Synthesis, molecular structure and anticancer activity of 1-allyl-3-amino-2-(4-chloro-2-mercaptobenzenesulphonyl)guanidine derivatives

Article abstract of DOI:10.1016/S0223-5234(02)01341-7

A series of 3-allylamino-6-chloro-7-R-1,1-dioxo-1,4,2-benzodithiazines (2a-e) was obtained by the reaction of 6-chloro-3-methylthio-1,4,2-benzodithiazine-1,1-dioxides (1a-e) with allylamine. Selective hydrazinolysis of the allylaminobenzodithiazines (2a-e) gave the appropriate 1-allyl-3-amino-2-(4-chloro-2-mercaptobenzenesulphonyl)guanidines (3a-e) in good yields. The reaction of 3a with dimethylsulphate under alkaline conditions provided the methylthio derivative 4. The structures of these compounds were confirmed on the basis of elemental analysis, spectral data (IR, ¹H- and ¹³C-NMR) and X-ray analysis. Screening data indicated that the compounds 3a and 3d exhibited significant in vitro activities against numerous human tumour cell lines, whereas compounds 3b and 3c showed a moderate activity.

Full text of DOI:10.1016/S0223-5234(02)01341-7

Eur. J. Med. Chem. 37 (2002) 285–293
www.elsevier.com/locate/ejmech
Original article
Synthesis, molecular structure and anticancer activity of
1-allyl-3-amino-2-(4-chloro-2-mercaptobenzenesulphonyl)guanidine
derivatives
Z. Brzozowski a, F. Sa˛czewski a,*, M. Gdaniec b
a Department of Chemical Technology of Drugs, Medical Uni6ersity of Gdan´sk, Gdan´sk, Poland
b Faculty of Chemistry, A. Mickiewicz Uni6ersity, 60-780 Poznan´, Poland
Received 27 August 2001; accepted 23 January 2002
Abstract
A series of 3-allylamino-6-chloro-7-R-1,1-dioxo-1,4,2-benzodithiazines (2a–e) was obtained by the reaction of 6-chloro-3-
methylthio-1,4,2-benzodithiazine-1,1-dioxides (1a–e) with allylamine. Selective hydrazinolysis of the allylaminobenzodithiazines
(2a–e) gave the appropriate 1-allyl-3-amino-2-(4-chloro-2-mercaptobenzenesulphonyl)guanidines (3a–e) in good yields. The
reaction of 3a with dimethylsulphate under alkaline conditions provided the methylthio derivative 4. The structures of these
compounds were confirmed on the basis of elemental analysis, spectral data (IR, 1H- and 13C-NMR) and X-ray analysis.
Screening data indicated that the compounds 3a and 3d exhibited significant in vitro activities against numerous human tumour
´
cell lines, whereas compounds 3b and 3c showed a moderate activity. © 2002 Editions scientifiques et me´dicales Elsevier SAS. All
rights reserved.
Keywords: 1-Allyl-3-amino-2-(2-mercaptobenzenesulphonyl)guanidines; X-ray structure analysis; Antitumour effect
1. Introduction
cancer activity prompted us to develop a method for
the synthesis of novel 1-allyl-3-amino-2-(4-chloro-2-
mercaptobenzenesulphonyl)guanidines (3a–e) for bio-
logical screenings.
Recently, arylsulphonamides have attracted attention
as anticancer [1–3] and anti-HIV [4–6] agents. We have
also described the syntheses of various 4-chloro-2-mer-
captobenzenesulphonamide derivatives with the nitro-
gen atom of the sulphonamide moiety attached to a
variety of heterocyclic ring systems. These compounds,
depending on structure, exhibited either anticancer [7–
13] or anti-HIV activities [7,9,12,14–16] and have been
described by Neamati et al. [16] as a novel class of
potent HIV-1 integrase inhibitors (MBSAs, Fig. 1). On
the other hand, aminoguanidine derivatives possess va-
rious biological activities but many of them are still
with unexplored pharmacological properties. Recent re-
ports on the syntheses of arylsulphonylaminoguanidi-
nes [17] as well as pyrimidinyl pyrazole derivatives
incorporating in their structure both aminoguanidine
and allylamino moieties [18] with pronounced anti-
2. Results
2.1. Synthesis
The syntheses of the target compounds 3a–e were
achieved by a convenient two step procedure starting
from methylthiobenzodithiazines 1a–e as shown in Fig.
2.
First, the reactions of 1a–e with allylamine carried
out in dry benzene at elevated temperature led to the
formation of 3-allylamino-1,4,2-benzodithiazines 2a–e
which could be separated either in high yields (91–98%)
for (2a–d) or in good yield (74%) for (2e). Then, upon
treatment of 2a–e with an excess of hydrazine hydrate
in methanolic solution at room temperature, the desired
1-allyl-3-amino-2-(4-chloro-2-mercaptobenzenesulpho-
* Correspondence and reprints.
E-mail address: saczew@amg.gda.pl (F. Sa˛czewski).
´
0223-5234/02/$ - see front matter © 2002 Editions scientifiques et me´dicales Elsevier SAS. All rights reserved.
