Synthesis and in vitro activities on anti-platelet aggregation of 4-methoxyisophthalamides

Article abstract of DOI:10.1007/s00044-018-2207-8

A series of 4-methoxyisophthalamides (1d–1w) were designed and synthesized and their chemical structures were confirmed by IR, MS, ¹H-NMR, and ¹³C-NMR. The in vitro on anti-platelet aggregation activities of these compounds were assessed by using Born method. Compounds with higher activities were selected to continue research via Cell Counting Kit-8 (CCK-8) assays of their cytotoxicities. Biological screening results revealed four compounds 1h, 1i, 1q, and 1v exhibited higher activities than the control drugs on against the platelet aggregation induced by adenosine triphosphate (ADP). Moreover, compounds 1p and 1q exhibited higher in vitro activities than picotamide induced by collagen at the concentration of 1.3 μM. Compound 1p also possessed anti-platelet aggregation activity superior to the control drug picotamide induced by arachidonic acid (AA) at the concentration of 1.3 μM. At the same time, the result of cytotoxicities exhibited that none of the compounds have significant cytotoxicities. Therefore, 4-methoxyisophthalamides are potential to become novel anti-platelet drugs with high activities and minimum toxicities.

Full text of DOI:10.1007/s00044-018-2207-8

MEDICINAL
CHEMISTRY
RESEARCH
Medicinal Chemistry Research (2018) 27:1971–1983
https://doi.org/10.1007/s00044-018-2207-8
ORIGINAL RESEARCH
Synthesis and in vitro activities on anti-platelet aggregation of 4-
methoxyisophthalamides
Xiujie Liu1,2 Yan Wang1,2 Lili Liu2 Guangling Chen2
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Received: 9 April 2018 / Accepted: 13 June 2018 / Published online: 26 June 2018
© Springer Science+Business Media, LLC, part of Springer Nature 2018
Abstract
A series of 4-methoxyisophthalamides (1d–1w) were designed and synthesized and their chemical structures were confirmed
1
by IR, MS, H-NMR, and 13C-NMR. The in vitro on anti-platelet aggregation activities of these compounds were assessed
by using Born method. Compounds with higher activities were selected to continue research via Cell Counting Kit-8 (CCK-
8) assays of their cytotoxicities. Biological screening results revealed four compounds 1h, 1i, 1q, and 1v exhibited higher
activities than the control drugs on against the platelet aggregation induced by adenosine triphosphate (ADP). Moreover,
compounds 1p and 1q exhibited higher in vitro activities than picotamide induced by collagen at the concentration of
1.3 μM. Compound 1p also possessed anti-platelet aggregation activity superior to the control drug picotamide induced by
arachidonic acid (AA) at the concentration of 1.3 μM. At the same time, the result of cytotoxicities exhibited that none of the
compounds have significant cytotoxicities. Therefore, 4-methoxyisophthalamides are potential to become novel anti-platelet
drugs with high activities and minimum toxicities.
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Keywords 4-Methoxyisophthalamides Anti-platelet aggregation Synthesis Picotamide Structure activity relationship
Cytotoxicity assay
Abbreviations
ADP
AA
Adenosine triphosphate
Arachidonic acid
CCK-8
DMSO
PAD
PGI2
Cell Counting Kit-8
Dimethyl sulfoxide
Peripheral arterial disease
Prostaglandin I2
Introduction
SAR
TLC
TXA2
Structure activity relationship
Thin layer chromatography
Thromboxane A2.
Anti-platelet drugs are typically used in the clinical treat-
ment of anti-thrombosis and prevention of thrombosis.
Thrombosis that is caused secondary to disrupted athero-
sclerotic plaques is the initiator of most cardiovascular
diseases including heart attacks and strokes. It is well-
known that platelets play a major role in the pathogenesis of
atherothrombosis (Brito et al. 2010; Eskandariyan et al.
2014). Picotamide (Figs. 1–1) is an anti-platelet drug with a
dual inhibitory action, which inhibits both thromboxane A2
(TXA2) receptors and TXA2 synthase (Balsano and Violi
1993; Berrettini et al. 1990; Violi et al. 1988; Yip and
Benavente 2011). Meanwhile, compared with aspirin,
picotamide has the benefit of not interfering with endothe-
lial prostacyclin (PGI2) production (Celestini and Violi
2007; Sartori et al. 1993). Recent research had indicated
that picotamide is more effective than aspirin in reducing
total mortality in patients with type II diabetes associated
Electronic supplementary material The online version of this article
(https://doi.org/10.1007/s00044-018-2207-8) contains supplementary
material, which is available to authorized users.
* Xiujie Liu
Liuxiujie@hotmail.com
* Yan Wang
chemwyan@163.com
1
Tianjin Key Laboratory of Organic Solar Cells and Photochemical
Conversion, School of Chemistry and Chemical Engineering,
Tianjin 300384, People’s Republic of China
2
Tianjin University of Technology, Tianjin 300384, People’s
Republic of China

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Medicinal Chemistry Research (2018) 27:1971–1983
Fig. 1 Structures of Picotamide
(1), target compounds (2),
compound 1l (3) and compound
1c (4)
Fig. 3 The Inhibition Rate and IC50 of Compounds for collagen and
Fig. 2 The Inhibition Rate and IC50 of Compounds
AA
with peripheral arterial disease (PAD) (Neri Serneri et al.
2005).
have the potential of becoming new anti-platelet drugs.
Nonetheless, researches of alkyl substituted 4-
methoxyisophthalamide are insufficient. And compound
1c (Figs. 1–4) with methyl groups in the p-position of the
side chain phenyl rings showed moderate anti-platelet
activity, which aroused our interest to continue to research.
To search for the new anti-platelet agents and perfect the
SAR of 4-methoxyisophthalamides, the depth and breadth
of research for the 4-methoxyisophthalamides have been
enhanced and expanded. Among which in the alkyl, the
original limited to methyl research extended to ethyl, iso-
propyl, n-/t-butyl and for unsaturated ethynyl and cyano
synthesis and research for the first time. Total 19 novel
compounds (1d–1w) were designed and synthesized.
Since 2000, we have concerned on the anti-platelet
aggregating activity studies of various series of 4-
methoxyisophthalamides (Figs. 1–2) and stated the rele-
vant report (Liu et al. 2006, 2011, 2012). Among those
compounds, nearly 30% compounds exhibited significant
anti-platelet aggregation activities than picotamide. In par-
ticularly, compound 1l (Figs. 1–3) with benzyl groups
attached to the phenyl rings in the parent compound showed
excellent activity against the platelet aggregation induced
by ADP, and its IC50 value is as low as 0.02 μM/L and its
LD50 is over 5000 mg/kg. It is 30 times as pictoamide’s
(IC50 = 0.60 μM/L, LD50 = 2500 mg/kg) (Liu et al. 2012,
2017a). All the data justify that 4-methoxyisophthalamides
Compounds 1d–1k with alkyl (–CH2CH3 –CH(CH3)2
–

Medicinal Chemistry Research (2018) 27:1971–1983
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Fig. 4 The cell morphology of L-929 following 48 h incubation
(CH2)3CH3 –C(CH3)3) in different positions were synthe-
sized to investigate the effect of increasing the steric hin-
drance of the substitutions on anti-platelet aggregation
activity. Compound 1m with ethynyl (–C≡CH) was syn-
thesized to investigate the effect of different hybridization
substituent having an unsaturated bond on anti-platelet
activity. We also try to increase the electron-withdrawing of
the substitutions, such as cyano (–C≡N) and trifluoromethyl
(–CF3) to synthesize the compounds 1n, 1o, 1p, 1q, 1r, 1s to
investigate the effect of their introducing on anti-platelet
aggregation. In order to investigate the effect of the number
of substitution on anti-platelet aggregation activity,

