synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age
Aim: To develop a rationally designed new organotin compound namely dimethyl tin 4-cyclohexyl thiosemicarbazone (D4-t) and evaluate its putative antitumor activity. Methods: Starting from 4-cyclohexyl thiosemicarbazone, a three step synthetic procedure was followed to obtain the title compound. In...
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2009
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irk-123456789-1351032018-06-15T03:10:10Z synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age Sen, A. Chaudhuri, T.K. Original contributions Aim: To develop a rationally designed new organotin compound namely dimethyl tin 4-cyclohexyl thiosemicarbazone (D4-t) and evaluate its putative antitumor activity. Methods: Starting from 4-cyclohexyl thiosemicarbazone, a three step synthetic procedure was followed to obtain the title compound. In vivo lymphocyte activation property of the compound at three different doses was assayed by measuring the blastogenesis. Concanavalin A (ConA) was used as standard mitogen for murine T cells stimulation in vivo. Also, the synthesis of DNA by the activated lymphocytes was measured after injecting the D4-t. The lymphocyte activation property and antitumor efficacy of D4-twere assessed inSarcoma-180 (S-180) bearing mice. The organization of lymphoid cells was studied in the histological preparations ofspleen and mesenteric lymph node. Tumor neutralization assay (Winn assay) was conducted to examine whether immune responses were associated with the manifestation of antitumor efficacies of this compound in S-180 in vivo. The DNA synthesis inhibitory effect of the compound in S-180 cells was studied in vitro, and was found significant (P < 0.001). Results: Different doses of the new compound caused differential response of blastogenesis and DNA synthesis. In comparison to ConA, the title compound showed a good number of blast cells at its optimum dose of 5 mg/kg. It caused maximum synthesis of DNA by the lymphoid cells. In histological preparations, the gradual transformation of lymphocytes into blasts was observed without any visible toxicity. Winn assay revealed that 5 mg/kg of D4-t was able to reduce tumor mass without severe toxicity. This organotin compound also inhibits the synthesis of DNA in S-180 tumor cells in comparison to Platin10 and ConA. Conclusion: The title compound has the lymphocyte activation property and stimulates immune response of the lymphoid cells, which in turn express the antitumor activity without any significant toxicity. Results indicate promising therapeutic potential of D4-t. 2009 Article synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age / A. Sen, T.K. Chaudhuri // Experimental Oncology. — 2009. — Т. 31, № 1. — С. 22-26. — Бібліогр.: 24 назв. — англ. 1812-9269 http://dspace.nbuv.gov.ua/handle/123456789/135103 en Experimental Oncology Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| collection |
DSpace DC |
| language |
English |
| topic |
Original contributions Original contributions |
| spellingShingle |
Original contributions Original contributions Sen, A. Chaudhuri, T.K. synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age Experimental Oncology |
| description |
Aim: To develop a rationally designed new organotin compound namely dimethyl tin 4-cyclohexyl thiosemicarbazone (D4-t) and
evaluate its putative antitumor activity. Methods: Starting from 4-cyclohexyl thiosemicarbazone, a three step synthetic procedure was
followed to obtain the title compound. In vivo lymphocyte activation property of the compound at three different doses was assayed
by measuring the blastogenesis. Concanavalin A (ConA) was used as standard mitogen for murine T cells stimulation in vivo. Also,
the synthesis of DNA by the activated lymphocytes was measured after injecting the D4-t. The lymphocyte activation property and
antitumor efficacy of D4-twere assessed inSarcoma-180 (S-180) bearing mice. The organization of lymphoid cells was studied in the
histological preparations ofspleen and mesenteric lymph node. Tumor neutralization assay (Winn assay) was conducted to examine
whether immune responses were associated with the manifestation of antitumor efficacies of this compound in S-180 in vivo. The
DNA synthesis inhibitory effect of the compound in S-180 cells was studied in vitro, and was found significant (P < 0.001). Results:
Different doses of the new compound caused differential response of blastogenesis and DNA synthesis. In comparison to ConA, the