PII: S0223-5234(02)01341-7

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Z. Brzozowski et al. / European Journal of Medicinal Chemistry 37 (2002) 285–293
nyl)guanidines (3a–e) were obtained in high yields (82–
92%).
the amino form [23] while others are so close to the
borderline that both imino and amino forms have been
isolated [24].
The mechanism of this reaction could be explained
by a regioselective nucleophilic attack of the hydrazine
nitrogen atom at the C-3 carbon atom of the ben-
zodithiazine 2 with simultaneous heterocyclic ring
opening. In this procedure an excess of hydrazine hy-
drate was used in order to avoid a plausible oxidation
of the mercapto group formed to the corresponding
disulphide.
The tautomeric form assumed by the sulphonyl-
guanidine 4 in the crystal (Fig. 3) is that observed for
sulphaguanidine in the solid state, i.e. with the
guanidine imino nitrogen joined to the sulphone S atom
[21,22]. This form has been confirmed by localisation of
the H atoms from the Fourier difference map and by
geometrical parameters characteristic of the sul-
phaguanidine molecule in this tautomeric form [21], i.e.
The structures of the compounds 2a–e and the final
products 3a–e were confirmed by elemental analyses as
,
S1ꢀN1 1.573(2) A, O1ꢀS1ꢀN1 114.7(1)°, O2ꢀS1ꢀN1
1
well as by IR, H-NMR, 13C-NMR spectrophotometry.
106.8(1)°. The three guanidine CꢀN bonds are of simi-
lar length [C9ꢀN1 1.344(3), C9ꢀN2 1.331(3), C9ꢀN3
1
For example, in H-NMR spectrum, the existence of an
,
intact N-allyl group in 3a was identified from the
signals at l 3.94 (NꢀCH2, d, JCHꢀNH=5.6 Hz), 5.11
(dd, NꢀCH2ꢀCHCꢁCHA, Jcis=6.1 Hz, Jgem=1.3 Hz),
1.329(3) A], however, like in sulphaguanidine [19,20],
the longest bond is formed to the N atom connected to
the sulphone group.
5.18 (NꢀCH2ꢀCHCꢁCHB, dd, Jtrans=13.5 Hz, Jgem
=
The overall molecular conformation can be described
as nearly perpendicular arrangement of the phenyl sub-
stituent and the allyl group relative to the guanidine
moiety with the best plane of the phenyl ring bisecting
the O2ꢀS1ꢀN1 angle. The hydrazine moiety is oriented
with the N2 hydrogen directed to N1 and N4 atom
directed to N3. This allows the intramolecular hydro-
gen bond to be formed between N2ꢀH and O1 of the
1.3 Hz) and 5.84 (m, NꢀCH2ꢀCHCꢁCH2). The 13C-
NMR spectrum exhibited the presence of three
carbons attributed to N-allyl group at
l 43.92
(NꢀCH2ꢀCHꢁCH2), 117.43 (NꢀCH2ꢀCHꢁCH2) and
138.92 (NꢀCH2ꢀCHꢁCH2). The presence of SH group
in 3a was confirmed in IR spectrum by the absorption
at 2543 cm−1
.
,
We were not able to obtain crystals of 3 suitable for
X-ray analysis. Therefore, in order to investigate in
sulphone group [N2ꢀH···O1 2.807(3) A, H···O1 2.20(3)
,
A, ꢂN2ꢀH···O1 135(3)°]. The N2ꢀH group participates
detail
a
tautomeric structure of the sulphony-
in the three-center hydrogen bond acting also as a
donor in a weak intermolecular hydrogen bond to O1
laminoguanidines obtained, we prepared the methylthio
derivative 4 by reacting 3a with dimethylsulphate under
alkaline conditions (Fig. 2).
of the molecule related by
a symmetry center
[N2ꢀH···O1i 3.004(3) A, H···O1i 2.43(3) A,
ꢂN2ꢀH···O1i 132(3)°; symmetry code (i): −x, −y,
−z].
,
,
2.2. Crystal and molecular structure of 4
The lone pair of the sp3 hybridised N4 atom is
pointing towards the hydrogen atom bonded to N3
giving rise to another weak intramolecular hydrogen
Previously, crystal structures were determined for a
number of guanidine derivatives containing substituents
such as cyano [19], nitro [20] and sulphonyl [21,22]. All
these showed the electron withdrawing group on imino
nitrogen. However, some acyl derivatives are firmly in
,
,
bond [N3ꢀH···N4 2.646(4) A, H···N(4) 2.27(3) A,
ꢂN3ꢀH···N4 112(3)°]. Only one hydrogen atom of the
NH2 group takes part in the intermolecular hydrogen
Fig. 1. Benzenesulphonylguanidines with anticancer and anti-HIV activities.