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Medicinal Chemistry Research (2018) 27:1971–1983
compounds 1t and 1u were designed and synthesized by
increasing the number of alkyl on the phenyl ring. In order
to investigate the effect of introducing chlorine on anti-
platelet aggregation, we try to introduce methyl and chlorine
on the phenyl ring to synthesize the compounds 1v and 1w.
The chemical structures of target compounds were con-
of compound 4’ was oxidized with potassium permanganate,
4-methoxyisophthalic acid 4 was obtained (Liu et al. 2017b).
At least, compound 4 reacted with sulfoxide chloride, a white
solid 4-methoxyisophthaloyl dichloride 5 was obtained.
Synthetic route of 4-methoxyisophthalamides
1
firmed by IR, MS, H-NMR, and 13C-NMR. Their inhibi-
tory effects on platelet aggregation were tested and assessed
by Born method for ADP inducer. Ten compounds that
were significantly more active in vitro than control drug
picotamide were selected and continued to test and assess
for two inducers (collagen and arachidonic acid). And the
compounds with higher activities on anti-platelet aggrega-
tion were selected to continue research of their
cytotoxicities.
N1,N3-bis(2-ethylphenyl)-4-methoxyisophthalamide (1d)
A round-bottomed flask (100 ml) equipped with a magnetic
stirrers was charged with 2-ethylaniline (10.0 mmol, 1.2 g)
in anhydrous tetrahydrofuran (15 ml). The fresh 4-
methoxyisophthaloyl dichloride 5 (5.0 mmol, 1.2 g) in dry
tetrahydrofuran (15 ml) was added drop-wise to the above
solution at room temperature. Ten hours after the addition
of compound 5, dry triethylamine (1 ml) was added drop-
wise to the reaction mixture. The resultant mixture reaction
was stirred for 24 h at room temperature. After the reaction
completed as monitored by thin layer chromatography
(TLC). The solvent was removed under reduced pressure to
obtain the crude product. The crude product was further
purified by recrystallization from anhydrous ethanol to yield
the desired product 1d as white solid. Yield: 55.00%, m.p.:
175.0~176.0 °C.
Materials and methods
General experimental techniques
All reagents for the synthesis of 4-methoxyisophthalamides
were purchased (purity > 99%) and used without further
purification. The melting points were determined with X-4
digital micro melting point apparatus (thermometer was
uncorrected); Mass spectra (MS) were measured on Xevo
GZQ-Tof VHRS mass spectrometer; IR spectra were
recorded on Avatar 370 spectrophotometer; 1H-NMR
spectra were determined with AVANCE III (400 MHz)
spectrometer (Tetramethyl silane as an internal standard);
13C-NMR spectra were determined with AVANCE III
(101 MHz) spectrometer.
Compounds 1e–1w were prepared in the similar manner.
N1,N3-bis(2-methylphenyl)-4-methoxyisophthalamide
(1a) White solid (The residue was recrystallized by acet-
one.) Yield: 41.00%; m.p.: 218.0–220.0 °C; IR (KBr) υmax
1
3347, 1677, 1605, 1551, 1319, 1091, 816 cm−1; H NMR
(400 MHz, CDCl3, TMS): δ = 9.77 (1H, s, CONH), 8.80
(1H, d, J = 2.2 Hz, 2-H), 8.24 (2H, m, Ar–H), 7.99 (1H, s
CONH), 7.82 (1H, s, 2a-H), 7.30 (1H, s, 2b-H), 7.19 (6H,
m, Ar–H), 4.16 (3H, s, OCH3), 2.36 (3H, s, Ar–CH3), 2.38
(3H, s, Ar–CH3); 13C NMR (101 MHz, CDCl3): δ = 162.2
(C, C = O), 159.5 (C, 4-C), 136.1 (C, 1a-C), 135.5 (C, 1b-
C), 134.8 (CH, 6-C), 133.9 (C, 2a-C), 133.8(C, 2b-C),
130.1 (C, 1-C), 128.6 (CH, 3a-C), 128.4 (CH, 3b-C), 128.0
(C, 3-C), 126.7 (CH, 4a-C), 126.5 (CH, 4b-C), 125.8 (CH,
2-C), 125.0(CH, 5a-C), 124.3 (CH, 5b-C), 122.9 (CH, 6a-
C), 121.4 (CH, 6b-C), 112.1 (CH, 5-C), 58.3 (CH3, OCH3),
14.1 (CH3, Ar–CH3), 13.8 (CH3, Ar–CH3); MS (m/z):
375.1063 [M+H]+ (Liu et al. 2012).
Chemistry
Synthetic routes of 4-methoxylisophthaloyl dichloride
Route a The reaction of phenol 1 with formaldehyde and
hydrochloric acid in benzene gave 2,4-bis (chloromethyl)
phenol 2, which reacted with dimethyl sulfate in water gave
2,4-bis (chloromethyl) -1-methoxybenzene 3. Then com-
pound 3 was oxidized with potassium permanganate, 4-
methoxyisophthalic acid 4 was obtained. Finally, using
compound 4 reacted with sulfoxide chloride, a white solid
4-methoxyisophthaloyl dichloride 5 was obtained.
N1,N3-bis(3-methylphenyl)-4-methoxyisophthalamide
(1b) White solid (The residue was recrystallized by acet-
one.) Yield: 51.00%; m.p.: 179.0–180.0 °C; IR (KBr) υmax
1
Route b 4-Methylphenol 1’ was used as starting material.
Acetylation of 1’ with acetic anhydride gave 4-methylphenyl
acetate 2’. And then compound 2’ was subjected to Fries
rearrangement to 1- (2-hydroxy-5-methylphenyl) ethan-1-one
3’ in the presence of AlCl3 (Shaikh et al. 2016). The reaction
of compound 3’ with dimethyl sulfate gave 2,4-bis (chlor-
omethyl) -1-methoxybenzene 4’. Then the methyl and acetyl
3264, 1618, 1593, 1521, 1487, 1236, 1070, 732 cm−1; H
NMR (400 MHz, CDCl3, TMS): δ = 9.72 (1H, s, CONH),
8.71 (1H, d, J = 1.9 Hz, 2-H), 8.24 (1H, dd, J = 1.0, 4.0 Hz,
6-H), 7.99 (1H, d J = 1.0 Hz, 2-H), 7.26 (1H, d, J = 4.0 Hz,
5-H), 7.55 (4H, m, 2a-H, 2b-H, 6a-H, 6b-H), 7.20 (4H, m,
3a-H, 3b-H, 5a-H, 5b-H), 4.14 (3H, s, OCH3), 2.35 (6H, s,
2 × Ar–CH3); 13C NMR (101 MHz, CDCl3): δ = 164.2 (C,

