Design, synthesis, and anticancer evaluation of some novel thiourea, carbamimidothioic acid, oxazole, oxazolidine, and 2-amino-1-phenylpropyl-2-chloroacetate derived from L-norephedrine

Article abstract of DOI:10.1134/S1068162016040026

A novel series of thiourea, carbamimidothioic acid, 4, 5-dihydrooxazole-2-thiol, oxazolidine-2thine, and 2-amino-1-phenylpropyl-2-chloroacetate derivatives was designed and synthesized using 2-amino-1-phenylpropan-1-ol (L-norephedrine) as a strategic starting material. The structures of the newly synthesized compounds were established by elemental analyses, IR, and ¹H NMR and ¹³C NMR spectral data. The compounds were evaluated for their in vitro anticancer activity against various cancer cell lines. The corresponding acetamide, carbamimidothioic acid, and 2-2-amino-1-phenylpropyl-2-chloroacetate derivatives showed almost the same activity as the standard drug doxorubicin against human breast cancer cell line (MCF-7). Also, the acetamide and 2-thioxoimidazolidin-4-one derivatives exhibited higher activity than the reference drug doxorubicin against human colon cancer cell line (HCT 116).

Full text of DOI:10.1134/S1068162016040026

ISSN 1068ꢀ1620, Russian Journal of Bioorganic Chemistry, 2016, Vol. 42, No. 4, pp. 434–440. © Pleiades Publishing, Ltd., 2016.
Design, Synthesis, and Anticancer Evaluation of Some Novel
Thiourea, Carbamimidothioic Acid, Oxazole, Oxazolidine,
and 2ꢀAminoꢀ1ꢀPhenylpropylꢀ2ꢀChloroacetate Derived
from LꢀNorephedrine1
Maged S. AbdelꢀKadera, b, Mostafa M. Ghorabc, d, 2, Mansour S. Alsaidc, and Saleh I. Alqasoumic
aDepartment of Pharmacognosy, College of Pharmacy, Prince Sattam Bin Abdulaziz University, Saudi Arabia
bDepartment of Pharmacognosy, College of Pharmacy, Alexandria University, Alexandria, Egypt
cDepartment of Pharmacognosy, College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh, 11451 Saudi Arabia
dDepartment of Drug Radiation Research, National Center for Radiation Research and Technology,
P.O. Box 29, Nasr City, Cairo, Egypt
Received February 8, 2016; in final form, March 6, 2016
Abstract—A novel series of thiourea, carbamimidothioic acid, 4, 5ꢀdihydrooxazoleꢀ2ꢀthiol, oxazolidineꢀ2ꢀ
thine, and 2ꢀaminoꢀ1ꢀphenylpropylꢀ2ꢀchloroacetate derivatives was designed and synthesized using
2ꢀaminoꢀ1ꢀphenylpropanꢀ1ꢀol (Lꢀnorephedrine) as a strategic starting material. The structures of the newly
1
synthesized compounds were established by elemental analyses, IR, and H NMR and 13C NMR spectral
data. The compounds were evaluated for their in vitro anticancer activity against various cancer cell lines. The
corresponding acetamide, carbamimidothioic acid, and 2ꢀ2ꢀaminoꢀ1ꢀphenylpropylꢀ2ꢀchloroacetate derivꢀ
atives showed almost the same activity as the standard drug doxorubicin against human breast cancer cell line
(MCFꢀ7). Also, the acetamide and 2ꢀthioxoimidazolidinꢀ4ꢀone derivatives exhibited higher activity than the
reference drug doxorubicin against human colon cancer cell line (HCT 116).
