazolidine-3-yl]propanamide 4h m.p., 154–157.7 °C.
HPLC tR (min.): 6.579, 7.608 (M1). IR (vmax, cm-1): 3261
(NH), 1713 (thiazolidinone, C=O), 1676 (amide, C=O).1H
NMR (400 MHz, DMSO-d6) d ppm: 1.27–1.35 (m, 3H,
CH3); 3.63, 3.68 (2d and each 1H, S–CH2, J = 15.8 Hz)
3.79–3.85 (m, 1H, CH); 6.05, 6.12 (2s, 1H, S–CH–N);
6.99–7.52 (m, 11H, Ar–H); 10.57,10.65 (2s, 1H, CONH).
13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 17.56 (CH–
CH3), 29.66 (S–CH2–), 42.46 (CH–CH3), 50.47 (S–CH–
N), 112.61, 115.11, 115.48, 123.75, 124.30, 127.05,
128.26, 129.15, 130.95, 132.02, 135.37, 143.06, 157.61,
160.76 (Ar–C), 168.43 (NH–C=O), 172.25 (thiazolidinone
C=O). Analysis for C24H19F3N2O2S (456.480). Calcd: C,
63.15; H, 4.20; N, 6.14; S, 7.02 %. Found: C, 63.03; H,
4.23; N, 6.01; S, 6.88 %.
2-(2-Fluoro-4-biphenylyl)-N-[2-(2-fluorophenyl)-4-oxo-1,3-thi-
azolidine-3-yl]propanamide 4e m.p., 158–166 °C. HPLC tR
(min.): 6.654, 7.275 (M1). IR (vmax, cm-1): 3267 (NH),
1711 (thiazolidinone, C=O), 1682(amide, C=O).1H NMR
(400 MHz, DMSO-d6) d ppm: 1.28–1.35 (m, 3H, CH3);
3.75, 3.78 (2d and each 1H, S–CH2, J = 15.9 Hz)
3.86–3.90 (m, 1H, CH); 5.96, 6.04 (2s, 1H, S–CH–N);
7.14–7.57 (m, 12H, Ar–H); 10.52 (s, 1H, CONH).
13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 17.91 (CH–
CH3), 29.60 (S–CH2–), 40.53 (CH–CH3), 55.94 (S–CH–
N), 115.28, 116.23, 123.90, 125.65, 127.04, 128.24,
129.16, 130.01, 130.89, 131.48, 135.37, 143.05, 157.52,
159.17, 162.46, 160.78 (Ar–C), 169.07 (NH–C=O), 172.18
(thiazolidinone C=O). Analysis for C24H20F2N2O2S
(438.489). Calcd: C, 65.74; H, 4.60; N, 6.96; S, 7.31 %.
Found: C, 65.26; H, 4.53; N, 6.21; S, 7.09 %.
2-(2-Fluoro-4-biphenylyl)-N-[2-(4-chlorophenyl)-4-oxo-1,3-
thiazolidine-3-yl]propanamide 4i m.p., 87–91 °C. HPLC
tR (min.): 6.261, 7.343 (M1)IR (vmax, cm-1): 3239 (NH),
1703 (thiazolidinone, C=O), 1682 (amide, C=O).1H NMR
(400 MHz, DMSO-d6) d ppm: 1.28–1.34 (m, 3H, CH3);
3.63, 3.76 (2d and each 1H, S–CH2, J = 15.8 Hz)
3.87–3.93 (m, 1H, CH); 5.75, 5.78 (2s, 1H, S–CH–N);
7.29–7.55 (m, 12H, Ar–H); 10.37 (s, 1H, CONH).
13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 17.93 (CH–
CH3), 29.74 (S–CH2–), 42.54 (CH–CH3), 61.44 (S–CH–
N), 115.11, 115.84, 124.35, 128.26, 129.11, 130.52,
130.64, 130.87, 134.51, 134.68, 135.35, 143.12, 157.52,
161.11 (Ar–C), 169.12 (NH–C=O), 171.93 (thiazolidinone
C=O). Analysis for C24H20ClFN2O2Sꢀ1/2 H2O (463.951).
Calcd: C, 62.13; H, 4.55; N, 6.03; S, 6.91 %. Found: C,
61.99; H, 4.38; N, 5.99; S, 6.81 %.
2-(2-Fluoro-4-biphenylyl)-N-[2-(3-fluorophenyl)-4-oxo-1,3-thi-
azolidine-3-yl]propanamide 4f m.p., 104 °C. HPLC tR
(min.): 5.979, 7.259 (M1). IR (vmax, cm-1): 3208 (NH),
1703 (thiazolidinone, C=O), 1661 (amide, C=O).1H NMR
(400 MHz, DMSO-d6) d ppm: 1.29–1.31 (m, 3H, CH3);
3.75, 3.91 (2d and each 1H, S–CH2, J = 15.9 Hz)
3.89–3.96 (m, 1H, CH); 5.77, 5.84 (2s, 1H, S–CH–N);
7.12–7.53 (m, 12H, Ar–H); 10.41 (s, 1H, CONH).
13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 18.01 (CH–
CH3), 29.60 (S–CH2–), 42.56 (CH–CH3), 60.90 (S–CH–
N), 114.80, 115.20, 116.43, 123.97, 124.31, 127.12,
128.04, 129.09, 131.05, 135.28, 141.76, 141.86, 143.17,
157.54, 160.87, 164.11 (Ar–C), 169.21 (NH–C=O), 172.00
(thiazolidinone C=O). Analysis for C24H20F2N2O2Sꢀ1/2
H2O (438.490). Calcd: C, 65.74; H, 4.60; N, 6.39; S,
7.31 %. Found: C, 65.80; H, 4.60; N, 6.35; S, 7.13 %.
2-(2-Fluoro-4-biphenylyl)-N-[2-(2-chlorophenyl)-4-oxo-1,3-
thiazolidine-3-yl]propanamide 4k m.p., 90–92 °C. HPLC
tR (min.): 6.034, 6.997 (M1). IR (vmax, cm-1): 3273 (NH),
2-(2-Fluoro-4-biphenylyl)-N-[2-(4-fluorophenyl)-4-oxo-1,3-thi-
azolidine-3-yl]propanamide 4g m.p., 65–70 °C. HPLC tR
(min.): 6.523, 7.798 (M1). IR (vmax, cm-1): 3208 (NH),
1
1713 (thiazolidinone, C=O), 1678 (amide, C=O). H NMR
(400 MHz, DMSO-d6) d ppm: 1.29–1.36 (m, 3H, CH3);
3.77, 3.81 (2d and each 1H, S–CH2, J = 15.9 Hz)
3.87–3.93 (m, 1H, CH); 6.09, 6.17 (2s, 1H, S–CH–N);
6.95–7.48 (m, 12H, Ar–H); 10.54 (s, 1H, CONH).
