M. Yousefi et al.
3
3
Complex 1 is the most active against stimulated PBMC (IC
.01 mg ml ) compared to activity on other cell lines used in
this study. The cytotoxic activity shown by these compounds
against all these cancer cell lines indicates that coupling of
d = 4.16 (t, H3,3
0
, 8 H, J = 10 Hz), 3.79 (t, H
0
, 8 H, J = 10 Hz),
5
0
H-H
2,2
H-H
ꢀ
1
3
13
0
2.11–1.45 (m, Hd ,b,g, 12 H), 0.97 (t, H , 6 H, J = 15 Hz ) ppm;
C
a
H-H
0 0
3
): d = 201 (C-1), 66.1 (C-3,3 ), 51.2 (C-2,2 ),
119
NMR (500 MHz, CDCl
26.3 (C ), 28.4 (C ), 34.19 (C
CDCl
280 (C
d
b
g a
), 13.7 (C ) ppm; Sn NMR (500 MHz,
+
ligand with R
2
Sn(IV) metal center results in metallic complexes
3
): d = ꢀ326 ppm; MS: m/z (70 eV) = 395 (C13
2
H26NOS Sn) ,
H
+
+
+
with important biological properties and remarkable cytotoxic
activity. Compounds 1 and 4 are considered agents with poten-
tial antitumor activity and can therefore be candidates for
further stages of screening in vitro and/or in vivo.
5
H
8
NOS
NO) , 77 (C
2
Sn) , 162 (C
5 8
H
NOS
2
) , 130 (C
5
8
NOS) , 86
+
+
(C H
4 8
H
6 5
) .
Synthesis of diphenyltin(IV)-bis(morpholine-1-carbodithioate) (3)
The preparation of compound 3 was carried out in an identical
Experimental
manner to compound 1. Ph
salt (0.21 g 0.87 mmol). M.p. 150 C; FT-IR (KBr): n = 995 (C-S), 1482
2
SnCl
2
(0.15 g, 0.43 mmol) and ligand
ꢁ
Materials and Methods
ꢀ1
1
(
C-N), 449 (Sn-S), 543 (Sn-C) cm ; H NMR (500 MHz, DMSO):
3
3
d = 4.03 (t, H3,3
.93 (H , SnC
500 MHz, DMSO): d = 196.7 (C-1), 65.3 (C-3,3 ), 51.7 (C-2,2 ), 150
0
, 8 H, JH-H = 10Hz), 3.77 (t, H2,2
0
, 8 H, JH-H = 10 Hz),
All chemicals and reagents purchased were of reagent grade
and used without further purification unless otherwise noted.
Dibenzyltin(IV) dichloride was prepared by a standard method
13
7
(
O
6 5
H , 4 H), 7.42 (m, Hm,p, SnC , 6 H) ppm; C NMR
6 5
H
0
0
1
19
[
32]
(C ), 133 (C ), 128.5 (C ), 128.2 (C ) ppm;
Sn NMR (500 MHz,
a
b
d
g
reported in the literature.
The solvents were purified and
+
DMSO): d = ꢀ225 ppm; MS: m/z (70 eV) = 520 (C16
21 2 2 4
H N O S Sn) ,
dried according to standard procedure. Melting points were
determined in open capillaries and were uncorrected. FT-IR spectra
were recorded on a Bomem MB-100 FT-IR spectrometer, using
+
+
+
4
8
43 (C10
H
16
N
2
O
2
S
4
Sn) , 358 (C11
H
13NOS
2
Sn) , 162 (C
H
5 8
NOS
2
) ,
+
+
6 (C NO) , 44 (C
H
4 8
2
H
4
O) .
ꢀ
1
1
13
119
KBr pellets (400–4000 cm ). The H, C and
Sn NMR were
Synthesis of dibenzyltin(IV)-bis(morpholine-1-carbodithioate) (4)
obtained using a Bruker AVANCE 500 spectrometer. The chemical
shifts are reported in ppm relative to the internal references,
The preparation of compound 4 was carried out in an identical
manner to compound 1. Bz SnCl (0.12 g, 0.32 mmol) and ligand
1
13
119
TMS for H, C and tetramethyltin for Sn shifts. The splitting
2
2
1
ꢁ
salt (0.16 g 0.64 mmol). M.p. 140 C ; FT-IR (KBr): n = 997.3 (C-S),
1 1
of proton resonances in the reported H NMR spectra are defined
ꢀ
as s = singlet, d = doublet, t = triplet, q = quadruplet, m = multiplet.
TGA was performed with a universal V3.8 B TA SDT Q500.
Mass spectroscopy was performed on a Hewlett-Packard 5973
instrument at 70 eV.
1474 (C-N), 417 (Sn-S), 544 (Sn-C) cm
;
H NMR (500 MHz,
, 8 H, JH-H = 10 Hz), 3.58 (t, H2,2 , 8 H,
JH-H = 10 Hz), 3.29 (s, Ha , 4 H), 7.15 ( H , SnCH C H ), 7.02
3
DMSO): d = 3.86 (t, H3
0
0
,3
3
O
2 6 5
13
(m, Hm,p, Sn-CH C H ) ppm; CNMR (500 MHz, DMSO): d = 199
2
6 5
0 0
C-1), 65.3 (C-3,3 ), 51 (C-2,2 ), 30.6 (C ) 139.3 (C ), 128.7 (C ),
a b g
(
Synthesis of morpholinium morpholine-1-carbodithioate (MCDT)
119
1
27.3(C ), 124.5 (C ) ppm; Sn NMR (500 MHz, DMSO): d = ꢀ484
e
d
+ +
Dropwise addition of CS
2
(in excess, 1.7 ml, 23mmol) in methanol
ppm. MS: m/z (70 eV) = 372 (C H NOS Sn) , 301 (C H Sn) ,
12 15 2 14 14
+
+
+
(30 ml) to morpholine (1ml, 11.5 mmol) in methanol (30 ml)
162 (C H NOS ) , 91 (C H ) , 77 (C H ) .
