Letters in Drug Design & Discovery, 2005, 2, 55-61 55

Synthesis and Biological Activity of C-8 Fluoroaryl Substituted Pyrimidine

Linked-Pyrrolobenzodiazepine Conjugates
Ahmed Kamal* , K. Laxma Reddy, V. Devaiah, N. Shankaraiah, M. Shiva Kumar and
G. Suresh Kumar Reddy

Biotransformation Laboratory, Division of Organic Chemistry, Indian Institute of Chemical Technology,

Hyderabad 500007, India
Received August 13, 2004: Accepted September 13, 2004

Abstract: Synthesis of C-8 fluoroaryl substituted pyrimidine linked-PBD conjugates are described. The
compounds are prepared with varying degrees of linker length in order to probe the structural requirements for
optimal in vitro antitumour activity. These compounds have been tested against a panel of 60 human cancer cell
lines, and it is demonstrated that compound 5b with four carbon spacer exhibited promising in vitro anticancer
activity in comparison to the other analogues (5a and 5c).
Keywords: Pyrrolobenzodiazepines, pyrimidines, cytotoxicity, antitumour activity.

INTRODUCTION engaged in the last few years in the structural modifications

Pyrrolo[2,1-c][1,4]benzodiazepines (PBDs) are of
considerable current interest due to their ability to recognize
and subsequently form covalent bonds to specific base
sequences of double stranded DNA. Such agents have
potential therapeutic targets, in the therapy of genetic based
diseases (eg. cancer) and validation of DNA sequences [1-3].
The pyrrolo[2,1-c][1,4]benzodiazepines (PBDs) are a family
of DNA interactive antitumour antibiotics derived from
Streptomyces species that include anthramycin, tomaymycin,
sibiromycin and DC-81 [4]. The cytotoxicity and
antitumour activity of these compounds are attributed to
their property of forming a covalent bond between the C11
position of DNA and the N2 amino group of a guanine base,
and they usually span approximately three base pairs with a
preference of Pu-G-Pu [5]. Recently, a number of structurally
modified PBDs have been synthesized and examined for their
biological properties, particularly their anticancer potential
[6] including the novel dimers of PBD [7]. We have recently
designed and synthesized non-cross-linking mixed imine-
amide PBD dimers (2) that have significant DNA-binding
ability and potent antitumour activity [8]. Pyrimidine ring
systems are reported to have shown a variety of biological
activities such as antitumour [9a], anti-allergic [9b],
antipyretic [9c], anti-inflammatory antiparasitic [9d] and
antibacterial [9e]. Substituted 2,4-bisanilino pyrimidines (3)
and 2,4-diamino-N4,6-diarylpyrimidines (4) have been
identified as potential agents for cancer treatment (Fig. 1)
[10,11]. Moreover, in literature a number of antiviral and
antitumour agents have been developed in which fluorine
substitution has played a key role in their biological
activity. The importance of fluorine substitution in
pharmaceutical development is evident in a large number of
fluorinated compounds that have been approved by food and
drug administration (FDA) as drugs [12]. We have been
*Address correspondence to this author at the Biotransformation
Laboratory, Division of Organic Chemistry, Indian Institute of Chemical
Technology, Hyderabad 500007, India; Tel: +91-40-27193157; Fax: +91-
40-27193189, 27160512; E-mail: ahmedkamal@iict.res.in,
ahmedkamal@ins.iictnet.com
[13] and the development of new synthetic strategies [14] for
the PBD based ring system. In view of this and as a
continuation of previous efforts carried out in our laboratory,
pyrimidine moiety bearing 2-methyl and 4-(p-fluoro)aryl
substituents have been linked at C-8 position of the PBD
ring system with different alkyl spacers to explore their in
vitro anticancer activity [15].
RESULTS AND DISCUSSION
The compounds 5a-c have been prepared from the (2S)-N-
(4-benzyloxy-5-methoxy-2-nitrobenzoyl)-pyrrolidine-2-carbo-
xaldehydediethyl thioacetal (8). This compound has been
prepared by literature method [16], which upon
debenzylation gives (2S)-N-(4-hydroxy-5-methoxy-2-
nitrobenzoyl)-pyrrolidine-2-carboxaldehydediethyl thioacetal
(9). The main precursors 10a-c have been obtained by
coupling of 9 and 4-(4-fluorophenyl)-6-bromoalkyloxy-2-
methylpyrimidine (7a-c). These upon nitro reduction
followed by deprotection of thioacetal group afford the target
compounds 5a-c (Scheme 2). The precursors 7a-c have been
prepared by monoalkylation of compound 6 with
dibromoalkanes (Scheme 1), and whereas compound 4-(4-
fluorophenyl)-6-hydroxy-2-methylpyrimidine (6) has been
prepared by literature method [17].
Cytotoxicity
Compounds 5a-c have been evaluated for in vitro
antitumour screening program of the National Cancer
Institute against sixty human tumour cell lines derived from
nine cancer types, leukemia, non-small lung, colon, CNS,
melanoma, ovarian, renal, prostate and breast cancer. For
each compound, dose response curves for each cell line were
measured at a minimum of live concentrations at 10 fold
dilutions in protocol of 48 h continuous drug exposure, and
a sulfurhodamine B (SRB) protein assay is used to estimate
cell viability or growth. The concentration causing 50% cell
growth inhibition (GI50), total cell growth inhibition (TGI,
0% growth), and 50% cell death (LC50 -50% growth)