title compound showed a good number of blast cells at its optimum dose of 5 mg/kg. It caused maximum synthesis of DNA by the
lymphoid cells. In histological preparations, the gradual transformation of lymphocytes into blasts was observed without any visible
toxicity. Winn assay revealed that 5 mg/kg of D4-t was able to reduce tumor mass without severe toxicity. This organotin compound
also inhibits the synthesis of DNA in S-180 tumor cells in comparison to Platin10 and ConA. Conclusion: The title compound has
the lymphocyte activation property and stimulates immune response of the lymphoid cells, which in turn express the antitumor
activity without any significant toxicity. Results indicate promising therapeutic potential of D4-t. |
| format |
Article |
| author |
Sen, A. Chaudhuri, T.K. |
| author_facet |
Sen, A. Chaudhuri, T.K. |
| author_sort |
Sen, A. |
| title |
synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age |
| title_short |
synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age |
| title_full |
synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age |
| title_fullStr |
synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age |
| title_full_unstemmed |
synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age |
| title_sort |
synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age |
| publisher |
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| publishDate |
2009 |
| topic_facet |
Original contributions |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/135103 |
| citation_txt |
synthesis and evaluation of dimethyl tin 4-cyclohexyl thiosemicarbazone as a novel antitumor age / A. Sen, T.K. Chaudhuri // Experimental Oncology. — 2009. — Т. 31, № 1. — С. 22-26. — Бібліогр.: 24 назв. — англ. |
| series |
Experimental Oncology |
| work_keys_str_mv |
AT sena synthesisandevaluationofdimethyltin4cyclohexylthiosemicarbazoneasanovelantitumorage AT chaudhuritk synthesisandevaluationofdimethyltin4cyclohexylthiosemicarbazoneasanovelantitumorage |
| first_indexed |
2025-07-09T22:42:47Z |
| last_indexed |
2025-07-09T22:42:47Z |
| _version_ |
1837211027574358016 |
| fulltext |
22 Experimental Oncology 31, 22–26, 2009 (March)
The earliest reports on the therapeutic use of the
metals or metal-containing compounds in cancer and
leukemia were dated back to 19th century. Until 1960s,
the knowledge about mechanism of their antitumor
activity was limited due to the technological restrictions
and lack of scientific approach for their exploration.
The discovery of the inorganic complex cis-diammine-
dichloroplatinum (II) (cisplatin) led to the develop-
ment of other types of non-organic cytostatic drugs
for the treatment of tumors. Numerous other metal
compounds that contained platinum, other platinum
metals, and even non-platinum main group metals, in-
cluding tin, were then shown to be effective against tu-
mors. In the field of organometallic chemistry organo-
tin compounds shares a very important prospective.
The organotin compounds or stannanes are chemical
compounds based on the tin with hydrocarbon sub-
stituents. In recent years, organotin compounds have
drawn much attention as antitumor agents [1–3]. The
R2Sn(IV)2+ compounds exhibit maximal antitumor ac-
tivity with low toxicity, are the type R2SnX2L2 (X = halo-
gen, pseudo halogen and L= O- or N-donor ligands)
[4–7]. Diorganotin(IV) compounds, R2SnCl2 are often
tetrahedral, and structurally similar with cisplatin.
A large number of such complexes have been tested
for antitumor activity [3, 8]. Investigations of thiosemi-
carbazone Sn(IV) complexes have been carried out
by many researchers [2, 9, 10]. It has been observed
that several di- and triorganotin(IV) species were ac-
tive against various types of cancer cells [5, 11]. The
compounds usually realize their effect by interacting
with the nucleic acid base [12]. A number of positive
results have been reported in this direction [3, 7, 13].
Organotin compound has antiproliferative ability and
antitumor activity on mammalian cells both in vitro and
in vivo [3, 7, 8, 13]. Also, in Japan works regarding the
immune response induced by implantation of tumor
cells was further stimulated by D-fraction of Maitake
mushroom have been carried out [14].
In the present study, we studied the potential anti-
tumor activity of a novel rationally designed compound
dimethyl tin 4-cyclohexyl thiosemicarbazone (D4-t),
which have ONS (oxygen/ nitrogen/sulphur) donors
in its structure to develop an appropriate “bite angles”.
The compound is expected to coordinate with DNA
base pairs. The synthesis, lymphocyte activation and
evaluation of D4-t anticancer activity were performed
and discussed.