Keywords: thiourea carbamimidothioic acid, oxazole, oxazolidine, 2ꢀaminoꢀ1ꢀphenylpropylꢀ2ꢀchloroacetate,
anticancer activity
DOI: 10.1134/S1068162016040026
INTRODUCTION
Thioureas can be used in control of plant pathogens,
such as Penicillum expansum and Fusarium oxysporum
[17]. 1,3ꢀDialkyl or diarylthioureas exhibit significant
antifungal activity against Pyricularia oryzae and
Drechslera oryzae [18]. It has been reported that thioꢀ
urea, oxazole, and oxazolidine derivatives show potent
anticancer activity [19, 20]. Crank et al. [21, 22]
reported for the first time the antifungal activity of 2ꢀ
aminooxazole thiourea derivatives against Botrytis
cinerea. In this work, a series of new thiourea, carbamꢀ
imidothioic acid, oxazole, oxazolidine, and 2ꢀaminoꢀ
1ꢀphenylpropylꢀ2ꢀchloroacetate derivatives bearing
biologically active moieties were prepared and evaluꢀ
ated for their anticancer activity against three human
cancer cell lines.
According to the World Health Organization, canꢀ
cer is a foremost health tricky. More than 7 million
deaths are reported worldwide annually [1, 2].
Because of limitation of surgery and radiotherapy in
curing cancer, chemotherapy is of increasing imporꢀ
tance [1, 2]. Therefore, identification of novel potent,
selective, and less toxic anticancer agents remains one
of the most pressing health problems. Thioureas are
sulfur and nitrogenꢀcontaining compounds that have
proved to be promising in drug research in recent years
[3–8]. Some carbamimidothioic acid derivatives posꢀ
sess valuable antibacterial and anticancer activities
[9–12]. Most of these compounds include heterocyꢀ
clic rings, such as oxazole and oxazolidine. It is well
known that oxazoles and oxazolidines find applicaꢀ
tions in the fields of medicine and industry [13–16].
Oxazole and oxazolidine nuclei have also been incorꢀ
porated into a wide variety of therapeutically imporꢀ
tant molecules to transform them into better drugs.
RESULTS AND DISCUSSION
2ꢀAminoꢀ1ꢀphenylpropanꢀ1ꢀol (Lꢀnorephedrine)
(I) was reacted with pꢀchlorophenylisothiocyanate(II)
to give three novel compounds: 1ꢀ(4ꢀchlorophenyl)ꢀ3ꢀ
(1ꢀhydroxyꢀ1ꢀphenylpropanꢀ2ꢀyl)thiourea (III),
methylꢀ5ꢀphenylꢀ4,5ꢀdihydrooxazoleꢀ2ꢀthiol
and its isomer 4ꢀmethylꢀ5ꢀphenyloxazolidineꢀ2ꢀ
1
4ꢀ
The article is published in the original.
2
(V),
Corresponding author: phone:
+ 966ꢀ534292860; fax:
+966014670560; eꢀmail: mmsghorab@yahoo.com.
434

DESIGN, SYNTHESIS, AND ANTICANCER EVALUATION
435
thione (VI) (Scheme 1). Compounds (
were formed via intermediate (IV) (Scheme 2). The
structures of compounds (III), ( ), and (VI) were
established on the basis of elemental analysis, IR, H
NMR, 13C NMR, and mass spectral data. IR spectra
of compounds ( ) and (VI) revealed the absence of
V
) and (VI
)
nature and the addition of ClCH2C=O to (III). IR
spectrum of compound (VIII) showed clear absence of
OH band. Besides 1H NMR signals of Lꢀnorephedrine
V
1
and 4ꢀchlorophenyl moieties, compound
(VII)
showed singlet at δH 3.73 assigned for CH2Cl group.
The disappearance of the NH signal of Lꢀnorepheꢀ
drine indicated the site of acylation at this position.
Signals of the CH2Cl–C=O moiety appeared at δC
31.83 and 155.72 ppm in the 13C NMR. The 1H NMR
spectrum of compounds (VIII) revealed the presence
of singlet at δH 3.76 assigned for CH2Cl group and the
disappearance of OH signal and the shift of the CH–
OH proton to δH 5.02 indicating that the acylation
occurred at the OH group. The signals for CH2Cl–
C=O moiety appeared at δC 33.97 and 166.91 ppm in
V
OH and C–Cl bands and the presence of bands at
3100, 3068 cm–1 (CH arom.) and 2942, 2835, 2956,
and 2876 cm–1 (CH aliph.), respectively. The main difꢀ
ference between the two compounds was the presence
of C=N band at 1612 cm–1 in compound (
V
) replaced
by C=S band at 1271 cm–1 in compound (VI). The 1H
NMR spectra of compounds ( ) and (VI) showed
V
closely related peaks for CH3, CH–N and CH–O proꢀ
tons and five aromatic protons. The main difference
was in the singlet at δH 7.61 assigned for SH together
with the 13C NMR signal at δC 155.52 assigned for
the 13C NMR spectrum. Another significant features
1
of the H and 13C NMR spectra of compound (VIII
)
C=N function in compound (V). Compound (VI)
were the singlet at δH 10.32 assigned for SH proton and
replacement of C=S signal with C=N signal at δC
164.79 ppm. The 1H NMR spectrum of compound (IX
showed singlet at δH 10.26 assigned for NH together
with the 13C NMR signal at δC 187.91 assigned for
C=S function. Both compounds showed the same M+ at
)
showed signals characteristic of Lꢀnorephedrine.