1570-1808/05 $50.00+.00 © 2005 Bentham Science Publishers Ltd.

56 Letters in Drug Design & Discovery, 2005, Vol. 2, No. 1 Kamal et al.

10
9 N 11
HO
H1
8 11a
H3CO 7 N 2
6 5 4
O 3

DC-81

O H
N O (CH2) n O N
H H

N OCH3 H3CO N

O O
n = 3-5, 8
Imine-aimde dimers (2)

NH2
N

R1 N N
HN N NH
NH
X
OH R3
R2
Me 2N O
3 X= H, F, Cl, CF 3 4

CH3

N N

O (CH2) n O N
H
F
H3CO N
n = 3-5 O
5a-c

Fig. (1). Synthesis of pyrrolobenzodiazepines and pyrimidine based analogues.

compared with the control was calculated. The GI50 values
concentration for killing 50% of the cells) for compounds 5a-
c against the 60 different cell lines are given in Table 1.
Compounds 5a-c possess cytotoxic potency against
many cell lines. Compound 5b exhibits a wide spectrum of
activity against 60 human cancer cell lines in nine cell line
panel with GI50 value of <1 µM. The average GI50 value of
compound 5b against leukemia cancer RFMI-8226, CCRF-
CEM and MOLT-4 are 0.01, 0.08 and 0.08 µM
respectively. In repeated testing of compound 5b (Fig. 2) the
cytotoxic activity is further confirmed in most of the cell
lines particularly GI50 value is 0.002 µM in case of colon
cancer HCC-2998. Compounds 5a and 5c exhibit cytotoxic
potency against number of cell lines in the nine panels, with
the GI50 value range of <20 µM. Compound 5c exhibits
cytotoxic potency in the leukemia which CCRF-CEM,

CH3 CH3

N N N N
i
OH O (CH2) n Br

F 6 F 7a-c n = 3-5

Scheme 1. Teagents and conditions: (i) dibromoalkanes, K2 CO 3 , acetone, 48h, reflux, 92-95%.
Synthesis and Biological Activity of C-8 Fluoroaryl Substituted Pyrimidine Letters in Drug Design & Discovery, 2005, Vol. 2, No. 1 57

BnO NO2 CH(SEt)2 HO NO2 CH(SEt)2

N
H3 CO N
H3 CO
O O
8 9

CH3 ii

N N

O (CH2) n O NO2 CH(SEt) 2

F N
H3CO
10a-c n = 3-5
O

CH3 iii

N N

O (CH2) n O NH2 CH(SEt) 2

F N
H3CO
11a-c n = 3-5
O
iv

5a-c

Scheme 2. Reagents and condition: (i) EtSH-BF3 OEt 2 , CH 2 Cl 2 , 12 h , rt, 75%; (ii) 7a-c, K2 CO 3 , acetone 48 h, reflux, 93-95%; (iii)
SnCl 2 . 2H 2 O, MeOH, reflux, 4 h 80-85%; (iv) HgCl2 -CaCO 3 , CH3 CN-H2 O, 12 h, rt, 68-70%.