Medicinal Chemistry Research (2018) 27:1971–1983
1975
C = O), 162.3 (C, C = O), 159.4 (C, 4-C), 140.7 (C, 3a-C),
140.4 (C, 3b-C), 135.6 (C, 1a-C), 135.5 (C, 1b-C), 133.7
(CH, 6-C), 130.0 (C, 1-C), 128.3 (CH, 5a-C), 128.2 (CH,
5b-C), 128.1 (CH, 2-C), 121.2 (CH, 2a-C, 2b-C), 120.7
(CH, 4a-C, 4b-C), 120.4 (CH, 6a-C, 6b-C), 112.1 (C, 3-C),
58.4 (CH3, OCH3), 15.6 (CH3, Ar–CH3), 15.6 (CH3, Ar–
CH3); MS (m/z): 375.1065 [M+H]+ (Liu et al. 2012).
3368.70, 2958.00, 2863.60, 1665.00, 1553.00, 1441.09,
1262.70, 1013.88, 785.75 cm−1
;
1H-NMR (400 MHz,
CDCl3, TMS): δ = 9.69 (1H, s, CONH), 8.70 (1H, d, J =
2.3 Hz, 2-H), 8.22 (1H, dd, J = 8.7, 2.3 Hz, 6-H), 8.17 (1H,
s, CONH), 7.57 (4H, dd, J = 8.2, 3.4 Hz, Ar–H), 7.21 (4H,
dd, J = 8.2, 3.2 Hz, Ar–H), 7.14 (1H, d, J = 8.7 Hz, 5-H),
4.12 (3H, s, OCH3), 2.69–2.61 (4H, m, 2 × CH2CH3), 1.25
(6H, td, J = 7.6, 2.7 Hz, 2 × CH2CH3); 13C-NMR
(101 MHz, CDCl3): δ = 164.3 (C, C = O), 162.5 (C, C =
O), 159.6 (C, 4-C), 140.8 (C, 1a-C), 140.6 (C, 1b-C), 135.8
(C, 3a-C), 135.7 (C, 3b-C), 133.9 (CH, 6-C), 130.1 (C, 1-
C), 128.5 (2 × CH, 5a-C, 5b-C), 128.4 (2 × CH, 2a-C, 2b-
C), 128.3 (CH, 2-C), 121.4 (2 × CH, 2a-C, 2b-C), 120.9
(2 × CH, 6a-C, 6b-C), 120.6 (C, 3-C), 112.3 (C,5-C), 56.7
(CH3, O-CH3), 28.4 (2 × CH2, ArCH2CH3), 15.7 (2 × CH3,
ArCH2CH3); MS: (m/z): 403.2031 [M+H]+.
N1,N3-bis(4-methylphenyl)-4-methoxyisophthalamide (1c)
White solid (The residue was recrystallized by acetone);
Yield: 30.00%; m.p.: 246.0–247.0 °C; IR (KBr) υmax 3264,
1
1618, 1593, 1521, 1487, 1236, 1070, 732 cm−1; H NMR
(400 MHz, CDCl3, TMS): δ = 9.68 (1H, s, CONH), 8.71
(1H, s, 2-H), 8.25 (1H, dd, J = 1.9, 3.8 Hz, 6-H), 7.99 (1H,
s CONH), 7.53 (2H, s, 2a-H, 2b-H), 7.45 (2H, m, 6a-H, 6b-
H), 7.30 (1H, d, J = 3.8 Hz, 5-H), 7.27 (1H, m, 5a-H), 7.18
(1H, m, 5b-H), 6.99 (2H, m, 4a-H, 4b-H), 4.16 (3H, s,
OCH3), 2.39 (6H, s, 2 × Ar–CH3); 13C NMR (101 MHz,
CDCl3): δ = 164.2 (C, C = O), 162.3 (C, C = O), 159.0 (C,
4-C), 140.5 (C, 4a-C), 140.4 (C, 4b-C), 135.8 (C, 1a-C),
135.6 (C, 1b-C), 133.7 (CH, 6-C), 129.9 (C, 1-C), 128.4
(2 × CH, 3a-C, 5a-C), 128.3 (2 × CH, 3b-C, 5b-C), 127.9
(CH, 2-C), 121.0 (C, 3-C), 120.4 (2 × CH, 2a-C, 6b-C),
120.2 (2 × CH, 2b-C, 6b-C), 112.9 (CH, 5-C), 65.5 (CH3,
OCH3), 15.6 (2 × CH3, Ar–CH3); MS (m/z): 375.1064 [M
+H]+ (Liu et al. 2012).
N1,N3-bis(4-ethylphenyl)-4-methoxyisophthalamide (1f)
Compound (1f): White solid (The residue was recrystallized
by anhydrous ethanol.) Yield: 65.30%; m.p.: 221.0–222.5 °
C; IR (KBr) υmax 3358.42, 2961.45, 2929.90, 2871.20,
1674.39, 1652.30, 1601.80, 1517.70, 1319.96, 1264.69,
1118.60 cm−1 1H-NMR (400 MHz, CDCl3, TMS): δ =
;
9.65 (1H, s, CONH), 8.65 (1H, d, J = 2.0 Hz, 2-H), 8.33
(1H, s, CONH), 8.15 (1H, dd, J = 8.6, 2.0 Hz, 6-H), 7.55
(4H, dd, J = 14.3, 8.3 Hz, 2a-H, 6a-H, 2b-H, 6b-H), 7.18
(4H, d, J = 8.2 Hz, 3a-H, 5a-H, 3b-H, 5b-H), 7.06 (1H, d, J
= 8.7 Hz, 5-H), 4.05 (3H, s, OCH3), 2.64 (4H, q, J =
7.4 Hz, 2 × Ar–CH2CH3), 1.24 (6H, t, J = 7.6 Hz, 2 ×
CH2CH3); 13C-NMR (101 MHz, CDCl3): δ = 164.4 (C,
C=O), 162.4 (C, C=O), 159.1 (C, 4-C), 140.6 (C, 1a-C),
140.6 (C, 1b-C), 135.9 (C, 4a-C), 135.8 (C, 4b-C), 133.9
(CH, 6-H), 130.1 (C, 1-C), 128.6 (2 × CH, 2a-C, 2b-C),
128.4 (2 × CH, 6a-C, 6b-C), 128.1 (CH, 2-H), 121.2 (C, 3-
C), 120.6 (2 × CH, 3a-C, 3b-C), 120.4 (2 × CH, 5a-C, 5b-
C), 113.0 (CH, 5-C), 65.7 (CH3, OCH3), 28.5 (CH2, Ar–
CH2CH3), 28.5 (CH2, Ar–CH2CH3), 15.7 (CH3,
Ar–CH2CH3), 14.9 (CH3, Ar–CH2CH3); MS (m/z):
403.2031 [M+H]+.
N1,N3-bis(2-ethylphenyl)-4-methoxyisophthalamide (1d)
White solid (The residue was recrystallized by anhydrous
ethanol.) Yield: 55.00%; m.p.: 175.0–176.0 °C; IR (KBr)
υmax 3354.80, 2960.69, 1659.50, 1597.94, 1530.14,
1463.90, 1256.00, 753.97, 627.80 cm−1
;
1H-NMR
(400 MHz, CDCl3, TMS): δ = 9.76 (1H, s, CONH), 8.80
(1H, d, J = 2.4 Hz, 2-H), 8.24 (1H, dd, J = 8.7, 2.4 Hz, 6-
H), 8.19 (1H, d, J = 8.0 Hz, 5-H), 7.95 (1H, s, CONH), 7.83
(1H, d, J = 7.8 Hz, Ar–H), 7.29–7.23 (3H, m, Ar–H),
7.21–7.10 (4H, m, Ar–H), 4.14 (3H, s, OCH3), 2.71 (4H, m,
2 × CH2CH3), 1.34–1.24 (6H, m, 2 × CH2CH3); 13C-NMR
(101 MHz, CDCl3): δ = 164.7 (C, C = O), 162.5 (C, C =
O), 159.8 (C, 4-C), 136.3 (C, 1a-C), 135.8 (C, 1b-C), 135.1
(CH, 6-C), 134.2 (2 × C, 2a-C, 2b-C), 130.4 (C, 1-C), 128.8
(CH, 4a-C), 128.2 (CH, 4b-C), 126.9 (CH, 5a-C), 126.1
(CH, 5b-C), 125.3 (2 × CH, 3a-C, 3b-C), 124.5 (CH, 2-C),
123.2 (CH, 5-C), 121.6 (C, 3-C), 112.4 (2 × CH, 6a-C, 6b-
C), 56.8 (CH, O-CH3), 25.0 (CH2, Ar–CH2CH3), 24.5
(CH2, Ar–CH2CH3), 14.4 (CH3, Ar–CH2CH3), 14.1 (CH3,
Ar–CH2CH3); MS (m/z): 403.2042 [M+H]+.
N1,N3-bis(2-isopropylphenyl)-4-methoxyisophthalamide
(1g)
White needles (The residue was recrystallized by anhydrous
ethanol.) Yield: 70.83%; m.p.: 154.8–156.0 °C; IR (KBr)
υmax 3376.10, 3270.50, 2961.18, 1656.57, 1527.50,
1467.14, 1265.48 cm−1
;
1H-NMR (400 MHz, CDCl3,
TMS): δ = 9.76 (1H, s, CONH), 8.82 (1H, d, J = 2.3 Hz, 2-
H), 8.29 (1H, dd, J = 8.7, 2.1 Hz, 6-H), 8.05 (1H, d, J =
7.8 Hz, 5-H), 7.97 (1H, s, CONH), 7.73 (1H, m, 6a-H), 7.34
(2H, m, 3a-H, 3b-H), 7.29 (1H, dd, J = 7.6, 1.4 Hz, 6b-H),
7.27–7.19 (4H, m, 4a-H, 5a-H, 4b-H, 5b-H), 4.16 (3H, s,
N1,N3-bis(3-ethylphenyl)-4-methoxyisophthalamide (1e)
White solid (The residue was recrystallized by 75% etha-
nol.) Yield: 82.85%; m.p.: 219.2–221.7 °C; IR (KBr) υmax

