Synthesis, Biological Activity of Pyrimidine Linked with Morpholinophenyl Derivatives

Article abstract of DOI:10.1002/jhet.2498

A new series of 5-fluoro-N⁴-(3-(4-substitutedbenzylamino)phenyl)-N²-(4-morpholinophenyl)pyrimidine-2,4-diamine derivatives (7a, 7b, 7c, 7d, 7e, 7f, 7g, 7h, 7i, 7j) are prepared from using an intermediate compound 5-fluoro-N⁴-(3-(aminophenyl)-N²-(4-morpholinophenyl)pyrimidine-2,4-diamine (5). The structures of the newly synthesized products are established from their spectral¹H-NMR,¹³C-NMR,¹⁹F-NMR, ESI-MS, and analytical data. Here we report the synthesized compounds and larvicidal activity. All the compounds are screened for their significant larvicidal activity against third instar larvae at 24, 48, and 78-h time exposure, and values were compared with standard drug Malathion. The Compounds 7i, 7a, 7c, 7f, and 7j exhibited significant activity. However the compounds 7b, 7e, 7d, and 7h showed excellent activity when compared to the above compounds and to standard drug malathion too because of the presence of mild electron withdrawing groups such as trifluoro, fluorine, hydroxy, nitro, and methoxy derivatives which are attached to the benzyl ring.

Full text of DOI:10.1002/jhet.2498

Month 2015
Synthesis, Biological Activity of Pyrimidine Linked with
Morpholinophenyl Derivatives
a
b
c
d
a
*
Sridevi Gorle, Suresh Maddila, Santosh Chokkakula, Palakondu Lavanya, Moganavelli Singh, and
Sreekanth B. Jonnalagaddab
aDiscipline of Biochemistry, School of Life Sciences, University of KwaZulu-Natal, Westville Campus, Chiltern Hills,
Durban 4000, South Africa
bSchool of Chemistry and Physics, University of KwaZulu-Natal, Westville Campus, Chilten Hills, Private Bag 54001,
Durban 4000, South Africa
cSchool of Life and Health sciences, Adikavi Nannaya University, Rajahmundry 533296, India
dDepartment of Chemistry, Annamacharya Institute of Technology and Sciences J.N.T. University,
Tirupati 517 502 Andhra Pradesh, India
*
E-mail: gajulapallilavanya@gmail.com
Received April 12, 2015
DOI 10.1002/jhet.2498
Published online 00 Month 2015 in Wiley Online Library (wileyonlinelibrary.com).
A new series of 5-fluoro-N4-(3-(4-substitutedbenzylamino)phenyl)-N2-(4-morpholinophenyl)pyrimidine-
2,4-diamine derivatives (7a–j) are prepared from using an intermediate compound 5-fluoro-N4-(3-
(aminophenyl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine (5). The structures of the newly synthesized
products are established from their spectral 1H-NMR, 13C-NMR, 19F-NMR, ESI-MS, and analytical data. Here
we report the synthesized compounds and larvicidal activity. All the compounds are screened for their
significant larvicidal activity against third instar larvae at 24, 48, and 78-h time exposure, and values were
compared with standard drug Malathion. The Compounds 7i, 7a, 7c, 7f, and 7j exhibited significant activity.
However the compounds 7b, 7e, 7d, and 7h showed excellent activity when compared to the above compounds
and to standard drug malathion too because of the presence of mild electron withdrawing groups such as
trifluoro, fluorine, hydroxy, nitro, and methoxy derivatives which are attached to the benzyl ring.
J. Heterocyclic Chem., 00, 00 (2015).
INTRODUCTION
most dangerous and case list is also increasing day by day
[4]. In the last five years most of the insecticides did not
show any effect on insects like mosquitoes because of
increased drug resistance [5,6].
Mosquito-borne diseases, such as dengue fever, malaria,
encephalitis, yellow fever, chikungunya, and filariasis (are
among the most common/(or) have become one of the most
dreadful) diseases, which are a major threat to millions of
people spread throughout the world. The common reasons
being the tropical, subtropical climate, poor drainage system
especially during rainy seasons, adversely maintained fish
ponds, irrigation ditches, and rice fields which provide abun-
dant mosquito breeding places. Although chemical vector
control programs have been carried out for long time, these
mosquito-vector diseases remain because of the refusal by
householders to house spraying with synthetic insecticides
[1,2]. The main vector of these diseases is the mosquito
Aedes aegypti (Ae. aegypti) that transmits dengue fever, dengue
hemorrhagic fever, and yellow fever over many areas of tro-
pics and subtropics [3]. Among them dengue fever is the
4-Morpholinophenyl derivatives represent an interesting
class of compounds possessing a wide spectrum of biological
activities. A large number of morpholinophenyl derivatives
exhibited antibacterial [7,8], antifungal [9], anticonvulsant
[10], analgesic [11], anticancer [12], and antiviral activity
[13]. A number of types of pyrimidines were proved to possess
valuable pharmacological properties, such as antiviral [14,15],
antioxidant [16,17], antimalarials [18], anticonvulsant [19,20],
Alzheimer’s disease [21], antidepressant [22], antimicrobial
[23,24], antitubercular [25,26], anti-inflammatory [27,28],
antiplatelet [29], antibacterial [30], antifungal [31,32],
anticancer agents [33,34], antiproliferative agent [35], and
anti-HIV agents [14,36,37]. Integration of these moieties may
show synergistic effect, and in the present study an attempt
© 2015 HeteroCorporation

S. Gorle, S. Maddila, S. Chokkakula, P. Lavanya, M. Singh, and S. B. Jonnalagadda
Vol 000
1
is made for the synthesis of some novel heterocyclic mole-
cules containing the 4-morpholinophenyl and pyrimidine
systems, with a hope that they may exhibit a synergistic
and larvicidal activity.