Z. Brzozowski et al. / European Journal of Medicinal Chemistry 37 (2002) 285–293
287
Fig. 2. Synthesis of arylsulphonylaminoguanidines from methylthiobenzodithiazines.
bond and is joined to the sulphone group of the
molecule related by a glide plane [N4ꢀH···O2ii 3.127(3)
ii
ii
,
,
A, H···O2 2.37(3) A, ꢂN4ꢀH···O2 135(3)°; symmetry
code (ii): x−0.5, −y+0.5, z−0.5].
We have also examined tautomers of 4 by quantum
chemical calculations using B3LYP hybrid density
functional model with 6-31G* polarisation basis set
[25]. The structures and relative energies obtained for
tautomers 4A, 4B and 4C are shown in Fig. 4. The
sulphonylimino tautomer 4A was calculated to be subs-
tantially lower in energy than sulphonylamino tau-
tomers 4B and 4C. According to the following equation
[26]:
N1
N2
=exp−1060(E1−E2)
N1,N2=number of molecules;
E1,E2=energy in a.u.
Fig. 3. ORTEP drawing of 4 with the atom labelling.

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Z. Brzozowski et al. / European Journal of Medicinal Chemistry 37 (2002) 285–293
Fig. 4. Tautomeric forms of benzenesulphonylguanidine 4.
at the temperature 296 K 4A is identified as the over-
whelmingly predominant tautomer by a factor of
7.22×105 over the less stable 4B which in turn is
favoured by a factor of 1.45×105 over 4C. Moreover,
based upon their calculated dipole moments, 4A (4.82
D) would be predicted to predominate over 4B (4.51 D)
and 4C (1.93 D) in polar solvents.
parameters: GI50, molar concentration of the com-
pound that inhibits 50% net cell growth; TGI, molar
concentration of the compound leading to total inhibi-
tion; and LC50, molar concentration of the compound
leading to 50% net cell death.
Compounds 3a–e exhibited reasonable antineoplastic
activity against most (72–93%) of the 61 human cancer
cell lines. Relatively highest sensitivity to the com-
pounds described here was found for lung cancer (NCI-
HS22), renal cancer (A 498) and melanoma (COX
IMVI, MALME-3M, SF-MEL-2 and SK-MEL-5). The
data of the most sensitive cell lines recorded in Table 1
indicate the following rank order of activity: 3e (R=p-
FꢀPhNHCO)B3b (R=CH3OCO)B3c (R=PHN-
HCO),B3a (R=CH3)53d (R=p-ClꢀPHNHCO).
It is noteworthy that the activity of the compound 3a
containing a small, neutral, lipophilic CH3 substituent
was similar to that of 3d with bulky, conjugative elec-
tron-withdrawing p-ClꢀPhNHCO group, while the p-
FꢀPhNHCO-derivative 3e was the least active in this
series.
2.3. Biology
All the 2-mercaptobenzenesulphonamides 3a–e syn-
thesised were tested in US National Cancer Institute
(Bethesda) for their in vitro anticancer activity.
The antitumour activities of the compounds were
evaluated by using total 61 human tumour. Cell Lines,
derived from nine different cancer types: leukemia,
lung, colon, CNS, melanoma, ovarian, renal, prostate
and breast. The compounds were tested at five concen-
tration at 10-fold dilution. A 48 h continuous drug
exposure protocol was used and sulphorhodamine B
(SRB) protein assay was used to estimate cell growth.
Details of this test system and the information which is
encoded by the activity pattern over all cell lines, have
been published [27–29]. The antitumour activity of a
test compound is reported for each cell line by three
These results are rather surprising from structure–ac-
tivity relationships point of view, and may suggest that
mechanisms by which compounds 3a and 3d exert their
antineoplastic effects are different.

Table 1
Inhibition of in vitro cancer cell lines by compounds 3a–e
Panel cell line
Response parameters a (mM) GI50 (A), TGI (B) and LC50 (C)
Compound 3a Compound 3b Compound 3c
Compound 3d
Compound 3e
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
Leukemia
K-562
RPMI-8226
SR
27.0
20.1
37.1
80.9
61.8
90.4
*
*
*
47.1
31.2
37.5