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Medicinal Chemistry Research (2018) 27:1971–1983
OCH3), 3.20 (2H, m, 2 × Ar-CH(CH3)2), 1.34 (6H, d, J =
6.8 Hz, Ar-CH(CH3)2), 1.29 (6H, d, J = 6.8 Hz, Ar-CH
(CH3)2); 13C-NMR (101 MHz, CDCl3) δ (ppm): 162.7 (2 ×
C, C=O), 159.8 (C, 4-C), 141.8 (C, 1a-C), 139.6 (C, 1b-C),
134.8 (C, 1-C), 134.2 (C, 2a-C), 134.2 (C, 2b-C), 130.4
(CH, 6-C), 128.2 (CH, 2-C), 126.6 (CH, 3a-C), 126.6 (CH,
3b-C), 125.9 (CH, 5a-C), 125.8 (CH, 5b-C), 125.6 (2 × CH,
4a-C, 4b-C), 124.3 (CH, 2-C), 121.6 (CH, 5-C), 112.4 (2 ×
CH, 6a-C, 6b-C), 56.8 (CH3, O-CH3), 28.4 (CH, Ar–CH
(CH3)2), 28.3 (CH, Ar–CH(CH3)2), 23.3 (2 × CH3, Ar–CH
(CH3)2), 23.2 (2 × CH3, Ar–CH(CH3)2); MS (m/z):
431.2354 [M+H]+.
136.1 (C, 1a-C), 135.8 (C, 1b-C), 134.0 (C, 6-C), 130.0 (C,
1-C), 128.1 (CH, 2-C), 127.2 (2 × CH, 2a-C, 6a-C), 127.1
(2 × CH, 2a-C, 6a-C), 121.2 (C, 3-C), 120.6 (2 × CH, 3a-C,
5a-C), 120.4 (2 × CH, 5b-C, 5b-C), 113.1 (CH, 5-C), 65.7
(CH3, O-CH3), 33.8 (2 × CH, Ar–CH(CH3)2), 24.2 (4 ×
CH3,Ar–CH(CH3)2); MS (m/z):431.2361 [M+H]+.
N1,N3-bis(4-butylphenyl)-4-methoxyisophthalamide (1j)
White solid (The residue was recrystallized by 75% etha-
nol.) Yield: 73.58%; m.p.: 254.0–257.0 °C; IR (KBr) υmax
3240.13, 2959.03, 1591.35, 1507.43, 1485.28, 1461.83,
2927.05 cm−1 1H-NMR (400 MHz, CDCl3, TMS): δ =
;
N1,N3-bis(3-isopropylphenyl)-4-methoxyisophthalamide
(1h)
9.64 (1H, s, CONH), 8.65 (1H, d, J = 1.7 Hz, 2-H), 8.35
(1H, s, CONH), 8.14 (1H, dd, J = 8.5, 1.7 Hz, 6-H), 7.54
(4H, dd, J = 16.2, 8.1 Hz, 2a-H, 6a-H, 2b-H, 6b-H), 7.15
(4H, d, J = 8.1 Hz, 3a-H, 5a-H, 3b-H, 5b-H), 7.05 (1H, d, J
= 8.7 Hz, 5-H), 4.04 (3H, s, OCH3), 2.59 (4H, t, J = 7.4 Hz,
Ar–CH2CH2CH2CH3), 1.59 (4H, dt, J = 15.2, 7.6 Hz,
Ar–CH2CH2CH2CH3), 1.36 (4H, dq, J = 14.5, 7.2 Hz,
White solid (The residue was recrystallized by cyclohex-
ane.) Yield: 61.58%; m.p.: 152.0–154.0 °C; IR (KBr) υmax
3248.30, 1647.45, 1610.23, 1548.99, 1487.90, 1463.93,
2958.72 cm−1 1H-NMR (400 MHz, CDCl3, TMS): δ =
;
9.74 (1H, s, CONH), 8.75 (1H, d, J = 2.1 Hz, 2-H), 8.26
(1H, dd, J = 2.2, 8.6 Hz, 6-H), 8.14 (1H, s, CONH), 7.61
(1H, s, Ar–H), 7.55 (2H, d, J = 11.0 Hz, Ar–H), 7.45 (1H,
d, J = 8.0 Hz, Ar–H), 7.31 (2H, td, J = 7.8, 4.5 Hz, Ar–H),
7.19 (1H, d, J = 8.7 Hz, 5-H), 7.05 (2H, t, J = 8.4 Hz,
Ar–H), 4.16 (3H, s, OCH3), 2.94 (2H, dq, J = 13.4, 6.8 Hz,
2 × Ar–CH(CH3)2), 1.32–1.26 (12H, m, 2 × Ar–CH(CH3)2);
13C-NMR (101 MHz, CDCl3): δ = 164.1 (C, C=O), 162.5
(C, C=O), 159.8 (C, 4-C), 150.2 (C, 3a-C), 150.1 (C, 3b-
C), 138.1 (C, 1a-C), 138.0 (C, 1b-C), 134.2 (CH, 5-C),
129.9 (C, 1-C), 129.1 (CH, 5a-C), 129.1 (CH, 5b-C), 128.3
(CH, 2-C), 122.9 (CH, 4a-C), 122.9 (CH, 4b-C), 121.5 (C,
3-C), 119.0 (CH, 6a-C), 118.4 (CH, 6b-C), 118.3 (CH, 2a-
C), 117.9 (CH, 2b-C), 112.5 (CH, 5-C), 56.8 (CH3, O-CH3),
34.4 (2 × CH, Ar–CH(CH3)2), 24.1 (2 × CH3, Ar–CH(CH3)
2); MS (m/z): 431.2332 [M+H]+.
Ar–CH2CH2CH2CH3),
0.94
(6H,
t,
J = 7.2 Hz,
Ar–CH2CH2CH2CH3); 13C-NMR (101 MHz, CDCl3): δ =
164.4 (C, C=O), 162.4 (C, C=O), 159.5 (C, 4-C), 139.4 (C,
4a-C), 139.2 (C, 4b-C), 135.7 (2 × C, 1a-C, 1b-C), 133.8
(CH, 6-C), 130.1 (C, 1-C), 130.0 (4 × CH, 2a-C, 6a-C, 2b-
C, 6b-C), 128.2 (CH, 2-C), 121.3 (C, 3-C), 120.7 (2 × CH,
3a-C, 5a-C), 120.4 (2 × CH, 3b-C, 5b-C), 112.1 (CH, 5-C),
56.6 (CH3, O-CH3), 35.2 (2 × CH2, Ar–CH2CH2CH2CH3),
33.8 (2 × CH2, Ar–CH2CH2CH2CH3), 22.4 (2 × CH2,
Ar–CH2CH2CH2CH3), 14.1 (2 × CH3, Ar–CH2CH2
CH2CH3); MS (m/z): 459.2656 [M+H]+.
N1,N3-bis(4-(tert-butyl)phenyl)-4-methoxyisophthalamide
(1k)
White solid (The residue was recrystallized by anhydrous
ethanol.) Yield: 55.43%; m.p.: 259.5–261.0 °C; IR (KBr)
υmax 3339.43, 2953.56, 1659.56, 1606.20, 1529.48,
N1,N3-bis(4-isopropylphenyl)-4-methoxyisophthalamide (1l)
1252.80, 1100.98, 952.2, 829.8 cm−1; H-NMR (400 MHz,
1
White flocculent (The residue was recrystallized by acet-
one.) Yield: 82.90%; m.p.: 221.0–224.0 °C; IR (KBr) υmax
3429.6, 2954.2, 1656.9, 1526.9, 1530.1, 1316.3, 1256.0,
1030.7 cm−1; 1H-NMR (400 MHz, CDCl3): δ = 9.67 (1H, s,
CONH), 8.69 (1H, d, J = 2.3 Hz, 2-H), 8.24 (1H, s,
CONH), 8.20 (1H, dd, J = 8.7, 2.3 Hz, 6-H), 7.58 (4H, t, J
= 8.0 Hz, 2a-H, 2b-H, 6a-H, 6b-H), 7.23 (4H, dd, J = 8.4,
3.2 Hz, 3a-H, 3b-H, 5a-H, 5b-H), 7.12 (1H, d, J = 8.7 Hz,
5-H), 4.09 (3H, s, OCH3), 2.91 (2H, qd, J = 10.8, 6.8 Hz,
Ar–CH(CH3)2), 1.27 (6H, d, J = 2.7 Hz, Ar–CH(CH3)2),
1.25 (6H, d, J = 2.7 Hz, Ar–CH(CH3)2); 13C-NMR
(100 MHz, CDCl3): δ = 164.3 (C, C=O), 162.4 (C,
C=O), 159.2 (C, 4-C), 145.3 (C, 4a-C), 145.3 (C, 4b-C),
CDCl3, TMS): δ = 9.66 (1H, s, CONH), 8.69 (1H, s,
CONH), 8.20 (2H, m, 6-H, 2-H), 7.58 (4H, m, 2a-H, 6a-H,
2b-H, 6b-H), 7.36 (4H, m, 3a-H, 5a-H, 3b-H, 5b-H), 7.13
(1H, s, 5-H), 4.10 (3H, s, OCH3), 1.32–1.33 (18H, s, 2 ×
Ar–C(CH3)3); 13C-NMR (101 MHz, CDCl3): δ = 164.3 (C,
C=O), 162.4 (C, C=O), 159.2 (C, 4-C), 147.6 (C, 4a-C),
147.5 (C, 4b-C), 135.8 (C, 1a-C), 135.5 (C, 1b-C), 133.9
(CH, 6-H), 130.0 (C, 1-C), 128.1 (CH, 2-C), 126.1 (2 × CH,
3a-C, 5a-C), 126.0 (2 × CH, 3b-C, 5b-C), 121.2 (C, 3-C),
120.1 (2 × CH, 2a-C, 6a-C), 113.1 (2 × CH, 2b-C, 6b-C),
112.9 (CH, 5-C), 65.7 (CH3, O-CH3), 34.5 (2 × C, Ar–C
(CH3)3), 31.5 (6 × CH3, Ar–C(CH3)3); MS (m/z): 459.2649
[M+H]+.

