Synthesis and antiprotozoal activity of mono- and bis-uracil isatin conjugates against the human pathogen Trichomonas vaginalis

Article abstract of DOI:10.1016/j.bmc.2015.04.075

A library of mono- and bis-uracil isatin conjugates were synthesized and subjected for the assessment of their in vitro activity against the protozoal pathogen Trichomonas vaginalis. The structure activity studies (SAR) revealed that the bis-uracil-isatin based conjugates were more effective than their corresponding mono conjugates in inhibiting the growth of T. vaginalis at approximately 10 μM with no visual effect on mammalian cells at the same concentration.

Full text of DOI:10.1016/j.bmc.2015.04.075

Bioorganic & Medicinal Chemistry xxx (2015) xxx–xxx
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com/locate/bmc
Synthesis and antiprotozoal activity of mono- and bis-uracil isatin
conjugates against the human pathogen Trichomonas vaginalis
Kewal Kumar a, Nicole Liu b, Donald Yang b, Daniel Na b, John Thompson b, Lisa A. Wrischnik b,
Kirkwood M. Land b, , Vipan Kumar a,
⇑
⇑
a Department of Chemistry, Guru Nanak Dev University, Amritsar 143005, India
b Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, USA
a r t i c l e i n f o
a b s t r a c t
Article history:
A library of mono- and bis-uracil isatin conjugates were synthesized and subjected for the assessment of
their in vitro activity against the protozoal pathogen Trichomonas vaginalis. The structure activity studies
(SAR) revealed that the bis-uracil-isatin based conjugates were more effective than their corresponding
Received 24 January 2015
Revised 22 April 2015
Accepted 24 April 2015
Available online xxxx
mono conjugates in inhibiting the growth of T. vaginalis at approximately 10
l
M with no visual effect on
mammalian cells at the same concentration.
Published by Elsevier Ltd.
Keywords:
Uracil-isatin conjugates
Trichomonas vaginalis
Cytotoxicity
Structure activity relationship
1. Introduction
development of novel, non-cytotoxic and efficient scaffolds against
trichomoniasis.12–16
Human trichomoniasis is caused by Trichomonas vaginalis. This
protozoal infection is the most common chronic non-viral infection
and is responsible for approximately 248 million cases each year,
being more prevalent than either chlamydia or gonorrheal infec-
tions.1,2 Trichomoniasis has a ubiquitous distribution especially
in under-developed countries with a combination of poor sanita-
tion, hygiene and tropical climate. Malodorous vaginal discharge,
vulval irritation, inflammation and cervical micro hemorrhages
are the common symptoms which may lead to serious health com-
plications such as infertility, preterm delivery, low birth weight
and cervical cancer.3,4 Despite its high prevalence, trichomoniasis
is neglected compared to other sexually transmitted diseases and
was long regarded as a self-clearing infection.5,6 The importance
of trichomoniasis has further increased dramatically because of
its strong association with the increased risk of acquisition and
transmission of human immunodeficiency virus (HIV).7–10
Metronidazole has been the main-stay for the treatment of tri-
chomoniasis for more than 40 years,11 however recent disclosures
of genotoxicity, gastric mucus irritation and developed of clinical
resistant isolates have provided strong impetus for the
Isatin (1H-indole-2,3-dione) is a chemical scaffold present
widely in both human and other mammalian tissues and is also
found to be a common structural motif in a variety of dyes, agro-
chemicals and pharmacologically active compounds.17 The syn-
thetic versatility of isatin makes it an ideal platform for
structural alteration and derivatization. This observation has been
confirmed by its enormous biological potential with myriad activ-
ities such as anticancer,18 antidepressant,19 anticonvulsant,20 anti-
fungal,21 anti-HIV,22 and anti-angiogenic properties.23 Further, it
can function as a potent inhibitor of DNA gyrase, the enzyme which
catalyzes the introduction of negative supercoils in a closed circu-
lar DNA.24 Note-worthy examples of 2-oxoindole analogues
include SU11248 (Sutent), a 5-fluoro-3-substituted isatin deriva-
tive, was approved by the FDA in 2006 for the treatment of
advanced renal carcinoma and gastrointestinal stromal tumors.25
C5- and C6-substituted isatin analogues were revealed to be selec-
tive inhibitors of monoamine oxidase B (MAOB) with 5-(4-phenyl-
butyl) isatin exhibiting the highest activity and being 18,500-fold
more potent than isatin.26
Uracils are also considered important scaffolds in drug discov-
ery with a wide range of biological activities, synthetic expediency
and ability to confer drug like properties to the compound libraries
appended on them at the N1, N3, C5 and C6 positions.27 Apart from
their antiviral and anti-tumor potential, uracil analogs also possess
herbicidal, insecticidal and bactericidal activities.28 Inhibition of a
⇑
Corresponding authors. Tel.: +1 209 946 7608; fax: +1 209 946 3022 (K.M.L.);
tel.: +91 183 2258802 09x3320; fax: +91 183 2258819 20 (V.K.).
E-mail addresses: KLAND@pacific.edu (K.M. Land), vipan_org@yahoo.com
(V. Kumar).
http://dx.doi.org/10.1016/j.bmc.2015.04.075
0968-0896/Published by Elsevier Ltd.
Please cite this article in press as: Kumar, K.; et al. Bioorg. Med. Chem. (2015), http://dx.doi.org/10.1016/j.bmc.2015.04.075

2
K. Kumar et al. / Bioorg. Med. Chem. xxx (2015) xxx–xxx
X
O
O
O
R
R
R
O
NaH, DMF
O
O
O
Br
Br
n
HN
NH
N
N
N
°
Br, 60 C
Br
H
H
n
1
O
n
Br
2
68-85%
X
O
Scheme 1. Synthesis of N-alkylbromo-isatins.
X
O
O
O
O
O
O
O
N
N
N
HN
N
N
N
R
n
n
Table 1
% age yields of synthesized N-alkylbromo-isatins 2
O
n
R
O
R
Compound
n
R
Yield (%)
Figure 1. General structures assigned to the target hybrids.
2a
2b
2c
2d
2e
2f
2g
2h
2i
2j
2k
2l
2m
2n
2o
2p
2q
2r
2
3
4
5
6
8
2
3
4
5
6
8
2
3
4
5
6
8
H
H
H
H
H
H
F
F
F
F
F
71
77
73
71
80
83
75
70
68
78
81
79
67
74
72
78
82
85
key step in viral replication pathways form the basis of their anti-
viral potential resulting in potent activities against HIV, hepatitis B
and C (HBV and HCV), the herpes viruses (HSV-1 and -2), Epstein
Barr virus (EBV), and human cytomegalovirus (HCMV).29 5-fluo-
rouracil is widely used as an anti-metabolism drug in the treat-
ment of malignancies including primarily colorectal, stomach and
breast cancer. Its poor selectivity, however, limits its therapeutic
relevance, resulting in high incidences of bone marrow, gastroin-
testinal tract and central nervous toxicity.30 Numerous alterations
of 5-FU structure have been accomplished to tackle these problems
ensuing in the development of 5-FU derivatives viz. floxuridine and
tegafur exhibiting better pharmacological and pharmacokinetic
properties.31 Molecular conjugate of 5-FU and Camptothecin
(CPT), a topoisomerase cytotoxic alkaloid, is another approach to
circumvent the problems associated with uracil analogues and
have shown to exhibit comparable or superior cytotoxicity to
irinotecan (semi-synthetic analogue of CPT and acts via topoiso-
merase 1 inhibition) with improved selectivity, efficacy and safety.
Recent studies have shown the synthesis of 1H-1,2,3-triazole-
tethered isatin-b-lactam conjugates and preliminary analysis of
their in vitro activity against T. vaginalis.32 The methodology was
further extended to the synthesis of b-amino alcohol tethered isa-
tin-b-lactam as well as piperazine-linked isatin-7-chloroquinoline
conjugates, along with their in vitro inhibitory activity against
T. vaginalis and Tritrichomonas foetus.33 In continuation of these
findings and in view of the reported anti-parasitic potential of ura-
cil, the present study expands the synthesis of mono- and bis-
uracil-isatin conjugates as shown in Figure 1 and their in vitro
activity against T. vaginalis. The chain length has been meticulously
altered so as to carefully analyze the structure–activity relation-
ship among the synthesized hybrids.
F
Cl
Cl
Cl
Cl
Cl
Cl
This chemical library of mono- and bis-uracil-isatin conjugates
was evaluated in a general inhibitory screen against the protozoal
pathogen Trichomonas vaginalis at 50 lM; and the results are sum-
marized in Table 3. As observed, the mono uracil-isatin conjugates
viz. 4a–r were not efficient in inhibiting the growth of T. vaginalis
and exhibited percentage growth inhibition in the range of
1.76–48.55%. The observed activity of the conjugates were found
to be independent of the nature of substituent at C-5 position of
isatin as well as the length of alkyl chain linker. Further, a closer
inspection of Table 3 revealed that bis-uracil-isatin conjugates
(5a–r) exhibited better percentage inhibition compared to mono-
uracil-isatin conjugates. Analyzing the effect of substituent at C-5
position of isatin ring among bis-conjugates revealed that the pres-
ence of electron withdrawing substituent viz. chloro and flouro
substantially improved the activity profiles compared to the un-
substituted isatins (R = H). Further, an increase in alkyl chain
length, introduced as a linker, improved the activity profile regard-
less to the nature of substituent at C-5 position of isatin ring as evi-
dent by conjugate 5f (R = H, n = 8), exhibiting 92.93% inhibition.
The scaffolds 5k (R = F, n = 6); 5l (R = F, n = 8); 5q (R = Cl, n = 6)
and 5r (R = Cl, n = 8) exhibited >90% inhibition confirming the role
of electron withdrawing halogen substituents and longer chain
length in improving the activity profiles of the synthesized conju-
gates. These conjugates were then serially diluted and re-tested on
parasite cultures.