13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 17.98 (CH–
CH3), 29.42 (S–CH2–), 42.57 (CH–CH3), 58.17 (S–CH–
N), 115.24, 123.88, 127.06, 128.22, 129.19, 129.04,
130.00, 130.83, 132.71, 135.38, 157.51, 136.17, 142.98,
135.84, 160.77 (Ar–C), 169.56 (NH–C=O), 172.18 (thia-
zolidinone C=O). Analysis for C24H20ClFN2O2S (438.49).
Calcd: C, 63.36; H, 4.43; N, 6.16; S, 7.05 %. Found: C,
63.22; H, 4.60; N, 6.13; S, 7.11 %.
1
1705 (thiazolidinone, C=O), 1663(amide, C=O). H NMR
(400 MHz, DMSO-d6) d ppm: 1.27–1.33 (m, 3H, CH3);
3.74, 3.76 (2d and each 1H, S–CH2, J = 15.8 Hz)
3.86–3.91 (m, 1H, CH); 5.77, 5.82 (2s, 1H, S–CH–N);
7.03–7.54 (m, 12H, Ar–H); 10.08–10.62 (bs, 1H, CONH).
13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 17.93 (CH–
CH3), 29.71 (S–CH2–), 42.54 (CH–CH3), 61.44 (S–CH–
N), 115.15, 115.66, 124.35, 127.07, 128.26, 129.11,
130.79, 134.51, 134.68, 135.38, 143.12, 157.56, 161.11,
164.36 (Ar–C), 169.12 (NH–C=O), 171.94 (thiazolidinone
C=O). Analysis for C24H20F2N2O2SꢀH2O (456.489). Calcd:
123

Med Chem Res
General procedure for the synthesis 2-[2-(2-
fluorobiphenyl-4-yl)propanoyl]-N-substituted hydrazine
carbothioamide 5a–h
143.84, 157.65 (Ar–C), 160.91 (C=O), 172.90 (C=S).
Analysis for C19H22FN3OꢀH2O (377.476). Calcd: C, 60.46;
H, 6.41; N, 11.13; S, 8.49 %. Found: C, 60.49; H, 6.28; N,
11.07; S, 8.30 %.
2-(2-Fluorobiphenyl-4-yl)propanehydrazide (2) (0.01 mol)
was dissolved in boiling absolute ethanol. Equimolar
amounts of appropriate isothiocyanate were added and
refluxed for 2 h. The flask content was allowed to cool and
the filtered and dried precipitates were recrystallized from
ethanol.
2-[2-(2-Fluorobiphenyl-4-yl)propanoyl]-N-n-butyl hydra-
zinecarbothioamide 5d
m.p., 157–160 °C. HPLC tR
(min.): 5.34 (M2). IR (vmax, cm-1): 3156 (NH), 1695
(C=O), 1226 (C=S).1H NMR (400 MHz, DMSO-d6) d
ppm: 0.86 (t, 3H, CH2CH2CH2CH3); 1.24 (m, 2H,
NHCH2CH2CH2CH3); 1.42 (d, 3H, CHCH3); 1.47 (m, 2H,
NHCH2CH2CH2CH3); 3.41 (m, 2H, NHCH2CH2CH2CH3);
3.72 (q, 1H, CHCH3); 7.26–7.54 (m, 8H, Ar–H); 7.68
(s, 1H, NH); 9.16 (s, 1H, NH); 9.94 (s, 1H, NH). 13C-NMR
(75 MHz) (DMSO-d6/TMS) d ppm: 14.24 (NH–CH2–
CH2–CH2–CH3), 18.52 (CH–CH3), 19.85 (NH–CH2–CH2–
CH2–CH3), 31.29 (NH–CH2–CH2–CH2–CH3), 43.20
(NH–CH2–CH2–CH2–CH3), 43.76 (CH–CH3), 115.43,
124.53, 126.92, 127.10, 128.25, 129.13, 131.00, 135.39,
143.84, 157.66 (Ar–C), 160.92 (C=O), 172.90 (C=S).
Analysis for C20H24FN3OS (373.487). Calcd: C, 64.32; H,
6.48; N, 11.25; S, 8.59 %. Found: C, 64.03; H, 6.26; N,
11.19; S, 8.77 %.
2-[2-(2-Fluorobiphenyl-4-yl)propanoyl]-N-methyl hydra-
zinecarbothioamide 5a m.p., 184–185 °C. HPLC tR
(min.): 7.634 (M2). IR (vmax, cm-1): 3160 (NH), 1676
(C=O), 1205 (C=S).1H NMR (400 MHz, DMSO-d6) d
ppm: 1.43 (d, 3H, CHCH3); 2.89 (d, 3H, NHCH3); 3.70 (q,
1H, CHCH3); 7.26–7.54 (m, 8H, Ar–H); 7.88 (s, 1H, NH);
9.21 (s, 1H, NH); 9.93 (s, 1H, NH). 13C-NMR (75 MHz)
(DMSO-d6/TMS) d ppm: 18.52 (CH–CH3), 22.41 (NH–
CH3), 45.74 (CH–CH3), 115.73, 124.53, 126.93, 127.10,
128.25, 129.14, 131.00, 135.39, 143.84, 157.66 (Ar–C),
160.91 (C=O), 172.89 (C=S). Analysis for C17H18FN3
OSꢀH2O (349.423). Calcd: C, 58.43; H, 5.77; N, 12.03;
S, 9.18 %. Found: C, 58.62; H, 5.84; N, 11.82; S,
8.94 %.
2-[2-(2-Fluorobiphenyl-4-yl)propanoyl]-N-allyl
hydra-
zinecarbothioamide 5e m.p., 133 °C. HPLC tR (min.):
5.921 (M2). IR (vmax, cm-1): 3169 (NH), 1680 (C=O),
1194 (C=S).1H NMR (400 MHz, DMSO-d6) d ppm: 1.40
(d, 3H, CHCH3); 3.72 (q, 1H, CHCH3); 4.10 (q, 2H,
NHCH2); 5.05 (d, 1H, CH=CH2, J = 10.4 Hz,cis); 5.12 (d,
1H, CH=CH2, J = 17.3 Hz, trans); 5.82 (m, 1H,
CH=CH2); 7.26–7.54 (m, 12H, Ar–H); 7.99 (s, 1H, NH);
9.28 (s, 1H, NH); 9.98 (s, 1H, NH). 13C-NMR (75 MHz)
(DMSO-d6/TMS) d ppm: 18.60 (CH–CH3), 43.23 (NH–
CH2–CH=CH2), 46.25 (CH–CH3), 115.45, 124.56, 126.91,
127.09, 128.25, 129.15, 130.99, 143.75, 143.85, 157.65
(Ar–C), 115.76 (NH–CH2–CH=CH2), 135.36 (NH–CH2–
CH=CH2), 160.91 (C=O), 172.96 (C=S). Analysis for
C19H20FN3OSꢀ1/2 H2O (366.452). Calcd: C, 62.27; H,
5.77; N, 11.46; S, 8.75 %. Found: C, 62.35; H, 5.75; N,
11.45; S, 8.651 %.