EPMI-8226 and SR cell lines are 2.86, 2.21 and 2.98 µM
respectively. In the non-small cell lung cancer, the growth of
HOP-92 cell line is affected by compound 5c with GI 50 value
0.37 µM. The in vitro cytotoxicity (IC50) for the naturally
occurring DC-81 [18] is 0.38, 0.33 and 0.1 µM in L1210,
PC6 and CH1 cell lines respectively.
binding has the potential to inhibit the BamH1 cleavage
activity. The study has been carried out to determine the
ability of 5a-c, which inhibits the DNA linearization by
BamH1. It is observed that for the compound 5b the
inhibition of BamH1 cleavage is in agreement with the DNA
binding affinity as determined by thermal denaturation.

DNA Interactions: Thermal Denaturation Studies CONCLUSIONS

The DNA binding ability of these compounds has also
been investigated by thermal denaturation studies using calf
thymus (CT) DNA at pH 7.0, incubated at 37 °C. It is
observed that compound 5b elevates the helix melting
temperature of the CT-DNA to 1.4 °C after incubation of 18
h while compounds 5a and 5c have not exhibited any
significant ∆T m value. In the same experiment the naturally
occurring DC-81 exhibits a ∆T m of 0.7 °C.
RED100-restriction Endonuclease Digestion assay
Many studies have employed restriction endonuclease
inhibition to confirm the relative binding affinity of DNA-
interactive small molecule ligands [19]. A quantitative
restriction enzyme digest (RED100) assay has been developed
in which the inhibition of DNA cleavage by BamH1 is used
to probe the DNA binding capability of PBD monomers
[20]. The BamH1 cleavage sequence G↓GATCC overlaps
with several favored PBD binding sites suggesting ligand
In conclusion, new fluoroaryl substituted pyrimidine-
PBD conjugates have been synthesized that exhibit in vitro
anticancer activity in many cancer cell lines and has potential
for further development.
EXPERIMENTAL
Reaction progress was monitored by thin-layer
chromatography (TLC) using GF254 silica gel with
fluorescent indicator on glass plates. Visualization was
achieved with UV light and iodine vapor unless otherwise
stated. Chromatography was performed using Acme silica
gel (100-200 and 60-120mesh). The majority of reaction
solvents were purified by distillation under nitrogen from the
indicated drying agent and used fresh: dichloromethane
(calcium hydride), tetrahydrofuran (sodium benzophenone
ketyl), methanol (magnesium methoxide), acetonitrile
(calcium hydride).
58 Letters in Drug Design & Discovery, 2005, Vol. 2, No. 1 Kamal et al.

Table 1. In Vitro Anticancer Activity of Compound 5a-c in Selected Cancer Cell Lines

Cancer panel/cell line GI50 ( M)

5a 5b 5b (repeated) 5c

Leukemia
CCRF-CEM 15.8 0.08 0.20 2.8
HL-60 (TB) 24.9 0.32 0.24 -
K-562 - 0.24 0.29 4.67
MOLT-4 14.3 0.08 0.13 3.23
RPMI-8226 26.0 0.01 0.19 2.21
SR 10.3 0.16 - 2.98

Non-small cell lung

HOP-62 - 0.26 0.78 25.0
HOP-92 27.3 0.11 - 0.3
NCI-H226 15.2 0.54 - -
NCI-H23 15.1 0.65 0.33 -
NCI-H322M 26.5 0.11 - 28.7
NCI-H460 - 0.20 0.85 12.9
NCI-H522 14.6 0.14 0.30 4.19

Colon
COLO 205 29.1 0.53 0.22 18.6
HCT-116 16.9 0.64 - 28.0
HCT-15 12.6 0.55 0.89 17.6
HT29 23.1 0.38 0.51 20.4
KM12 19.3 0.66 0.79 23.3
SW-620 22.8 0.85 0.85 10.3
HCC-2998 - - 0.002 -

CNS
SF-268 17.2 0.56 - 10.5
SF-295 16.4 037 0.24 25.9
SF-539 16.6 0.94 0.91 -
SNB-19 20.1 0.86 0.55 21.4
SNB-75 19.1 0.45 - -
U251 14.8 0.19 0.20 12.1