*
*
*
*
*
*
35.2
31.4
41.8
*
*
*
*
*
*
31.1
28.9
27.2
*
70.8
*
*
*
*
32.8
34.9
43.4
*
*
*
*
*
*
Non-small cell lung cancer
NCI-H522
22.3
47.0
99.0
19.5
47.5
*
23.4
76.1
*
28.2
64.1
*
33.4
94.2
*
/
Colon cancer
HCC-2998
KM-12
3.73
61.8
*
*
*
*
42.2
40.4
*
*
*
*
46.1
33.3
*
*
*
*
19.1
28.0
49.1
97.0
*
*
40.5
49.3
*
*
*
*
CNS cancer
SF-268
SF-539
15.9
43.8
93.5
73.0
*
*
*
*
*
13.5
40.5
24.7
60.7
*
75.3
*
*
*
26.1
38.4
*
*
*
*
*
*
*
11.5
30.7
NT
52.1
81.8
NT
*
*
NT
50.4
39.5
NT
*
*
NT
*
*
NT
SNB-75
Melanoma
LOX IMVI
MALME-3M
M 14
SK-MEL-2
SK-MEL-28
SK-MEL-5
UACC-62
18.4
16.7
27.8
21.0
41.6
35.0
51.2
37.7
69.3
*
84.3
*
77.4
37.5
24.0
31.5
26.0
36.7
26.4
47.0
*
97.5
*
84.5
*
82.3
*
*
*
*
*
*
*
*
21.0
31.2
26.5
22.2
45.3
23.2
43.1
*
77.0
*
71.6
*
*
*
*
*
*
*
*
*
27.4
17.9
21.7
28.9
22.3
14.0
29.2
*
*
85.3
*
*
*
54.5
29.1
42.3
28.3
56.2
26.3
57.3
*
70.1
*
76.7
*
*
*
*
*
*
*
*
*
*
*
*
*
*
*
39.1
69.5
71.2
83.9
28.8
84.1
*
*
59.2
*
*
*
3
O6arian cancer
IGRV 1
OVCAR-8
23.9
32.1
53.3
*
*
*
29.6
34.8
*
*
*
*
6.2
30.8
83.3
*
*
*
36.2
25.4
*
59.3
*
*
71.0
73.8
*
*
*
*
(
Renal cancer
285
A 498
21.6
63.6
69.8
*
*
*
34.5
42.8
72.8
*
*
*
17.7
29.0
57.4
*
*
*
18.1
30.0
37.1
94.0
75.8
*
17.2
30.3
64.1
*
*
*
293
Prostate cancer
PC-3
Breast cancer
MDA-MB-435
MDA-N
32.2
39.7
*
*
*
*
40.3
35.3
*
*
*
*
17.7
24.9
52.5
71.5
*
*
23.4
27.3
73.1
*
*
*
35.7
54.7
*
*
*
*
Percent of the cell lines giving positive GI50, TGI and LC50 testing result
91.4
17.2
5.2
77.6
12.1
0
81.0
12.1
0
93.1
29.3
5.2
72.4
6.9
0
*The values TGI or LC50\100 mM. NT, not tested.
a GI50, concentration giving 50% inhibition; TGI, concentration giving total growth inhibition; LC50, concentration having 50% lethal effect.

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Z. Brzozowski et al. / European Journal of Medicinal Chemistry 37 (2002) 285–293
3. Experimental
127.98, 128.18, 131.63, 132.02, 138.51, 139.00 (aromatic
carbons); 144.62 (NꢀCH2ꢀCHꢁCH2); 163.67 (CꢁN).
3.1. Synthesis
3.2.2. 3-Allylamino-6-chloro-7-methoxycarbonyl-
1,4,2-benzodithiazine-1,1-dioxide (2b)
M.p.s were taken on a Bu¨chi 535 apparatus and are
reported uncorrected. IR spectra in KBr were recorded
on a Perkin–Elmer 1600 FTIR spectrophotometer. H-
IR: 3330 (NH), 1730 (CꢁO); 1642, 1587, 1555 (CꢁN
and aromatic ring); 1360, 1305, 1160, 1140, (SO2).
1H-NMR (DMSO-d6): 3.89 (s, 3H, CH3OCO); 4.01
(d, 2H, NꢀCH2CHꢁCH2); 5.15–5.27 (m, 2H,
CH2CHꢁCH2); 5.79–5.92 (m, 1H, NꢀCH2ꢀCHꢁCH2);
8.12 (s, 1H, aromat.); 8.33 (s, 1H, aromat.).
1
and 13C-NMR spectra were recorded on a Varian XL
200 spectrometer 200 MHz PMR using TMS as inter-
nal standard (l values in ppm). The results of elemental
analyses for C, H and N were within 90.4% of the
theoretical values. The starting 6-chloro-3-metylthio-
1,4,2-benzodithiazine-1,1-dioxide derivatives were ob-
tained according to methods described previously: 1a
[30]; 1b [31] and 1c–e [32].
3.2.3. 3-Allylamino-6-chloro-7-phenylcarbamoyl-
1,4,2-benzodithiazine-1,1-dioxide (2c)
IR: 3300, 3215 (NH); 1660 (CꢁO); 1615, 1595, 1555;
1
(CꢁN and aromatic ring), 1320, 1140 (SO2). H-NMR
(DMSO-d6): 4.03 (d, 2H, NꢀCH2CHꢁCH2); 5.16–5.27
(m, 2H, NꢀCH2CHꢁCH2); 5.77–5.96 (m, 1H,
NꢀCH2CHꢁCH2); 7.10–7.71 (m, 5H, aromat.); 8.09 (s,
1H, aromat.); 8.12 (s, 1H, aromat.); 10.02 (br.s, 1H,
NH); 10.69 (s, 1H, NHCO).
3.2. Preparation of 3-allylamino-6-chloro-1,4-2-
benzodithiazine-1,1-dioxides (2a–e)
A solution of the corresponding methylthioben-
zodithiazine (1a), (1b), (1c), (1d) or (1e) (0.05 mol) and
2.9 g (0.05 mol) of allylamine in dry C6H6 (120–180
mL) was stirred for 3 h at room temperature (r.t.). The
suspension obtained was refluxed until the evolution of
CH3SH had ceased (20–28 h). The precipitate was
filtered off, washed successively with C6H6 (3×10 mL)
and CH3OH (3×5 mL). Experimental data: see Table
2.