In the pursuit of design of new drugs, the development
of hybrid molecules through the combination of different
pharmacophores in one frame could lead to compounds
with interesting biological properties. Prompted by these
observations, in the present study, the synthesis and
anti-larvicidal screening of new derivatives (7a–j) with
different pyrimidines and morpholinophenyl moiety
pharmacophores as hybrid molecules possessing anti-
larvicidal activity are investigated.
compounds were determined from their spectral H-NMR,
13C-NMR, 19F-NMR, and ESIMS and analytical data.
All the synthesized compounds gave satisfactory anal-
yses for the proposed structures, which were confirmed
on the basis of their spectral data. The H-NMR spectra
of compound (3) showed peak at δ 7.43 ppm as a singlet
for pyrimidine –CH proton and a broad singlet at δ
1
1
8.24 ppm for –NH proton. The H-NMR spectra of the
compound (4) exhibited two triplets for the –NCH2, –
OCH2 groups at δ 3.27, δ 3.90 ppm, a singlet at δ
7.36 ppm because of pyrimidine –CH proton and two
broad singlets appeared at δ 8.20, δ 8.23 ppm because
of two –NH protons respectively. Similarly compound
(5) showed two triplets at δ 3.15, δ 3.86 ppm because
of –NCH2, –OCH2 groups, while the –NH2 group
protons appeared as a singlet at δ 6.20 ppm, two broad
singlets appeared at δ 8.20, δ 8.26 ppm because of two
–NH group protons respectively.
The structures of title compounds (7a–j) were assigned
on the basis of their spectral studies. The physical data,
1H-NMR, 13C-NMR, 19F-NMR, ESIMS, and analytical
data for all the synthesized compounds are reported in
experimental protocols.
Biological results. Insecticidal resistance is the main
problem to control the mosquito population in the world.
This is mainly achieved by synthesis of new compounds
having high potential activity. The present study of
anti-larvicidal activity indicated mortality of larvae varies
from different concentrations of compounds. The
percentages of mortality increased when increasing
concentrations of the compounds. The mortality of third
instar larvae was indicated in Table 1. Among all the
newly synthesized compounds, 7b showed highest
RESULTS AND DISCUSSION
Chemistry.
Compound 1 was treated with 3-
nitrobenzenamine 2 in the presence of K2CO3 and
dimethyl sulfoxide medium to give 2-chloro-5-fluoro-N-
(3-nitrophenyl)pyrimidin-4-amine (3). The compound
nitro
pyrimidine
(3)
was
reacted
with
4-
morpholinobenzenamine in the presence of K2CO3 as a
basic media to afford compound (4) respectively. The
aminopyrimidine compound (5) was performed by the
treatment of compound 4 with Pd-Carbon in methanol-
ethyl acetate solvent condition. Interestingly, the resultant
amino pyrimidine (5) was further converted into 5-
fluoro-N4-(3-(4-substitutedbenzylamino)phenyl)-N2-(4-
morpholinophenyl)pyrimidine-2,4-diamine
derivatives
(7a–j) through one-pot reaction by dehydrohalogenation
with various substituted benzyl halides 6a–j in the
presence of K2CO3 which is observed to give a very good
yield (Scheme 1). The structures of the newly synthesized
Scheme 1. Synthesis pathway for the pyrimidine linked with morpholinophenyl derivatives.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2015
Synthesis, Biological Activity of Pyrimidine Linked with Morpholinophenyl
Derivatives
Table 1
Results of anti-larvicidal activity of fluoro pyrimidine linked with morpholinophenyl, benzyl derivatives (7a–j).