Five potent synthesized conjugates viz. 5f, 5j, 5k, 5m and 5q
were further evaluated for their IC50 (lM) and compared with
the standard drug metronidazole, the only and current FDA
approved drug used for the treatment of Trichomonas; shown in
Table 4. The synthesized conjugates were not as active as the stan-
dard drug metronidazole. However, 5f and 5m represented the
most potent inhibitory activity in serial dilution. The IC50 values
of 5f and 5m were determined to be 9.86 and 9.79 lM respectively.
These IC50’s were then re-confirmed using the same assays and
standard error was determined to be within 10%. Mammalian cells
2. Results and discussion
C-5 substituted N-alkylbromo-isatins
2 (Scheme 1) were
synthesized via our previously reported protocol involving NaH
promoted reaction of isatin with dibromoalkanes at 60 °C. The pre-
cursors 2 were purified via column chromatography using 15:85
EtOH/Hexane as eluent and the % age yields of each substituent
have been shown in Table 1. The N-alkylated precursors were then
employed in the synthesis of desired mono- and bis-uracil-isatin
hybrids 4 and 5, respectively as shown in Scheme 2.34 It was
observed that the use of 1.0 mmol of uracil and 1.1 mmol of
N-alkylbromo-isatins 2 resulted in the formation of mono- and
bis-uracil-isatin hybrids 4 and 5 (8:2) respectively, while the use
of 1.0 mmol of uracil and the 2.2 mmol of C-5 substituted N-alkyl-
bromo-isatins exclusively resulted in the formation of bis-uracil-
isatin conjugates 5 (Table 2).
The mono- and di-alkylation of uracil in the present case could
be rationalized in terms of the acidity of the ionizable protons.35
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K. Kumar et al. / Bioorg. Med. Chem. xxx (2015) xxx–xxx
3
O
O
O
N
O
O
O
O
O
N
2
(1.1 mol)
N
N
n
+
HN
N
N
NaH (0.7 mol)
n
DMF, 60 C, 1h
°
O
n
R
R
O
O
R
O
14-20%
69-78%
4
5
HN
O
N
H
O
O
O
N
O
2
(2.0 mol)
NaH (2.0 mol)
3
N
N
DMF, 60 C, 2h
°
N
n
n
O
R
R
79-87%
5
Scheme 2. Synthesis of mono- and bis-uracil-isatin conjugates 4 and 5.
Table 2
% age yields of mono- and bis-uracil-isatin conjugates 4 and 5
were then tested with 5f and 5m at the same concentration and no
Compound
n
R
Yield (%)
Compound
n
R
Yield (%)
visual effects on morphology or cell count was observed.
4a
4b
4c
4d
4e
4f
4g
4h
4i
4j
4k
4l
4m
4n
4o
4p
4q
4r
2
3
4
5
6
8
2
3
4
5
6
8
2
3
4
5
6
8
H
H
H
H
H
H
F
F
F
F
F
73
78
71
75
69
72
72
76
75
74
78
73
71
73
75
75
74
76
5a
5b
5c
5d
5e
5f
5g
5h
5i
5j
5k
5l
5m
5n
5o
5p
5q
5r
2
3
4
5
6
8
2
3
4
5
6
8
2
3
4
5
6
8
H
H
H
H
H
H
F
F
F
F
F
80
79
86
84
81
79
84
87
83
86
87
80
82
85
81
84
80
82
3. Conclusion
A series of mono- and bis-uracil-isatin conjugates were synthe-
sized and assessed for their in vitro activity against the protozoal
pathogen T. vaginalis. SAR studies revealed that the bis-uracil-isatin
conjugates viz. 5f, 5k, 5l, 5q and 5r were more effective than their
corresponding mono-uracil-isatin counterparts with more than
90% inhibition at 50 lM. The most potent and non-cytotoxic conju-
gates of this group, 5f and 5m, had an optimum combination of
longer alkyl chain length (n = 2 for 5f and, 6 for 5m) and no or elec-
tron withdrawing-substituent at C-5 position of isatin, exhibited
F
F
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
an IC50 values of 9.86 and 9.79 lM respectively. These conjugates
may represent a novel scaffold for development of chemotherapy
for resistant cases of human trichomoniasis, and may represent
an effective strategy for new anti-protozoals against related anaer-
obic pathogens.
Table 3
Antiprotozoal inhibition of 4a–r and 5a–r against T. vaginalis at 50
4. Material and methods
4.1. Biological evaluation
l
M
Compound
Percentage inhibition
Compound
Percentage inhibition
4a
4b
4c
4d
4e
4f
4g
4h
4i
4j
4k
4l
4m
4n
4o
4p
4q
4r
37.77 11.04
30.13 16.27
41.48 17.72
26.56 18.64
19.58 15.62
32.23 17.37
32.38 7.62
26.68 2.92
20.95 11.05
35.80 3.80
37.01 18.19
22.05 13.15
14.04 14.03
4.39 4.38
5a
5b
5c
5d
5e
5f
5g
5h
5i
5j
5k
5l
5m
5n
5o
5p
5q
5r
10.46 1.37
0.54 0.54
18.28 18.28
4.30 4.30
67.45 8.30
92.93 7.08
47.23 5.29
44.54 7.98
37.00 0.64
86.69 9.27
100
97.47 2.53
82.46 0.40
78.07 0.70
57.89 2.72
74.56 1.45
90.35 0.18
96.49 0.92
4.1.1. In vitro protozoal parasite susceptibility assay
Protozoal parasites were cultured for 24 h at 37 °C. To perform
the initial susceptibility screens on T. vaginalis, compounds were
suspended in DMSO to obtain concentrations of 100
quots of these suspensions were diluted in 5 mL of TYM diamond’s
media to obtain a final concentration of 100 M. After 24 h, cells
lM; 5 lL ali-
l
were counted using a hemacytometer. Cell counts were normal-
ized to the DMSO controls, in order to allow direct comparison
and averaging of the various trials. These data sets were then trans-
formed using Prism Software, GraphPad, by taking the log of the
drug concentrations for the trials, and inputting this transform into
a log (inhibitor) versus response-variable slope regression option.
Within this non-linear regression, constraints were set to force
the maximum value (top) to 1 and the minimum value (bottom)
to 0. The slope was left variable, and then determined through
which regression was performed. The sample size consists of 4
independent trials carried out on four different days (to account
for possible variation in parasite culture). The assays were per-
formed in 15 mL culture tubes, with both wildtype parasites and
0.1% DMSO-only treated parasites serving as control tubes to nor-
malize for the effects of the solvent and in vitro conditions. After
24 h, cells were counted using a hemacytometer. The IC50 value
for compound 5q was determined by running assays of increasing
48.55 0.57
1.76 1.75
25.38 23.74
41.84 23.95
Table 4
Antiprotozoal inhibition of 4a–r and 5a–r against T. vaginalis
Compound
IC50 value (mM)
5f
5j
5k
5m
9.86
10.41
19.50
9.79
drug concentrations, 5–40
sis using Prism software from GraphPad. Calculated IC50 value of
lM, and performing a regression analy-
5q
10.26
0.72
Metronidazole
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4
K. Kumar et al. / Bioorg. Med. Chem. xxx (2015) xxx–xxx
5q was then re-confirmed by testing again using the same assay
described above.
150.8, 151.3, 158.7, 164.2, 183.8; HRMS: Calcd for C15H13N3O4
[M]+ 299.0906, found 299.0901; Anal. Calcd (%) for: C, 60.20; H,
4.38; N, 14.04, found: C, 60.12; H, 4.30; N, 14.16.
4.1.2. Mammalian cell cytotoxicity studies
HeLa cells were cultured in DMEM/penicillin streptomycin/10%
FBS, 5%CO2, 37 °C, and 60% confluency. Cells were treated with 5q
to a final concentration of 10 lM. DMSO was also used as a nega-
tive control. Cells were then incubated for 24 h, trypsinized with
0.25% Trypsin-EDTA, then counted using light microscopy and a
corpuscle counting chamber. Controls included untreated cells
and cells treated with 0.1% DMSO.
4.2.1.3. 1-[4-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-butyl]-
1H-indole-2,3-dione (4c).
Yield 71%; orange solid; mp
171–172 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.73–1.79 (m, 4H,
2ꢀCH2), 3.89–3.92 (m, 2H, –NCH2), 3.96–3.98 (m, 2H, –NCH2),
5.58 (d, J = 7.4 Hz, 1H, olefinic H), 7.11–7.15 (m, 1H, –ArH), 7.26
(d, J = 8.1 Hz, 1H, –ArH), 7.63 (d, J = 7.4 Hz, 1H, olefinic H),
7.66–7.69 (m, 2H, –ArH), 11.21 (s, 1H, –NH–, exchangeable with
D2O); 13C NMR (DMSO-d6, 125 MHz): 25.5, 26.1, 37.8, 44.2, 101.0,
111.3, 118.1, 123.3, 124.6, 138.3, 146.1, 150.2, 151.5, 158.2,
164.6, 183.7; HRMS: Calcd for C16H15N3O4 [M]+ 313.1063, found
313.1054; Anal. Calcd (%) for: C, 61.34; H, 4.83; N, 13.41, found:
C, 61.39; H, 4.88; N, 13.32.
4.2. General
Melting points were determined by open capillary using Veego
Precision Digital Melting Point apparatus (MP-D) and are uncor-
rected. 1H NMR spectra were recorded in DMSO-d6 with BRUKER
AVANCE II (500 MHz) spectrometer using TMS as internal stan-
dard. Chemical shift values are expressed as parts per million
downfield from TMS and J values are in hertz. Splitting patterns
are indicated as s: singlet, d: doublet, t: triplet, m: multiplet, dd:
double doublet, ddd: doublet of a doublet of a doublet, and br:
broad peak. 13C NMR spectra were recorded in DMSO-d6 with
BRUKER AVANCE II (125 MHz) using TMS as internal standard.