Melanoma
LOX EMVI 13.6 0.09 0.27 11.1
MALME-3M 12.8 0.67 1.00 23.3
M14 15.5 0.74 0.61 20.7
SK-MEL-2 17.2 0.76 0.94 23.4
SK-MEL-5 15.9 0.62 0.56 1.24
UACC-257 17.2 0.68 0.69 -
UACC-62 14.5 0.21 0.67 14.5

Ovarian
IGROVI 19.2 0.29 - 7.74
OVCAR-3 15.2 0.43 0.26 14.6
OVCAR-4 33.5 0.59 0.11 16.5
OVCAR-5 24.5 0.17 0.42 -
OVCAR-8 24.6 0.86 0.11 13.9

Renal
786-0 16.8 0.27 0.86 20.1
ACHN 16.1 0.17 0.21 14.2
CAKI-1 17.5 0.25 0.38 17.4
RXF393 20.2 0.36 0.50 10.8
SN12C 16.5 0.37 - 19.6
TK-10 18.1 0.14 0.34 24.9
UO-31 17.7 0.26 0.94 8.0

Prostate
PC-3 - 0.62 0.91 11.1
DU-145 9.0 0.20 0.25 15.9

Breast
MCF7 29.4 0.33 0.75 5.58
MDA-MB-231/ATCC 13.8 0.66 - 15.6
HS 57BT
MDA-MB-435 24.7 0.84 0.67 22.6
BT-549 15.2 0.63 0.54 20.5
T-47D - 0.61 0.28 10.8
25.8 0.30 - 6.27
Synthesis and Biological Activity of C-8 Fluoroaryl Substituted Pyrimidine
Fig. (2). Log 10 GI50 data from the NCI 60 cell line screen for compound 5b and its repeat testing data.
1H NMR spectra were recorded on Varian Gemini 200
MHz spectrometer using tetramethyl silane (TMS) as an
internal standard. Chemical shifts are reported in parts per
million (ppm) down field from tetramethyl silane. Spin
multiplicities are described as s (singlet), d (doublet), t
(triplet), q (quartet), m (multiplet). Coupling constants are
reported in Hertz (Hz). Low resolution mass spectra were
recorded on VG-7070H Micromass mass spectrometer at 200
oC, 70 eV with trap current of 200 A and 4 KV
acceleration voltage. Optical rotations are measured on
Horiba, High Sensitive Polarimeter, SEPA-300.
3-Bromopropyl-6-(4-fluorophenyl)-2-methyl-4-pyrimidyl
ether (7a)
A solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol (6)
(1.0 g, 4.90 mmol), 1,3-dibromopropane (2.475 g, 12.25
mmol) and K2CO3 (2.03 g, 14.7 mmol) in dry acetone (40
Letters in Drug Design & Discovery, 2005, Vol. 2, No. 1 59
mL) was refluxed for 48 h. After the completion of reaction
as indicated by TLC, ethyl acetate-hexane (2:8), the reaction
mixture was poured on to the water and then extracted with
ethyl acetate. Evaporation of the organic layer gave the crude
product, which was further purified by column
chromatography on silica gel eluting with ethyl acetate-
hexane (5:95) gave the pure 7a (1.465g, 92%): 1H NMR
(CDCl 3) δ 2.20-2.35 (m, 2H), 2.65 (s, 3H), 4.25 (t, 2H, J =
4.8 Hz), 4.50 (t, 2H, J = 4.5 Hz), 6.70 (s, 1H), 7.08-7.16
(m, 2H), 7.90-8.05 (m, 2H); MS (EI) m/z 325. Anal. calcd
for C14H14BrFN2O: C, 51.71; H, 4.33; N, 8.61. Found: C,
51.67; H, 4.23; N, 8.49.
4-Bromobutyl-6-(4-fluorophenyl)-2-methyl-4-pyrimidyl
ether (7b)
The compound 7b was prepared according to the method
described for the compound 7a employing compound 6 (1.0
60 Letters in Drug Design & Discovery, 2005, Vol. 2, No. 1
g, 4.90 mmol), 1,4-dibromobutane (2.65 g, 12.25 mmol)