Period of exposure
24
S.E
48
S.E
72
S.E
Compound
Concentration (in ppm)
M
M
M
100
75
50
100
75
50
100
75
50
100
75
50
100
75
50
100
75
50
100
75
50
100
75
50
72 3.39
70 3.75
65 1.61
78 1.45
75 1.11
70 1.56
71 1.49
69 1.76
64 1.46
76 3.33
73 2.68
68 1.55
77 2.22
74 1.89
69 2.71
69 1.27
68 1.65
63 3.49
65 1.12
63 1.27
59 2.26
74 2.42
72 1.61
67 3.62
73 1.52
71 1.64
66 2.86
68 3.36
66 2.49
62 2.54
100 0.00
74 2.65
71 1.72
66 2.83
84 1.53
79 2.12
72 1.56
73 2.42
70 1.63
65 2.32
78 1.22
74 3.23
69 26
81 2.77
76 1.43
70 2.57
71 2.21
69 3.74
64 3.42
67 3.24
64 2.68
51 3.48
77 3.42
73 2.32
68 2.33
75 3.43
72 2.54
67 2.59
70 2.62
68 2.28
63 1.47
100 0.00
81 3.12
74 3.54
71 1.74
91 2.12
88 1.91
82 2.23
80 3.41
74 3.41
71 1.43
87 1.84
78 1.49
75 1.63
89 2.67
84 2.86
79 1.93
78 2.61
71 2.68
70 3.42
70 2.78
68 2.57
65 1.46
84 3.46
77 2.72
74 2.21
82 1.48
75 2.81
72 1.53
75 3.35
70 2.27
68 2.46
100 0.00
7a
7b
7c
7d
7e
7f
7g
7h
7i
100
75
50
100
75
50
7j
Malathion
25
M = mortality; SE = standard error
mortality (91 2.12) and reached to standard value at
100ppm concentration at 72-h exposures, followed by 7e,
7d, and 7h compounds. Moderate mortality was observed
by 7i, 7a, 7c, 7f, and 7j compounds. 65 1.12, 67 3.24,
70 2.78 mortality rates were considered as a least at
100ppm concentration in 24, 48, and 72-h exposure of
compound 7g respectively.
The structure–activity relationship (SAR) of compounds
7a–j was determined using the data presented in Table 1.
SAR studies revealed that the presence of a strong electron-
withdrawing group on the benzyl ring increased the anti-
larvicidal activity, and activity decreased in the presence of
weak electron-withdrawing group or electron-releasing
atoms or groups. Specifically, compounds with a trifluoro,
fluoro groups in either position 2 or position 4 of the benzyl
ring significantly increased potency against different panel of
microbial strains. The dependence of compounds efficacy
(biological activity) on the position of the fluoro group did
not appear to be important. In this case, the use of electron
withdrawing group on the benzyl ring in basic structures
was worthy. Compounds bearing 4-F, 2-F, 4-CF3, and
2-NO2 exhibited more pronounced anti-larvicidal activity.
CONCLUSION
In conclusion, we have described efficient and benign
synthesis of 5-fluoro benzylamino pyrimidine systems con-
taining morpholinophenyl rings with excellent yields. All
the synthesized compounds have been investigated for
their anti-larvicidal activity. With our newly synthesized
compounds, it is evident that, 7b 7e, 7d, and 7h com-
pounds have exhibited high activity. Compounds 7i, 7a,
7c, 7f, and 7j have revealed excellent larvicidal activity.
Accordingly, this novel class of new 5-fluoro-(4-
substitutedbenzylamino)-(4-morpholinophenyl)pyrimidine
derivatives which were reported from our laboratory,
emerged as a valuable lead series with great potential as
anti-larvicidal agents. This area of research may have a
broad scope for the upcoming and very much promising
researchers for further efficacy of evaluation.
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. Gorle, S. Maddila, S. Chokkakula, P. Lavanya, M. Singh, and S. B. Jonnalagadda
Vol 000
EXPERIMENTAL
Melting points were
2H, J = 7.34Hz), 3.90 (t, 4H, J =4.95 Hz), 3.27 (t, 4H,
J = 4.83Hz); 13C NMR (CDCl3, 75MHz): δ 165.6, 154.4,
151.8, 148.5, 143.2, 140.4, 132.6, 132.2, 130.3, 123.2,
119.2, 115.3, 114.6, 110.8, 69.4, 51.8; 19F NMR (CDCl3,
376 MHz): δ ꢀ118.31 (s, 1F); ESIMS: m/z 411 (M+H)+.
Anal. Cald. for C20H19FN6O3: C, 58.54; H, 3.65; N,
General experimental protocols.
determined in open capillaries on a Mel-Temp apparatus
and are uncorrected. All the reactions were monitored by
thin layer chromatography (TLC). The silica gel F254
plates were used for thin layer chromatography (TLC) in
which the spots were examined under UV light and then
developed by an iodine vapor. Column chromatography
was performed with silica gel (BDH 100–200 mesh).
Solvents were purified according to standard procedures.