3.2.4. 3-Allylamino-6-chloro-7-(4-chlorophenyl-
carbamoyl)-1,4,2-benzodithiazine-1,1-dioxide (2d)
IR: 3310, 3285 (NH); 1640 (CꢁO), 1625, 1598, 1540
1
(CꢁN and aromatic ring), 1345, 1310, 1160 (SO2). H-
NMR (DMSO-d6): 4.02 (d, 2H, NꢀCH2ꢀCHꢁCH2);
5.16–5.27 (m, 2H, NꢀCH2CHꢁCH2); 5.71–5.96 (m, 1H,
NꢀCH2ꢀCHꢁCH2); 7.43 (d, 2H, aromat.); 7.72 (d, 2H,
aromat.); 8.12 (s, 1H, aromat.); 8.13 (s, 1H, aromat.);
10.02 (br.s, 1H, NH); 10.83 (s, 1H, NHCO).
In this manner the following products were obtained.
3.2.1. 3-Allylamino-6-chloro-7-methyl-1,4,2-
benzodithiazine-1,1-dioxide (2a)
3.2.5. 3-Allylamino-6-chloro-7-(4-fluorophenyl-
carbamoyl)-1,4,2-benzodithiazine-1,1-dioxide (2e)
IR: 3300, 3235, (NH); 1655 (CꢁO), 1615, 1570, 1555
IR: 3260 (NH), 1642; 1565 (CꢁN and aromatic ring);
1
1
1345, 1305, 1152 (SO2). H-NMR (CDCl3): 2.45 (s, 3H,
(CꢁN and aromatic ring), 1320, 1155 (SO2). H-NMR
CH3); 4.12 (d, 2H, NꢀCH2ꢀCHꢁCH2); 5.18–5.30 (m,
2H, NCH2CHꢁCH2); 5.76–5.95 (m, 1H, NCH2ꢀ
CHꢁCH2); 6.76 (br.s, 1H, NH); 7.34 (s, 1H, aromat.);
7.97 (s, 1H, aromat.). 13C-NMR (CDCl3): 20.62 (CH3);
47.01 (N CH2ꢀCHꢁCH2); 119.54 (NꢀCH2ꢀCHꢁCH2);
(DMSO-d6): 4.03 (d, 2H, NꢀCH2ꢀCHꢁCH2); 5.16–5.27
(d, 2H, NꢀCH2CHꢁCH2); 5.80–5.94 (m, 1H,
NꢀCH2ꢀCHꢁCH2); 7.22 (t, 2H, aromat.); 7.67–7.75 (m,
2H, aromat.); 8.11 (s, 2H, aromat.); 10.03 (s, NH);
10.75 (s, 1H, NHCO).
Table 2
Experimental data of compounds 2a–e, 3a–e and 4
Compound
R
Yield (%)
m.p. (°C)
Molecular formula (molecular weight)
2a
2b
2c
2d
2e
3a
3b
3c
3d
3e
4
CH3
CH3OCO
C6H5NHCO
4-ClꢀC6H4NHCO
4-FꢀC6H4NHCO
CH3
95
96
98
91
74
92
84
88
82
83
83
161–163
154–156
169–171
185–187
188–189
130–131
129–131
142–144
145–147
171–173
154–155
C11H11ClN2O2S2(302.8)
C12H11ClN2O4S (346.8)
C17H14ClN3O3S2 (407.9)
C17H13Cl2N3O3S2 (442.3)
C
17H13ClFN3O3S2 (425.9)
C11H15ClN4O2S2 (334.8)
C12H15ClN4O4S2 (378.8)
C17H18ClN5O3S2 (439.9)
C17H17Cl2N5O3S2 (474.4)
C17H17ClFN5O3S2 (457.9)
C12H17ClN4O2S2 (348.87)
CH3OCO
C6H5NHCO
4-ClꢀC6H4NHCO
4-FꢀC6H4NHCO
CH3

Z. Brzozowski et al. / European Journal of Medicinal Chemistry 37 (2002) 285–293
291
3.3. Preparation of 1-allyl-3-amino-2-(4-chloro-2-
mercaptobenzenesulphonyl)guanidines (3a–e)
7.77(s, 1H, aromat.); 8.02 (s, 1H, aromat.); 8.14 (s, 1H,
NH); 10.52 (s, 1H, CONH). 13C-NMR (DMSO-d6):
42.41 (NꢀCH2ꢀCHꢁCH2); 78.93 (NꢀCH2ꢀCHꢁCH2);
115.28, 119.56, 119.75, 123.88, 128.40, 128.74, 130.76,
131.87, 132.50, 135.25 (aromatic carbons); 138.76
(NꢀCH2ꢀCHꢁCH2); 156.52 (CꢁN); 163.56 (CꢁO).