Medicinal Chemistry Research (2018) 27:1971–1983
1977
N1,N3-bis(2-benzylphenyl)-4-methoxyisophthalamide (1l)
= 8.0 Hz, 4a-H), 7.78 (1H, d, J = 7.8 Hz, 4b-H), 7.44 (2H,
t, J = 6.7 Hz, 5a-H, 5b-H), 7.38 (2H, t, J = 7.5 Hz, 6a-H,
6b-H), 7.16 (1H, d, J = 8.8 Hz, 5-H), 4.14 (3H, s, OCH3);
13C-NMR (101 MHz, DMSO-d6) δ = 166.4 (C, C=O),
164.5 (C, C=O), 164.3 (C, 4-C), 159.9 (C, 1a-C), 159.2 (C,
1b-C), 140.0 (CH, 4a-C), 139.6 (CH, 4b-C), 133.6 (CH, 2a-
C), 132.2 (CH, 2b-C), 131.0 (CH, 6-C), 130.2 (CH, 5a-C),
129.3 (CH, 5b-C), 127.2 (C, 1-C), 126.1 (CH, 2-C), 124.8
(CH, 6a-H), 124.5 (CH, 6b-H), 124.4 (CH, 3-C), 123.0 (C,
Ar–C≡N), 122.5 (C, Ar–C≡N), 118.6 (CH, 5-C), 112.1 (C,
3a-C), 111.6 (C, 3b-C), 56.4 (CH3, O–CH3); MS (m/z):
419.1131 [M + Na]+.
White solid (The residue was recrystallized by acetone.)
Yield: 59.00%; m.p.: 238.0–240.0 °C; IR (KBr) υmax 3301,
3025, 2922, 1653, 1606, 1587, 1493, 1451, 1300, 1262,
1
1027, 825 cm−1; H-NMR (400 MHz, DMSO-d6, TMS): δ
= 9.96 (1H, s, CONH), 9.79 (1H, s, CONH), 8.40 (1H, dd,
J = 4.0, 12.0 Hz, 6-H), 8.04 (1H, d, J = 12.0 Hz, 2-H), 7.75
(1H, d, J = 4.0 Hz, 5-H), 7.11–7.36 (18H, m, Ar–H), 4.07
(2H, s, Ar–CH2–Ph), 4.00 (2H, s, Ar–CH2–Ph), 3.85 (3H, s,
OCH3); 13C-NMR (101 MHz, DMSO-d6): δ = 164.7 (C,
C=O), 163.5 (C, C=O), 160.8 (C, 4-C), 140.2 (C, 1a-C),
140.0 (C, 1b-C), 138.3 (C, 2a-C), 138.2 (C, 2b-C), 131.9
(CH, 6-H), 131.0 (2 × C, 1a’-C, 1b’-C), 128.5 (2 × CH, 3a’-
C, 5a’-C), 128.4 (2 × CH, 3b’-C, 5b’-C), 128.2 (2 × CH,
2a’-C, 6a’-C), 128.0 (4 × CH, 2b’-C, 6b’-C, 4a-C, 4b-C),
126.6 (C, 1-C), 126.0 (C, 2-C), 125.5 (2 × CH, 3a-C, 3b-C),
125.4 (2 × CH, 4a’-C, 4b’-C), 120.4 (2 × CH, 6a-C, 6b-C),
116.5 (C, 3-C), 112.3 (C, 5-C), 60.3 (CH3, OCH3), 34.5
(CH2, Ar–CH2–Ph), 32.7 (CH2, Ar–CH2–Ph); MS (m/z):
527.6063 [M+H]+ (Liu et al. 2012).
N1,N3-bis(4-cyanophenyl)-4-methoxyisophthalamide (1o)
White solid (The residue was recrystallized by ethanol.)
Yield: 62.51%; m.p.: 271.5–273.3 °C; IR (KBr) υmax
3339.20, 2997.36, 1678.58, 1592.21, 1493.40, 2224.40,
1176.00, 1014.90, 823, 753 cm−1 1H-NMR (400 MHz,
;
DMSO-d6, TMS): δ = 10.70 (1H, s, CONH), 10.62 (1H, s,
CONH), 8.26 (1H, d, J = 2.2 Hz, 2-H), 8.18 (1H, dd, J =
8.7, 2.2 Hz, 6-H), 8.00 (2H, d, J = 8.7 Hz, 2a-H, 2b-H),
7.94 (2H, d, J = 8.7 Hz, 6a-H, 6b-H), 7.83 (4H, dd, J = 8.7,
4.7 Hz, 3a-H, 5a-H, 3b-H, 5b-H), 7.36 (1H, d, J = 8.9 Hz,
5-H), 3.96 (3H, s, OCH3); 13C-NMR (101 MHz, DMSO-
d6): δ = 164.8 (C, C=O), 164.6 (C, C=O), 159.3 (C, 4-C),
143.5 (C, 1a-C), 143.1 (C, 1b-C), 133.3 (2 × CH, 3a-C, 5a-
C), 133.1 (2 × CH, 3b-C, 5b-C), 132.3 (CH, 6-C), 129.2 (C,
1-C), 126.0 (CH, 2-C), 124.8 (C, 3-C), 120.2 (2 × CH, 2a-C,
6a-C), 119.6 (2 × CH, 2b-C, 6b-C), 119.0 (2 × C, Ar–C≡N),
112.0 (CH, 5-C), 105.4 (C, 4a-C), 105.3 (C, 4b-C), 56.4
(CH3, O-CH3); MS (m/z): 419.1129 [M + Na]+.
N1,N3-bis(3-ethynylphenyl)-4-methoxyisophthalamide (1m)
White needles (The residue was recrystallized by ethanol.)
Yield: 67.40%; m.p.: 174.1–175.7 °C; IR (KBr) υmax
3346.23, 32837, 2955.33, 1671.50, 649.38, 1582.54,
1554.22, 1431.70, 2104.20, 1185.81, 1017.99, 840, 790 cm
1
−1; H-NMR (400 MHz, CDCl3, TMS): δ = 9.69 (1H, s,
CONH), 8.63 (1H, d, J = 2.3 Hz, 2-H), 8.58 (1H, s,
CONH), 8.16 (1H, dd, J = 8.7, 2.3 Hz, 6-H), 7.86 (1H, s,
Ar–H), 7.73–7.63 (3H, m, Ar–H), 7.32–7.28 (2H, m,
Ar–H), 7.25 (2H, d, J = 8.6 Hz, 5a-H, 5b-H), 7.08 (1H, d, J
= 8.8 Hz, 5-H), 4.09 (3H, s, OCH3), 3.11 (1H, s, C≡CH),
3.09 (1H, s, C≡CH); 13C-NMR (101 MHz, CDCl3): δ =
164.4 (C, C = O), 162.5 (C, C = O), 159.8 (C, 4-C), 138.2
(C, 1a-C), 138.1 (C, 1b-C), 134.3 (CH, 6-C), 130.3 (C, 1-
C), 129.3 (CH, 5a-C), 129.2 (CH, 5b-C), 128.4 (CH, 4a-C),
128.3 (CH, 4b-C), 128.0 (CH, 2-C), 124.2 (CH, 2a-C),
123.9 (CH, 2b-C), 123.0 (C, 4a-C), 130.0 (C, 4b-C), 121.4
(CH, 6a-C), 121.1 (CH, 6b-C), 120.9 (C, 3-C), 112.4 (C, 5-
C), 83.4 (2 × C, Ar–C≡CH), 77.7 (2 × CH, Ar–C≡CH),
56.8 (CH3, O–CH3); MS (m/z): 395.1388 [M+H]+.
N1,N3-bis(2-(trifluoromethyl)phenyl)-4-
methoxyisophthalamide (1p)
White solid (The residue was recrystallized by acetonitrile.)
Yield: 74.71%; m.p.: 172.6–174.3 °C; IR (KBr) υmax
3354.20, 2957.38, 1679.53, 1645.22, 1535.50, 1134.10,
1058.90, 824, 764 cm−1
;
1H-NMR (400 MHz, CDCl3,
TMS): δ = 10.27 (2H, s, CONH), 8.63 (1H, d, J = 1.7 Hz,
2-H), 8.23–8.14 (2H, m, Ar–H), 7.80 (2H, dd, J = 2.4,
10.8 Hz, 3a-H, 3b-H), 7.75 (2H, t, J = 7.7 Hz, 4a-H, 4b-H),
7.55 (2H, t, J = 6.5 Hz, 5a-H, 5b-H), 7.43 (2H, t, J =
8.5 Hz, 6a-H, 6b-H), 4.09 (3H, s, OCH3); 13C-NMR
(101 MHz, DMSO-d6): δ = 165.2 (C, C=O), 162.9 (C,
C=O), 159.6 (C, 4-C), 135.8 (CH, 5a-C), 135.3 (CH, 5b-
C), 133.3 (C, 1a-C), 133.1 (C, 1b-C), 131.3 (CH, 6-C),
127.4 (2 × CH, 3a-C, 3b-C), 126.7 (C, 1-C), 126.6 (2 × C,
Ar–CF3), 126.3 (2 × CH, 4a-C, 4b-C), 126.2 (C, 2-C), 125.4
(2 × C, 2a-C, 2b-C), 122.3 (C, 3-C), 121.0 (CH, 5-C), 112.6
(2 × CH, 6a-C, 6b-C), 56.7 (CH3, O-CH3); MS (m/z):
483.1153 [M+H]+.
N1,N3-bis(3-cyanophenyl)-4-methoxyisophthalamide (1n)
Brown solid (The residue was recrystallized by DMF: H2O
= 1.5: 1); Yield: 56.70%; m.p.: 263.6–265.2 °C; IR (KBr)
υmax 3356.13, 2952.32, 1668.50, 1546.26, 1420.47,
1
2228.49, 1179.80, 1086.95, 830, 792, 680 cm−1; H-NMR
(400 MHz, CDCl3, TMS): δ = 9.85 (1H, s, CONH), 8.73
(1H, s, 2-H), 8.65 (1H, s, CONH), 8.23 (1H, d, J = 8.7 Hz,
6-H), 8.02 (1H, s, 3a-H), 7.91 (1H, s, 3b-H), 7.84 (1H, d, J