The spectra were recorded with the following instruments;
NMR: Varian Gemini 400 MHz (1H), 75 MHz (13C), and
376MHz (19F) spectrometer; ESIMS: VG-Autospec
micromass, analyses of all the compounds were recorded
in LCQ Fleet, Thermo Fisher Instruments Limited.
Organic extracts were dried over anhydrous Na2SO4.
20.32%: Found: C, 58.50; H, 3.67; N, 20.36%.
General procedure for the synthesis of 5-fluoro-N4-(3-
(aminophenyl)-N2-(4-morpholino--phenylpyrimidine-2,4-diamine
(5). The compound 4 (500mg, 1mmol) in a mixture of
methanol-ethyl acetate, (1:2, 20mL) was treated with 10%
Pd-carbon (5% w/w). The reaction was subjected to
hydrogenation under 50 psi hydrogen gas pressures at
room temperature for 2.5h, and the reaction was monitored
by TLC. After completion of the reaction, the mixture was
filtered through a celite pad and concentrated under
reduced pressure to afford pure product in a good yield.
Yellow solid (0.410 g, 79%) m.p. 205–206°C (ethanol
solvent used for crystallization); 1H NMR (CDCl3,
400 MHz): δ 8.26 (brs, 1H), 8.20 (brs, 1H), 7.56–7.44
(m, 4H), 7.36 (t, 1H, J = 7.2 Hz), 7.30 (s, 1H), 7.10 (dd,
1H, J = 7.0, 8.20Hz), 6.89 (d, 2H, J = 7.30Hz), 6.20 (s,
2H), 3.86 (t, 4H, J= 4.89 Hz), 3.15 (t, 4H, J =4.78 Hz);
13C NMR (CDCl3, 75MHz): δ 165.8, 154.6, 151.4,
149.0, 143.2, 139.8, 132.6, 132.3, 130.4, 119.4, 115.0,
106.5, 102.4, 69.4, 51.8; 19F NMR (CDCl3, 376MHz): δ
ꢀ118.33 (s, 1F); ESIMS: m/z 403 (M+Na)+. Anal. Cald.
for C20H21FN6O: C, 63.27; H, 5.38; N, 22.82%; Found:
C, 63.30; H, 5.35; N, 22.85%.
General procedure for the synthesis of 2-chloro-5-fluoro-N-(3-
nitrophenyl)pyrimidin-4-amine (3).
3-Nitrobenzenamine 2
(832 mg, 2mmol) was added to a mixture of 2,4-dichloro-5-
fluoropyrimidine 1 (500mg, 1mmol) and K2CO3 (625mg,
1.5 mmol) in dimethyl sulfoxide (10mL). The reaction
mixture was stirred at 90°C for 3 h and followed by TLC.
After completion of the reaction (monitored by TLC), the
mixture was diluted with ethyl acetate and washed with
water. The ethyl acetate fraction was separated, dried over
Na2SO4, and was evaporated. The obtained compound was
recrystallized with ethanol solvent to get 2-chloro-5-fluoro-
N-(3-nitrophenyl)pyrimidin-4-amine as solid.
Yellow solid (0.432 g, 86%) m.p. 167–169°C (ethanol sol-
vent used for crystallization); 1H NMR (CDCl3, 400MHz): δ
8.24 (brs, 1H), 7.62 (d, 1H, J=8.70Hz), 7.43 (s, 1H), 7.40
(t, 1H, J=8.2Hz), 7.32–7.28 (m, 1H), 7.20 (d, 1H,
J=7.0Hz); 13C NMR (CDCl3, 75MHz): δ 156.6, 155.5,
153.4, 147.6, 143.8, 141.8, 130.2, 122.8, 114.8, 111.7; 19F
NMR (CDCl3, 376MHz): δ ꢀ118.34 (s, 1F); ESIMS: m/z
291 (M+Na)+. Anal. Cald. for C10H6ClFN4O2: C, 45.82; H,
3.20; N, 21.78%; Found: C, 45.84; H, 3.22; N, 21.76%.
General procedure for the synthesis of 5-fluoro-N4-(3-(4-
substitutedbenzylamino)--phenyl)-N2-(4-morpholinophenyl)
pyrimidine-2,4-diamine derivatives (7a–j). To a solution
of compound 5 (100 mg, 1 mmol) in acetone 20 mL was
added K2CO3 (155 mg, 2.5 mmol) at room temperature.