2-[2-(2-Fluorobiphenyl-4-yl)propanoyl]-N-ethyl
hydra-
zinecarbothioamide 5b m.p., 190–192 °C. HPLC tR
(min.): 6.00 (M2). IR (vmax, cm-1): 3169 (NH), 1695
(C=O), 1221 (C=S). 1H NMR (400 MHz, DMSO-d6) d
ppm: 1.05 (t, 3H, CH2CH3); 1.43 (d, 3H, CHCH3); 3.54 (m,
2H, CH2CH3); 3.71 (q, 1H, CHCH3); 7.26–7.54 (m, 8H,
Ar–H); 7.78 (s, 1H, NH); 9.17 (s, 1H, NH); 9.94 (s, 1H,
NH). 13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 14.76
(NH–CH2–CH3), 18.45 (CH–CH3), 38.95 (NH–CH2–CH3),
43.23 (CH–CH3), 115.38, 124.32, 126.95, 127.99, 128.85,
129.00, 130.75, 135.42, 143.66, 157.68 (Ar–C), 160.95
(C=O), 172.89 (C=S). Analysis for C18H20FN3OS
(345.434). Calcd: C, 62.59; H, 5.84; N, 12.16; S, 9.28 %.
Found: C, 62.38; H, 5.64; N, 12.09; S, 9.58 %.
2-[2-(2-Fluorobiphenyl-4-yl)propanoyl]-N–n-propyl hydra-
zinecarbothioamide 5c m.p., 155 °C. HPLC tR (min.):
5.669 (M2). IR (vmax, cm-1): 3160 (NH),1676 (C=O), 1223
(C=S).1H NMR (400 MHz, DMSO-d6) d ppm: 0.81 (t, 3H,
CH2CH2CH3); 1.34 (d, 3H, CHCH3); 1.47 (m, 2H,
CH2CH2CH3); 3.73 (q, 1H, CHCH3); 3.63–3.82 (m,
2H, NCH2CH2CH3); 7.26–7.54 (m, 8H, Ar–H); 7.72 (s, 1H,
NH); 9.17 (s, 1H, NH); 9.95 (s, 1H, NH). 13C-NMR
(75 MHz) (DMSO-d6/TMS) d ppm: 11.58 (NH–CH2–
CH2–CH3), 18.52 (CH–CH3), 22.41 (NH–CH2–CH2–CH3),
43.20 (CH–CH3), 45.73 (NH–CH2–CH2–CH3), 115.73,
124.53, 126.93, 127.10, 128.25, 129.14, 131.00, 135.39,
2-[2-(2-Fluorobiphenyl-4-yl)propanoyl]-N-phenyl hydra-
zinecarbothioamide 5f m.p., 197–201 °C. HPLC tR
(min.): 5.785 (M2). IR (vmax, cm-1): 3121 (NH), 1672
(C=O), 1192 (C=S).1H NMR (400 MHz, DMSO-d6) d
ppm: 1.45 (d, 3H, CHCH3); 3.79 (q, 1H, CHCH3);
7.29–7.55 (m, 13H, Ar–H); 9.79 (s, 2H, NH); 10.18 (s, 1H,
NH). 13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 18.58
(CH–CH3), 43.25 (CH–CH3), 115.52, 115.83, 124.52,
125.54, 126.93, 127.10, 128.26, 128.65, 129.16, 131.00,
139.55, 143.73, 143.83, 157.66 (Ar–C), 160.92 (C=O),
172.91 (C=S). Analysis for C22H20FN3OS (393.477).
123

Med Chem Res
Calcd: C, 67.15; H, 5.12; N, 10.68; S, 8.15 %. Found: C,
66.76; H, 5.01; N, 10.59; S, 8.13 %.
13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 18.66 (CH–
CH3), 29.04 (NH–CH3), 33.25 (–CH2–S), 43.18 (CH–
CH3), 115.49, 124.33, 127.03, 128.22, 129.16, 131.00,
135.42, 144.21, 157.64, 160.90 (Ar–C), 159.66 (C=N),
169.61 (C=O), 171.87 (thiazolidinone C=O). Analysis for
C19H18FN3O2S (371.428). Calcd: C, 61.44; H, 4.88; N,
11.31; S, 8.63 %. Found: C, 61.04; H, 4.82; N, 11.17; S,
8.54 %. HR-MS (Calculated [MH?]/Found [MH?] m/
z) 372.11765/372.11775.
2-[2-(2-Fluorobiphenyl-4-yl)propanoyl]-N-benzyl hydra-
zinecarbothioamide 5g m.p., 153–154 °C. HPLC tR
(min.): 5.539 (M2). IR (vmax,cm-1): 3161 (NH), 1705
(C0O), 1172 (C=S). 1H NMR (400 MHz, DMSO-d6) d
ppm: 1.43 (d, 3H, CHCH3); 3.72 (q, 1H, CHCH3); 4.75 (m,
2H, NHCH2C6H5); 7.23–7.54 (m, 13H, Ar–H); 8.41 (bs,
1H, NH); 9.36 (s, 1H, NH); 9.93 (b, 1H, NH). 13C-NMR
(75 MHz) (DMSO-d6/TMS) d ppm: 18.65 (CH–CH3),
43.26 (CH–CH3), 47.17 (–NH–CH2–), 115.48, 115.79,
124.56, 126.91, 127.10, 127.40, 127.80, 128.54, 129.15,
130.97, 135.41, 139.71, 143.84, 157.66 (Ar–C), 160.91
(C=O), 173.04 (C=S). Analysis for C23H22FN3OS
(407.504). Calcd: C, 67.79; H, 5.44; N, 10.31; S, 7.87 %.
Found: C, 67.50; H, 5.37; N, 10.21; S, 7.77 %.