Mass spectra were recorded on a BRUCKER high resolution mass
spectrometer (micrOTOF-QII). Elemental analyses were performed
on Heraus CHN-O-Rapid Elemental Analyzer. Column chromatog-
raphy was performed on a silica gel (60–120 mesh) using ethyl
acetate–hexane mixture as eluent.
4.2.1.4.
1-[5-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-pen-
Yield 75%; orange solid; mp
tyl]-1H-indole-2,3-dione (4d).
166–167 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.33–1.37 (m, 2H,
–CH2), 1.81–1.87 (m, 4H, 2ꢀCH2), 3.82–3.84 (m, 2H, –NCH2),
3.89–3.91 (m, 2H, –NCH2), 5.62 (d, J = 7.2 Hz, 1H, olefinic H),
7.12–7.17 (m, 1H, –ArH), 7.22 (d, J = 7.8 Hz, 1H, –ArH), 7.60 (d,
J = 7.2 Hz, 1H, olefinic H), 7.64–7.66 (m, 2H, –ArH), 11.25 (s, 1H,
–NH–, exchangeable with D2O); 13C NMR (DMSO-d6, 125 MHz):
24.7, 25.1, 26.0, 37.7, 44.1, 101.4, 111.5, 118.5, 123.2, 124.4,
138.6, 146.0, 150.3, 151.2, 158.7, 164.4, 183.8; HRMS: Calcd for
C
17H17N3O4 [M]+ 327.1219, found 327.1208; Anal. Calcd (%) for:
C, 62.38; H, 5.23; N, 12.84, found: C, 62.49; H, 5.36; N, 12.71.
4.2.1. Typical procedure for the synthesis of hybrids 4a–r
To a stirred suspension of NaH (0.7 mmol) in dry DMF (5 mL) at
0 °C was added uracil 3 (1.0 mmol) and the mixture was allowed to
stir for 5 min. followed by the dropwise addition of a solution of
C-5 substituted N-alkylbromo-isatins 2 (1.1 mmol) in dry DMF.
The reaction mixture was stirred at 60 °C for 1 h and the progress
was monitored by using TLC. On completion of the reaction, the
solvent was evaporated under vacco followed by the extraction
with ethyl acetate (2 ꢀ 25 ml) and water. The combined organic
layers were dried over anhydrous Na2SO4 and concentrated under
reduced pressure. The crude product was purified via column chro-
matography using silica gel (60–120 mesh) yielding 4 in good
yields.
4.2.1.5. 1-[6-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-hexyl]-
1H-indole-2,3-dione (4e).
Yield 69%; orange solid; mp 173–
1.33–137 (m, 4H,
174 °C; 1H NMR (DMSO-d6, 500 MHz):
d
2ꢀCH2), 1.55–1.61 (m, 4H, 2ꢀCH2), 3.69–3.71 (m, 4H, 2ꢀNCH2),
5.58 (d, J = 7.4 Hz, 1H, olefinic H), 7.13–7.17 (m, 1H, –ArH), 7.25
(d, J = 7.7 Hz, 1H, –ArH), 7.64 (d, J = 7.4 Hz, 1H, olefinic H), 7.68–
7.73 (m, 2H, –ArH), 11.20 (s, 1H, –NH–, exchangeable with D2O);
13C NMR (DMSO-d6, 125 MHz): 25.8, 26.4, 26.8, 28.9, 47.5, 101.2,
111.4, 118.7, 123.1, 124.5, 138.8, 146.4, 150.6, 151.8, 158.4,
164.3, 183.3; HRMS: Calcd for C18H19N3O4 [M]+ 341.1376, found
341.1367; Anal. Calcd (%) for: C, 63.33; H, 5.61; N, 12.31, found:
C, 63.24; H, 5.52; N, 12.43.
4.2.1.6. 1-[8-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-octyl]-
4.2.1.1. 1-[2-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-ethyl]-
1H-indole-2,3-dione (4f).
Yield 72%; orange solid; mp 163–
1.28–133 (m, 8H,
1H-indole-2,3-dione (4a).
Yield 73%; orange solid; mp 168–
3.84 (t, J = 6.3 Hz,
164 °C; 1H NMR (DMSO-d6, 500 MHz):
d
169 °C; 1H NMR (DMSO-d6, 500 MHz):
d
4ꢀCH2), 1.56–1.60 (m, 4H, 2ꢀCH2), 3.68–3.71 (m, 4H, 2ꢀNCH2),
5.57 (d, J = 7.8 Hz, 1H, olefinic H), 7.14–7.19 (m, 1H, –ArH), 7.29
(d, J = 7.7 Hz, 1H, –ArH), 7.61 (d, J = 7.8 Hz, 1H, olefinic H), 7.64–
7.68 (m, 2H, –ArH), 11.24 (s, 1H, –NH–, exchangeable with D2O);
13C NMR (DMSO-d6, 125 MHz): 26.0, 26.2, 27.3, 28.6, 28.8, 29.3,
47.5, 101.4, 111.1, 118.6, 123.3, 124.8, 138.2, 146.2, 150.1, 151.2,
158.6, 164.7, 183.8; HRMS: Calcd for C20H23N3O4 [M]+ 369.1689,
found 389.1680; Anal. Calcd (%) for: C, 65.03; H, 6.28; N, 11.37,
found: C, 65.15; H, 6.36; N, 11.29.
2H, –NCH2), 3.95 (t, J = 6.3 Hz, 2H, –NCH2), 5.59 (d, J = 7.4 Hz, 1H,
olefinic H), 7.12–7.16 (m, 1H, –ArH), 7.20 (d, J = 8.0 Hz, 1H,
–ArH), 7.63 (d, J = 7.4 Hz, 1H, olefinic H), 7.69–7.72 (m, 2H, –
ArH), 11.23 (s, 1H, –NH–, exchangeable with D2O); 13C NMR
(DMSO-d6, 125 MHz): 43.1, 45.5, 101.4, 111.2, 118.0, 123.3,
124.7, 138.1, 146.3, 150.5, 151.2, 158.4, 164.1, 183.5; HRMS:
Calcd for C14H11N3O4 [M]+ 285.0750, found 285.0742; Anal. Calcd
(%) for: C, 58.95; H, 3.89; N, 14.73, found: C, 58.86; H, 3.94; N,
14.66.
4.2.1.2.
1-[3-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-pro-
Yield 78%; orange solid; mp
4.2.1.7. 1-[2-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-ethyl]-
pyl]-1H-indole-2,3-dione (4b).
5-fluoro-1H-indole-2,3-dione (4g).
Yield 72%; orange solid;
3.81 (t,
164–165 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.97 (t, J = 7.0 Hz,
2H, –CH2), 3.72–3.78 (m, 4H, 2ꢀNCH2), 5.57 (d, J = 7.8 Hz, 1H, ole-
finic H), 7.15–7.18 (m, 1H, –ArH), 7.23 (d, J = 7.8 Hz, 1H, –ArH), 7.59
(d, J = 7.2 Hz, 1H, olefinic H), 7.67–7.70 (m, 2H, –ArH), 11.25 (s, 1H,
–NH–, exchangeable with D2O); 13C NMR (DMSO-d6, 125 MHz):
26.7, 37.3, 45.9, 101.3, 111.0, 118.1, 123.6, 124.8, 138.5, 146.0,
mp 158–159 °C; 1H NMR (DMSO-d6, 500 MHz):
d
J = 6.1 Hz, 2H, –NCH2), 3.97 (t, J = 6.1 Hz, 2H, –NCH2), 5.57 (d,
J = 7.4 Hz, 1H, olefinic H), 7.22 (d, J = 7.8 Hz, 1H, –ArH), 7.44 (d,
J = 7.8 Hz, 1H, –ArH), 7.61 (s, 1H, –ArH), 7.67 (d, J = 7.4 Hz, 1H, ole-
finic H), 11.20 (s, 1H, –NH–, exchangeable with D2O); 13C NMR
(DMSO-d6, 125 MHz): 42.8, 44.6, 101.6, 112.1, 118.3, 124.3,
Please cite this article in press as: Kumar, K.; et al. Bioorg. Med. Chem. (2015), http://dx.doi.org/10.1016/j.bmc.2015.04.075

K. Kumar et al. / Bioorg. Med. Chem. xxx (2015) xxx–xxx
5
124.9, 146.7, 147.8, 151.6, 157.3, 158.8, 164.4, 183.2; HRMS: Calcd
for C14H10FN3O4 [M]+ 303.0655, found 303.0647; Anal. Calcd (%)
for: C, 55.45; H, 3.32; N, 13.86, found: C, 55.41; H, 3.26; N, 13.93.
387.1594, found 387.1585; Anal. Calcd (%) for: C, 62.01; H, 5.72;
N, 10.85, found: C, 61.97; H, 5.65; N, 10.94.
4.2.1.13. 5-Chloro-1-[2-(2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-
4.2.1.8.
1-[3-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-pro-
Yield 76%; orange
yl)-ethyl]-1H-indole-2,3-dione (4m).