This was followed by the addition of substituted benzyl
halides 6 (180 mg, 1.2 mmol). The reaction mixture was
heated under reflux for 3 h, and the completion of
reaction was checked by TLC. The reaction mixture was
cooled to room temperature and poured into ice-water
followed by extraction with ethyl acetate. The combined
organic layer was evaporated, recrystallized with ethanol
General procedure for the synthesis of 5-fluoro-N4-(3-
(nitrophenyl)-N2-(4-morpholinophenyl)pyrimidine-2,4-diamine
(4). To a stirred solution of compound 3 (500 mg, 1mmol),
4-morpholinobenzenamine (665 mg, 2 mmol) and K2CO3
(390 mg, 1.5mmol) in dimethyl sulfoxide (10 mL). The
reaction mixture was stirred for 3 h at 90°C. After
completion of the reaction, the mixture was diluted and
extracted with ethyl acetate solution. The organic layer
was dried over anhydrous Na2SO4. Finally the compound
was concentrated and recrystallized by ethanol to get pure
solid compound of 5-fluoro-N4-(3-(nitrophenyl)-N2-(4-
morpholinophenyl)pyrimidine-2,4-diamine.
Yellow solid (0.448 g, 85%) m.p. 191–193°C (ethanol
solvent used for crystallization); 1H NMR (CDCl3,
400MHz): δ 8.23 (brs, 1H), 8.20 (brs, 1H), 7.76–7.68
(m, 2H), 7.40 (t, 1H, J = 8.2 Hz), 7.36 (s, 1H), 7.21 (dd,
1H, J = 7.05, 8.0 Hz), 6.86 (d, 2H, J = 7.32Hz), 6.78 (d,
to get pure compounds (7a–j) as solids.
5-Fluoro-N4-(3-(4-fluorobenzylamino)phenyl)-N2-(4-morpholi
nophenyl)pyrimidine-2,4-diamine (7a). Yellow solid (0.091g,
91%) m.p. 238–240°C (ethanol solvent used for
1
crystallization); H NMR (DMSO–d6, 400MHz): δ 8.24
(brs, 1H), 8.22 (brs, 1H), 7.58–7.46 (m, 3H), 7.43 (s, 1H),
7.34 (d, 2H, J=8.54Hz), 7.28–7.14 (m, 4H), 7.10 (t, 1H,
J=10.0Hz), 6.94 (d, 2H, J= 7.84Hz), 6.26 (brs, 1H), 4.42
(d, 2H, J= 5.61 Hz), 3.82 (t, 4H, J=4.88Hz), 3.28 (t, 4H,
J=4.79Hz); 13C NMR (DMSO–d6, 75MHz): δ 165.5,
161.4, 154.5, 151.2, 148.4, 143.8, 140.2, 137.4, 132.6,
132.3, 130.5, 128.8, 118.8, 115.2, 104.9, 103.7, 99.4, 69.2,
51.7, 46.3; 19F NMR (DMSO–d6, 376MHz): δ ꢀ118.30
(s, 1F); ESIMS: m/z 489 (M+H)+. Anal. Cald. for
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

Month 2015
Synthesis, Biological Activity of Pyrimidine Linked with Morpholinophenyl
Derivatives
C27H26F2N6O: C, 66.42; H, 5.36; N, 18.40%; Found: C,
66.38; H, 5.33; N, 18.38%.
J=7.0 Hz), 7.36–7.28 (m, 2H), 6.97 (d, 2H, J=10.2Hz),
6.35 (brs, 1H), 4.52 (d, 2H, J=5.84 Hz), 3.95 (t, 4H,
J=4.82Hz), 3.39 (t, 4H, J=4.90Hz); 13C NMR (DMSO–
d6, 75MHz): δ 164.8, 159.7, 155.3, 151.2, 148.7, 143.8,
139.6, 132.4, 132.2, 131.6, 130.3, 124.6, 124.4, 118.9,
118.5, 117.7, 115.1, 104.6, 103.3, 99.8, 69.5, 51.6, 44.6;
19F NMR (DMSO–d6, 376 MHz): δ ꢀ118.29 (s, 1F),
ꢀ122.86 (m, 3F); ESIMS: m/z 557 (M+H)+. Anal. Cald. for
C28H25F5N6O: C, 58.60; H, 3.75; N, 14.86%; Found: C,
5-Fluoro-N4-(3-(4-fluoro-2-(trifluoromethyl)benzylamino)
phenyl)-N2-(4-morpholinophenyl)-pyrimidine-2,4-diamine
(7b). Light yellow solid (0.074g, 76%) m.p. 255–257°C
(ethanol solvent used for crystallization); 1H NMR
(DMSO–d6, 400 MHz): δ 8.34 (brs, 1H), 8.28 (brs, 1H),
7.62–7.56 (m, 4H), 7.42 (t, 1H, J = 8.58Hz), 7.36 (d, 2H,
J =4.58 Hz), 7.28 (s, 1H), 7.20–7.12 (m, 2H), 6.98 (d,
2H, J= 8.12 Hz), 6.28 (brs, 1H) 4.45 (d, 2H, J= 5.58Hz),
3.84 (t, 4H, J = 4.86Hz), 3.29 (t, 4H, J =4.82 Hz); 13C
NMR (DMSO–d6, 75 MHz) δ: 165.7, 161.2, 155.0,
151.4, 148.6, 144.0, 140.3, 132.6, 132.3, 130.5, 128.9,
127.5, 121.8, 119.2, 118.8, 115.1, 112.2, 104.8, 103.5,
99.8, 69.4, 51.8, 46.4; 19F NMR (DMSO–d6, 376MHz):
δ ꢀ118.27 (s, 1F), ꢀ122.39 (m, 3F); ESIMS: m/z 579 (M
+Na)+. Anal. Cald. for C28H25F5N6O: C, 58.63; H, 3.78;
N, 14.98%; Found: C, 58.65; H, 3.75; N, 14.96%.