2-(2-Fluorobiphenyl-4-yl)-N0-[(3-ethyl-4-oxo-1,3-thiazolidine-
2-ylidene]propanehydrazide 6b m.p., 164 °C. HPLC tR
(min.): 8.83 (M2). IR (vmax, cm-1): 3196 (NH), 1720
(thiazolidinone, C=O), 1637 (amide, C=O). 1H NMR
(400 MHz, DMSO-d6) d ppm: 1.12 (t, 3H, CH2CH3); 1.41
(d, 3H, CHCH3); 3.67 (q, 2H, CH2CH3); 3.86 (q, 1H,
CHCH3); 4.00 (s, 2H, SCH2); 7.19–7.55 (m, 8H, Ar–H);
10.48 (s, 1H, CO–NH). 13C-NMR (75 MHz) (DMSO-d6/
TMS) d ppm: 12.55 (NH–CH2-CH3), 18.64 (CH–CH3),
33.14 (–CH2–S), 37.95 (NH–CH2–CH3), 43.23 (CH–CH3),
115.51, 124.35, 127.03, 128.22, 129.16, 130.99, 135.42,
144.20, 157.64, 160.90 (Ar–C), 159.43 (C=N), 169.57
(C=O), 171.68 (thiazolidinone C=O). Analysis for
C20H20FN3O2S (385.455). Calcd: C, 62.32; H, 5.23; N,
10.90; S, 8.32 %. Found: C, 62.18; H, 5.35; N, 10.79;
S, 8.20 %. HR-MS (Calculated [MH?]/Found [MH?]
m/z) C20H20FN3O2S 386.13330/386.13348.
2-[2-(2-Fluorobiphenyl-4-yl)propanoyl]-N-cyclohexyl hydra-
zinecarbothioamide 5h
m.p., 150–152 °C. HPLC tR
(min.): 5.067 (M2). IR (vmax, cm-1): 3171 (NH), 1692
(C=O), 1175 (C=S).1H NMR (400 MHz, DMSO-d6) d
ppm: 0.81–1.14 (m, 5H, cyclohexyl CH2); 1.42 (d, 3H,
CHCH3); 1.53–1.61 (m, 5H, cyclohexyl CH2); 3.24–3.30
(s, 1H, cyclohexyl CH); 3.73 (q, 1H, CHCH3); 7.27–7.58
(m, 9H, Ar–H and N–H); 9.16 (bs, 1H, NH); 9.96 (s, 1H,
NH). 13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm: 18.48
(CH–CH3), 25.86, 30.70, 37.42, 50.15 (C6H11), 43.18 (CH–
CH3), 115.72, 124.50, 126.93, 127.10, 128.25, 129.12,
131.00, 135.40, 143.83, 157.58 (Ar–C), 160.94 (C=O),
172.88 (C=S). Analysis for C22H26FN3OS (399.525).
Calcd: C, 66.14; H, 6.56; N, 10.52; S, 8.03 %. Found: C,
66.33; H, 6.70; N, 9.99; S, 7.54 %.
2-(2-Fluorobiphenyl-4-yl)-N0-[(3-allyl-4-oxo-1,3-thiazol-
idine-2-ylidene]propanehydrazide 6e m.p., 165 °C.
HPLC tR (min.): 9.106 (M2). IR (vmaks, cm-1): 3180 (NH),
1720 (thiazolidinone, C=O), 1658 (amide, C=O).
1H NMR (300 MHz, DMSO-d6) d ppm: 1.41 (d, 3H,
CHCH3); 3.84 (q, 1H, CHCH3); 4.12 (s, 2H, SCH2); 4.24
(d, 2H, NCH2); 5.12 (d, 1H,CH=CH2, J = 17.0 Hz, trans);
5.14 (d, 1H, CH=CH2, J = 10.0 Hz, cis); 5.75–5.88 (m,
1H, CH=CH2); 7.15–7.55 (m, 12H, Ar–H); 10.48 (s, 1H,
CO–NH).13C-NMR (75 MHz) (DMSO-d6/TMS) d ppm:
18.65 (CH–CH3), 33.08 (–CH2–S), 43.23 (CH–CH3), 44.71
(N–CH2–CH=CH2), 117.19 (N–CH2–CH=CH2), 115.51,
124.36, 126.87, 128.22, 129.16, 131.00, 131.52, 144.18,
157.64, 160.89 (Ar–C), 135.43 (N–CH2–CH=CH2), 159.11
(C=N), 169.47 (C=O), 171.61 (thiazolidinone C=O).
Analysis for C21H20FN3O2Sꢀ1/2 H2O (406.473). Calcd: C,
62.05; H, 5.20; N, 10.34; S, 7.89 %. Found: C, 62.61; H,
5.09; N, 10.40; S, 7.90 %. HR-MS (Calculated [MH?]/
Found [MH?] m/z) C21H20FN3O2S 398.13330/398.13349.
General procedure for the synthesis 2-(2-fluorobiphenyl-4-
yl)-N0-[(3-substituted-4-oxo-1,3-thiazolidine-2-
ylidene]propanehydrazide 6a–b, 6e and 6g
2-[2-(2-Fluorobiphenyl-4-yl)propanoyl]-N-substituted hy-
drazinecarbothioamide (5a–e and 5g) (0.01 mol) was dis-
solved in boiling absolute ethanol. Ethyl a-bromoacetate
(0.011 mol) and anhydrous sodium acetate (0.04 mol) were
added and refluxed for 2–8 h. The flask content was
allowed to cool and the filtered and dried precipitates were
recrystallized from ethanol/methanol (for 6g).
2-(2-Fluorobiphenyl-4-yl)-N0-[(3-methyl-4-oxo-1,3-thiazol-
idine-2-ylidene]propanehydrazide 6a m.p., 199–200 °C.
HPLC tR (min.): 11.49 (M2). IR (vmax, cm-1): 3144 (NH),
2-(2-Fluorobiphenyl-4-yl)-N0-[(3-benzyl-4-oxo-1,3-thiazol-
idine-2-ylidene]propanehydrazide 6g m.p., 168–170 °C.
HPLC tR (min.): 8.894 (M2). IR (vmax, cm-1): 3192 (NH),
1722 (thiazolidinone, C=O), 1637 (amide, C=O).
1
1726 (thiazolidinone, C=O), 1637 (amide, C=O). H NMR
(400 MHz, DMSO-d6) d ppm: 1.41 (d, 3H, CHCH3);
3.08 (s, 3H, NHCH3); 3.86 (q, 1H, CHCH3); 4.02 (s, 2H,
SCH2); 7.27–7.55 (m, 8H, Ar–H); 10.45 (s, 1H, CO–NH).