A mixture of the corresponding allylaminoben-
zodithiazine 2a–e (0.01 mol) and 1.3 g (0.025 mol) of
hydrazine hydrate (99–100%) in 15 mL of CH3OH was
stirred at r.t. for 20–24 h. The solvent was removed
under vacuum (rotary evaporator) to give a dry residue,
which was further purified by dissolving in water (350
mL, pH 8.5–9). After stirring for 0.5 h, a small amount
of insoluble side-products (0.1–0.3 g) was filtered off
and the filtrate was acidified with 1% HCl to pH 3. The
precipitate thus obtained was collected by filtration,
washed with water and dried initially at r.t. and then at
100 °C. Experimental data: see Table 1.
3.3.4. 1-Allyl-3-amino-2-[4-chloro-2-mercapto-5-(4-
chlorophenylcarbamoyl)-benzenesulphonyl]guanidine (3d)
IR: 3340, 3290 (NH); 2552 (SH); 1670 (NCꢁO); 1650
1
(CꢁN); 1350, 1130 (SO2). H-NMR (DMSO-d6): 3.83
(d, 2H, NꢀCH2ꢀCHꢁCH2); 4.30–4.80 (br.s, 2H, SH and
NH); 4.97–5.13 (m, 2H, NꢀCH2ꢀCHꢁCH2); 5.73–5.92
(m, 1H, (NꢀCH2ꢀCHꢁCH2); 7.43 (d, 2H, aromat.); 7.74
(d, 2H, aromat.); 7.62 (s, 1H, NH); 7.80 (s, 1H, aro-
mat.); 8.03 (s, 1H, aromat.); 8.15 (s, 1H, SO2NH); 10.69
(s, 1H, HNCO).
In this manner the following products were obtained.
3.3.1. 1-Allyl-3-amino-2-(4-chloro-2-mercapto-5-
methylbenzenesulphonyl)guanidine (3a)
IR: 3345, 3320, 3245 (NH); 2540 (SH); 1650 (CꢁN);
1350, 1330 (SO2). 1H-NMR (CDCl3): 2.34 (s, 3H,
CH3Ph); 3.91 (s, 2H, NH2); 3.94 (d, 2H,
NꢀCH2ꢀCHꢁCH2); 5.04 (br.s, 1H, NH); 5.11 (dd, 1H,
NꢀCH2ꢀCHꢁCHA); 5.18 (dd, 1H, NꢀCH2ꢀCHꢁCHB);
5.84 (m, 1H, NꢀCH2ꢀCHꢁCH2); 6.70 (br.s, 1H, NH);
7.34 (s, 1H, aromat.); 7.87 (s, 1H, aromat.); 8.28 (s, 1H,
NH). 13C-NMR (CDCl3): 20.08 (CH3ꢀPh); 43.92
(NꢀCH2ꢀCHꢁCH2); 117.75 (NꢀCH2ꢀCHꢁCH2); 131.1,
131.4, 133.4, 134.3, 138.1 (aromatic carbons); 138.93
(NꢀCH2ꢀCHꢁCH2); 157.34 (CꢁN).
3.3.5. 1-Allyl-3-amino-2-[(4-chloro-2-mercapto-5-(4-
fluorophenylcarbamoyl)-benzenesulphonyl]guanidine (3e)
IR: 3340, 3260 (NH); 2550 (SH); 1665 (NCꢁO); 1640
1
(CꢁN); 1350, 1130 (SO2). H-NMR (DMSO-d6): 3.82
(d, 2H, NꢀCH2ꢀCHꢁCH2); 4.4–4.6 (br.s, 2H, NH and
SH); 4.97–5.13 (m, 2H, NꢀCH2ꢀCHꢁCH2); 5.73–5.92
(m, 1H, NꢀCH2ꢀCHꢁCH3); 7.14–7.28 (m, 2H, aro-
mat.) and 7.69–7.76 (m, 2H, aromat.); 7.61 (s, 1H,
NH); 7.79 (s, 1H, aromat.); 8.02 (s, 1H, aromat.); 8.15
(s, 1H, NH); 10.60 (s, 1H, HNCO).
3.4. Preparation of 1-allyl-3-amino-2-(4-chloro-5-
methyl-2-methylthiobenzenesulphonyl)guanidine (4)
3.3.2. 1-Allyl-3-amino-2-(4-chloro-2-mercapto-5-
methoxycarbonylbenzenesulphonyl)guanidine (3b)
IR: 3375, 3345, 3325, 3271 (NH); 2550 (SH); 1720
(CꢁO); 1640 (CꢁN); 1360, 1140 (SO2). 1H-NMR
(DMSO-d6): 3.76 (s, 1H, SH); 3.82 (d, 2H,
NꢀCH2ꢀCHꢁCH2); 3.86 (s, 3H, CH3O); 5.02 (s, 1H,
NH); 4.97–5.10 (m, 2H, NꢀCH2ꢀCHꢁCH2); 5.8 (m,
1H, NHꢀCH2ꢀCHꢁCH2); 7.63 (br.s, 1H, NH); 7.81 (s,
1H, aromat.); 8.05 (s, 1H, NH); 8.30 (s, 1H, aromat.).