MATERIALS AND METHODS
Chemical synthesis. The ligand 4-cylohexyl
thiosemicarbazone was prepared from cyclohexyl
thiosemicarbazide and salicyl aldehyde as described
[15]. Then methanol solution of dimethyltin dichloride
and sodium hydroxide solution (50% excess) were
mixed to prepare dimethyltin oxide [10, 16]. After that,
condensation of dimethyltin oxide (1 M) and the ligand
SyNTHESIS AND EvALuATION Of DIMETHyL TIN 4-CyCLOHExyL
THIOSEMICARbAzONE AS A NOvEL ANTITuMOR AgENT
A. Sen1, 2, T.K. Chaudhuri1, *
1Cellular Immunology Laboratory, Department of Zoology, University of North Bengal, Siliguri, West Bengal
734013, India
2Department of Chemistry, University of North Bengal, Siliguri, West Bengal 734013, India
Aim: To develop a rationally designed new organotin compound namely dimethyl tin 4-cyclohexyl thiosemicarbazone (D4-t) and
evaluate its putative antitumor activity. Methods: Starting from 4-cyclohexyl thiosemicarbazone, a three step synthetic procedure was
followed to obtain the title compound. In vivo lymphocyte activation property of the compound at three different doses was assayed
by measuring the blastogenesis. Concanavalin A (ConA) was used as standard mitogen for murine T cells stimulation in vivo. Also,
the synthesis of DNA by the activated lymphocytes was measured after injecting the D4-t. The lymphocyte activation property and
antitumor efficacy of D4-t were assessed in Sarcoma-180 (S-180) bearing mice. The organization of lymphoid cells was studied in the
histological preparations of spleen and mesenteric lymph node. Tumor neutralization assay (Winn assay) was conducted to examine
whether immune responses were associated with the manifestation of antitumor efficacies of this compound in S-180 in vivo. The
DNA synthesis inhibitory effect of the compound in S-180 cells was studied in vitro, and was found significant (P < 0.001). Results:
Different doses of the new compound caused differential response of blastogenesis and DNA synthesis. In comparison to ConA, the
title compound showed a good number of blast cells at its optimum dose of 5 mg/kg. It caused maximum synthesis of DNA by the
lymphoid cells. In histological preparations, the gradual transformation of lymphocytes into blasts was observed without any visible
toxicity. Winn assay revealed that 5 mg/kg of D4-t was able to reduce tumor mass without severe toxicity. This organotin compound
also inhibits the synthesis of DNA in S-180 tumor cells in comparison to Platin10 and ConA. Conclusion: The title compound has
the lymphocyte activation property and stimulates immune response of the lymphoid cells, which in turn express the antitumor
activity without any significant toxicity. Results indicate promising therapeutic potential of D4-t.
Key Words: new organotin compound, synthesis, lymphocyte activation, antitumor activity.
Received: July 25, 2008.
*Correspondence: Fax: +91353 2699001
E-mail: dr_tkc_nbu@rediffmail.com
Abbreviations used: CHN — carbon hydrogen nitrogen; D4-t —
dimethyl tin 4-cyclohexyl thiosemicarbazone; LN — lymph nodes;
MLN — mesenteric lymph node; ONS — oxygen nitrogen sulphur;
S-180 — sarcoma180; SP — spleen.
Exp Oncol 2009
31, 1, 22–26
Experimental Oncology 31, 22–26, 2009 (March) 23
(1 M) in benzene (100 ml) was performed by Dean
Stark method for 8 h [10, 16]. The prepared D4-t was
purified from pet ether and benzene.
IR spectroscopy (FTIR-8300-Shimadzu, KBR op-
tics using nujol mull) and 1H NMR spectra (1H NMR:
Bruker 300-DPX, δ values, TMS as internal standard)
of D4-t were measured. Release of radioactivity was
counted in liquid scintillation counter (Model no. LKB
1209 Rack-Beta).
In vivo screening. All in vivo experiments were
conducted according to the rules and regulations
of Committee for the Purpose of Control and Super-
vision of Experiments on Animals (CPCSEA), India.
Sarcoma-180 (S-180) cells were obtained from the
National Center for Cell Sciences (Pune, India) and
maintained in Swiss albino male mice (6–7 weeks
old, 24 ± 2 g) by intra peritoneal (i. p.) transplantation
of 1.0 × 106 cells/0.2 ml physio logical saline/animal
[17]. Mice were divided in groups as control (untreated),
D4-t treated and ConA / Platin 10 treated. At least 6 ani-
mals were used for a particular group in an experiment.
Physiological saline containing 0.2 ml of Freund’s in-
complete adjuvant (Sigma-Aldrich Chemicals Pvt.