However, the absence of OH signal and the downfield
shift of the CH–OH to δH 5.02, together with the
CH2Cl singlet at δH 3.57, indicated that the OH group
193 m/z, however, in compound (VI) the molecular ion
was the base peak due to the stability of the ion thioxo
group at position 2 comparing with the much less stable
ketoꢀenol thiol (mercapto) group. Reaction of compound
was acylated. The 13C NMR spectrum of compound (IX
)
(
III) with chloroacetyl chloride in chloroform in the presꢀ
ence of triethylamine as a catalyst afforded four novel comꢀ
pounds: 2ꢀchloroꢀ ꢀ(4ꢀchlorophenylcarbamothioyl)ꢀ
(1ꢀhydroxyꢀ1ꢀphenylpropanꢀ2ꢀyl)acetamide (VII), ꢀ(1ꢀ
(2ꢀchloroacetoxy)ꢀ1ꢀphenylpropanꢀ2ꢀyl)ꢀ 'ꢀ(4ꢀchloꢀ
rophenyl)carbamimidothioic acid VIII),
aminoꢀ1ꢀphenylpropylꢀ2ꢀchloroacetate (IX), and 3ꢀ
(4ꢀchlorophenyl)ꢀ2ꢀthioxoimidazolidinꢀ4ꢀone XI
(Schemes 3–5). The structures of compounds (VII),
showed all signals of Lꢀnorephedrine and CH2Cl sigꢀ
nal at δC 31.62 ppm. The M+ at 227
m/z
was in comꢀ
N
Nꢀ
1
plete agreement with the proposed structure. The H
NMR spectrum of compound (XI) indicated the comꢀ
plete absence of Lꢀnorephedrine signals. The two douꢀ
N
N
(
2ꢀ
blets at δH 7.33 and 7.59,
J = 8, each integrated for 2H,
were assigned for the ꢀsubstituted aromatic ring. The
p
(
)
singlet at δH 3.69 integrated for 2H, together with corꢀ
responding 13C NMR signal at δC 44.45 ppm was
assigned to the NH–CH2–C=O moiety. The signals at
δC 168.51 and 178.78 ppm were assigned ftoor C=O
(
VIII), (IX), and (XI) were established on the basis of
1
microanalyses, IR, H NMR, 13C NMR, and mass
spectral data. Compounds (VII) and (VIII) showed
the same M+ at 397
m/z indicating their isomeric and C=S carbons, respectively.
Cl
S
NH
HN
H2N
CHCl
TEA
3
+
N=C=S
Cl
OH
III
OH
(I
)
(II
)
(
)
+
N
NH
+
O
O
S
S H
(V)
(VI)
Scheme 1. Formation of compounds (III), (
V
), and (VI).
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 42
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2016

436
MAGED S. ABDELꢀKADER et al.
Cl
S
HN
Cl
NH
N
HN
S−H
O−H
OH
III
(IV)
(
)
NH2
–
Cl
N
NH
O
O
S H
S
(VI
)
(V)
Scheme 2. Postulated mechanisms of the formation of compounds (
V
) and (VI).
Cl
Cl
O
Cl
S
Cl
O
S
Cl
NH
NH
N
HN
CHCl
TEA
3
OH
III
OH
VII
(
)
(
)
+
H
S
N
Cl
S
NH2
O
NH
HN
+
+
Cl
Cl
Cl
N
O
O
O
O
(
XI
)
(IX)
(VIII
)
Scheme 3. Formation of compounds (VII)–(IX), and (XI).