123

Med Chem Res
1H NMR (300 MHz, DMSO-d6) d ppm: 1.27 (d,
CH3OH, recrystalize solvent); 1.41 (d, 3H, CHCH3); 1.65
(s, CH3OH, recrystalize solvent); 3.80–3.87 (q, 1H,
CHCH3); 4.14 (s, 2H, SCH2); 4.84 (s, 2H, CH2C6H5);
7.29–7.55 (m, 13H, Ar–H); 10.56 (s, 1H, CO–NH). 13C-
NMR (75 MHz) (DMSO-d6/TMS) d ppm: 18.91 (CH–
CH3), 33.55 (–CH2–S), 43.25 (CH–CH3), 45.93 (N–CH2),
115.55, 124.39, 126.85, 127.72, 127.96, 128.29, 128.91,
129.16, 130.98, 135.45, 136.36, 144.24, 157.65, 160.90
(Ar–C), 159.27 (C=N), 169.58 (C=O), 171.02 (thiazolidi-
none C=O). Analysis for C25H22FN3O2S. 2CH3OH
(511.608). Cald: C, 63.89; H, 5.91; N, 8.21; S, 6.27 %.
Found: C, 63.40; H, 4.95; N, 8.79; S, 6.62 %. HR-MS
(Calculated [MH?]/Found [MH?] m/z) C25H22FN3O2S
448.14895/448.14931.
Engineering and Architecture, Yeditepe University, Istan-
bul, Turkey. The microorganisms used were Micrococcus
luteus, Staphylococcus cohnii, Staphylococcus aureus,
Escherichia coli, Proteus vulgaris, Pseudomonas aeru-
ginosa, Micrococcus lylae, Salmonella paratyphi, Acine-
tobacter
radioresistens,
Enterobacter
aerogenes,
Photorhabdus luminescens, Candida albicans, Candida
parapsilosis, Candida tropicalis, Candida glabrata,
Schwanniomyces occidentalis, Kluyveromyces marxianus,
Pichia membranaefaciens and Aspergillus sp. Ampisillin,
Cefepime and Amfoterisin B were used as the positive
sensitivity reference standard for bacteria and fungi, yeast.
Disc-diffusion assay
The compounds were dissolved in DMSO to a final con-
centration of 10 mg/mL. Antimicrobial tests were deter-
mined by the disc-diffusion method using 100 lL of
suspension containing 108 CFU/mL of bacteria, 106 CFU/
mL of yeast and 104 spore/mL of fungi spread on nutrient
agar (NA) for bacterial species, and potato dextrose agar
(PDA) for yeast and fungi species. The discs (6 mm in
diameter) were impregnated with 20 lL of dissolved
compounds and placed on the inoculated agar. DMSO
served as the negative control. 200 lg/disc of Ampicillin
(Bio Basic Inc) was used as the positive sensitivity refer-
ence standard for bacteria. 200 lg/disc of Amphotericin B
(Riedel–de Haen) was used as a positive reference standard
for yeast and fungi. The inoculated plates were incubated at
37 °C for 24 h for bacterial strains, 48 h for yeast and 72 h
for fungal isolates. Antimicrobial activity was evaluated by
measuring the zone of inhibition against the test organisms.
Biological activity
HCV NS5B polymerase ınhibitory activity
Inhibition of HCV NS5B RNA-dependent RNA polymer-
ase activity by flubiprofen compounds was evaluated in the
Department of Biochemistry and Molecular Biology, New
Jersey Medical School, UMDNJ. All synthesized com-
pounds (3a–k, 4a–b, 4d–k, 5a–h and 6a–b, 6e, 6g) were
evaluated for their ability to inhibit HCV NS5B polymer-
ase by the standard primer-dependent elongation reactions
employing poly rA/U12 template primers (TP) as previ-
ously described (Kaushik-Basu et al., 2008a, b). Recom-
binant NS5B carrying the N-terminal histidine-tag was
expressed from the plasmid pTh NS5BCD21in Escherichia
coli strain DH5a and purified using Ni–NTA chromatog-
raphy (Kaushik-Basu et al., 2008a, b). The compounds
were screened at 200-lM concentration. Wedelolactone, a
previously characterized NS5B inhibitor, was employed as
a positive control and DMSO served as negative control.
Reactions were incubated at 30 °C for 60 min, RNA
product was precipated and radioactive incorporation was
quantified on a liquid scintillation counter. Inhibition of the
NS5B RdRp activity was determined relative to the DMSO
control.
Micro-well dilution assay
The sensitivity of the bacterial strains towards the com-
pounds was quantitively evaluated from the minimal
inhibitory concentration (MIC) values obtained by the
micro-well dilution method. The inocula of the bacterial
strains were prepared from 12-h broth cultures and sus-
pensions were adjusted to 0.5 McFarland standard turbid-
ity. Compounds dissolved in DMSO were first prepared at
the highest concentration to be tested (200 lg/mL), and
then serial two-fold dilutions were made in order to obtain
a concentration range from 6.25 to 200 lg/mL, in 15-mL
sterile test tubes containing nutrient broth. The 96-well
plates were prepared by dispensing into each well 95 lL of
nutrient broth and 5 lL of the inoculum. 200 lL of nutrient
broth without inoculum was transferred into the first wells
as positive control. Aliquots, (100 lL) taken from the
200-lg/mL stock solution, were added to the second well.
100 lL from the respective serial dilutions was transferred
into 5 consecutive wells. The last well containing 195 lL
Antimicrobial activity
The antimicrobial activities of all compounds were evalu-
ated in the Department of Genetics and Bioengineering,
Faculty of Engineering and Architecture, Yeditepe Uni-
versity. The antibacterial and antifungal activities of the
compounds were evaluated against 20 microbial cultures
isolates of 11 bacteria, 1 fungi and 7 yeast species by disc-
diffusion assay and micro-well dilution assay as described
below (Gu¨l et al., 2005). Microorganisms were provided by
the Department of Genetics and Bioengineering, Faculty of
123

Med Chem Res
of nutrient broth without compound and 5 lL of the
inoculum on each strip was used as negative control.
Contents of each well were mixed on plate shaker at
300 rpm for 20 s and then incubated at appropriate tem-
peratures for 24 h. Microbial growth in each medium was
determined by reading the absorbance (Abs) at 630 nm
using the ELx 800 universal microplate reader (Biotek
Instrument inc, Highland Park, Vermont, USA) and con-
firmed by plating 5-lL samples from clear wells on
nutrient agar medium. The MIC was defined as the lowest
concentration of the compounds to inhibit the growth of
microorganisms. Cefepime (MaxipimeÒ, Bristol-Myers
Squibb, Istanbul, Turkey) at the concentration range of
500–7.8 lg/mL was prepared in nutrient broth and used as
standard drug for positive control.