Yield 71%; orange
pyl]-5-fluoro-1H-indole-2,3-dione (4h).
solid; mp 156–157 °C;; 1H NMR (DMSO-d6, 500 MHz): d 3.81 (t,
J = 6.1 Hz, 2H, –NCH2), 3.94 (t, J = 6.1 Hz, 2H, –NCH2), 5.59 (d,
J = 7.7 Hz, 1H, olefinic H), 7.22 (d, J = 7.2 Hz, 1H, –ArH), 7.42–7.68
(m, 3H, 2ArH+1olefinic H), 11.23 (s, 1H, –NH–, exchangeable with
D2O); 13C NMR (DMSO-d6, 125 MHz): 43.8, 45.4, 101.0, 112.5,
119.8, 124.2, 127.5, 137.3, 146.4, 149.2, 151.8, 158.0, 164.6,
solid; mp 152–153 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.95 (t,
J = 6.9 Hz, 2H, –CH2), 3.71–3.75 (m, 4H, 2ꢀNCH2), 5.58 (d,
J = 7.2 Hz, 1H, olefinic H), 7.25 (d, J = 7.7 Hz, 1H, –ArH), 7.48 (d,
J = 7.7 Hz, 1H, –ArH), 7.63 (s, 1H, –ArH), 7.68 (d, J = 7.2 Hz, 1H, ole-
finic H), 11.22 (s, 1H, –NH–, exchangeable with D2O); 13C NMR
(DMSO-d6, 125 MHz): 26.6, 37.1, 45.8, 101.1, 112.4, 118.0, 124.3,
124.7, 146.8, 147.6, 151.4, 157.2, 158.3, 164.0, 183.7; HRMS:
Calcd for C15H12FN3O4 [M]+ 317.0812, found 318.0803; Anal.
Calcd (%) for: C, 56.78; H, 3.81; N, 13.24, found: C, 56.87; H, 3.89;
N, 13.15.
182.3; HRMS: Calcd for
C
14H10ClN3O4 [M]+ 319.0360, found
319.0369; Anal. Calcd (%) for: C, 52.60; H, 3.15; N, 13.14, found:
C, 52.69; H, 3.20; N, 13.03.
4.2.1.14. 5-Chloro-1-[3-(2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-
yl)-propyl]-1H-indole-2,3-dione (4n).
Yield 73%; orange
4.2.1.9. 1-[4-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-butyl]-
solid; mp 150–151 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.91 (t,
J = 6.6 Hz, 2H, –CH2), 3.72–3.78 (m, 4H, 2ꢀNCH2), 5.58 (d,
J = 7.8 Hz, 1H, olefinic H), 7.20 (d, J = 7.5 Hz, 1H, –ArH), 7.46–7.64
(m, 3H, 2ArH+1olefinic H), 11.26 (s, 1H, –NH–, exchangeable with
D2O); 13C NMR (DMSO-d6, 125 MHz): 26.9, 38.2, 45.1, 101.6, 112.4,
119.7, 124.5, 127.7, 137.2, 146.6, 149.0, 151.7, 158.3, 164.2, 182.7;
HRMS: Calcd for C15H12ClN3O4 [M]+ 333.0516, found 333.0508;
Anal. Calcd (%) for: C, 53.98; H, 3.62; N, 12.59, found: C, 54.07; H,
3.68; N, 12.54.
5-fluoro-1H-indole-2,3-dione (4i).
Yield 75%; orange solid;
mp 154–155 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.70–1.74 (m,
4H, 2ꢀCH2), 3.88–3.93 (m, 2H, –NCH2), 3.94–3.99 (m, 2H,
–NCH2), 5.60 (d, J = 7.4 Hz, 1H, olefinic H), 7.29 (d, J = 7.8 Hz, 1H,
–ArH), 7.49 (d, J = 7.8 Hz, 1H, –ArH), 7.60 (s, 1H, –ArH), 7.66 (d,
J = 7.4 Hz, 1H, olefinic H), 11.21 (s, 1H, –NH–, exchangeable with
D2O); 13C NMR (DMSO-d6, 125 MHz): 25.2, 26.8, 37.0, 44.4, 101.0,
112.2, 118.2, 124.5, 124.6, 146.4, 147.5, 151.7, 157.3, 158.2,
164.1, 183.4; HRMS: Calcd for C16H14FN3O4 [M]+ 331.0968, found
331.0959; Anal. Calcd (%) for: C, 58.01; H, 4.26; N, 12.68, found:
C, 57.92; H, 4.17; N, 12.79.
4.2.1.15. 5-Chloro-1-[4-(2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-
yl)-butyl]-1H-indole-2,3-dione (4o).
Yield 75%; orange solid;
mp 155–156 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.75–1.78 (m, 4H,
2ꢀCH2), 3.83–3.91 (m, 2H, –NCH2), 3.95–3.98 (m, 2H, –NCH2), 5.61
(d, J = 7.4 Hz, 1H, olefinic H), 7.25 (d, J = 7.8 Hz, 1H, –ArH), 7.42–
7.61 (m, 3H, 2ArH+1olefinic H), 11.24 (s, 1H, –NH–, exchangeable
with D2O); 13C NMR (DMSO-d6, 125 MHz): 25.1, 26.2, 37.6, 44.9,
101.4, 112.6, 119.5, 124.8, 127.9, 137.0, 146.3, 149.1, 151.3,
158.5, 164.3, 182.5; HRMS: Calcd for
347.0673, found 347.0677; Anal. Calcd (%) for: C, 55.26; H, 4.06;
N, 12.08, found: C, 55.35; H, 4.11; N, 11.98.
4.2.1.10. 1-[5-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-pen-
tyl]-5-fluoro-1H-indole-2,3-dione (4j).
Yield 74%; orange
solid; mp 151–152 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.31–1.34
(m, 2H, –CH2), 1.77–1.81 (m, 4H, 2ꢀCH2), 3.80–3.85 (m, 2H, –
NCH2), 3.87–3.90 (m, 2H, –NCH2), 5.57 (d, J = 7.7 Hz, 1H, olefinic
H), 7.24 (d, J = 8.0 Hz, 1H, –ArH), 7.42 (d, J = 8.0 Hz, 1H, –ArH),
7.62 (s, 1H, –ArH), 7.63 (d, J = 7.7 Hz, 1H, olefinic H), 11.27 (s, 1H,
–NH–, exchangeable with D2O); 13C NMR (DMSO-d6, 125 MHz):
24.7, 25.7, 26.3, 37.5, 44.2, 101.7, 112.1, 118.3, 124.3, 124.8,
146.1, 147.8, 151.6, 157.4, 158.5, 164.4, 183.6; HRMS: Calcd for
C
16H14ClN3O4 [M]+
4.2.1.16. 5-Chloro-1-[5-(2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-
C
17H16FN3O4 [M]+ 345.1125, found 345.1114; Anal. Calcd (%) for:
C, 59.13; H, 4.67; N, 12.17, found: C, 59.19; H, 4.63; N, 12.22.
yl)-pentyl]-1H-indole-2,3-dione (4p).
Yield 75%; orange
solid; mp 152–153 °C;; 1H NMR (DMSO-d6, 500 MHz): d 1.37–
1.42 (m, 2H, –CH2), 1.76–1.82 (m, 4H, 2ꢀCH2), 3.83–3.86 (m, 2H,
–NCH2), 3.89–3.94 (m, 2H, –NCH2), 5.58 (d, J = 7.7 Hz, 1H, olefinic
H), 7.23 (d, J = 7.2 Hz, 1H, –ArH), 7.47–7.68 (m, 3H, 2ArH+1 olefinic
H), 11.23 (s, 1H, –NH–, exchangeable with D2O); 13C NMR (DMSO-
d6, 125 MHz): 24.9, 25.0, 26.6, 37.3, 44.7, 101.5, 112.5, 119.7, 124.7,
127.3, 137.2, 146.4, 149.4, 151.0, 158.6, 164.5, 182.2; HRMS: Calcd
for C17H16ClN3O4 [M]+ 361.0829, found 361.0820; Anal. Calcd (%)
for: C, 56.44; H, 4.46; N, 11.61, found: C, 56.38; H, 4.37; N, 11.66.
4.2.1.11.
1-[6-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-
Yield 78%; orange
hexyl]-5-fluoro-1H-indole-2,3-dione (4k).
solid; mp 157–158 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.31–138
(m, 4H, 2ꢀCH2), 1.54–1.62 (m, 4H, 2ꢀCH2), 3.64–3.70 (m, 4H,
2ꢀNCH2), 5.58 (d, J = 7.4 Hz, 1H, olefinic H), 7.21 (d, J = 7.8 Hz,
1H, –ArH), 7.43 (d, J = 7.8 Hz, 1H, –ArH), 7.60 (s, 1H, –ArH), 7.65
(d, J = 7.4 Hz, 1H, olefinic H), 11.20 (s, 1H, –NH–, exchangeable with
D2O); 13C NMR (DMSO-d6, 125 MHz): 26.1, 26.4, 27.3, 28.6, 28.9,
30.3, 46.7, 101.3, 112.2, 118.1, 124.5, 124.9, 146.0, 147.6, 151.5,
157.7, 158.2, 164.6, 183.5; HRMS: Calcd for C18H18FN3O4 [M]+
359.1281, found 359.1294; Anal. Calcd (%) for: C, 60.16; H, 5.05;
N, 11.69, found: C, 60.10; H, 4.99; N, 11.76.
4.2.1.17. 5-Chloro-1-[6-(2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-
yl)-hexyl]-1H-indole-2,3-dione (4q).
Yield 74%; orange solid;
mp 158–159 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.31–138 (m,
4H, 2ꢀCH2), 1.59–1.62 (m, 4H, 2ꢀCH2), 3.67–3.68 (m, 4H,
2ꢀNCH2), 5.57 (d, J = 7.4 Hz, 1H, olefinic H), 7.25 (d, J = 7.7 Hz,
1H, –ArH), 7.48–7.74 (m, 3H, 2ArH+1olefinic H), 11.24 (s, 1H,
–NH–, exchangeable with D2O); 13C NMR (DMSO-d6, 125 MHz):
25.9, 26.1, 26.9, 28.7, 47.8, 101.2, 112.8, 119.3, 124.3, 127.7,
137.4, 146.1, 149.6, 151.3, 158.3, 164.1, 182.8; HRMS: Calcd for
4.2.1.12.