H
–
NCS
Cl
NH2
S
Cl
N
Cl
O
NH
O
Cl
O
O
(VIII
)
(IX)
Scheme 4. Postulated mechanism for the formation of compound (IX).
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 42
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DESIGN, SYNTHESIS, AND ANTICANCER EVALUATION
437
Cl
Cl
S
N
O
Cl
S
S
O
H N
NH
HN
Cl
–HCl
HN
Cl
Cl
N
H
CHCl
TEA
3
OH
–
O
OH
OH
III
(
)
(
X)
(XI)
Scheme 5. Postulated mechanism for the formation of compound (XI).
In Vitro Anticancer Activity
itive control doxorubicin against the liver cancer cell
line HepG2. Compounds (VII) and (IX) exhibited no
activity against the liver cancer cell line. The structure–
activity relationship indicated that the addition of thioꢀ
urea to 4ꢀchloroaniline with free OH group was proved to
be effective in the case of compound (VII) showing a slight
The newly synthesized compounds were evaluated
for their in vitro cytotoxic activity against human
breast (MCFꢀ7), liver (HepG2), and colon (HCT 116)
cancer cell lines. The clinically used drug doxorubicin,
one of the most effective anticancer agents, was used
as a reference drug in the current study. The obtained
IC50 values are presented in the table indicating the in
vitro anticancer activity of the tested compounds
compared to the reference drug. It was found in the
negative control that the solvent had no effect on the
cells since the surviving fraction was 100%. Comꢀ
pounds (VII) and (IX) were the most potent with IC50
increase in the activity (IC50 5.50
trary, the activity of compound (VIII) decreased to
5.60 g/mL when the free OH group was substituted by
the ꢀacetyl chloride group. However, additional in vitro
µ
g/mL). On the conꢀ
µ
O
and in vivo biological experiments are required in order
to explore the possibility of utilization of these promising
compounds in practice.
of 3.60 μg/mL and exhibited higher cytotoxic activiꢀ
ties as compared with effect of the reference drug doxꢀ
orubicin on HCT116 cells. The thiourea derivatives
EXPERIMENTAL
General
carrying
Nꢀacetyl chloride moiety (VII), carbamimꢀ
idothioic acid (VIII) having a biologically active thiol
(SH) group, 2ꢀaminoꢀ1ꢀphenylpropylꢀ2ꢀchloroaceꢀ
tate (IX) were nearly as active as the reference drug
doxorubicin against the breast cancer cell line MCFꢀ7. In
addition, compounds (V) and (VI) revealed a moderꢀ
ate activity against breast cancer cell line. All the
tested compounds showed lower activity than the posꢀ
Melting points (uncorrected) were determined in
an open capillary on a Gallen Kamp melting point
apparatus (Sanyo Gallen Kamp, UK). Precoated silꢀ
ica gel plates (Kieselgel 0.25 mm, 60 F254, Merck,
Germany) were used for thin layer chromatography. A
developing solvent system of chloroform–methanol
(8 : 2) was used; the spots were detected by ultraviolet
light. IR spectra (
, cm–1; KBr disc) were recorded
ν
using a Perkin Elmer FTꢀIR spectrophotometer
(USA). NMR spectra were recorded on a NMR specꢀ
trophotometer (Bruker AXS Inc., Switzerland) operꢀ
In vitro anticancer screening of the newly synthesized comꢀ
pounds against human breast (MCFꢀ7), liver (HEPG2),
and colon (HCT 116) cancer cell lines
1
ating at 500 MHz for H and 125.76 MHz for 13C
NMR. Chemical shifts are expressed in ppm relative
to TMS as an internal standard using DMSOꢀd6 as a
solvent. Elemental analyses were done on a model
2400 CHNSO analyzer (Perkin Elmer); all the values
were within 0.4% of theoretical values. All reagents
used were of AR grade. The starting 2ꢀaminoꢀ1ꢀpheꢀ
nylpropanꢀ1ꢀol (I) was purchased from Sigma (USA)
and was directly used for preparation of the target
compounds.