1978
Medicinal Chemistry Research (2018) 27:1971–1983
N1,N3-bis(3-(trifluoromethyl)phenyl)-4-
methoxyisophthalamide (1q)
(400 MHz, DMSO-d6, TMS): δ = 10.76 (1H, s, CONH),
10.43 (1H, s, CONH), 8.74 (1H, d, J = 2.1 Hz, 2-H), 8.71
(1H, d, J = 8.8 Hz, 6a-H), 8.26 (1H, dd, J = 8.7, 2.2 Hz, 6-
H), 8.00 (2H, d, J = 6.0 Hz, 3a-H, 3b-H), 7.90 (1H, d, J =
8.4 Hz, 6b-H), 7.79 (2H, t, J = 7.3 Hz, 5a-H, 5b-H), 7.48
(1H, d, J = 8.8 Hz, 5-H), 4.18 (3H, s, OCH3); 13C-NMR
(101 MHz, DMSO-d6): δ = 164.2 (C, C=O), 162.5 (C,
C=O), 160.0 (C, 4-C), 139.1 (C, 1a-C), 138.7 (C, 1b-C),
133.8 (2 × C, 4a-C, 4b-C), 131.7 (CH, 6-C), 129.5 (2 × C,
2a-C, 2b-C), 128.3 (C, 1-C), 126.7 (CH, 3a-C), 126.4 (CH,
3b-C), 125.3 (C, 1-C), 124.8 (2 × CH, 6a-C, 6b-C), 124.6
(2 × CH, 5a-C, 5b-C), 123.3 (CH, 2-C), 122.0 (2 × C, Ar–
CF3), 120.4 (C, 3-C), 113.0 (CH, 5-H), 57.4(CH3, O–CH3);
MS (m/z): 551.0373 [M+H]+.
White solid (The residue was recrystallized by ethanol.)
Yield: 81.60%; m.p.: 201.1–202.6 °C; IR (KBr) υmax
3335.26, 2961.30, 1686.50, 1568.21, 1496.48, 1441.10,
1337.38, 1226.16, 1014.30, 829, 790, 698 cm−1; H-NMR
1
(400 MHz, DMSO-d6, TMS): δ = 10.60 (1H, s, CONH),
10.54 (1H, s, CONH), 8.31 (1H, d, J = 2.1 Hz, 2-H), 8.25
(2H, s, 2a-H, 2b-H), 8.19 (1H, dd, J = 8.8, 2.1 Hz, 6-H),
8.09 (1H, d, J = 8.4 Hz, 4a-H), 7.97 (1H, d, J = 8.1 Hz, 4b-
H), 7.60 (2H, td, J = 8.0, 2.7 Hz, 5a-H, 5b-H), 7.46 (2H, t,
J = 7.4 Hz, 6a-H, 6b-H), 7.36 (1H, d, J = 8.8 Hz, 5-H), 3.97
(3H, s, OCH3); 13C-NMR (101 MHz, DMSO-d6): δ = 164.7
(C, C = O), 164.5 (C, C=O), 159.1 (C, 4-C), 140.0 (C, 1a-
C), 139.7 (1, 1b-C), 132.1 (CH, 6-C), 130.0 (C, 3a-C),
129.8 (C, 3b-C), 129.5 (CH, 5a-C), 129.3 (CH, 5b-C),
129.1 (C, 1-C), 126.1 (CH, 2-C), 125.5 (2 × CH, 2a-C, 2b-
C), 124.8 (2 × CH, 6a-C, 6b-C), 123.8 (2 × C, Ar–CF3),
123.2 (C, 3-C), 120.0 (CH, 4a-C), 116.4 (CH, 4b-C), 112.0
(CH, 5-H), 56.4 (CH3, O-CH3); MS (m/z): 483.1141 [M
+H]+.
N1,N3-bis(2,3-dimethylphenyl)-4-methoxyisophthalamide
(1t)
White solid (The residue was recrystallized by acetone.)
Yield: 61.58%; m.p.: 173.0–174.0 °C; IR (KBr) υmax
3369.52, 3273.61, 1665.90, 1637.69, 1607.32, 1526.80,
2821.76 cm−1 1H-NMR (400 MHz, CDCl3, TMS): δ =
;
9.68 (1H, s, CONH), 8.81 (1H, d, J = 1.8 Hz, 2-H), 8.26
(1H, dd, J = 8.5, 1.4 Hz, 6-H), 7.98 (1H, s, CONH), 7.89
(1H, d, J = 8.0 Hz, 5-H), 7.49 (1H, d, J = 7.9 Hz, Ar–H),
7.17 (1H, ddd, J = 19.7, 10.2, 5.5 Hz, Ar–H), 7.06 (2H, t, J
= 8.1 Hz, Ar–H), 4.15 (3H, s, OCH3), 2.35 (3H, s, Ar–
CH3), 2.33 (3H, s, Ar–CH3), 2.27 (3H, s, Ar–CH3), 2.22
(3H, s, Ar–CH3); 13C-NMR (101 MHz, CDCl3): δ = 162.6
(2 × C, C=O), 159.8 (C, 4-C), 137.7 (C, 3a-C), 137.4 (C,
3b-C), 136.2 (C, 1a-C), 135.4 (C, 1b-C), 134.1 (CH, 6-C),
130.4 (2 × C, 2a-C, 2b-C), 128.1 (C, 1-C), 128.0 (CH, 2-C),
127.9 (CH, 4a-C), 127.1 (CH, 4b-C), 126.2 (CH, 5a-C),
126.0 (CH, 5b-C), 122.8 (C, 3-C), 121.6 (CH, 6a-C), 121.4
(CH, 6b-C), 112.3 (CH, 5-C), 56.8 (CH3, O–CH3), 20.8
(CH3, 3a-CH3), 20.7 (CH3, 3b-CH3), 14.2 (CH3, 2a-CH3),
13.9 (CH3, 2b-CH3); MS (m/z): 403.2015 [M+H]+.
N1,N3-bis(4-(trifluoromethyl)phenyl)-4-
methoxyisophthalamide (1r)
White solid (The residue was recrystallized by ethanol.)
Yield: 45.37%; m.p.: 244.7–245.0 °C; IR (KBr) υmax
3339.20, 1661.96, 1605.88, 1549.44, 1493.29, 1408.30,
1321.85, 1260.93, 1067.12 cm−1
;
1H-NMR (400 MHz,
DMSO-d6, TMS): δ = 10.64 (1H, s, CONH), 10.58 (1H, s,
CONH), 8.28 (1H, d, J = 1.2 Hz, 2-H), 8.19 (1H, d, J =
8.7 Hz, 6-H), 8.03 (2H, d, J = 8.5 Hz, 3a-H, 3b-H), 7.97
(2H, d, J = 8.3 Hz, 3b-H, 5b-H), 7.73 (4H, dd, J = 8.4,
2.6 Hz, 2a-H, 6a-H, 2b-H, 6b-H), 7.36 (1H, d, J = 8.8 Hz,
5-H), 3.96 (3H, s, OCH3); 13C-NMR (101 MHz, DMSO-
d6): δ = 164.8 (C, C = O), 164.6 (C, C = O), 159.2 (C, 4-
C), 142.9 (C, 1a-C), 142.6 (C. 1b-C), 132.3 (2 × CH, 4a-C,
4b-C), 129.2 (CH, 6-H), 126.2 (C, 1-C), 126.0 (2 × CH, 3a-
C, 5a-C), 125.9 (2 × CH, 3b-C, 5b-C), 125.8 (CH, 2-C),
125.8 (C, Ar–CF3), 125.0 (C, Ar–CF3), 123.9 (C, 3-C),
123.6 (CH, 2a-C), 123.1 (CH, 2b-C), 120.2 (CH, 6a-C),
119.6 (CH, 6b-C), 112.0 (CH, 5-C), 56.4 (CH3, O-CH3);
MS (m/z): 483.1152 [M+H]+.
N1,N3-bis(3,4-dimethylphenyl)-4-methoxyisophthalamide
(1u)
White floccule (The residue was recrystallized by 70%
ethanol.) Yield: 58.00%; m.p.: 172.0–173.0 °C; IR (KBr)
υmax 3133.96, 1666.73, 1543.65, 1402.17, 923.32, 840.95,
1
727.79 cm−1; H-NMR (400 MHz, CDCl3, TMS): δ = 9.52
N1,N3-bis(2-chloro-4-(trifluoromethyl)phenyl)-4-
methoxyisophthalamide (1s)
(1H, s, CONH), 8.60 (1H, s, CONH), 8.55 (1H, s, 2-H),
8.06 (1H, d, J = 7.9 Hz, 6-H), 7.44 (1H, s, Ar-H), 7.41–7.33
(2H, m, 2a-H, 2b-H), 7.30 (1H, d, J = 8.1 Hz, Ar–H), 7.04
(2H, dd, J = 8.0, 2.6 Hz, 5a-H, 5b-H), 6.95 (1H, d, J =
8.7 Hz, 5-H), 3.97 (3H, s, OCH3), 2.20 (12H, s, 4 ×
Ar–CH3); 13C-NMR (101 MHz, CDCl3): δ = 164.5 (2 × C,
C=O), 159.3 (C, 4-C), 137.2 (C, 3a-C), 137.0 (C, 3b-C),
White solid (The residue was recrystallized by acetone.)
Yield: 63.21%; m.p.: 226.0–229.0 °C; IR (KBr) υmax
3421.20, 3312.13, 1685.96, 1595.88, 1542.74, 1500.79,
1477.30, 1324.85, 1261.93, 1081.12 cm−1
;
1H-NMR