and K2CO3 (2.03 g, 14.70 mmol) to afford the crude
product, which was purified by column chromatography (4%
ethyl acetate-hexane) to afford the compound 7b (1.578g,
95%): 1H NMR (CDCl 3) δ 1.90-2.15 (m, 4H), 2.65 (s, 3H),
3.50 (t, 2H, J = 4.7 Hz), 4.5 (t, 2H, J = 4.9 Hz), 6.85 (s,
1H), 7.1-7.2 (m, 2H), 8.0-8.1 (m, 2H); MS (EI) m/z 339.
Anal. calcd for C15H16BrFN2O: C, 53.11; H, 4.75; N, 8.26.
Found: C, 53.02; H, 4.65; N, 8.07.
5-Bromopentyl-6-(4-fluorophenyl)-2-methyl-4-pyrimidyl
ether (7c)
The compound 7c was prepared according to the method
described for the compound 7a employing compound 6 (1.0
g, 4.90 mmol), 1,5-dibromopentane (2.82 g, 12.25 mmol)
and K2CO3 (2.03 g, 14.70 mmol) to afford the crude
product, which was purified by column chromatography (3%
ethyl acetate-hexane) to afford the compound 7c (1.608 g,
93%): 1H NMR (CDCl3) δ 1.50-1.60 (m, 2H), 1.70-1.80
(m, 2H), 1.90-2.00 (m, 2H), 2.60 (s, 3H), 3.40 (t, 2H, J =
5.1 Hz), 4.40 (t, 3H, J = 5.5 Hz), 6.7 (s, 1H), 7.10-7.12 (m,
2H), 7.95-8.05 (m, 2H); MS (EI) m/z 353. Anal. calcd for
C 16H18BrFN2O: C, 54.40; H, 5.14; N, 7.93. Found: C,
54.22; H, 5.05; N, 7.67.
(2S)-N-[{4-[6'-(4" -Fluorophenyl)-2'-methylpyrimidine-4'-
yloxy]-propoxy}-5-methoxy-2-nitrobenzoyl]pyrrolidine-
2-carboxaldehyde diethyl thioacetal (10a)
A solution of 7a (1.2 g, 3.69 mmol), A solution of (2S)-
N-(4-hydroxy-5-methoxy-2-nitrobenzoyl)pyrrolidine-2-
carboxaldehydediethylthioacetal (9) (1.472 g, 3.69 mmol)
and K 2CO3 (1.53 g, 11.07 mmol) in dry acetone (20 mL)
was refluxed for 48 h. After the completion of reaction as
indicated by TLC, ethyl acetate, the reaction mixture was
poured on to the water and then extracted with ethyl acetate.
Evaporation of the organic layer gave the crude product,
which was further purified by column chromatography on
silica gel eluting with ethyl acetate-hexane (1:3) gave the
pure 10a (2.186g, 92%): 1H NMR (CDCl3) δ 1.20-1.40 (m,
6H), 1.70-2.42 (m, 6H), 2.65 (s, 3H), 2.68-2.90 (m, 4H),
3.15- 3.30 (m, 2H), 3.95 (s, 3H), 4.25-4.35 (m, 2H), 4.58-
4.78 (m, 2H), 4.65-4.80 (m, 1H), 4.85 (d, 1H, J = 4.2 Hz),
6.78 (s, 1H), 6.90 (s, 1H), 7.10-7.20 (m, 2H), 7.70 (s, 1H),
8.00-8.10 (m, 2H); FABMS 645 (M+H)+. . Anal. calcd for
C 31H37FN4O6S 2: C, 57.75; H, 5.78; N, 8.69. Found: C,
57.63; H, 5.65; N, 8.48.
(2S)-N-[{4-[6'-(4" -Fluorophenyl)-2'-methylpyrimidine-4'-
yloxy]butoxy}-5-methoxy-2-nitrobenzoyl]pyrrolidine-2-
carboxaldehyde diethyl thioacetal (10b)
The compound 10b was prepared according to the
method described for the compound 10a employing
compound 9 (1.532 g, 3.83 mmol), 7b (1.3 g, 3.83 mmol)
and K2CO3 (1.585 g, 11.47 mmol) to afford the crude
product, which was purified by column chromatography
(30% ethyl acetate-hexane) to afford the compound 10b
(2.394g, 95%): 1H NMR (CDCl3) δ 1.20-1.50 (m, 8H),
