D. Bartulewicz et al. / European Journal of Medicinal Chemistry 36 (2001) 461–467
465
netropsin and distamycin. Molecular modelling calcu-
6.1.1.1. N-[3-(3-[4-Bis(2-chloroethyl)aminophenyl-
butyramido]benzamido)-benzoyl]-N%,N%-dimethyl-1,
lations made it possible to understand the reason for
the observed reduced affinity to AT pairs and in-
creased affinity towards GC sequences of the carbo-
cyclic lexitropsins with chlorambucil moiety in
comparison with netropsin and distamycin. Further
investigations on the mechanisms of the cytotoxicity
carried out by these compounds are in progress.
3-propanediamine hydrochloride (5)
1
Yield: 0.14 g (45.8%). H NMR (DMSO-d , l ppm):
6
1.20 (m, 2H, CH ); 1.83 (m, 4H, CH ); 2.40 (t, 2H,
2
2
CH ); 2.72 (s, 6H, N(CH ) ); 3.09 (m, 2H, CH ); 3.35
2
3 2
2
(m, 2H, CH ); 3.88 (s, 8H, ClCH CH N); 6.63–8.78 (m,
2
2
2
10H, Ar-H); 10.36, 10.49 2× (s, 1H, CONH); 10.83 (bs,
13
1
H, CONH). C NMR (DMSO-d , l ppm): 24.15
6
(
CH ); 27.12 (CH ); 33.61 (CH ); 35.83 (CH ); 36.45
2
2
2
2
(
CONHCH2);
41.87
(N(CH3)2);
39.80,
52.23
6
. Experimental protocols
(ClCH CH N); 54.40 (CH N(CH ) ); 111.91; 118.66;
2 2 2 3 2
1
1
1
19.89; 121.96; 122.18; 123.15; 127.32; 128.46; 128.63;
29.31; 129.84; 134.89; 135.33; 139.25; 139.58; 141.70;
44.40; 146.17 (Ar); 165.74; 166.36; 171.46 (3×CONH).
6
.1. Chemistry
Melting points were determined on a Buchi 535 ap-
Anal. Found: C, 59.97; H, 6.56; Cl, 16.24; N, 10.65.
Calc. for C H N O Cl HCl: C, 59.77; H, 6.38; Cl,
1
13
paratus and are uncorrected. H NMR and C NMR
spectra were recorded on a Bruker AC 200 F spectrom-
eter using tetramethylsilane as an internal standard.
Chemical shifts are expressed in l value (ppm). Multi-
plicity of resonance peaks is indicated as singlet (s),
doublet (d), triplet (t), quartet (q), broad singlet (bs) and
multiplet (m).
Thin-layer chromatograms were prepared on pre-
coated plates (Merck, silica gel 60F-254) using the sol-
vent system (all proportions by volume) methanol–25%
ammonia, 99:1. Silica gel 60 (230–400 mesh ASTM) was
used for column chromatography.
33
41
5
3
2
1
6.04; N, 10.56%.
6.1.1.2. N-[3-(3-(3-[4-Bis(2-chloroethyl)aminophenyl-
butyramido]benzamido)benzamido)benzoyl]-N%,N%-
dimethyl-1,3-propanediamine hydrochloride (6)
1
Yield: 0.17 g (46.2%). H NMR (DMSO-d , l ppm):
6
1
.84–1.94 (m, 4H, CH ); 2.37 (t, 2H, CH ); 2.43 (s, 6H,
2 2
N(CH ) ); 2.64 (m, 4H, CH ); 3.44 (t, 2H, CH ); 3.65
3
2
2
2
13
(m, 8H, ClCH CH N); 6.58–8.16 (m, 10H, Ar-H).
C
2
2
NMR (DMSO-d , l ppm): 25.99 (CH ); 27.41 (CH );
3
6
2
2
3.61 (CH ); 34.40 (CH ); 36.40 (CONHCH ); 40.54
2 2 2
(
N(CH ) ); 39.18; 53.53 (ClCH CH NH ); 56.72
3 2 2 2 2
6.1.1. General procedure
(CH N(CH ) ); 112.23; 112.27; 119.11; 119.60; 120.06;
2 3 2
4
-[p-[Bis(2-chloroethylamino)phenyl]butyric chloride
123.21; 123.36; 123.68; 123.81; 124.12; 124.53; 129.13;
129.21; 129.48; 129.64; 130.56; 134.87; 135.21; 135.46;
138.61; 138.70; 144.51 (Ar); 167.30; 167.39; 168.70;
173.31 (4×CONH). Anal. Found: C, 61.31; H, 6.06; Cl,
12.58; N, 10.07. Calc. for C H N O Cl HCl: C, 61.92;
was prepared by dissolving chlorambucil (140 mg, 0.46
mmol) in anhydrous tetrahydrofuran (20 mL) and oxa-
lyl chloride (3 mL), and warming to mild reflux under a
drying tube for 2 h. The excess oxalyl chloride and
solvent were removed under reduced pressure and the
residue co-evaporated with a dry dichloromethane (5
mL, twice). The above acid chloride dissolved in a dry
dichloromethane (20 mL) was added dropwise to a
chilled (0°C) solution of the aminoamide 1–4 (0.42
mmol) in dry pyridine (20 mL) with 4-(dimethyl-
amino)pyridine (DMAP) under stirring. The reaction
mixture was kept at 0°C for an additional 15 min, then
stirred at room temperature (20 h). The reaction mixture
was concentrated under a reduced pressure to a brown
foam. The crude product was purified by column chro-
matography (silica gel) with a methanol gradient (1%,
then 1% increase every 100 mL) in chloroform as eluent.
After removal of the solvent, the pure chlorambucil
derivatives 5–8 were obtained as glaze solids.
41
47
6
4
2
H, 6.08; Cl, 13.38; N, 10.57%.
6.1.1.3. 5-{5-[4-[Bis(2-chloroethyl)aminophenyl]-
butyramido]-2-methoxybenzamido}-N-[3-
(dimethvlamino)propylo]-2-methoxybenzeno-1-
carboxamide hydrochloride (7)
1
Yield: 0.14 g (42.7%). H NMR (CDCl /CD OD, l
3
3
ppm): 1.87 (m, 2H, CH ); 2.08 (m, 2H, CH ); 2.26 (t,
2
2
2H, CH ); 2.48 (t, 2H, CH ); 2.75 (s, 6H, N(CH ) ); 3.18
2
2
3 2
(t, 2H, CH ); 3.54 (t, 2H, CH ); 3.84 (s, 8H,
2
2
ClCH CH N); 3.92 2× (s, 3H, OCH ); 6.65–8.00 (m,
2
2
3
13
10H, Ar-H). C NMR (CDCl /CD OD, l ppm): 24.57
3
3
(CH ); 27.10 (CH ); 33.86 (CH ); 36.13 (CH ); 36.22
2
2
2
2
(CONHCH2);
42.56
(N(CH3)2);
40.29;
53.29
(ClCH CH N); 55.22 (CH N(CH ) ); 56.01; 56.24 (2×
2
2
2
3 2