IC50, µg/mL
Compound
MCFꢀ7
HepG2
HCT116
Doxorubicin
5.40
34.00
19.00
19.00
5.50
2.97
17.20
9.60
35.00
NA
5.26
10.40
7.60
29.2
3.60
7.20
3.60
(
(
(
(
(
(
III)
V)
VI
VII
VIII
IX
)
)
1ꢀ(4ꢀChlorophenyl)ꢀ3ꢀ(1ꢀhydroxyꢀ1ꢀphenylproꢀ
panꢀ2ꢀyl) thiourea (III), 4ꢀmethylꢀ5ꢀphenylꢀ4, 5ꢀdihyꢀ
drooxazoleꢀ2ꢀthiol (V) and isomeric 4ꢀmethylꢀ5ꢀpheꢀ
)
5.60
6.20
NA
)
5.60
NA: Not active.
nyloxazolidineꢀ2ꢀthione (VI). A mixture of 2ꢀaminoꢀ
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 42
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2016

438
MAGED S. ABDELꢀKADER et al.
1ꢀphenylpropanꢀ1ꢀol (Lꢀnorephedrine) (
1 mmol) and 4ꢀchlorophenyl isothiocyanate (II
(170 mg, 1 mmol) in chloroform (10 mL) containing 3
drops of triethylamine was stirred at room temperature yield 40%; mp 131.6
for 3 h. The mixture was fractionated over silica gel 3088 (CH arom.), 2966, 2863 (CH aliph.), 1687
I
) (151 mg, tion 2 afforded 11 mg of (XI). Fractions 5–7 afforded
48 mg of (VII). Fractions 10–14 afforded 50 mg of
VIII). Fractions 19–20 afforded 12 mg of (IX). (VII):
C; IR: 3427 (OH), 3244 (NH),
)
(
°
(C=O), 1257 (C=S), 733 (C–Cl); 1H NMR: H 1.61
(d, = 6.9 Hz, 3H, CH3 of Lꢀnorephedrine), 3.73
(s, 2H, CH2Cl), 4.64 (t, = 7.0 Hz, NH–CH–CH3
of Lꢀnorephedrine), 5.11 (bs, 1H, CH–OH), 5.79
(d, = 6.5 Hz, 1H, CH–OH), 6.72 (bs, 1H aromatic),
column (2 mm i.d., 30 g) using dichloromethane as an
eluent. Fractions of 50 mL were collected and
screened by TLC and similar fractions were pooled.
Fractions 3–7 afforded 230 mg of compound (III).
Fractions 11–17 were further purified on silica gel by
PTLC using CH2Cl2–MeOH (95 : 5) as a developing
J
J
J
7.26–7.42 (m, 8H, aromatic), 9.61 (s, NH of 4ꢀchloꢀ
system to afford 23 mg of compound (V) and 29 mg of
13
rophenyl); C NMR: C 13.95, 31.83, 56.17, 72.42,
122.39–129.19 (10), 142.52, 147.03, 155.72, 171.98;
compound (VI). (III): yield 82%; mp semiꢀsolid. IR:
3473 (OH), 3374, 3265 (2NH), 3096 (CH arom.),
2946, 2853 (CH aliph.), 1268 (C=S), 754 (C–Cl);
MS, m/z
(%): 411 (8, M+), 334 (31), 284 (7), 275 (19),
259 (24), 176 (54), 77 (100). Calcd. for
C19H20Cl2N2O2S (411): C, 55.48; H, 4.90; N, 6.81.