58.63; H, 3.78; N, 14.88%.
5-Fluoro-N4-(3-(3-nitrobenzylamino)phenyl)-N2-(4-morphol
inophenyl)pyrimidine-2,4-diamine (7f).
Pale yellow solid
(0.085 g, 85%) m.p. 234–235°C (ethanol solvent used for
1
crystallization); H NMR (CDCl3, 400MHz): δ 8.20 (brs,
1H), 8.17 (brs, 1H), 7.58–7.45 (m, 4H), 7.40 (d, 2H,
J = 10.0Hz), 7.35 (s, 1H), 7.32–7.25 (m, 3H), 7.10 (t, 1H,
J = 8.28Hz), 6.96 (d, 2H, J =8.64 Hz), 6.25 (brs, 1H),
4.42 (d, 2H, J= 5.64Hz), 3.76 (t, 4H, J = 4.52Hz), 3.32
(t, 4H, J =4.75 Hz); 13C NMR (CDCl3, 75MHz): δ 164.5,
155.2, 151.0, 148.6, 147.5, 143.9, 142.6, 139.8, 133.6,
132.4, 130.5, 123.4, 121.9, 119.1, 115.0, 104.5, 103.4,
99.8, 69.2, 51.5, 45.2; 19F NMR (CDCl3, 376 MHz): δ
ꢀ118.27 (s, 1F); ESIMS: m/z 538 (M+Na)+. Anal. Cald.
for C27H26FN7O3: C, 62.80; H, 4.65; N, 18.78%; Found:
5-Fluoro-N4-(3-(3-fluorobenzylamino)phenyl)-N2-
(4-morpholinophenyl)pyrimidine-2,4-diamine (7c).
Yellow
solid (0.077 g, 78%) m.p. 223–224°C (ethanol solvent
1
used for crystallization); H NMR (CDCl3, 400 MHz): δ
8.22 (brs, 1H), 8.19 (brs, 1H), 7.60–7.48 (m, 4H), 7.42
(d, 2H, J= 8.60 Hz), 7.38 (s, 1H), 7.34–7.28 (m, 3H),
7.12 (t, 1H, J =8.20 Hz), 6.98 (d, 2H, J= 10.0 Hz), 6.27
(brs, 1H), 4.45 (d, 2H, J =5.62 Hz), 3.79 (t, 4H,
J =4.76 Hz), 3.26 (t, 4H, J = 4.81Hz); 13C NMR (CDCl3,
75MHz): δ 165.5, 162.9, 154.4, 151.3, 148.6, 143.6,
139.8, 132.3, 132.1, 130.4, 122.7, 119.2, 115.1, 113.8,
113.5, 104.8, 103.6, 99.8, 69.3, 51.8, 46.3; 19F NMR
(CDCl3, 376MHz): δ ꢀ118.19 (s, 1F); ESIMS: m/z 511
(M + Na)+. Anal. Cald. for C27H26F2N6O: C, 66.36; H,
5.38; N, 18.38%; Found: C, 66.39; H, 5.36; N, 18.40%.
C, 62.78; H, 4.62; N, 18.80%.