1.90-2.30 (m, 8H), 2.65 (s, 3H), 2.70-2.90(m, 4H), 3.20-
3.35 (m, 2H). 3.92 (s, 3H), 4.05-4.25 (m, 4H), 4.45-4.55
(m, 2H), 4.65-4.75 (m, 1H), 4.85 (d, 1H. J = 4.85 Hz),
Kamal et al.
6.78 (s, 1H), 6.82 (s, 1H), 7.10-7.20 (m, 2H), 7.65 (s, 1H),
7.95-8.10 ( m, 2H); FABMS 659 (M+H)+. . Anal. calcd for
C 32H39FN4O6S 2: C, 58.34; H, 5.97; N, 8.50. Found: C,
58.23; H, 5.69; N, 8.49.
(2S)-N-[{4[6'-(4" -Fluorophenyl)-2'-methylpyrimidine-4'-
yloxy]pentoxy}-5-methoxy-2'-nitrobenzoyl]pyrrolidine-2-
carboxaldehydediethylthioacetal (10c)
The compound 10c was prepared according to the
method described for the compound 10a employing
compound 9 (1.59 g, 3. 97 mmol), 7c (1.4 g, 3.97 mmol)
and K2CO3 (1.64 g, 11.898 mmol) to afford the crude
product, which was purified by column chromatography
(27% ethyl acetate-hexane) to afford the compound 10c
(2.507g, 94%): 1H NMR (CDCl3) δ 1.20-1.50 (m, 6H),
1.60-2.40 (m, 10H), 2.64 (s, 3H), 2.70-2.90 (m, 4H), 3.20-
3.30 (m, 2H), 3.95 (s, 3H), 4.50 (t, 2H, J = 6.25 Hz), 4.45
(t, 2H, J = 6.5 Hz), 4.62-4.75 (m, H), 4.85 (d, 1H, J = 4.2
Hz), 6.78 (s, 1H), 6.82 (s, 1H), 7.05-7.20 (m, 2H), 7.65 (s,
1H), 7.95-8.1 (m, 2H); FABMS 939 (M+H)+. . Anal. calcd
for C33H41FN4O6S 2: C, 58.91; H, 6.14; N, 8.32. Found: C,
58.83; H, 5.98; N, 8.22.
(2S)-N-[{4-[6'-(4" -Fluorophenyl)-2'-methylpyrimidine-4'-
yloxy]propoxy}-5-methoxy-2-aminobenzoyl]pyrrolidine
-2-carboxaldehyde diethyl thioacetal (11a)
The 10a (0.5 g, 0.78 mmol) was dissolved in methanol
(10 mL) and added SnCl2.2H2O (0.873 g, 3.88 mmol) was
refluxed for 1.5 h. The reaction mixture was then carefully
adjusted to pH 8 with saturated NaHCO3 solution and then
extracted with ethyl acetate (3x30 mL). The combined
organic phase was dried over Na2SO4 and evaporated under
vacuum to afford the crude 11a
(2S)-N-[{4-[6'-(4" -Fluorophenyl)-2'-methylpyrimidine-4'-
yloxy]butoxy}-5-methoxy-2-aminobenzoyl]pyrrolidine-2-
carboxaldehyde diethyl thioacetal (11b)
The compound 11b was prepared according to the
method described for the compound 11a employing the
compound 10b (0.5 g, 0.76 mmol) and SnCl2.2H2O (0.855
g, 3.80 mmol) to afford the amino diethyl thioacetal 11b.
(2S)-N-[{4-[6'-(4" -Fluorophenyl)-2'-methylpyrimidine-4'-
yloxy]pentoxy}-5-methoxy-2-aminobenzoyl]pyrrolidine-
2-carboxaldehyde diethyl thioacetal (11c)
The compound 11c was prepared according to the
method described for the compound 11a employing the
compound 10c (0.5 g, 0.74 mmol) and SnCl2.2H2O (0.84
g, 3.72 mmol) to afford the amino diethyl thioacetal 11c.
7-Methoxy-8-[6'-(4" -fluorophenyl)-2 '-methylpyrimidine-
4'-yloxy]propoxy-(11aS)-1,2,3,11a-tetrahydro-5H-
pyrrolo[2,1-c][1,4]benzodiazepin-5-one (5a)
A solution of 11a (0.3 g, 0.50 mmol), HgCl2 (0.3 g,
1.10 mmol) and CaCO3 (0.12 g, 1.20 mmol) in
CH3CN/H2O (3:1, 15 mL) was stirred at room temperature