Ltd., India) was used as vehicle for drug administra-
tion by i.p. injection. Compounds in respective doses
were administered to each mouse in treated groups
as per the experiment performed. Group 1: The control
groups received equal volumes of vehicle. Group 2:
Mice treated with D4-t in respective doses through i.p.
route. Group 3: Concanavalin A (ConA) (Sigma-Aldrich
Chemicals Pvt. Ltd., India) dissolved in sterilized dis-
tilled water before intravenous (i. v.) injection was used
as positive control. For evaluation of antitumor activity
cisplatin injection (Platin10) (Cadila Pharmaceuticals
Ltd., India) was used as positive control.
In vivo lymphocyte activation. The degree
of activation by ConA of lymphocytes of secondary
lymphoid organs and peripheral blood is considered
as the measure of in vivo lymphocyte activation [18,
19]. Mice were injected with three different doses
of D4-t on day 0: 1 mg/kg, 5 mg/kg and 10 mg/kg
of body wt of mice. Cell suspensions from different
lymphoid organs such as spleen (SP), mesenteric
lymph node (MLN) and other lymph nodes (LN) were
prepared. The peripheral blood (PB) was on Ficoll and
Hypaque solution (Sigma-Aldrich Chemicals Pvt. Ltd.,
India) for separation of lymphocytes [18, 19]. Then
D4-t-stimulated blast transformation was measured.
The dose of 50 µg of ConA in 0.1 ml/animal was chosen
for lymphocyte activation. The rate of blast transforma-
tion of lymphocytes of lymphoid organs was recorded
at 24 h up to 96 h [18].
The process of activation and blast transformation
of lymphocytes were evaluated by the rate of DNA syn-
thesis, which was measured by the rate of 3H-thymidine
incorporation into DNA. The optimal dose of ConA for
DNA synthesis was 10 µg in 0.1 ml/animal [18]. So this
dose was used in this case apart from using three dif-
ferent doses of D4-t injected on day 0. The assay was
made at 24 h up to 96 h after injection. Cell suspen-
sions of SP were adjusted to 1.0 × 106 / 0.1 ml after
the addition of 3H-thymidine (activity 10 µCi each)
dissolved in 0.01 ml sterile saline. Using the standard
method of incubation for 8 h followed by precipita-
tion with 10% chilled trichloroacetic acid, the release
of radioactivity was recorded with the help of liquid
scintillation counter [18, 20].
Evaluation of antitumor activity. S-180 tumor
cells (1.0 × 106 cells/animal) were s. c. implanted
in normal Swiss albino mice on day 0. At least 6 animals
were used for a particular group, divided as cont-
rol (untreated) and treated (D4-t, ConA / Platin 10)
in an experiment.
S-180 bearing animals treated with D4-t on day
1 by i. p. route for this experiment (optimal dose
of 5 mg/kg) were killed, their SP and MLN were iso-
lated at different time points after day 1 (24, 48, 72 and
96 h), and fixed in Bouin’s fixative. Histological sec-
tions were made and stained with Delafield’s haema-
toxylin and eosin. The histopathological changes were
evaluated by microscopy. In control, the experiments
were performed without any drug injection and ConA
was used as standard [21].
In Winn or tumor neutralization assay, D4-t (5 mg/kg
dose by i. p.) and ConA (50 µg in 0.1 ml/ani mal by i. v.)
were injected on day 1 after injecting S-180 on day
0. Then on day 5, SP cell suspension from ConA and
D4-t treated mice or S-180 bea ring control mice was
mixed with S-180 tumor cells (1 : 1) and inoculated into
normal mice. ConA treated, D4-t treated and control
mice were weighed daily as described in [14].
To study the inhibitory effects of D4-t on DNA
synthesis in S-180 in vitro 3H-thymidine incorporation
assay was used as described in [22]. Tumor cells were
removed at the 7th day after tumor transplantation,
washed twice in Hank’s balanced salt solutions, re-
suspended in RPMI-1640 medium supplemented with
10% heat inactivated fetal calf serum, streptomycin
(100 µg/ml) and penicillin (100 units/ml), followed
by viable cell count. To the cells (1.0 × 106/0.1 ml) with
3H-thymidine (10 µCi each) D4-t was added at concen-
tration of 0.0016 µM and cells were incubated at 37 °C
for 30 min and 60 min. As controls, the same concen-
trations of ConA and Platin10 were used.
Statistical analysis. Values were expressed
as the mean ± SD. Experimental results were analyzed
by Student’s t-test. P < 0.05 was considered as the
level of significance for values obtained for treated
groups, compared with the control group.
RESuLTS
The pathway of D4-t synthesis is presented
on Fig. 1. Melting point (M. pt.) of D4-t was 97–98 °C.