HR-MS). Purity of the synthesized compounds have been
demonstrated by HPLC analysis using normal phase
chromatography. As thiazolidinones synthesized starting
from racemic flurbiprofen have two stereocentres and four
possible optical isomers; diastereoisomeric pairs of these
compounds existed. Our chromatographic system was able
to separate these isomeric pairs with well-resolved peaks.
All thiazolidinones 4a–b and 4d–k were observed to have
two peaks whose UV spectra recorded by PDA detector
were completely identical.
In the 1H-NMR spectra of 2-(2-Fluorobiphenyl-4-yl)-N0-
[(substituted phenyl/5-nitro-2-furyl)methylene] propa-
nehydrazide (3a–k) generated by the condensation of
appropriate aromatic aldehydes with 2-(4-fluorobiphenyl-
3-yl)propanoic acid hydrazide (2) (El-Sadek et al., 1996;
Amir and Kumar, 2005), N–H and azomethine protons
showed two singlet as is the case with -CH- protons of
flurbiprofen skeleton (CH–CH3) demonstrating two quar-
tets acquiring the integration of one proton. The conse-
quence is that the hydrazide-hydrazones (3a–k) occurred as
E and Z forms (Rutavichyus et al., 1995, 2000). Com-
pounds (3i) and (3j), carrying 4-chloro and 4-methoxy
groups on the phenyl ring simultaneously, have been pre-
viously described (El-Sadek et al., 1996). The purities of
(3i) and (3j) were determined by elemental analysis.
Anticancer activity
The cytotoxicity and anticancer activities of the synthe-
sized compounds were elucidated at the National Institutes
of Health-National Cancer Institute, Bethesda (NIH-NCI).
A total of 60 human tumour cell lines derived from nine
cancer types (non-small cell lung, colon, breast, ovarian,
leukaemia, renal, prostate, CNS and melanoma) formed the
basis of this test. The tumour cells were cultured in
RPMI1640 medium supplemented with 5 % foetal calf
serum and 2-mM L-glutamine. The tumour cells were
inoculated over a series of standard 96-well microtiter
plates in 100 lL of medium (Grever et al., 1992; Boyd and
Paull, 1995). Density of the inoculum depended on the type
of tumour cells and their growth characteristics (Weinstein
et al., 1997). These cells were then preincubated on the
microtiter plate for 24 h before adding the compounds at a
single concentration of 10-5 M. After incubation with the
chemical agent for 48 h with the tumour cell lines, the
sulforhodamine B (SRB) protein assay was used to esti-
mate cell viability and growth (Skehan et al., 1990). The
cytotoxic effects were evaluated and assay results and
dose–response parameters were calculated as described
previously (Grever et al., 1992). Details of this test system
and the information derived from the activity pattern over
all cell lines have been previously described (Shoemaker,
2006; Weinstein et al., 1997; Monks et al., 1991).
1
FT-IR and H-NMR spectral findings of 2-(2-fluoro-4-
biphenylyl)-N-(2-substituted-4-oxo-1,3-thiazolidine-3-yl)
propanamide (4a–b, 4d–k) supported the formation of the
target compounds which were synthesized by the reaction
of hydrazide-hydrazones (3a–b, 3d–k) with thioglycolic
acid in benzene with Dean-Stark water trap. In the
1H-NMR spectra of 1,3-thiazolidine-4-ones (4a–b, 4d–k);
lack of azomethine proton’s signals of hydrazide-hydra-
zones at 8.00–9.00 ppm (Ku¨c¸u¨kgu¨zel et al., 2003) and
existence of new signals attributable to S–CH2, N–CH–S
and CONHN = protons in accordance with literature
(Ku¨c¸u¨kgu¨zel et al., 2002, 2006; Tatar et al., 2010) revealed
that the cyclization through hydrazide-hydrazones
(3a–b and 3d–k) had occured. Chiral centre (N–CH–S) and
amide (CONH) protons of the mentioned ring were deter-
mined between 3.79–3.97 ppm and 10.08–10.65 ppm,
respectively. In the 1H-NMR spectra, methylene protons of
the 4-thiazolidinone ring displayed two signals appearing
as dublets at 3.77, 3.79 ppm for 4a, 3.73, 3.75 ppm for 4b,
3.64, 3.79 ppm for 4d, 3.75, 3.78 ppm for 4e, 3.75,
3.91 ppm for 4f, 3.74, 3.76 ppm for 4g, 3.63, 3.68 ppm for
4h, 3.63, 3.76 ppm for 4i and 3.77, 3.81 ppm for 4k,
respectively, due to the non-equivalent, geminal methylene
protons interacting with the chiral centre at position 2
(Ku¨c¸u¨kgu¨zel et al., 2006).
Results and discussion
Chemistry
In this study, thirty-two novel compounds were synthesized
as per the described Fig. 1. The structures of these com-
pounds were characterized using elemental analysis
(CHNS) and spectral data (FT-IR, 1H-NMR, 13C-NMR and
Existence of singlet peaks attributable to N–H protons of
thiosemicarbazide moiety (CO–NH–NH–CS–NH) between
1
7.68–8.41; 9.16–9.79 and 9.93–10.18 ppm in the H-NMR
123

Med Chem Res
CH3
CH3
CH3
NH
OCH3
OH
NH2
i
O
ii
O
O
F
F
F
2
1
Flurbiprofen
v
iii
CH3
S
CH3
NH
NH
NH
NH
R
N
R
O
O
F
5a-h
F
3a-k
vi
iv
CH3
S
O
NH
CH3
R
N
N
NH
R
O
N
S
O
F
O
F
6a-b, 6e, 6g
4a-b, 4d - k
(i)
CH3OH / H2SO4, reflux ; (ii) NH2NH2.H2O, reflux ; (iii) R–CH=O / EtOH, reflux ; (iv)
Toluene, HS-CH2-COOH (v) R-N=C=S, (vi) Br-CH2-COOC2H5 / CH3COONa.
Compound
Compound
R
R (Ar)
5-nitro-2-furyl
C6H4NO2 (4)
3a
3b
4a
4b
6a
6b
5a
5b
-CH3
-CH2CH3
n-C3H7
n-C4H9
C6H2(CH3)3 (2,4,6)
C6H4(CF3) (4)
3c
5c
3d
3e
4d
4e
5d
-CH2CH=
CH2
C6H4F (2)
6e
6g
5e
5g
C6H4F (3)
C6H4F (4)
3f
3g
4f
5f
-C6H5
-CH2C6H5
-C6H11
4g
C6H3F2 (2,6)
C6H4Cl (4)
3h
3i*
4h
4i
5h
C6H4(OCH3) (4)
C6H4Cl (2)
3j*
3k
4k
(*) El-Sadek et al., 1996.