1-[8-(2,4-Dioxo-3,4-dihydro-2H-pyrimidin-1-yl)-
octyl]-5-fluoro-1H-indole-2,3-dione (4l). Yield 73%; orange
solid; mp 153–154 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.31 (s,
8H, 4ꢀCH2), 1.58–1.61 (m, 4H, 2ꢀCH2), 3.66–3.67 (m, 4H,
2ꢀNCH2), 5.56 (d, J = 7.2 Hz, 1H, olefinic H), 7.22 (d, J = 7.7 Hz,
1H, –ArH), 7.43 (d, J = 7.7 Hz, 1H, –ArH), 7.61 (s, 1H, –ArH), 7.66
(d, J = 7.4 Hz, 1H, olefinic H), 11.23 (s, 1H, –NH–, exchangeable with
D2O); 13C NMR (DMSO-d6, 125 MHz): 26.1, 26.5, 27.0, 28.8, 28.9,
29.0, 47.8, 101.1, 112.5, 118.8, 124.3, 124.5, 146.1, 147.4, 151.3,
157.9, 158.5, 164.1, 183.4; HRMS: Calcd for C20H22FN3O4 [M]+
C
18H18ClN3O4 [M]+ 375.0986, found 375.0979; Anal. Calcd (%) for:
C, 57.53; H, 4.83; N, 11.18, found: C, 57.60; H, 4.91; N, 11.08.
4.2.1.18. 5-Chloro-1-[8-(2,4-dioxo-3,4-dihydro-2H-pyrimidin-1-
yl)-octyl]-1H-indole-2,3-dione (4r).
Yield 76%; orange solid;
mp 152–153 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.35 (s, 8H,
Please cite this article in press as: Kumar, K.; et al. Bioorg. Med. Chem. (2015), http://dx.doi.org/10.1016/j.bmc.2015.04.075

6
K. Kumar et al. / Bioorg. Med. Chem. xxx (2015) xxx–xxx
4ꢀCH2), 1.54–1.60 (m, 4H, 2ꢀCH2), 3.64–3.68 (m, 4H, 2ꢀNCH2),
5.58 (d, J = 7.2 Hz, 1H, olefinic H), 7.20 (d, J = 7.8 Hz, 1H, –ArH),
7.43–7.71 (m, 3H, 2ArH+1olefinic H), 11.26 (s, 1H, –NH–,
exchangeable with D2O); 13C NMR (DMSO-d6, 125 MHz): 26.0,
26.5, 27.4, 28.2, 28.7, 29.8, 47.3, 101.0, 112.3, 119.1, 124.7, 127.4,
137.5, 146.6, 149.3, 151.1, 158.0, 164.4, 182.7; HRMS: Calcd for
(m, 4H, 2ꢀCH2), 1.83–1.89 (m, 8H, 4ꢀCH2), 3.85–3.89 (m, 4H,
2ꢀNCH2), 3.92–3.96 (m, 4H, 2ꢀNCH2), 5.65 (d, J = 7.4 Hz, 1H, olefi-
nic H), 7.17–7.22 (m, 4H, –ArH), 7.58 (d, J = 7.7 Hz, 2H, –ArH), 7.61–
7.69 (m, 3H, 2ArH+1olefinic H); 13C NMR (DMSO-d6, 125 MHz):
25.6, 25.9, 26.4, 27.1, 27.7, 28.6, 36.8, 37.9, 44.2, 47.4, 100.7,
111.1, 111.4, 117.4, 118.7, 123.3, 123.5, 124.5, 124.7, 138.1,
138.6, 144.1, 150.2, 150.4, 151.2, 158.7, 158.9, 162.2, 183.6,
C
20H22ClN3O4 [M]+ 403.1299, found 403.1291; Anal. Calcd (%) for:
C, 59.48; H, 5.49; N, 10.40, found: C, 59.57; H, 5.56; N, 10.28.
183.8; HRMS: Calcd for C
30H30N4O6 [M]+ 542.2165, found
542.2161; Anal. Calcd (%) for: C, 66.41; H, 5.57; N, 10.33, found:
C, 66.49; H, 5.61; N, 10.26.
4.2.2. Typical procedure for the synthesis of hybrids 5a–r
To a stirred suspension of NaH (2.0 mmol) in dry DMF (5 mL) at
0 °C was added uracil 3 (1.0 mmol) and the mixture was allowed to
stir for 10 min. followed by the dropwise addition of a solution of
C-5 substituted N-alkylbromo-isatins 2 (2.0 mmol) in dry DMF. The
reaction mixture was stirred at 60 °C for 2 h and the progress was
monitored by using TLC. On completion of the reaction, the solvent
was evaporated under vacco followed by the extraction with ethyl
acetate (2ꢀ25 ml) and water. The combined organic layers were
dried over anhydrous Na2SO4 and concentrated under reduced
pressure. The crude product was purified via column chromatogra-
phy using silica gel (60–120 mesh) yielding 5 in good yields.
4.2.2.5. 1,10-(6,60-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis(hex-
ane-6,1-diyl))diindoline-2,3-dione (5e).
Yield 81%; orange
solid; mp 198–199 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.29–1.37
(m, 8H, 4ꢀCH2), 1.55–1.63 (m, 8H, 4ꢀCH2), 3.61–3.68 (m, 4H,
2ꢀNCH2), 3.75–3.79 (m, 4H, 2ꢀNCH2), 5.69 (d, J = 7.2 Hz, 1H, olefi-
nic H), 7.19–7.27 (m, 4H, –ArH), 7.59 (d, J = 7.7 Hz, 2H, –ArH), 7.64–
7.69 (m, 3H, 2ArH+1olefinic H); 13C NMR (DMSO-d6, 125 MHz):
25.3, 25.5, 26.4, 27.0, 27.3, 28.8, 28.9, 29.3, 37.8, 38.1, 44.1, 47.3,
100.2, 111.4, 111.6, 117.3, 118.5, 123.1, 123.2, 124.7, 124.9,
138.4, 138.8, 144.3, 150.0, 150.5, 151.6, 158.5, 158.7, 162.4,
183.3, 183.5; HRMS: Calcd for C32H34N4O6 [M]+ 570.2478, found
570.2469; Anal. Calcd (%) for: C, 67.35; H, 6.01; N, 9.82, found: C,
67.30; H, 5.94; N, 9.92.
4.2.2.1.
(ethane-2,1-diyl))diindoline-2,3-dione
1,10-(2,2’-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis
(5a). Yield 80%;
orange solid; mp 195–196 °C; 1H NMR (DMSO-d6, 500 MHz): d
3.81 (t, J = 6.3 Hz, 2H, –NCH2), 3.93 (t, J = 6.3 Hz, 2H, –NCH2),
3.97–4.01 (m, 4H, 2ꢀNCH2), 5.63 (d, J = 7.8 Hz, 1H, olefinic H),
7.14–7.20 (m, 4H, –ArH), 7.56–7.57 (m, 2H, –ArH), 7.66–7.72 (m,
3H, 2ArH+1olefinic H); 13C NMR (DMSO-d6, 125 MHz): 37.4, 38.0,
38.7, 46.9, 100.8, 110.7, 110.8, 117.8, 117.9, 123.7, 123.8, 125.0,
125.1, 138.6, 138.7, 145.0, 150.7, 151.0, 151.9, 158.7, 158.9,
162.8, 183.4, 183.6; HRMS: Calcd for C24H18N4O6 [M]+ 458.1226,
found 458.1221; Anal. Calcd (%) for: C, 62.88; H, 3.96; N, 12.22,
found: C, 62.94; H, 4.04; N, 12.16.
4.2.2.6. 1,10-(8,80-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis(oc-
tane-8,1-diyl))diindoline-2,3-dione (5f).
Yield 79%; orange
solid; mp 193–194 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.21–1.34
(m, 16H, 8ꢀCH2), 1.64–1.69 (m, 8H, 4ꢀCH2), 3.63–3.68 (m, 6H,
3ꢀNCH2), 3.89–3.93 (m, 2H, –NCH2), 5.60 (d, J = 7.7 Hz, 1H, olefinic
H), 7.23–7.29 (m, 4H, –ArH), 7.55 (d, J = 7.8 Hz, 2H, –ArH),
7.62–7.68 (m, 3H, 2ArH+1olefinic H); 13C NMR (DMSO-d6,
125 MHz): 26.3, 26.6, 27.6, 27.7, 27.9, 28.1, 28.4, 28.5, 28.7, 28.8,
29.0, 29.2, 37.8, 37.9, 48.0, 48.4, 100.4, 111.1, 111.2, 117.2, 118.6,
123.0, 123.4, 124.1, 124.4, 138.7, 138.9, 144.2, 150.4, 150.7,
151.3, 158.3, 158.5, 162.9, 183.8, 183.9; HRMS: Calcd for
4.2.2.2. 1,10-(3,30-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis(pro-
pane-3,1-diyl))diindoline-2,3-dione (5b).
Yield 79%; orange
solid; mp 197–198 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.91 (t,
J = 7.0 Hz, 2H, –CH2), 1.99 (t, J = 6.9 Hz, 2H, –CH2), 3.70–3.74 (m,
4H, 2ꢀNCH2), 3.80 (t, J = 6.9 Hz, 2H, –NCH2), 3.87 (t, J = 7.0 Hz,
2H, –NCH2), 5.69 (d, J = 7.8 Hz, 1H, olefinic H), 7.14–7.21 (m, 4H,
–ArH), 7.58 (d, J = 7.4 Hz, 2H, –ArH), 7.68–7.71 (m, 3H,
2ArH+1olefinic H); 13C NMR (DMSO-d6, 125 MHz): 25.6, 26.5,
36.4, 37.5, 38.3, 47.0, 100.6, 111.0, 111.1, 117.9, 118.0, 123.5,
123.6, 124.8, 124.9, 138.5, 138.6, 144.5, 150.8, 150.9, 151.4,
158.5, 158.7, 162.8, 183.7, 183.8; HRMS: Calcd for C26H22N4O6
[M]+ 486.1539, found 486.1534; Anal. Calcd (%) for: C, 64.19; H,
4.56; N, 11.52, found: C, 64.11; H, 4.44; N, 11.61.