5 8 2 7 7 6 5
ꢁ
followed by stirring for 4 h at 0 C gave the white product. This
was filtered off and washed with diethyl ether. M.p. 195 C; FT-IR
ꢁ
ꢀ
1
1
In Vitro Studies
(KBr): n = 1019 (C-S), 1450 (C-N) cm ; H NMR (500 MHz, DMSO):
3
3
d = 4.35 (t, H3,3
0
, 4H, JH-H = 10Hz ), 3.77 (t, H3a,3
0
a
, 4 H, JH-H = 10Hz),
Preparation of drug solutions
0
3
0
3
3
.60 (t, H2,2 , 4H, JH-H =10Hz), 3.07 (t, H2a,2 a, 4H, JH-H = 10Hz)
13
0
Stock solutions of the studied compounds were prepared in DMSO
ppm; C NMR (500 MHz, DMSO): d = 213.6 (C-1), 66.94 (C-3,3 ),
ꢀ
1
0 0
4.23 (C-3a,3’a), 50.68 (C-2,2 ), 43.71 (C-2a,2 a) ppm.
(Sigma Aldrich) at a concentration of 1000mg ml , sterilized by
filtration through Millipore filter (0.22 mm) before use, and diluted
by cell culture medium to various working concentrations. DMSO
was used owing to solubility problems. Nutrient medium was
RPMI-1640 (Gibco BRL, UK) supplemented with 10% fetal bovine se-
6
Synthesis of dimethyltin(IV)-bis(morpholine-1-carbodithioate) (1)
A solution of Me SnCl (0.1 g, 0.45 mmol) in methanol (15 ml) was
2
2
added dropwise to a solution of ligand (0.22 g, 0.9 mmol) in
methanol (30 ml) at 60 C. The reaction mixture was refluxed for
ꢀ
1
ꢁ
rum (FBS; Gibco BRL). MTT was dissolved (5mg ml ) in phosphate
buffered saline (PBS; pH 7.2) and filtered through Millipore filter
6
h with constant stirring and the solvent was evaporated; then
(0.22 mm) before use.
the colorless crystals were separated out 5 days later. M.p.
ꢁ
HeLa cells were split using 0.25% TRED (trypsin–
ethylenediaminetetraacetic acid; Gibco BRL) medium prior to
reaching 80% confluence.
1
62 C; FT-IR (KBr): n = 1000 (C-S), 1468 (C-N), 413 (Sn-S), 540
ꢀ
1
1
(
Sn-C) cm
Sn-H = 82 Hz ), 4.13 (t, H3,3
H-H = 10 Hz) ppm; C NMR (500 MHz, DMSO): d = 199.2 (C-1), 66
C-3,3 ), 51.25 (C-2,2 ), 12.77 (C-a, JC-Sn = 700 Hz ) ppm; Sn NMR
;
H NMR (500 MHz, DMSO): d = 1.57 (s, H
a
,6 H,
, 8 H,
2
3
J
0
, 8 H, JH-H = 10 Hz), 3.79 (t, H2,2
0
3
13
J
Cell line and cell culture
0
0
1 119
(
(
(
(
500 MHz, DMSO): d = ꢀ322 ppm; MS: m/z (70 eV)= 209.7
HeLa (human cervix carcinoma) cells (NCBI C115, National Cell
Bank of Iran) and K562 (human myelogenous leukemia) cells
(NCBI C122, National Cell Bank of Iran) were obtained from
the Pasteur Institute of Iran. Cells were cultured in RPMI-1640
medium supplemented with 10% heat-inactivated fetal calf
serum (FCS; Gibco BRL) 2 mM L-glutamine (Gibco BRL) and
+
+
+
+
C
C
2
S
2
H
3
Sn) , 162 (C
5
H
8
+
NOS
2
) , 133 (CH
3
Sn) , 130 (C
5
H
8
NOS) , 86
+
+
4
H
8
NO) , 72 (C
H
4 8
O) , 58 (NCS) .
Synthesis of dibutyltin(IV)-bis(morpholine-1-carbodithioate) (2)
The preparation of compound 2 was carried out in an identical
manner to compound 1. Bu SnCl (0.1 g, 0.33 mmol) and ligand
salt (0.16 g, 0.66 mmol). M.p. 140 C; FT-IR(KBr): n = 996 (C-S),
ꢀ
1
antibiotics, including streptomycin (100 mg ml ) and penicillin
2
2
ꢁ
ꢀ1
ꢁ
(100 IU ml ) (Sigma, USA) and incubated at 37 C in a humidified
1
1
465 (C-N), 418 (Sn-S), 543 (Sn-C); H NMR (500 MHz, CDCl3):
5% CO atmosphere.
2
wileyonlinelibrary.com/journal/aoc
Copyright © 2012 John Wiley & Sons, Ltd.
Appl. Organometal. Chem. 2012, 26, 438–444