13C-NMR (DMSO-d6): 42.41 (NꢀCH2ꢀCHꢁCH2); 52.55
(CH3O);115.22 (NHꢀCH2ꢀCꢁCH2); 130.78, 130.81,
132.12, 132.19, 134.54, 135.24 (aromatic carbons);
139.03 (NꢀCH2ꢀCHꢁCH2); 156.59 (CꢁN); 163.98
(CꢁO).
Compound 3a (1.7 g, 5 mmol) was dissolved in a
solution of NaOH (0.32 g, 8 mmol) in water (30 mL).
The resulting solution was stirred and cooled on an ice
bath, and then treated dropwise with Me2SO4 (0.76 g, 6
mmol). Stirring was continued at 0–3 °C for 1 h and at
r.t. for 2 h. Then, the reaction mixture was acidified
with 1% HCl to pH 6. The precipitate thus obtained
was filtered off, washed thoroughly with water, dried
(1.7 g, m.p. 152–154 °C) and purified by recrystallisa-
tion from 2-propanol. Yield: 1.4 g (83%), m.p. 154–
155 °C. IR (KBr): 3385, 3340, 3280, 3245 (NH2, NH);
2950, 2915, 2850 (CH3, CH2); 1635, 1570 (CꢁN, CꢁC);
1
1360, 1135 (SO2). H-NMR (DMSO-d6): 2.31 (s, 3H,
3.3.3. 1-Allyl-3-amino-2-(4-chloro-2-mercapto-5-
phenylcarbamoylbenzenesulphonyl)guanidine (3c)
PhCH3); 2.46 (s, 3H, SCH3); 3.79 (t, 2H, CH2); 4.57 (s,
2H, NH2); 4.98–5.11 (m, 2H, CꢁCH2); 5.71–5.87 (m,
1H, CHꢁC); 7.29 (s, 1H, H-3); 7.48 (t, 1H, NHꢀCH2);
7.81 (s, 1H, H-6); 7.99 (s, 1H, NH). 13C-NMR (DMSO-
IR: 3340, 3250 (NH); 2548 (SH); 1670 (NCꢁO); 1645
(CꢁN); 1350, 1130 (SO2). 1H-NMR (DMSO-d6): 3.48 (s,
1H, SH); 3.83 (d, 2H, NꢀCH2ꢀCHꢁCH2); 4.60 (br.s,
1H, NH); 4.97–5.13 (m, 2H, NꢀCH2ꢀCHꢁCH2); 5.72–
5.91 (m, 1H, NꢀCH2ꢀCHꢁCH2); 7.07–7.14, 7.31–7.38,
7.68–7.72 (m, 5H, aromat.); 7.57 (br.s, 1H, NH);
d6):
15.48
(CH3ꢀS);
19.16
(PhCH3);
42.60
(NꢀCH2ꢀCHꢁCH2); 115.39 (NꢀCH2ꢀCHꢁCH2); 125.69,
130.47, 130.94, 135.75, 136.74, 137.65 (aromatic car-
bons); 139.40 (NꢀCH2ꢀCHꢁCH2); 157.00 (CꢁN).

292
Z. Brzozowski et al. / European Journal of Medicinal Chemistry 37 (2002) 285–293
Table 3
Table 4
Atomic coordinates (×104) and equivalent isotropic displacement
,
Bond lengths (A) and angles (°) for 4
2
3
,
parameters (A ×10 ) for 4
Bond lengths
S(1)ꢀO(2)
S(1)ꢀO(1)
S(1)ꢀN(1)
S(1)ꢀC(1)
S(2)ꢀC(6)
S(2)ꢀC(8)
Cl(1)ꢀC(4)
N(1)ꢀC(9)
N(2)ꢀC(9)
N(2)ꢀN(4)
N(3)ꢀC(9)
N(3)ꢀC(10)
C(1)ꢀC(2)
C(1)ꢀC(6)
C(2)ꢀC(3)
C(3)ꢀC(4)
C(3)ꢀC(7)
C(4)ꢀC(5)
C(5)ꢀC(6)
C(10)ꢀC(11)
C(11)ꢀC(12)
x
y
z
Ueq
50(1)
1.434(2)
1.4498(18)
1.573(2)
1.783(2)
1.764(3)
1.778(4)
1.746(3)
1.344(3)
1.331(3)
1.406(3)
1.329(3)
1.441(3)
1.385(3)
1.406(3)
1.391(4)
1.377(4)
1.506(4)
1.381(4)
1.392(4)
1.474(5)
1.278(6)
S(1)
S(2)
895(1)
1648(1)
−3072(1)
273(2)
1964(2)
1271(2)
−173(2)
792(2)
−723(3)
−191(2)
−1400(2)
−2330(2)
−1977(2)
−788(2)
134(2)
−3637(4)
1563(4)
620(2)
1748(1)
4137(1)
4405(1)
677(1)
1794(1)
3392(1)
2683(1)
1193(2)
2891(2)
1063(2)
−813(2)
−549(2)
85(1)
81(1)
65(1)
67(1)
50(1)
59(1)
55(1)
66(1)
44(1)
54(1)
57(1)
55(1)
56(1)
51(1)
97(1)
76(1)
44(1)
58(1)
75(1)
106(1)
Cl(1)
O(1)
O(2)
N(1)
N(2)
N(3)
N(4)
C(1)
C(2)
C(3)
C(4)
C(5)
C(6)
C(7)
C(8)
C(9)
C(10)
C(11)