Found: C, 55.18; H, 5.17; N, 6.51. (VIII): yield 41%;
1H NMR: H 0.94 (d,
J= 6.5 Hz, 3H, CH3 of Lꢀnorepheꢀ
drine), 4.78 (bs, NH–CH–CH3 of Lꢀnorephedrine),
4.93 (bs, 1H, CH–OH), 7.26–7.42 (m, 9H, aroꢀ
mp 168.9
(CH aliph.), 1694 (C=O), 1599 (C=N), 733 (C–Cl); H
NMR: H 0.96 (d, = 6.5 Hz, 3H, CH3 of Lꢀnorepheꢀ
drine), 3.76 (s, 2H, CH2Cl), 4.32 (bq, = 5.4 Hz,
NH–CH–CH3 of Lꢀnorephedrine), 5.02 (d,
= 6.4 Hz, 1H, CH–OH), 7.34–7.63 (m, 9H, aromatꢀ
ic), 8.56 (d, = 6.9 Hz, 1H, NH of Lꢀnorephedrine),
°C; IR: 3286 (NH), 3065 (CH arom.), 2937, 2853
matic), 7.83 (d,
J = 7.8 Hz, 1H, NH of Lꢀnorepheꢀ
1
drine), 9.74 (s, NH of 4ꢀchlorophenyl); 13C NMR: C
12.47, 54.98, 73.03, 124.04–128.28 (10), 138.49,
J
143.22, 179.38; MS, m/z
(%): 320 (10, M+), 303 (11),
J
193 (24), 148 (18), 128 (100). Calcd. for
C16H17ClN2OS (320): C, 59.90; H, 5.34; N, 8.73.
J
J
Found: C, 59.67; H, 5.09; N, 8.46. (
semiꢀsolid. IR: 3100 (CH arom.), 2942, 2835 (CH 10.32 (s, 1H, SH); C NMR: C 17.53, 33.97, 52.52,
aliph.), 1612 (C=N); 1H NMR: H 0.68 (d,
= 6.7 Hz, 65.83, 120.53–128.65 (10), 137.91, 138.47, 164.79,
(%): 397 (7, M+), 396 (10), 271 (18),
3H, CH3 of Lꢀnorephedrine), 4.41 (bs, NHꢀCHꢀCH3
166.91; MS, m/z
V): yield 8%; mp
13
J
of Lꢀnorephedrine), 5.67 (d, J = 8.7 Hz, 1H, CH–O),
201 (55), 153 (12), 127 (100), 125 (62), 99 (16),
91 (27). Calcd. for C18H18Cl2N2O2S (411): C, 54.41;
H, 4.57; N, 7.05. Found: C, 54.73; H, 4.19; N, 6.79.
7.27–7.40 (m, 5H, aromatic), 7.61 (s, SH); 13C NMR:
C 17.60, 51.62, 82.07, 126.41–128.75 (5), 137.58,
155.52; MS, m/z
(%): 193 (4, M+), 180 (56), 140 (33),
(
IX): yield 10%; mp 151
3072 (CH arom.), 2959, 2845 (CH aliph.), 1688
(C=O), 851 (C–Cl); 1H NMR: H 1.26 (d,
= 6.4 Hz,
°C; IR: 3370, 3295 (NH2),
138 (99), 132 (88), 117 (37), 105 (47), 91(100). Calcd.
for C10H11NOS (193): C, 62.15; H, 5.74; N, 7.25.
Found: C, 62.43; H, 5.49; N, 7.66. (VI): yield 10%;
J
3H, CH3 of Lꢀnorephedrine), 3.57 (s, 2H, CH2Cl),
4.78 (bs, 1H, NH–CH–CH3 of Lꢀnorephedrine),
5.02 (bs, 1H, CH–OH), 6.75 (s, 2H, NH2), 7.16–7.44
mp semiꢀsolid. IR: 3186 (NH) 3068 (CH arom.),
1
2956, 2876 (CH aliph.), 1271 (C=S); H NMR: H
0.65 (d,
J = 6.5 Hz, 3H, CH3 of Lꢀnorephedrine), 4.42
13
(m, 5H, aromatic); C NMR: C 9.90, 31.62, 57.14,
(q, = 2 Hz, NH–CH–NH–CH–CH3 of Lꢀ
J
73.77, 121.50–128.38 (5), 140.49, 170.94; MS,
m/z
norephedrine), 6.00 (d, J = 9 Hz, 1H, CH–O), 7.26–
(%): 227 (100, M+), 153 (7), 133 (10), 110 (7), 77 (11).