The yield of the dark yellow coloured solid compound
was 80%. The calculated Carbon Hydrogen Nitro-
gen (CHN) percentage (%) was 45.3, 5.4, 9.9 and
the observed CHN % was 44.8, 5.2, 9.1 respec-
tively. IR (cm–1) values was 1600 (CH=N), 968 (C-S),
530 (Sn-O), 478 (Sn-C). 1H NMR (δ ppm, CDCl3)
of D4-t was 0.85 (s, 6H, 2×CH3), 1.35 (m,11H,C6H11),
24 Experimental Oncology 31, 22–26, 2009 (March)
4.7 (d,1H,NH), 6.77 (m,2H, aromH), 7.1 (d,1H,aromH),
7.26 (m,1H,aromH) and 8.6 (s,1H, CHN).
R = H, R1 = Cyclohexyl
R´ = Me
+ H2NNHC(S)NHR1
R
R
R
R1
R1
R´ R´
R´2SnO
S
SSn
O
O
N
N
N
OH
OH
NH
NH
HN
R
R1
S
N
OH
NH
HN
fig. 1. The pathway of D4-t synthesis
In vivo activation of lymphocytes with different
do ses of D4-t at different time intervals was exami-
ned. The dose wise effect of ConA is presented
on Fig. 2, a. The dose wise effects of D4-t are presen-
ted on Fig. 2, b–d. The peak of blastogenesis of cells
from all sources (SP, LN, MLN and PB) irrespective
of D4-t dose was effectively reached at 72 h. The
responses were almost at the same level in 24 h for
all three studied D4-t doses. The blast cells became
more vacuolated or exhausted at 72 h up to 96 h. The
increase in the percentage of vacuolated cells corre-
sponded to the 10 mg/kg of D4-t (see Fig. 2, d). The
significance of the difference between the groups
(P < 0.01) was found.
Patterns of DNA synthesis by SP cells upon treat-
ment by D4-t and ConA are presented on Fig. 3. With
all three studied D4-t doses, maximum incorpora-
tion of 3H-thymidine was observed at 48 h, and the
index decreased between 48 h up to 96 h time points
(P < 0.001). The rate of DNA synthesis was higher for
5 mg/kg dose and almost reached the ConA value.
0
20
40
60
80
100
0 20 40 60 80 100
Hours
In
co
rp
or
at
io
n
of
p
re
cu
rs
or
, %
c
on
tro
l
ConA
1 mg/kg
5 mg/kg
10 mg/kg
fig. 3. Pattern of 3H-Thymidine incorporation by splenic lym-
phocytes at different time points after in vivo stimulation by ConA
(10 µg) and D4-t (1 mg/kg, 5 mg/ kg, and 10 mg/ kg)
The in vivo antitumor activity of D4-t in S-180 was
determined following the treatment of animals at the
dose of 5 mg/kg.
In histological preparations of SP and MLN the
gradual transformation of the lymphocytes into blasts
was observed. Within 24 h of the treatment, SP cells
of D4-t treated animals were loosely packed in com-
parison to control mice (S-180 bearing untreated
mice). A higher percentage of blasts in MLN than
in SP were recorded within 24 h. By 48 h most of the
MLN cells differentiated into blast cells (data not
presented).
In Winn assay the weight of tumor bearing mice,
treated by D4-t increased from the day 1 to day 14,
but after day 15 decreased compared with the weight
of control animals and reached the value shown for
ConA treated animals (Fig. 4). Significant regression
(P < 0.01) was observed in mice inoculated with the
0
20
40
60
80
24 48 72 96
Hours
%
b
la
st
c
el
ls
0
20
40
60
80
24 48 72 96
Hours
%
b
la
st
c
el
ls
0
20
40
60
80
24 48 72 96
Hours
%
b
la
st
c
el
ls
0
20
40
60
80
24 48 72 96
Hours
%
b
la
st
c
el
ls
a
c
b
d
SP
LN
MLN
PB
fig. 2. Kinetics of blastogenesis of lymphocytes treated by ConA at a dose of 50 µg in 0.1 ml/animal (a), by D4-t at a doses
of 1 mg/kg body weight (b), of 5 mg/kg (c) and 10 mg/kg (d)
Experimental Oncology 31, 22–26, 2009 (March) 25
mixture of tumor cells and SP cells obtained from
D4-t pre-treated mice.