Medicinal Chemistry Research (2018) 27:1971–1983
1979
136.0 (C, 1a-C), 135.7 (C, 1b-C), 133.4 (CH, 6-C), 132.8
(C, 4a-C), 132.6 (4b-C), 130.3 (C, 1-C), 129.9 (CH, 5a-C),
129.9 (CH, 5b-C), 128.2 (CH, 2-C), 122.0 (CH, 2a-C),
121.8 (CH, 2b-C), 121.3 (C, 3-C), 118.2 (CH, 6a-C), 118.0
(CH, 6b-C), 111.8 (CH, 5-C), 56.5 (CH3, O–CH3), 19.9
(2 × CH3, 3a-CH3, 3b-CH3), 19.2 (2 × CH3, 4a-CH3, 4b-
CH3); MS (m/z): 403.2039 [M+H]+.
Pharmacology
In vitro anti-platelet aggregation activity
The in vitro activity studies on anti-platelet aggregation of
the target compounds have been done by using turbidi-
metric test (Born 1962): fresh venous blood was taken from
the ear vein of rabbits, with 3.8% sodium citrate as antic-
oagulant (blood in proportion to anticoagulant agent were 9:
1), and then centrifuged at room temperature for 10 min
(500–800 r/min) to obtain the platelet-rich plasma (PRP).
The platelet-poor plasma (PPP) was obtained by centrifu-
ging of PRP at room temperature for 15 min (3000 r/min).
The target compounds were dissolved in DMSO (5 μl), and
then, the solution was added into PRP (20 μl), and the same
volume of DMSO without target compounds was added as a
control group. After 2 min incubation, using ADP as the
active inducer, platelet aggregation rate was measured.
Using the same test method, the platelet aggregation rates of
eight compounds induced by collagen and AA were also
tested. The inhibition rates for the compounds were calcu-
lated according to the following formulas:
N1,N3-bis(3-chloro-2-methylphenyl)-4-
methoxyisophthalamide (1v)
White solid (The residue was recrystallized by dioxane.)
Yield: 85.10%; m.p.: 232.7–234.3 °C; IR (KBr) υmax
3375.13, 3255.68, 1667.20, 1524.26, 1437.48, 1180.20,
1047.42, 2948.29, 821, 779 cm−1 1H-NMR (400 MHz,
;
DMSO-d6, TMS): δ = 10.27 (1H, s, CONH), 10.03 (1H, s,
CONH), 8.45 (1H, d, J = 2.0 Hz, 2-H), 8.19 (1H, dd, J =
9.2, 1.5 Hz, 6-H), 7.68 (1H, d, J = 7.9 Hz, 4a-H), 7.55 (1H,
d, J = 8.0 Hz, 4b-H), 7.50 (1H, d, J = 7.9 Hz, 5-H), 7.38
(1H, d, J = 8.8 Hz, 5a-H), 7.33 (1H, d, J = 8.2 Hz, 5b-H),
7.20 (2H, td, J = 7.8, 4.0 Hz, 6a-H, 6b-H), 4.05 (3H, s,
OCH3), 2.40 (3H, s, 2a-CH3), 2.28 (3H, s, 2b-CH3); 13C-
NMR (101 MHz, DMSO-d6): δ = 164.4 (C, C=O), 163.4
(C, C=O), 159.3 (C, 4-C), 138.0 (C, 1a-C), 137.8 (C, 1b-
C), 134.0 (C, 2a-C), 132.2 (C, 2b-C), 131.3 (CH, 6-C),
130.0 (C, 3a-C), 129.2 (C, 3b-C), 127.4 (CH, 5a-C), 127.3
(CH, 5b-C), 126.6 (C, 1-C), 126.3 (CH, 2-C), 124.7 (CH,
4a-C), 124.6 (CH, 4b-C) 124.4 (C, 3-C), 123.3 (CH, 5-C),
112.1 (2 × CH, 6a-C, 6b-C), 56.6 (CH3, O–CH3), 18.4
(CH3, 2a-CH3), 18.0 (CH3, 2b-CH3); MS (m/z): 443.0937
[M+H]+.
PPPÀPRP
Aggregation% ¼
 100%
PPP
SÀD
Inhibition%
¼
 100%
S
S: the platelet aggregation in the presence of solvent.
D: the platelet aggregation in the presence of test compounds.
Cytotoxicity assay
Mouse fibroblast cells (L-929) was chosen to evaluate the
in vitro cytotoxicity of the target compounds via Cell
Counting Kit-8 (CCK-8) assays. Experiment is mainly
divided into the following steps:
N1,N3-bis(3-chloro-4-methylphenyl)-4-
methoxyisophthalamide (1w)
White solid (The residue was recrystallized by ethanol.)
Yield: 84.30%; m.p.: 195.8–197.6 °C; IR (KBr) υmax
3361.19, 3327.30, 1645.22, 1592.26, 1400.48, 1258.40,
1. Preparation for experiment: through cell thawing, cell
culture and cell passage cultivation to obtain healthy
cells. Target compounds were diluted into 10 μM/L
and 100 μM/L with DMSO, respectively.
1181.27, 2947.25, 817, 578 cm−1 1H-NMR (400 MHz,
;
DMSO-d6, TMS): δ = 10.32 (2H, s, 2 × CONH), 8.25 (1H,
d, J = 2.2 Hz, 2-H), 8.26 (1H, dd, J = 2.4, 8.8 Hz, 6-H),
8.18–8.11 (1H, m, Ar-H), 7.99–7.90 (1H, m, Ar-H), 7.63
(1H, dd, J = 8.3, 1.9 Hz, Ar-H), 7.58–7.51 (1H, m, Ar-H),
7.32 (3H, dd, J = 8.3, 1.9 Hz, Ar-H), 7.19–7.24 (3H, m,
Ar–H), 3.96 (3H, s, OCH3), 2.29 (6H, s, 2 × Ar-CH3); 13C-
NMR (101 MHz, DMSO-d6): δ = 164.42 (2 × C, C = O),
159.0 (C, 4-C), 138.3 (C, 1a-C), 138.1 (C, 1b-C), 133.0 (C,
3a-C), 132.8 (C, 3b-C), 131.8 (C, 6-C), 131.2 (C, 4a-C),
131.0 (C, 4b-C), 130.3 (C, 5a-C), 130.1 (C, 5b-C), 129.0
(C, 1-C), 126.3 (CH, 2-C), 124.8 (C, 3-C), 120.2 (CH, 2a-
C), 119.5 (CH, 2b-C), 118.8 (CH, 6a-C), 118.3 (CH, 6b-C),
111.9 (CH, 5-C), 56.3 (CH3, O–CH3), 19.0 (2 × CH3,
Ar–CH3); MS (m/z): 443.0924 [M+H]+.
2. Cell inoculation: L-929 was added into 96-well
microplates (1 × 104 cell/well), cultivated in a humi-
dified 5% CO2 atmosohere at 37 °C for 24 h to allow
cells to attach.
3. Cell to medicine: cells were exposed to target
compounds at the concentrations of 10 μM/L and
100 μM/L and incubated at 37 °C for 48 h.
4. Measuring absorbance: After incubation at 37 °C for
48 h, the medium was removed and replaced with
fresh complete medium of RPMI-1640, then CCK-8
solution was added to the 96-well plates at 10 μL per
well and the absorbance of the solution was monitored
at 450 nm using a microplate reader (Bio-Tek Flx800