5-Fluoro-N4-(3-(3-methyl-4-nitrobenzylamino)phenyl)-N2-
(4-morpholinophenyl)pyrimidine-2,4--diamine (7g). White
solid (0.087 g, 88%) m.p. 217–219°C (ethanol solvent used
1
for crystallization); H NMR (CDCl3, 400 MHz): δ 8.22
(brs, 1H), 8.14 (brs, 1H), 7.50–7.42 (m, 3H), 7.36 (d, 2H,
J=8.0 Hz), 7.32 (s, 1H), 7.30–7.27 (m, 3H), 7.04 (t, 1H,
J=8.0 Hz), 6.90 (d, 2H, J=8.20 Hz), 6.28 (brs, 1H), 4.40
(d, 2H, J= 5.60 Hz), 3.72 (t, 4H, J= 4.52 Hz), 3.28 (t, 4H,
J=4.80Hz), 2.19 (s, 3H); 13C NMR (CDCl3, 75MHz): δ
164.3, 155.0, 149.8, 148.4, 147.5, 143.7, 142.2, 139.6,
132.2, 132.0, 130.4, 129.8, 124.9, 123.6, 119.0, 114.8,
104.3, 103.2, 99.7, 69.1, 51.3, 45.0, 19.8; 19F NMR
(CDCl3, 376MHz): δ ꢀ118.28 (s, 1F); ESIMS: m/z 552 (M
+Na)+. Anal. Cald. for C28H28FN7O3: C, 62.84; H, 4.68; N,
5-Fluoro-N4-(3-(4-(trifluoromethoxy)benzylamino)phenyl)-
N2-(4-morpholinophenyl)pyrimidine-2,4-diamine (7d). Light
yellow solid (0.079g, 80%) m.p. 260–262°C (ethanol
solvent used for crystallization); 1H NMR (CDCl3,
400MHz): δ 8.45 (brs, 1H), 8.38 (brs, 1H), 7.82–7.74
(m, 3H), 7.63 (s, 1H), 7.51 (d, 2H, J= 10.0Hz), 7.44–
7.32 (m, 4H), 7.20 (t, 1H, J= 8.62Hz), 7.10 (d, 2H,
J =8.86Hz), 6.32 (brs, 1H), 4.48 (d, 2H, J= 5.80Hz),
3.92 (t, 4H, J= 4.72Hz), 3.38 (t, 4H, J = 4.89Hz); 13C
NMR (CDCl3, 75MHz): δ 165.5, 161.4, 155.2, 151.3,
148.5, 143.8, 139.8, 133.9, 132.5, 130.7, 128.0, 121.7,
119.3, 115.2, 114.0, 104.7, 103.5, 99.8, 69.6, 51.7, 46.2;
19F NMR (CDCl3, 376MHz): δ ꢀ118.32 (s, 1F),
ꢀ123.84 (m, 3F); ESIMS: m/z 577 (M+Na)+. Anal. Cald.
for C28H26F4N6O2: C, 58.80; H, 3.78; N, 16.08%; Found:
C, 58.78; H, 3.75; N, 16.06%.
20.50%; Found: C, 62.86; H, 4.65; N, 20.48%.
5-Fluoro-N4-(3-(4-fluoro-2-nitrobenzylamino)phenyl)-N2-(4-
morpholinophenyl)pyrimidine--2,4-diamine (7h). Yellow solid
(0.077 g, 78%) m.p. 280–282°C (ethanol solvent used for
1
crystallization); H NMR (CDCl3, 400MHz): δ 8.23 (brs,
1H), 8.19 (brs, 1H), 7.59–7.48 (m, 3H), 7.40 (s, 1H), 7.36
(d, 2H, J=8.56Hz), 7.30–7.22 (m, 3H), 7.14 (t, 1H,
J=10.0Hz), 6.98 (d, 2H, J=8.0Hz), 6.28 (brs, 1H), 4.46
(d, 2H, J= 5.74 Hz), 3.85 (t, 4H, J=4.80Hz), 3.32 (t, 4H,
J=4.46Hz); 13C NMR (CDCl3, 75MHz): δ 164.5, 162.0,
155.5, 151.3, 148.9, 143.8, 139.6, 132.5, 132.3, 130.0,
129.4, 121.7, 121.5, 119.2, 115.1, 111.2, 104.5, 103.6,
99.9, 69.4, 51.8, 45.5; 19F NMR (CDCl3, 376MHz): δ
ꢀ118.31 (s, 1F); ESIMS: m/z 556 (M+Na)+. Anal. Cald.
for C27H25F2N7O3: C, 60.76; H, 3.82; N, 18.56%; Found:
C, 60.78; H, 3.84; N, 18.54%.
5-Fluoro-N4-(3-(2-fluoro-3-(trifluoromethyl)benzylamino)p
henyl)-N2-(4-morpholinophenyl)--pyrimidine-2,4-diamine
(7e). White solid (0.091 g, 92%) m.p. 245–247°C (ethanol
solvent used for crystallization); 1H NMR (DMSO–d6,
400 MHz): δ 8.42 (brs, 1H), 8.33 (brs, 1H), 7.90–7.82 (m,
4H), 7.71 (s, 1H), 7.63 (d, 2H, J= 8.02 Hz), 7.40 (t, 1H,
Journal of Heterocyclic Chemistry
DOI 10.1002/jhet

S. Gorle, S. Maddila, S. Chokkakula, P. Lavanya, M. Singh, and S. B. Jonnalagadda
Vol 000
2-((3-(5-Fluoro-2-(4-morpholinophenylamino)pyrimidin-4-
ylamino)phenylamino)methyl)-4-nitrophenol (7i). Pale yellow
solid (0.089g, 90%) m.p. 210–212°C (ethanol solvent used
for crystallization); 1H NMR (CDCl3, 400MHz): δ 8.26
(brs, 1H), 8.20 (brs, 1H), 7.62–7.58 (m, 3H), 7.48 (d, 2H,
J=10.0Hz), 7.42 (s, 1H), 7.38–7.29 (m, 4H), 7.08 (t, 1H,
J=8.0Hz), 6.94 (d, 2H, J= 8.0Hz), 6.28 (brs, 1H), 4.46 (d,
2H, J= 5.68Hz), 3.82 (t, 4H, J=4.80 Hz), 3.38 (t, 4H,
J=4.50Hz); 13C NMR (CDCl3, 75MHz): δ 165.2, 160.5,
155.3, 151.6, 148.5, 143.9, 140.5, 139.7, 132.6, 132.3,
130.4, 129.8, 124.7, 123.5, 119.4, 116.7, 115.3, 104.8,
103.5, 99.8, 69.5, 51.8, 45.6; 19F NMR (CDCl3, 376MHz):
δ ꢀ118.27 (s, 1F); ESIMS: m/z 554 (M+Na)+. Anal. Cald.