ConA
Control
D4-t
0
10
20
30
40
50
0 5 10 15 20
Days
%
b
od
y
we
ig
ht
c
ha
ng
es
fig. 4. Body weight changes of mice treated by D4-t, ConA and
control mice in tumor neutralization assay (Winn assay)
To ascertain whether drug-induced tumor growth
inhibition was achieved due to the inhibitory effect
of D4-t on DNA synthesis, we performed DNA syn-
thesis assay. The untreated S-180 cells demonstrated
an almost linear pattern of 3H-thymidine uptake over
a period of 60 min (Fig. 5). Simultaneous exposure
of tumor cells to 0.0016 µM D4-t resulted in gradual and
marked inhibition of 3H-thymidine uptake (P < 0.001).
The effect was almost equal to that in Platin10-treated
cells, but greater than for ConA-treated cells. Thus,
at the end of 60 min incubation Platin10, ConA and
D4-t exhibited 95%, 66% and 80% of DNA synthesis
inhibition compared with control untreated cells, re-
spectively (see Fig. 5).
Incubation time, min
In
co
rp
or
at
io
n
of
p
re
cu
rs
or
, %
c
on
tro
l
0
20
40
60
80
100
0 10 20 30 40 50 60
ConA
Platin 10
D4-t
fig. 5. Effect of Platin10, D4-t and ConA on DNA synthesis
by S-180 tumor cells
DISCuSSION
The evaluation of D4-t induced blastogenesis was
the first criterion to get an idea whether D4-t could
effectively induce blast transformation, and what
are the optimal time interval and D4-t concentra-
tions. Blastogenesis is the culmination of several
biochemical events, and affects almost every meta-
bolic pathway [23]. The blast cell count in the present
investigation shows that D4-t can stimulate lymphoid
system of mice in vivo and cause blast differen-
tiation of lymphocytes. As it was shown previously
by others, ConA seems to be the best stimulating
agent of the lymphocytes in vivo, with maximal effect
at 48 h [18]. In comparison with ConA, and different
examined D4-t doses (see Fig. 2, a–d), we found that
5 mg/kg of D4-t showed greater degree of stimula-
tion of lymphocytes, as documented by high number
of healthy blast cells without any sign of vacuolation
or exhaustion. The optimal D4-t dose of 5 mg/kg was
confirmed by duplicate screening experiments.
The DNA synthesis assay is also widely used for
evaluation of lymphocytes activation in vivo [24]. The
rate of DNA synthesis by the SP lymphocytes has been
followed as the pattern of blastogenesis in current
study. By conventional criteria the difference was con-
sidered to be very statistically significant (P < 0.001).
It was previously shown that the optimal dose of ConA
for the blastogenic responses of the lymphocytes did
not correlate with DNA synthesis [18]. But as we have
shown, in the case of D4-t the maximum rate of blas-
togenesis corresponded to the optimal D4-t dose and
correlated with the rate of DNA synthesis.
Next, SP and MLN were isolated from the S-180 tu-
mor bearing mice, treated by 5 mg/kg dose of D4-t,
and histological sections were studied. The loosely
packed cells of SP possibly reflected the gradual trans-
formation of lymphocytes into blast cells. MLN cells
are more sensitive to ConA treatment than SP cells,
as it was observed on the histological sections of MLN
and SP that most of the MLN cells differentiated into
blasts by 48 h [21]. In current study the same effect
was observed for the optimal dose of D4-t (data not
presented). We didn’t observe any abnormalities
of cells treated by D4-t at its optimal dose.
It was found that significant (P < 0.01) inhibition
of tumor growth was in full agreement with the Winn as-
say data’s. In Winn assay tumor mass regression allow
us to suggest the antitumor efficacy of D4-t without
severe toxicity (see Fig. 4).
As D4-t showed 80% inhibition of 3H-thymidine
uptake in comparison to Platin10 and ConA it can
be concluded that D4-t could inhibit DNA synthesis
of S-180 tumor cells (highly significant P < 0.001).
In conclusion, the above results indicate promi-
sing therapeutic potential of dimethyl tin 4-cyclohexyl
thiosemicarbazone (D4-t).
ACKNOWLEDgEMENTS
We sincerely thank the Department of Science &
Technology (DST), Women Scientists Scheme (WOS-A),
New Delhi, India, (No.SR/WOS-A/CS-07/2004) for
financial assistance to Dr. A. Sen. Also we thank
to Dr. U. Sanyal, A. Mukherjee and S. Dutta of Dept.
of ACDD & Chemotherapy, Chittaranjan National Cancer
Institute, Kolkata for their immense help and support
in performing all the radioactive assays for the project.
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