C
36H42N4O6 [M]+ 626.3104, found 626.3113; Anal. Calcd (%) for:
C, 68.99; H, 6.75; N, 8.94, found: C, 69.04; H, 6.83; N, 8.88.
4.2.2.7.
(ethane-2,1-diyl))bis(5-fluoroindoline-2,3 dione) (5g).
1,10-(2,20-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis
Yield
84%; orange solid; mp 183–184 °C; 1H NMR (DMSO-d6,
500 MHz): d 3.83 (t, J = 6.1 Hz, 2H, –NCH2), 3.92–3.95 (m, 2H,
–NCH2), 3.99–4.04 (m, 4H, 2ꢀNCH2), 5.65 (d, J = 7.4 Hz, 1H, olefinic
H), 7.23–7.29 (m, 2H, –ArH), 7.44–7.49 (m, 2H, –ArH), 7.50–7.55
(m, 2H, –ArH), 7.65 (d, J = 7.4 Hz, 1H, olefinic H); 13C NMR
(DMSO-d6, 125 MHz): 37.9, 38.2, 38.5, 48.4, 100.3, 111.3, 112.2,
112.3, 112.7, 112.9, 118.5, 118.8, 124.2, 124.6, 144.4, 147.6,
147.9, 151.4, 157.6, 158.2, 158.5, 159.6, 162.5, 183.7; HRMS:
Calcd for C24H16F2N4O6 [M]+ 494.1038, found 494.1030; Anal.
Calcd (%) for: C, 58.30; H, 3.26; N, 11.33, found: C, 58.23; H, 3.21;
N, 11.40.
4.2.2.3. 1,10-(4,40-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis(bu-
tane-4,1-diyl))diindoline-2,3-dione (5c).
Yield 86%; orange
solid; mp 190–191 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.61–165
(m, 6H, 3ꢀCH2), 1.67–1.71 (m, 2H, –CH2), 3.65–3.72 (m, 4H,
2ꢀNCH2), 3.79–3.83 (m, 2H, –NCH2), 3.87 (s, 2H, –NCH2), 5.69 (d,
J = 7.7 Hz, 1H, olefinic H), 7.12–7.20 (m, 4H, –ArH), 7.55 (d,
J = 7.4 Hz, 2H, –ArH), 7.63–7.70 (m, 3H, 2ArH+1olefinic H); 13C
NMR (DMSO-d6, 125 MHz): 24.3, 24.7, 25.4, 25.8, 36.5, 37.5, 45.4,
47.2, 100.2, 111.5, 111.8, 117.5, 118.6, 123.8, 123.9, 124.3, 124.6,
138.2, 138.8, 144.3, 150.4, 150.7, 151.1, 158.0, 158.3, 162.5,
183.5, 183.7; HRMS: Calcd for C28H26N4O6 [M]+ 514.1852, found
514.1845; Anal. Calcd (%) for: C, 65.36; H, 5.09; N, 10.89, found:
C, 65.33; H, 5.03; N, 10.95.
4.2.2.8. 1,10-(3,30-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis(pro-
pane-3,1-diyl))bis(5-fluoroindoline-2,3-dione) (5h).
Yield
87%; orange solid; mp 180–181 °C; 1H NMR (DMSO-d6,
500 MHz): d 1.94 (t, J = 6.8 Hz, 2H, –CH2), 1.97 (t, J = 6.9 Hz, 2H,
–CH2), 3.73–3.78 (m, 4H, 2ꢀNCH2), 3.82 (t, J = 6.9 Hz, 2H, –NCH2),
3.88 (t, J = 6.9 Hz, 2H, –NCH2), 5.67 (d, J = 7.8 Hz, 1H, olefinic H),
7.21–7.27 (m, 2H, –ArH), 7.43–7.47 (m, 2H, –ArH), 7.56–7.59 (m,
2H, –ArH), 7.62 (d, J = 7.8 Hz, 1H, olefinic H); 13C NMR (DMSO-d6,
125 MHz): 25.2, 26.6, 36.3, 37.7, 38.1, 47.4, 100.4, 111.1, 112.1,
112.2, 112.5, 112.6, 118.1, 118.5, 124.6, 124.9, 144.8, 147.3,
147.5, 151.6, 157.3, 158.8, 158.9, 159.4, 162.3, 183.5; HRMS:
4.2.2.4. 1,10-(5,50-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis(pen-
tane-5,1-diyl))diindoline-2,3-dione (5d).
Yield 84%; orange
solid; mp 194–195 °C; 1H NMR (DMSO-d6, 500 MHz): d 1.30–1.36
Please cite this article in press as: Kumar, K.; et al. Bioorg. Med. Chem. (2015), http://dx.doi.org/10.1016/j.bmc.2015.04.075

K. Kumar et al. / Bioorg. Med. Chem. xxx (2015) xxx–xxx
7
Calcd for C26H20F2N4O6 [M]+ 522.1351, found 522.1343; Anal.
Calcd (%) for: C, 59.77; H, 3.86; N, 10.72, found: C, 59.69; H, 3.75;
N, 10.78.
500 MHz): d 3.80 (t, J = 6.3 Hz, 2H, –NCH2), 3.96–3.99 (m, 2H,
–NCH2), 4.01–4.05 (m, 4H, 2ꢀNCH2), 5.67 (d, J = 7.4 Hz, 1H, olefinic
H), 7.29–7.33 (m, 2H, –ArH), 7.35–7.40 (m, 2H, –ArH), 7.45–7.49
(m, 2H, –ArH), 7.64 (d, J = 7.4 Hz, 1H, olefinic H); 13C NMR
(DMSO-d6, 125 MHz): 37.3, 38.1, 38.3, 48.8, 100.7, 111.8, 112.3,
112.5, 112.8, 118.3, 118.5, 124.2, 124.4, 124.8, 144.7, 146.4,
146.6, 148.5, 157.2, 158.6, 158.8, 159.7, 162.2, 183.6; HRMS:
Calcd for C24H16Cl2N4O6 [M]+ 526.0447, found 526.0443; Anal.
Calcd (%) for: C, 54.67; H, 3.06; N, 10.62, found: C, 54.60; H, 2.98;
N, 10.58.
4.2.2.9. 1,10-(4,40-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis(bu-
tane-4,1-diyl))bis(5-fluoroindoline-2,3-dione)
83%; orange solid; mp 187–188 °C; 1H NMR (DMSO-d6,
500 MHz): 1.61 (s, 6H, 3xCH2), 1.68–1.69 (m, 2H, CH2),
(5i).
Yield
d
3.70–3.72 (m, 4H, 2ꢀNCH2), 3.75–3.77 (m, 2H, –NCH2), 3.83 (s,
2H, –NCH2), 5.69 (d, J = 7.7 Hz, 1H, olefinic H), 7.24–7.28 (m, 2H,
–ArH), 7.46–7.47 (m, 2H, –ArH), 7.53–7.56 (m, 2H, –ArH), 7.69 (d,
J = 7.7 Hz, 1H, olefinic H); 13C NMR (DMSO-d6, 125 MHz): 24.0,
24.5, 24.9, 26.1, 48.4, 100.6, 111.8, 112.0, 112.4, 112.5, 112.6,
118.8, 118.9, 124.3, 124.4, 144.5, 147.2, 147.3, 151.5, 157.9,
158.6, 158.7, 159.8, 162.8, 183.2; HRMS: Calcd for C28H24F2N4O6
[M]+ 550.1664, found 550.1669; Anal. Calcd (%) for: C, 61.09; H,
4.39; N, 10.18, found: C, 60.97; H, 4.34; N, 10.23.
4.2.2.14.
1,10-(3,30-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis
(propane-3,1-diyl))bis(5-chloroindoline-2,3-dione) (5n). Yield
85%; orange solid; mp 186–187 °C; 1H NMR (DMSO-d6,
500 MHz): d 1.93 (t, J = 6.7 Hz, 2H, –CH2), 1.95 (t, J = 6.7 Hz, 2H, –
CH2), 3.70–3.77 (m, 4H, 2ꢀNCH2), 3.85 (t, J = 6.6 Hz, 2H, –NCH2),
3.88 (t, J = 6.6 Hz, 2H, –NCH2), 5.69 (d, J = 7.8 Hz, 1H, olefinic H),
7.24–7.29 (m, 2H, –ArH), 7.31–7.37 (m, 2H, –ArH), 7.47–7.50 (m,
2H, –ArH), 7.61 (d, J = 7.8 Hz, 1H, olefinic H); 13C NMR (DMSO-d6,
125 MHz): 25.0, 26.3, 36.7, 37.8, 38.4, 47.6, 100.2, 111.5, 112.5,
112.8, 112.9, 118.2, 118.3, 124.5, 124.6, 124.9, 144.4, 146.6,
146.8, 148.1, 157.8, 158.5, 158.6, 159.4, 162.7, 183.3; HRMS:
Calcd for C26H20Cl2N4O6 [M]+ 554.0760, found 554.0769; Anal.
Calcd (%) for: C, 56.23; H, 3.63; N, 10.09, found: C, 56.28; H, 3.68;
N, 9.98.
4.2.2.10.
diyl)bis(pentane-5,1-diyl))bis(5-fluoroindoline-2,3-dione)
(5j).