Calcd. for C11H14ClNO2 (227): C, 58.03; H, 6.20; N,
6.15. Found: C, 58.31; H, 6.47; N, 5.78. (XI): yield
9%; mp semiꢀsolid; IR: 3389 (NH), 3099 (CH arom.),
2951, 2836 (CH aliph.), 1699 (C=O), 1266 (C=S),
7.45 (m, 5H, aromatic), 10.26 (s, NH); 13C NMR: C
16.56, 55.37, 85.34, 126.60–128.92 (5), 135.33, 187.91;
MS, m/z
(%): 193 (100, M+), 150 (36), 132 (70), 117
(51), 105 (40), 91(100). Calcd. for C10H11NOS (193):
C, 62.15; H, 5.74; N, 7.25. Found: C, 61.82; H, 6.07;
N, 7.55.
1
773 (C–Cl); H NMR: H 3.69 (s, 2H, NH–CH2–
C=O), 7.33 (d,
NMR: C 44.45, 122.52 (2X), 129.86 (3X), 132.85,
168.51, 178.78; MS,
(%): 226 (10, M+), 201 (26),
J = 8, 2H), 7.59 (d, J
= 8, 2H); –13C
2ꢀChloroꢀ
(1ꢀhydroxyꢀ1ꢀphenylprꢀopanꢀ2ꢀyl)acetamide (VII),
ꢀ(1ꢀ(2ꢀchloroacetoxy)ꢀ1ꢀphenylpropanꢀ2ꢀyl)ꢀ 'ꢀ
Nꢀ(4ꢀchlorophenylcarbamothioyl)ꢀNꢀ
m/z
N
N
154 (8), 129 (32), 127 (100), 99 (23). Calcd. for
C9H7ClN2OS (226): C, 47.69; H, 3.11; N, 12.36.
Found: C, 47.44; H, 2.84; N, 12.69.
(4ꢀchlorophenyl)carbamimidothioic acid (VIII), 2ꢀamiꢀ
noꢀ1ꢀphenylpropylꢀ2ꢀchloroꢀacetate (IX), and 3ꢀ(4ꢀ
chlorophenyl)ꢀ2ꢀthioxoimidazolidinꢀ4ꢀone (XI). A mixꢀ
ture of (III) (160 mg, 0.5 mmol) and chloroacetylꢀ
chloride (56.5 mg, 0.5 mmol) in chloroform (10 mL)
containing 3 drops of triethylamine was stirred for
10 min. The mixture was fractionated on a silica gel
In Vitro Anticancer Activity
In vitro cytotoxic activity of the novel synthesized
column (2 mm i.d., 30 g) eluted with dichloꢀ compounds was measured using the sulforhodamine B
romethane. Fractions of 50 mL were collected and stain (SRB) assay and the method of Skehan et al. [23].
screened by TLC; similar fractions were pooled. Fracꢀ The human tumor breast cancer cell line MCFꢀ7,
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 42
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2016

DESIGN, SYNTHESIS, AND ANTICANCER EVALUATION
439
human colon cancer cell line HCT 116, and human compounds (VII) and (IX) exhibited higher activity
liver cancer cell line HEPG2 (obtained from National than the reference drug doxorubicin against human
Cancer Institute, Cairo, Egypt) were maintained at colon cancer cell line HCT 116. The active comꢀ
37°C in 5%ꢀCO2 as subconfluent monolayers in pounds could be considered as promising templates
80 cm3 culture flasks (Nunclon) and were subcultured for further development of more potent anticancer
once or twice weekly in Dulbecco’s modification of agents.
Eagle’s medium (Flow) supplemented with 5% heatꢀinacꢀ
tivated fetal calf serum (FCS) and 1 mM Lꢀglutamine.
During experiments, 50 g/mL gentamicin was added
ACKNOWLEDGMENTS
μ
to the culture medium. Passage levels were in the range
of 5–20 according to the original receipt. Cells were
harvested from exponential phase cultures by
trypsinization, counted, and plated in 96ꢀwell flatꢀ
bottomed microliter plates (Greiner Labortechnik,
The authors would like to extend their sincere appreꢀ
ciation to the Deanship of Scientific Research at King
Saud University for the funding of this research through
the Research Group Project No. RGPꢀVPPꢀ 302.
Germany) (100
μ
L cell suspension containing
104 cells per well). Following plating and a 24ꢀh recovꢀ
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