C(12)
1592(2)
2611(2)
1955(2)
3734(2)
2151(3)
2563(2)
2176(2)
2725(2)
3684(2)
4116(3)
3578(2)
2287(5)
5480(4)
2744(2)
4669(2)
4577(4)
3760(5)
−2015(2)
2062(2)
1519(2)
1668(3)
2407(2)
2939(2)
2769(2)
1040(6)
4065(4)
−91(2)
98(3)
1630(3)
2851(3)
3210(5)
148(3)
−304(5)
Bond angles
O(2)ꢀS(1)ꢀO(1)
O(2)ꢀS(1)ꢀN(1)
O(1)ꢀS(1)ꢀN(1)
O(2)ꢀS(1)ꢀC(1)
O(1)ꢀS(1)ꢀC(1)
N(1)ꢀS(1)ꢀC(1)
C(6)ꢀS(2)ꢀC(8)
C(9)ꢀN(1)ꢀS(1)
C(9)ꢀN(2)ꢀN(4)
C(9)ꢀN(3)ꢀC(10)
C(2)ꢀC(1)ꢀC(6)
C(2)ꢀC(1)ꢀS(1)
C(6)ꢀC(1)ꢀS(1)
C(1)ꢀC(2)ꢀC(3)
C(4)ꢀC(3)ꢀC(2)
C(4)ꢀC(3)ꢀC(7)
C(2)ꢀC(3)ꢀC(7)
C(3)ꢀC(4)ꢀC(5)
C(3)ꢀC(4)ꢀCl(1)
C(5)ꢀC(4)ꢀCl(1)
C(4)ꢀC(5)ꢀC(6)
C(5)ꢀC(6)ꢀC(1)
C(5)ꢀC(6)ꢀS(2)
C(1)ꢀC(6)ꢀS(2)
N(3)ꢀC(9)ꢀN(2)
N(3)ꢀC(9)ꢀN(1)
N(2)ꢀC(9)ꢀN(1)
N(3)ꢀC(10)ꢀC(11)
C(12)ꢀC(11)ꢀC(10)
116.39(12)
106.80(12)
114.73(11)
106.87(12)
106.33(11)
104.85(11)
103.94(17)
123.81(17)
119.8(2)
124.9(2)
120.2(2)
117.33(19)
122.48(19)
122.9(2)
115.8(2)
123.2(3)
121.0(3)
123.0(2)
119.1(2)
117.9(2)
121.0(2)
117.1(2)
Ueq is defined as one-third of the trace of the orthogonalised Uij
tensor.
4. X-Ray structure analysis
The data were collected on a KumaCCD diffrac-
tometer using graphite monochromatised Mo Ka radia-
tion with detector distance of 6 cm. More than
hemisphere of reciprocal space was covered by combi-
nation of four sets of exposures; each set had a different
ƒ-angle (0, 90, 180, 270) and each exposure of 30 s
covered 0.75° in ꢀ. Coverage of the unique set is over
99% complete. Out of 8871 reflections measured up to
(sin q/u)max=0.625, 3257 were symmetry independent
(Rint=0.0263). The collected data were reduced using
the program CRYSALIS RED [33]. The structure was
solved by direct methods with the program SHELXS-97
[34] and refined by full-matrix least-squares method on
F2 with SHELXL-97 [35]. Hydrogen atoms have been
located on DF maps and refined with isotropic displace-
ment parameters.
122.2(2)
120.7(2)
117.5(2)
117.4(2)
125.1(2)
114.1(3)
126.4(4)
Crystal data for 4: C12H17ClN4O2S2, monoclinic,
,
P21/n, a=12.2808(9), b=11.2825(7), c=13.0269(9) A,
3
,
i=117.668(7)°, V=1598.6(2) A , Z=4, T=293 K,
Dx=1.450 g cm−3, v=0.509 mm−1. The structure
was refined on 3257 reflections; 258 refined parameters;
R1=0.0500, wR2=0.1221, Goodness-of-fit=1.087 for
2575 reflections with F\4|(F) [R1=0.0686, wR2=
0.1366 for all 3257 independent reflections]. Final
atomic coordinates, bond lengths and angles are listed
in Tables 3 and 4, respectively. Atom labelling is shown
in Fig. 3.
5. Supplementary data
Further details of the crystal structure investigation
may be obtained from The Director of the Cambridge
Crystallographic Data Centre, 12 Union Road, Cam-
bridge CB2 1EZ, UK, in quoting the full journal
citation.

Z. Brzozowski et al. / European Journal of Medicinal Chemistry 37 (2002) 285–293
293
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