Yield 86%; orange solid; mp 182–183 °C; 1H NMR
1,10-(5,50-(2,4-Dioxopyrimidine-1,3(2H,4H)-
(DMSO-d6, 500 MHz): d 1.32–1.35 (m, 4H, 2ꢀCH2), 1.80–1.88 (m,
8H, 4ꢀCH2), 3.83–3.87 (m, 4H, 2ꢀNCH2), 3.94–3.98 (m, 4H,
2ꢀCH2), 5.64 (d, J = 7.4 Hz, 1H, olefinic H), 7.23–7.29 (m, 2H, –
ArH), 7.44–7.49 (m, 2H, –ArH), 7.52–7.59 (m, 2H, –ArH), 7.68 (d,
J = 7.4 Hz, 1H, olefinic H); 13C NMR (DMSO-d6, 125 MHz): 25.2,
25.4, 26.3, 27.5, 27.8, 28.3, 36.5, 37.7, 44.5, 47.7, 100.8, 111.5,
112.1, 112.3, 112.6, 112.9, 118.5, 118.6, 124.6, 124.9, 144.4,
147.6, 147.8, 151.3, 157.8, 158.4, 158.7, 159.5, 162.6, 183.7;
HRMS: Calcd for C30H28F2N4O6 [M]+ 578.1977, found 578.1971;
Anal. Calcd (%) for: C, 62.28; H, 4.88; N, 9.68, found: C, 62.44; H,
4.91; N, 9.76.
4.2.2.15. 1,10-(4,40-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis(bu-
tane-4,1-diyl))bis(5-chloroindoline-2,3-dione) (5o).
81%; orange solid; mp 184–185 °C; 1H NMR (DMSO-d6,
500 MHz): 1.60 (s, 6H, 3xCH2), 1.63–1.68 (m, 2H, –CH2),
Yield
d
3.71–3.75 (m, 4H, 2ꢀNCH2), 3.77–3.81 (m, 2H, –NCH2), 3.87 (s,
2H, –NCH2), 5.65 (d, J = 7.2 Hz, 1H, olefinic H), 7.29–7.33 (m, 2H,
–ArH), 7.35–7.38 (m, 2H, –ArH), 7.44–7.46 (m, 2H, –ArH), 7.66 (d,
J = 7.2 Hz, 1H, olefinic H); 13C NMR (DMSO-d6, 125 MHz): 24.3,
24.6, 24.8, 26.0, 48.1, 100.1, 111.4, 112.2, 112.7, 112.8, 118.4,
118.6, 124.2, 124.7, 124.8, 144.6, 146.7, 146.8, 148.0, 157.3,
158.6, 158.7, 159.5, 162.5, 183.2; HRMS: Calcd for C28H24Cl2N4O6
[M]+ 582.1073, found 582.1068; Anal. Calcd (%) for: C, 57.64; H,
4.15; N, 9.60, found: C, 57.58; H, 4.08; N, 9.67.
4.2.2.11.
(hexane-6,1-diyl))bis(5-fluoroindoline-2,3-dione) (5k).
87%; orange solid; mp 186–187 °C; 1H NMR (DMSO-d6,
500 MHz):
1.25–1.31 (m, 8H, 4ꢀCH2), 1.53–1.61 (m, 8H,
1,10-(6,60-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis
Yield
d
4ꢀCH2), 3.63–3.66 (m, 4H, 2ꢀNCH2), 3.74–3.77 (m, 4H, 2ꢀNCH2),
5.67 (d, J = 7.8 Hz, 1H, olefinic H), 7.20–7.28 (m, 2H, –ArH), 7.43–
7.47 (m, 2H, –ArH), 7.51–7.56 (m, 2H, –ArH), 7.64 (d, J = 7.8 Hz,
1H, olefinic H); 13C NMR (DMSO-d6, 125 MHz): 25.1, 25.3, 26.2,
27.5, 27.6, 28.4, 28.7, 29.6, 37.4, 38.3, 44.0, 47.2,100.4, 111.4,
112.0, 112.5, 112.8, 112.9, 118.2, 118.7, 124.6, 124.8, 144.2,
147.5, 147.9, 151.3, 157.6, 158.2, 158.5, 159.8, 162.8, 183.4;
HRMS: Calcd for C32H32F2N4O6 [M]+ 606.2290, found 606.2298;
Anal. Calcd (%) for: C, 63.36; H, 5.32; N, 9.24, found: C, 63.27; H,
5.27; N, 9.33.
4.2.2.16.
1,10-(5,50-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis
(pentane-5,1-diyl))bis(5-chloroindoline-2,3-dione) (5p). Yield
84%; orange solid; mp 180–181 °C; 1H NMR (DMSO-d6,
500 MHz):
d
1.34–1.38 (m, 4H, 2ꢀCH2), 1.82–1.85 (m, 8H,
4ꢀCH2), 3.81–3.84 (m, 4H, 2ꢀNCH2), 3.92–3.97 (m, 4H, 2ꢀNCH2),
5.61 (d, J = 7.4 Hz, 1H, olefinic H), 7.26–7.31 (m, 2H, –ArH), 7.33–
7.37 (m, 2H, –ArH), 7.41–7.44 (m, 2H, –ArH), 7.62 (d, J = 7.4 Hz,
1H, olefinic H); 13C NMR (DMSO-d6, 125 MHz): 25.4, 25.7, 26.2,
27.4, 27.6, 28.5, 36.7, 37.9, 44.3, 47.5, 100.4, 111.6, 112.0, 112.3,
112.7, 118.5, 118.8, 124.0, 124.2, 124.5, 144.1, 146.3, 146.6,
148.4, 157.1, 158.5, 158.8, 159.9, 162.4, 183.8; HRMS: Calcd for
4.2.2.12.
(octane-8,1-diyl))bis(5-fluoroindoline-2,3-dione) (5l).
1,10-(8,80-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis
Yield
80%; orange solid; mp 184–185 °C; 1H NMR (DMSO-d6,
500 MHz): d 1.26–1.33 (m, 16H, 8ꢀCH2), 1.62–1.67 (m, 8H,
4ꢀCH2), 3.66–3.69 (m, 6H, 3ꢀNCH2), 3.84–3.90 (m, 2H, –NCH2),
5.68 (d, J = 7.7 Hz, 1H, olefinic H), 7.22–7.25 (m, 2H, –ArH),
7.41–7.46 (m, 2H, –ArH), 7.55–7.58 (m, 2H, –ArH), 7.67 (d,
J = 7.7 Hz, 1H, olefinic H); 13C NMR (DMSO-d6, 125 MHz): 26.2,
26.7, 27.2, 27.5, 27.8, 28.0, 28.2, 28.7, 28.8, 28.9, 29.3, 29.6, 37.5,
37.7, 48.4, 48.6, 100.1, 111.3, 112.4, 112.5, 112.7, 112.9, 118.6,
118.8, 124.4, 124.6, 144.1, 147.4, 147.6, 151.2, 157.7, 158.7,
158.9, 159.4, 162.6, 183.3; HRMS: Calcd for C36H40F2N4O6 [M]+
662.2916, found 662.2907; Anal. Calcd (%) for: C, 65.24; H, 6.08;
N, 8.45, found: C, 65.16; H, 5.96; N, 8.57.
C
30H28Cl2N4O6 [M]+ 610.1386, found 610.1379; Anal. Calcd (%)
for: C, 58.93; H, 4.62; N, 9.16, found: C, 58.86; H, 4.54; N, 9.27.
4.2.2.17.
1,10-(6,60-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis
(hexane-6,1-diyl))bis(5-chloroindoline-2,3-dione) (5q). Yield
80%; orange solid; mp 186–187 °C; 1H NMR (DMSO-d6,
500 MHz):
d
1.21–1.28 (m, 8H, 4ꢀCH2), 1.51–1.57 (m, 8H,
4ꢀCH2), 3.65–3.68 (m, 4H, 2ꢀNCH2), 3.72–3.78 (m, 4H, 2ꢀNCH2),
5.65 (d, J = 7.8 Hz, 1H, olefinic H), 7.28–7.33 (m, 2H, –ArH),
7.35–7.40 (m, 2H, –ArH), 7.43–7.46 (m, 2H, –ArH), 7.60 (d,
J = 7.8 Hz, 1H, olefinic H); 13C NMR (DMSO-d6, 125 MHz): 25.3,
25.6, 26.5, 27.7, 27.8, 28.2, 28.5, 29.4, 37.7, 38.2, 44.3, 47.5,
100.2, 111.4, 112.2, 112.4, 112.8, 118.2, 118.4, 124.4, 124.5,
124.7, 144.6, 146.7, 146.8, 148.1, 157.3, 158.7, 158.9, 159.5,
162.7, 183.4; HRMS: Calcd for C32H32Cl2N4O6 [M]+ 638.1699, found
4.2.2.13.
1,10-(2,20-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis
(ethane-2,1-diyl))bis(5-chloroindoline-2,3-dione) (5m). Yield
82%; orange solid; mp 180–181 °C; 1H NMR (DMSO-d6,
Please cite this article in press as: Kumar, K.; et al. Bioorg. Med. Chem. (2015), http://dx.doi.org/10.1016/j.bmc.2015.04.075

8
K. Kumar et al. / Bioorg. Med. Chem. xxx (2015) xxx–xxx
14. Blaha, C.; Duchene, M.; Aspock, H.; Walochnik, J. J. Antimicrob. Chemother. 2006,
57, 273.
15. Upcroft, J. A.; Dunn, L. A.; Wal, T.; Tabrizi, S.; Delgadillo-Correa, M.; Johnson, G.
P. J.; Garland, S.; Siba, P.; Upcroft, P. Sex Health 2009, 6, 334.
16. Wright, J. M.; Webb, R. I.; O’donoghue, P.; Upcroft, P.; Upcroft, J. A. J. Eukaryot.
Microbiol. 2010, 57, 171.
17. (a) Pandeya, S. N.; Smitha, S.; Jyoti, M.; Sridhar, S. K. Acta Pharm. 2005, 55, 27;
(b) Vine, K. L.; Matesic, L.; Locke, J. M.; Ranson, M.; Skropeta, D. Anti-Cancer
Agents Med. Chem. 2009, 9, 397.
18. (a) Cane, A.; Tournaire, M. C.; Barritault, D.; Crumeyrolle-Arias, M. Biochem.