for 12 h until TLC (ethyl acetate), indicates complete loss of
starting material. Then organic layer is evaporated in
Synthesis and Biological Activity of C-8 Fluoroaryl Substituted Pyrimidine
vacuum and the residue is diluted with ethyl acetate. To this
saturated NaHCO 3 was added slowly at room temperature
and the mixture is filtered through celite and washed with
ethyl acetate. The filterate is evaporated in vacuum to get
crude 5a, which was further purified by column
chromatography on silica gel eluting first with ethyl acetate
to remove traces of mercuric salts and further eluted with
ethyl acetate-methanol (9:1) (0.166g, 68%):1H NMR
(CDCl 3) δ 1.90-2.15 (m, 4H), 2.25-2.4 (m, 2H), 2.65 (s,
3H), 3.5-3.8 (m, 3H), 3.95 (s, 3H), 4.00-4.20 (m, 2H),
4.45-4.55 (m, 2H), 6.78 (s, 1H), 6.8 (s, 1H), 7.10-7.20 (m,
2H), 7.50 (s, 1H), 7.50 (d, 1H, J = 4.3 Hz), 7.95-8.20 (m,
2H); FABMS: 491 (M+H)+. . Anal. calcd for C 27H27FN4O4:
C, 66.11; H, 5.55; N, 11.42. Found: C, 65.94; H, 5.38; N,
11.34.; [α]26D +213 (c 0.0075, MeOH)
7-Methoxy-8-[6'-(4" -fluorophenyl)-2 '-methylpyrimidine-
4'-yloxy]butoxy-(11aS)-1,2,3,11a-tetrahydro-5H-
pyrrolo[2,1-c][1,4]benzodiazepin-5-one (5b)
The compound 5b was prepared according to the method
described for the compound 5a employing 11b (0.3 g, 0.50
mmol), HgCl2 (0.3 g, 1.10 mmol) and CaCO3 (0.12 g, 1.20
mmol) to afford 5b (0.176g, 70%): 1HNMR (CDCl3) δ
1.92-2.42 (m, 8H), 2.63 (s, 3H), 3.502-3.80 (m, 2H), 3.92
(s, 3H), 4.14-4.28 (m, 3H), 4.45-4.55 (m, 2H), 6.76 (s,
1H), 6.80 (s, 1H) 7.10-7.30 (m, 2H), 7.58 (s, 1H), 7.65 (d,
1H), 7.95-8.10 (m, 2H); FABMS: 505 (M+H)+. . Anal.
calcd for C28H29FN4O4: C, 66.65; H, 5.79; N, 11.10.
Found: C, 66.54; H, 5.67; N, 11.08.; [α]26D +86 (c 0.001,
MeOH)
7-Methoxy-8-[6'-(4" -fluorophenyl)-2 '-methylpyrimidine-
4'-yloxy]pentoxy-(11aS)-1,2,3,11a-tetrahydro-5H-
pyrrolo[2,1-c][1,4]benzodiazepin-5-one (5c)
The compound 5c was prepared according to the method
described for the compound 5a employing 11b (0.3 g, 0.47
mmol), HgCl2 (0.280 g, 1.03 mmol) and CaCO3 (0.11 g,
1.10 mmol) to afford 5c (0.170g, 70%): 1HNMR (CDCl3) δ
1.6-1.76 (m, 3H), 1.82.2.20 (m, 5H), 2.30-2.40 (m, 2H),
2.65 (s, 3H), 3.60-3.80 (m, 3H), 3.97 (s, 3H), 4.10-4.20
(m, 3H) 4.40-4.50 (m, 2H), 6.84 (s, 1H), 6.86 (s, H), 7.10-
7.25 (m, 2H), 7.55 (s, 1H), 7.70 (d, 1H, J = 4.4 Hz), 8.0-
8.1 (m, 2H); FABMS: 519 (M+H)+. .; Anal. calcd for
C 29H31FN4O4: C, 67.17; H, 6.03; N, 9.26. Found: C,
67.05; H, 5.59; N, 9.09.; [α]26D +259 (c 0.07, MeOH).
ACKNOWLEDGEMENTS
We thank the National Cancer Institute, Maryland for the
in vitro anticancer assay in human cancer cell lines. We are
also grateful to CSIR, New Delhi for the award of research
fellowships to K. L. R., V. D., N. S. and G. S. K. R.
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