Fig. 1 Synthetic route of compounds 3a–k, 4a–b, 4d–k, 5a–h and 6a–b, 6e, 6g
123

Med Chem Res
CH3
Fig. 2 Synthetic route for
1,3,4-oxadiazoline derivatives
CH3
O
NH
N
(CH3CO)2O
N
Ar
N
O
O
CH3
Ar
F
F
spectra of 2-[2-(2-fluorobiphenyl-4-yl)propanoyl]-N-substi-
tuted hydrazine carbothioamide (5a–h), that were gained by
the condensation of substituted isothiocyanates with 2-(4-
fluorobiphenyl-4-yl)propanehydrazide (2), proved the for-
mation of thiosemicarbazide structure.
appropriate number of resonances that exactly resembled the
number of carbon atoms.
IR spectra of hydrazide-hydrazones (3a–k) showed char-
acteristic bands at 3227–3171, 1682–1647 and 1645–
1601 cm-1 corresponding to hydrazone N–H, C=O and C=N
groups, respectively (Ku¨c¸u¨kgu¨zel et al., 2002, 2003, 2006).
Thiosemicarbazides (5a–h) were also characterized by IR
spectra displaying N–H at 3,275–3,115 and C=S bands at
1,174–1,227 cm-1 (Rollas, 1983). N–H and C=O bands of
4-thiazolidinones obtained from hydrazones or thiosemicar-
bazides were detected within the 3273–3140, 3277–3144 and
1720–1703, 1726–1716 cm-1 region, respectively.
The thiosemicarbazides were then reacted with ethyl
a-bromoacetate in the presence of anhydrous sodium ace-
tate in absolute ethanol to yield 2-(2-fluorobiphenyl-4-yl)-
N0-[(3-methyl-4-oxo-1,3-thiazolidine-2-ylidene]propanehyd-
razide 6a–b, 6e and 6g. By the reaction of thiosemicar-
bazides 5a–h with ethyl a-bromoacetate in the presence of
anhydrous sodium acetate in absolute ethanol (Fig. 1), four
compounds 6a–b, 6e and 6g instead of eight were gained.
The presence of SCH2 signals between 4.00 and 4.10 ppm
An additional goal of the present study was to synthesize
1,3,4-oxadiazoline compounds (Fig. 2). However, all efforts
for the synthesis of 1,3,4-oxadiazoline derivatives from flur-
biprofen hydrazones failed despite many attempted reaction
conditions (steam bath, heating mantle and oil bath). It has
been described in previous literature that in structures with
biphenyl moiety the cyclization of hydrazones to 1,3,4-ox-
adiazoline ring have not been achieved (Joshi et al., 2008).
1
in the H-NMR spectra of compounds 6a–b, 6e and 6g)
supported the formation of the thiazolidinone ring. Lack of
N–H signals of thiosemicarbazides (5a–h) between
7.68–8.41; 9.16–9.79 and 9.93–10.18 ppm are also other
evidences for the formation of 4-thiazolidinones. High-
resolution mass spectra (HR-MS) confirmed the molecular
weights and empirical formulae of compounds 6a–b, 6e
and 6g, with less than 5-mmu bias between calculated and
experimental m/z values of molecular ions.
Pharmacology
Additional support for the structures of the synthesized
compounds was provided by 13C-NMR spectra. The signals
belonging to –C=O group and N=CH group derived from 3a–
h and 3k were recorded at 174.53–175.28 ppm and
160.92–161.91 ppm, respectively (Gu¨rsoy and Gu¨zeldemirci,
2007). The peaks resonated at 29.42–29.71 ppm, 50.47–
61.44 ppm, 168.43–169.56 ppm and 171.93–172.25 ppm in
the 13C-NMR spectrum of 4-thiazolidinones, 4a–b and 4d–
k obtained from hydrazones, assigned for S–CH2, S–CH–N,
amide C=O and thiazolidinone C=O moieties, confirmed the
carbon skeleton of 4-thiazolidinone ring (Ku¨c¸u¨kgu¨zel et al.,
2006; Patel and Shaikh, 2010). On the other hand, the 13C-
NMR spectra exhibited resonated at 160.91–160.95 ppm and
172.88–173.04 ppm assigned for flubiprofen thiosemicarbaz-
ides 5a–h, C=O and C=S, respectively (Gu¨rsoy and Karali,
1995, Abdel-Aziz and Abdel-Rahman, 2010). Another series of
4-thiazolidinones (6a–b, 6e and 6g), prepared from flubiprofen
thiosemicarbazides, were characterized by 13C-NMR spec-
trum at 33.08–33.55, 159.11–159.66, 169.47–169.61 and
171.61–172.01 ppm due to S–CH2, C=N, amide C=O and
4-thiazolidinone C=O peaks, respectively (Tatar et al., 2008,
2010) The 13C-NMR spectra of all compounds displayed the
HCV NS5B polymerase ınhibitory activity
2-(2-Fluorobiphenyl-4-yl)-N0-[(substituted phenyl/5-nitro-2-
furyl)methylene]propane hydrazide (3a–k) and 2-(2-Fluoro-
4-biphenylyl)-N-(2-substituted-4-oxo-1,3-thiazolidine-3-yl)
propanamide (4a–b, 4d–k), 2-[2-(2-fluorobiphenyl-4-yl)
propanoyl]-N-substituted hydrazinecarbothioamide (5a–h)
and novel 2-(2-fluorobiphenyl-4-yl)-N0-[(3-methyl-4-oxo-
1,3-thiazolidine-2-ylidene]propanehydrazides (6a–b, 6e,
6g) were evaluated for their activities against HCV NS5B
RdRp employing the in vitro NS5B RdRp assay (Kaushik-
Basu et al., 2008a, b). For these experiments, wedelolactone,
a previously characterized NS5B inhibitor (Kaushik-Basu
et al., 2008b) served as a positive control and yielded NS5B
inhibition consistent with previously reported values (data
not shown). Hydrazide-hydrazones (3a–k), thiosemicar-
bazides (5a–h) and 1,3-thiazolidine-4-ones (4a–b, 4d–k and
6a–b, 6e, 6g) exhibited a wide range of activity ranging from
8 to 70 % inhibition at 200 lM compound concentration.