Biophys. Res. Commun. 2000, 276, 379; (b) Vine, K. L.; Locke, J. M.; Ranson, M.;
Pyne, S. G.; Bremner, J. B. Bioorg. Med. Chem. 2007, 15, 931.
19. Singh, G. S.; Singh, T.; Lakhan, R. Indian J. Chem. 1997, 36, 951.
20. Verma, M.; Pandeya, S. N.; Singh, K. N.; James, P. S. Acta Pharm. 2004, 54, 49.
21. Pandeya, S. N.; Yogeeshwari, P.; Sriram, D.; Nath, G. Indian J. Pharm. Sci. 2002,
64, 209.
22. Selvam, P.; Murugesh, N.; Chandramohan, M.; Debyser; Witvroum, Z. M. Indian
J. Pharm. Sci. 2008, 70, 779.
23. Maskell, L.; Blanche, E. A.; Colucci, M. A.; Whatmore, J. L.; Moody, C. J. Bioorg.
Med. Chem. Lett. 2007, 17, 1575.
24. (a) Maxwell, A. Biochem. Soc. Trans. 1999, 27, 48; (b) Oblak, M.; Grdadolnik, S.
G.; Kotnik, M.; Jerala, R.; Filipic, M.; Solmajer, T. Bioorg. Med. Chem. Lett. 2005,
15, 5207.
638.1691; Anal. Calcd (%) for: C, 60.10; H, 5.04; N, 8.76, found: C,
59.98; H, 4.96; N, 8.82.
4.2.2.18.
(octane-8,1-diyl))bis(5-chloroindoline-2,3-dione) (5r).
1,10-(8,80-(2,4-Dioxopyrimidine-1,3(2H,4H)-diyl)bis
Yield
82%; orange solid; mp 188–189 °C; 1H NMR (DMSO-d6,
500 MHz): d 1.22–1.31 (m, 16H, 8ꢀCH2), 1.64–1.69 (m, 8H,
4ꢀCH2), 3.63–3.68 (m, 6H, 3xNCH2), 3.82–3.88 (m, 2H, –NCH2),
5.61 (d, J = 7.4 Hz, 1H, olefinic H), 7.24–7.31 (m, 2H, –ArH), 7.34–
7.39 (m, 2H, –ArH), 7.45–7.48 (m, 2H, –ArH), 7.63 (d, J = 7.4 Hz,
1H, olefinic H); 13C NMR (DMSO-d6, 125 MHz): 26.4, 26.8, 27.5,
27.6, 27.9, 28.3, 28.5, 28.6, 28.8, 28.9, 29.2, 29.5, 37.7, 37.9, 48.1,
48.5, 100.3, 111.3, 112.5, 112.6, 112.8, 118.6, 118.7, 124.2, 124.4,
124.8, 144.4, 146.8, 146.9, 148.0, 157.0, 158.5, 158.7, 159.8,
162.8, 183.8; HRMS: Calcd for C36H40Cl2N4O6 [M]+ 694.2325, found
694.2321; Anal. Calcd (%) for: C, 62.16; H, 5.80; N, 8.05, found: C,
62.24; H, 5.86; N, 7.97.
25. (a) Motzer, R. J.; Michaelson, M. D.; Redman, B. G.; Hudes, G. R.; Wilding, G.;
Figlin, R. A.; Ginsberg, M. S.; Kim, S. T.; Baum, C. M.; DePrimo, S. E.; Li, J. Z.;
Bello, C. L.; Theuer, C. P.; George, D. J.; Rini, B. I. J. Clin. Oncol. 2006, 24, 16; (b)
Prenen, H.; Cools, J.; Mentens, N.; Folens, C.; Sciot, R.; Schoffski, P.;
VanOosterom, A.; Marynen, P.; Debiec-Rychter, M. Clin. Cancer Res. 2006, 12,
2622.
26. Clarina, I. M.; Jacobus, J. B.; Jacobus, P. P. Bioorg. Med. Chem. 2011, 19, 261.
27. Newkome, G. R.; Pandler, W. W. Contemporary Heterocyclic Chemistry; Wiley:
New York, 1982.
28. (a) Saladino, R.; Crestini, C.; Palamara, A. T.; Danti, M. C.; Manetti, F.; Corelli, F.;
Garaci, E.; Botta, M. J. Med. Chem. 2001, 44, 4554; (b) Zhi, C.; Long, Z.;
Manikowski, A.; Brown, N. C.; Tarantino, P. M.; Holm, K.; Dix, E. J.; Wright, G. E.;
Foster, K. A.; Butler, M. M.; LaMarr, W. A.; Skow, D. J.; Motorina, I.; Lamothe, S.;
Storer, R. J. Med. Chem. 2005, 48, 7063; (c) Fustero, S.; Piera, J.; Sanz-Cervera, J.
F.; Catalan, S.; Ramirez, A. C. Org. Lett. 2004, 6, 1417; (d) Jain, K. S.; Chitre, T. S.;
Minutosiyar, P. B.; Kathiravan, M. K.; Bendre, V. S.; Veer, V. S.; Shahane, R. S.;
Shishoo, C. J. Curr. Sci. 2006, 793.
Acknowledgements
Financial assistance from Department of Science and
Technology, New Delhi under Innovation in Science Pursuit for
Inspired Research (INSPIRE) Fellowship (K.K.) is gratefully
acknowledged. We also thank the Department of Biological
Sciences, University of the Pacific, for support of this project.
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.bmc.2015.04.075.
29. (a) Schinazi, R. F.; Bassit, L.; Gavegnano, C. J. Viral Hepat. 2010, 17, 77; (b) Wen,
Y. M.; Lin, X.; Ma, Z. M. Curr. Drug Targets Infect. Disord. 2003, 3, 241; (c) De
Clercq, E.; Andrei, G.; Snoeck, R.; De Bolle, L.; Naesens, L.; Degreve, B.; Balzarini,
J.; Zhang, Y.; Schols, D.; Leyssen, P.; Ying, C.; Neyts, J. Nucleosides Nucleotides
Nucleic Acids 2001, 20, 271.
References and notes
1. World Health Organization. Global incidence and prevalence of selected
curable sexually transmitted infections-2008. Geneva, Switzerland, 2012.
http://apps.who.int/iris/handle/10665/75181.
30. Pan, X.; Wang, C.; Wang, F.; Li, P.; Hu, Z.; Shan, Y.; Zhang, J. Curr. Med. Chem.
2011, 18, 4538.
2. Soper, D. Am. J. Obstet. Gynecol. 2004, 190, 281.
31. (a) Kemeny, N.; Gonen, M.; Sullivan, D.; Schwartz, L.; Benedetti, F.; Saltz, L.;
Stockman, J.; Fong, Y.; Jarnagin, W.; Bertino, J.; Tong, W.; Paty, P. J. Clin. Oncol.
2001, 19, 2687; (b) Jones, R.; Twelves, C. Expert Opin. Pharmacother. 2001, 2,
1495; (c) Ohwada, S.; Ikeya, T.; Yokomori, T.; Kusaba, T.; Roppongi, T.;
Takahashi, T.; Nakamura, S.; Kakinuma, S.; Iwazaki, S.; Ishikawa, H.; Kawate, S.;
Nakajima, T.; Morishita, Y. Br. J. Cancer 2004, 90, 1003.
32. Nisha, N.; Mehra, V.; Hopper, M.; Patel, N.; Hall, D.; Wrischnik, L. A.; Land, K.
M.; Kumar, V. Med. Chem. Commun. 2013, 4, 1018.
33. Nisha; Kumar, K.; Bhargava, G.; Land, K. M.; Chang, K. H.; Arora, R.; Sen, S.;
Kumar, V. Eur. J. Med. Chem. 2014, 74, 657.
3. (a) Singh, B. N.; Lucas, J. J.; Fichorova, R. N. Trichomonasvaginalis: Pathobiology
and Pathogenesis; Taylor & Francis Group: London, UK, 2007. 411; (b) Smith, L.
M.; Wang, M.; Zangwill, K.; Yeh, S. J. Perinatol. 2002, 22, 502.
4. Conrad, M. D.; Bradic, M.; Warring, S. D.; Gorman, A. W.; Carlton, J. M. Trends
Parasitol. 2013, 29, 17.
5. Van Der Pol, B. Clin. Infect. Dis. 2007, 44, 23.
6. McCelland, R. S. J. Infect. Dis. 2007, 195, 698.
7. Sorvillo, F.; Smith, L.; Kerndt, P.; Ash, L. Emerg. Infect. Dis. 2001, 7, 927.
8. Guenthner, P. C.; Secor, W. E.; Dezzutti, C. S. Infect. Immun. 2005, 73, 4155.
9. McClelland, R. S.; Sangare, L.; Hassan, W. M. J. Infect. Dis. 2007, 195, 698.
10. van der Pol, B.; Kwok, C.; Pierre-Louis, B. J. Infect. Dis. 2008, 197, 548.
11. Sobel, J. D.; Nagappan, V.; Nyirjesy, P. N. Engl. J. Med. 1999, 341, 292.
12. Kim, D. W.; Park, J. M.; Yoon, B. W.; Baek, M. J.; Kim, J. E.; Kim, S. J. Neurol. Sci.
2004, 224, 107.
34. Piens, T. N.; Vreese, R. D.; Neve, N. D.; Hecke, K. V.; Balzarini, J.; Kimpe, N. D.;
D’hooghe, M. Synthesis 2014, 2436.
35. (a) Nakanishi, K.; Suzuki, N.; Yamazaki, F. Bull. Chem. Soc. Jpn. 1961, 34, 53; (b)
Wittenburg, E. Chem. Ber. 1966, 99, 2391.
13. El-Nahas, A. F.; El-Ashmawy, I. M. Basic Clin. Pharmacol. Toxicol. 2004, 94, 226.
Please cite this article in press as: Kumar, K.; et al. Bioorg. Med. Chem. (2015), http://dx.doi.org/10.1016/j.bmc.2015.04.075