for C27H26FN7O4: C, 60.04; H, 3.96; N, 18.84%; Found: C,
Adult female mosquitos were moved to feed on rabbit for
blood (normally exposed site is dorsal side and 1 rabbit
for day) continued up to 5 days for sufficient blood intake
for development and growth. After completion of blood
meal, that enamel water tray were moved to cage for
oviposition. By following the method of Kamaraj et al.
[38], all were maintained and reared in the lab.
Larvicidal bioassay. The larvicidal assay was done by
the standard protocol of WHO with slight modifications
[39]. Different concentrations (50, 75, and 100ppm) of
newly synthesized compounds (7a–j) were transferred in
to glass petridish. In every batch 25 larvae of 3rd instar
were transferred to the petri dish, and 25 ppm
concentration of Malathion was also transferred to
petridish and exposed with larvae forms. Results were
observed in every 24, 48, and 72h. The dead larvae were
identified when they are failed to move after probing with
needle in the siphon or cervical region. In this same way
60.07; H, 3.94; N, 18.86%.
5-Fluoro-N4-(3-(4-methoxybenzylamino)phenyl)-N2-(4-
morpholinophenyl)pyrimidine-2,4--diamine (7j). Yellow solid
(0.080 g, 81%) m.p. 218–220°C (ethanol solvent used
for crystallization); 1H NMR (CDCl3, 400 MHz): δ
8.21 (brs, 1H), 8.18 (brs, 1H), 7.50–7.48 (m, 3H),
7.45 (s, 1H), 7.38 (d, 2H, J = 8.0 Hz), 7.34–7.28 (m,
4H), 7.06 (t, 1H, J = 10.0 Hz), 6.85 (d, 2H, J = 8.0 Hz),
6.25 (brs, 1H), 4.38 (d, 2H, J = 5.80 Hz), 3.90 (s, 3H),
3.70 (t, 4H, J = 4.80 Hz), 3.20 (t, 4H, J = 4.88 Hz); 13C
NMR (CDCl3, 75 MHz): δ 164.5, 158.3, 155.2, 151.4,
148.6, 143.5, 139.7, 134.1, 132.5, 132.3, 130.6, 128.0,
118.8, 114.9, 114.3, 104.5, 103.3, 99.9, 69.2, 58.8,
46.2; 19F NMR (CDCl3, 376 MHz): δ ꢀ118.34 (s, 1F);
ESIMS: m/z 523 (M+Na)+. Anal. Cald. for
C28H29FN6O2: C, 68.42; H, 4.98; N, 16.86%; Found:
C, 68.40; H, 4.96; N, 16.88%.
the experiment was carried out for
4 replicates
simultaneously to all the concentrations. During the entire
experimental period the larvae were unfed. The mortality
results were identified as mortality in % standard error,
and p value less than 0.05 was considered as significant.
Corrected mortality was calculated with the Abbott’s
formula [40].
% of mortality ¼ ½Number of dead larvae=
Number of larva introducedꢁꢂ100:
Corrected mortality ¼ Observed mortality in treatment
ꢀobserved mortality in control
Biological assay. Preparation of different concentrations of
compound. One gram of each newly synthesized series
of compounds 7a–j was taken in different tubes and
mixed with 100mL of distilled water to each tube, and
these were considered as the blank. Then different
concentrations ranging from 50, 75, and 100ppm were
prepared from the above stock. In contrast, 25ppm of
Malathion was prepared, and it was considered as
standard. All chemicals and reagents used were of
analytical grade and obtained from Sigma.
ꢂ100=100 ꢀ control mortality:
Acknowledgments. The authors greatly acknowledge the National
Research Foundation, South Africa, College of AES, University of
KwaZulu-Natal, Durban, South Africa, and Department of
Chemistry, Annamacharya Institute of Technology and Sciences,
Tirupati, India for facilities.
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Journal of Heterocyclic Chemistry
DOI 10.1002/jhet