Compound 4k bearing 2-chlorophenyl moiety at C-2 of
1,3-thiazolidine-4-one ring and compound 5d bearing butyl
moiety at N-1 position of thiosemicarbazide were identified
123

Med Chem Res
Fig. 3 Anticancer result of compound 4d
as the most active compounds with *70 % inhibition of
NS5B activity. Among the ten 1,3-thiazolidine-4-one
derivatives (4a–b and 4d–k), six (4a, 4d–f, 4i and 4k)
exhibited higher inhibitory activity against NS5B when
compared to the corresponding flurbiprofen hydrazide-
hydrazones used as the starting compounds. For compounds
(4b) and (4j), closure of the corresponding hydrazide-
hydrazones to yield corresponding 1,3-thiazolidine-4-one
123

Med Chem Res
derivatives caused a decrease in terms of their inhibitory
activity. The inhibitory activity of the hydrazide-hydrazone
derivatives (3g and 3h) and their corresponding 1,3-thia-
zolidine-4-one derivatives (4g and 4h) exhibited near equal
inhibition of NS5B. As this inhibition was relatively weak
compared to other NS5B inhibitors reported in literature,
none of the compounds were further pursued for IC50 value
evaluation.
hydrazide-hydrazones used as the starting compounds. The
other compounds, exhibited no inhibitory effect on any
other bacterial strains, fungi or yeast isolates.
Anticancer activity
The cytotoxicity and anticancer activities of synthesized
compounds (3i), (3j), (3h), (4d), (4i) and (6b) were evaluated
at NIH-NCI. A primary anticancer assay was performed
according to the protocolofthe DrugEvaluationBranchofthe
NationalCancerInstitute, Bethesda(Alleyetal., 1988;Monks
et al., 1991; Grever et al., 1992; Boyd and Paull, 1995;
Shoemaker, 2006). Selection of the compounds for anticancer
screening was made according to the criteria which are
available online at the DTP web site (http://www.dtp.
nci.nih.gov/docs/misc/common_files/guidelines.html). The
cytotoxic and/or growth inhibitory effects of the compounds
were tested in vitro at a single concentration (10-5 M) against
the full panel of 60 human tumour cell lines derived from nine
neoplastic diseases. The growth percentage was evaluated
spectrophotometrically and compared to controls which were
not treated with test agents. A 48-h continuous drug exposure
protocol was followed and a sulforhodamine B (SRB) protein
assay was used to estimate cell viability or growth. The cell
lines used in the National Cancer Institute screen were leu-
kaemia, non-small cell lung cancer (NSCL), colon cancer,
central nervous system (CNS) cancer, melanoma, ovarian
Antimicrobial activity
The antimicrobial activities of the compounds against the
laboratory strains of bacterial species, isolates of seven
fungi and a species of yeast were tested using disc-diffu-
sion and microdilution assays (Gu¨l et al., 2005). Of the
compounds tested, only compounds (4a) and (4i) exhibited
activity against M. luteus, S. cohnii and S. aureus between
100 and 200 lg/mL concentrations. From the data it has
been shown that, compound (4a) containing 5-nitro-2-furyl
moiety presented inhibition zone against S. cohnii and
S. aureus (10 mm), whereas ampicillin has not shown
inhibition against same microorganisms in the in vitro disc-
diffusion test assays. Hence, the preliminary experimental
data revealed that some of the 4-thiazolidinones of flurbi-
profen showed a very interesting activity profile against
M. luteus, S. cohnii and S. aureus when compared with the
standard ampicillin and the corresponding flurbiprofen
Table 1 Anticancer screening
data of selected compounds at
10-5
Compound
NCI code
Mean growth
The most sensitive cell line
Growth percent % of the
most sensitive cell line
M
3h
3j
749789/1
749725/1
89.43
90.58
Breast cancer
T-47D
55.26
Leukaemia SR
Breast cancer
T-47D
46.04
57.41
4d
749726/1
66.73
Leukaemia SR
Leukaemia HL (TB)
Leukaemia K562
Leukaemia MOLT-4
Leukaemia RPMI-8226
Breast cancer
T-47D
20.80
36.88
39.48
43.50
43.34
33.42
4i
749727/1
749728/1
77.39
86.59
Renal cancer
CAK-1
29.58
50.90
Breast cancer
T-47D
Prostate cancer PC-3
Breast cancer
T-47D
49.18
55.24
6b
Breast cancer
MDA-MB-468
43.60
123

Med Chem Res
NS5B inhibition studies were supported by the National Institute of
Health research Grants DK066837 and CA153147 to N.K-B (Neerja
Kaushik-Basu). The authors are grateful to Dr. Ju¨rgen Gross from the
Institute of Organic Chemistry, University of Heidelberg, for his gener-
ous help on obtaining HR-EI/FAB mass spectra of the synthesized
compounds. We thank the Division of Cancer Research, National Cancer
Institute, Bethesda, MD, for the anticancer activity screening.
cancer, renal cancer, prostate cancer and breast cancer cell
lines. The one-dose data will be reported as a mean graph of
the percent growth of treated cells. Mean Graphs were con-
structed for each effect, with bars depicting the deviation of
individualtumourcelllinesfromthe overallmeanvaluefor all
the cells tested. In the Mean Graph, the centre point is the
mean of the growth inhibition (GI) percentage over all cell
lines. Bars that point to the right are cell lines where the
inhibition is greater than the average, while bars that point to
the left are cell lines where the inhibition is less than the
average. Compounds withcell lines appearing onpositiveside
ofmeangraph exhibit highgrowthinhibitionofcancercellsof
that particular cancer. The One-dose assay allows detection of
both growth inhibition (values between 0 and 100) and
lethality (values less than 0). It means that a value of 100
means no growth inhibition. A value of 40 would mean 60 %
growth inhibition. A value of 100[means cytotoxicity at this
cell line. Tested compounds have moderate growth inhibition
effects, so they have not been passed the 5-dose assay. Despite
these results, with compound (4d), containing thiazolidinone
ring, growth percent was found to be 20.80 and 33.42 % for a
leukaemia cancer cell line SR and breast cancer cell line
T-47D, respectively (Fig. 3). There are also several reports in
the literature on 4-thiazolidinones which were observed to
inhibit leukaemia cells (Grasso et al., 1988, Mosula et al.,
2009; Havrylyuk et al., 2009). The results of in vitro anti-
cancer screening are summarized in Table 1. Moreover,
compounds 3h, 3i, 3j, 4d, 4i and 6b had a strong cytotoxicity
against melanoma SK-MEL-2, colon cancer COLO-205 and
colon cancer HCC-2998.
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