1980
Medicinal Chemistry Research (2018) 27:1971–1983
fluorescence microplate reader) after 2 h of incubation
(Xu et al. 1991).
At the onset of the experiments, the laboratory’s tradi-
tional method (Route a) was used to synthesize the inter-
mediates. The drawback of this method is the use of volatile
hydrogen chloride, it is not only difficult to handle but it
also corrode the equipment. Moreover, it is dangerous for
an industrial process. Considering the above question, a
safer method (Route b) was used to synthesize the inter-
mediates in subsequent experiments. Route b use Fries
rearrangement as the key step, avoiding the use of large
doses of hydrochloric acid, which is beneficial for the
environment and equipment.
Their cytotoxicity effect on L-929 cells were calculated
by the following formulas:
AbsðtestcellÞ À AbsðblankcellÞ
Cellviabilityð%Þ ¼
 100%
AbsðcontrolledcellÞ À AbsðblankcellÞ
Results and discussion
Pharmacology
Chemistry
In vitro anti-platelet aggregation activity
The synthetic pathway was disclosed in Scheme 1 and the
structure of target compounds were shown in Table 1. Key
intermediate 4-methoxylisophthaloyl dichloride was synthe-
sized in two routes (Route a and Route b). The intermediate
of compounds (1m, 1n, 1o, 1p, 1q, 1r, 1s, 1v, 1w) were
prepared by Route a and the intermediate of compounds (1d,
1e, 1f, 1g, 1h, 1i, 1j, 1k, 1t, 1u) were synthesized by Route b.
Taking picotamide and aspirin as positive control drugs,
in vitro activities on anti-platelet aggregation were tested
and assessed by using Born test for ADP inducer (Born
1962). After that, eight compounds (1c, 1h, 1i, 1k, 1p, 1q,
1v, and 1w), they were significantly more active in vitro
than control drug picotamide and their IC50 values for ADP
Scheme 1 Synthesis routes of intermediate (Route a and b) and target compounds (1d–1w)

Medicinal Chemistry Research (2018) 27:1971–1983
1981
Table 1 Structures of 4-
Compound
R1
R2
R3
Compound
R1
R2
R3
Methoxylisophthalamides (1a–
1w)
1aa
CH3
H
H
1m
1n
1o
1p
1q
1r
H
C≡CH
C≡N
H
H
1ba
1ca
1d
1e
1f
H
CH3
H
H
H
H
H
CH3
H
C≡N
H
CH2CH3
H
H
CF3
H
H
H
CH2CH3
H
CF3
H
H
H
H
CH2CH3
H
H
CF3
CF3
H
1g
1h
1i
CH(CH3)2
H
1s
Cl
CH3
H
H
H
CH(CH3)2
H
1t
CH3
CH3
Cl
H
H
H
H
H
CH(CH3)2
(CH2)3CH3
C(CH3)3
H
1u
1v
1w
CH3
H
1j
H
CH3
H
1k
1la
H
Cl
CH3
Bn
aReported compound
were calculated, they were selected and continued to testing
and assessing for two inducers both collagen and AA. The
anti-platelet aggregation activity of the target compounds
was listed in Table 2.
These results analysis for ADP and for collagen and for
AA were shown in Figs. 2 and 3, respectively.
Table 2 The inhibition rate and IC50 of tested compounds
Compound
Does
Inhibition IC50 Inhibition Inhibition
(μmol/ rate (%) in (μM/ rate (%) in rate (%) in
L)
ADP
L) in collagen
ADP
AA
Control group
–
–
–
–
–
As were shown in Fig. 2, compared with positive control
drugs picotamide and aspirin, 9 compounds 1h, 1i, 1n, 1o,
1p, 1q, 1s, 1v, and 1w had superior platelet aggregation
inhibition rate induced by ADP. Among them, compounds
1d, 1j, and 1k were equivalent to the positive control drug
picotamide, slightly lower than aspirin. Simultaneously, the
IC50 values of compounds 1h, 1i, 1k, 1p, 1q, 1v, 1w were
lower than the positive control drug picotamide and the
in vitro activities of four compounds 1h, 1i, 1q, and 1v with
IC50 values of 0.24 μM/L, 0.16 μM/L, 0.09 μM/L and
0.09 μM/L, respectively, they were higher than that of two
control drugs picotamide and aspirin with IC50 values of
0.60 μM/L and 0.26 μM/L induced by ADP (1.3 μM). Of
course, among them, compound 1q, that was a N1,N3-bis (3-
(trifluoromethyl)phenyl) -4-methoxyisophthalamide, had
the highest platelet aggregation inhibition rate with lowest
IC50 value of 0.09 μM/L. The inhibition rate of compound
1v was not high, but its IC50 value was the same as that of
compound 1q, which indicated that 1v had high platelet
inhibition rate at lower concentration.
Eight compounds (1h, 1i, 1k, 1p, 1q, 1u, 1v, and 1w)
which were significantly more active in vitro than control
drug picotamide were selected and continued to test and
assess for two inducers both collagen and AA. The analysis
in Fig. 3 showed two compounds 1p and 1q showed higher
in vitro activities of inhibition rates of 51.25% and 50.07%
than that of the 46.67% of picotamide and 46.67% of
aspirin induced by collagen, while 1p showed higher
in vitro activity of inhibition rate of 31.57% than that of the
29.67% of picotamide induced by AA at the concentration
Picotamide
Aspirin
1aa
1ba
1ca
1d
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
1.3
44.38
54.67
0.00
0.60 46.67
0.26 42.82
29.67
40.22
–
–
–
–
0.00
–
–
63.50
46.96
16.12
0.00
0.50
0.78
–
1e
–
–
–
–
1f
–
–
1g
43.58
91.27
58.45**
49.02
52.22*
89.30
22.34
62.60
67.63
57.64*
96.73**
10.27
56.71
27.61
11.62
86.24**
75.11**
–
–
1h
0.24 32.25
0.16 47.13**
–
13.56
28.47*
–
1i
1j
1k
1la
0.34 22.76
0.02
13.10
1m
1n
–
–
–
–
–
–
–
–
1o
–
1p
0.41 51.25**
0.09 50.07**
31.57*
24.55
–
1q
1r
–
–
–
–
–
–
–
–
1s
–
1t
–
1u
11.9
0.00
0.38
1v
0.09 0.00
0.31 0.00
1w
aReported compound
*p < 0.05 VS Control group
**p < 0.01 VS Control group

1982
Medicinal Chemistry Research (2018) 27:1971–1983
Table 3 Cytotoxicity effect on L-929
of 1.3 μM. Compound 1p, that was a N1,N3-bis (2-(tri-
fluoromethyl)phenyl)
-4-methoxyisophthalamide,
had
Compound
Dose (μM/L)
Absorbance
Survival rate (%)
similar structure with compound 1q and both of them had
shown good results in the pharmacological tests. which is
worth further researching.
Blank group
Control group
Picotamide
–
0.191
0.318
0.275
0.246
0.311
0.285
0.268
0.242
0.303
0.231
0.290
0.223
0.311
0.287
0.259
0.256
0.284
0.254
0.245
0.206
–
–
–
10
66.14
43.31
94.49
74.02
60.63
40.16
88.19
31.50
77.95
25.20
94.49
75.59
53.54
51.18
73.23
49.61
42.52
11.81
100
10
Cytotoxicity assay
Compound 1h
Compound 1i
Compound 1n
Compound 1o
Compound 1p
Compound 1q
Compound 1v
Compound 1w
100
10
The effect of the compounds with higher activities on
cytotoxicities was evaluated on mouse fibroblast cell (L-
929) via CCK-8 assays. The survival rate of cells was set
out in Table 3 and the cell morphology after exposed to
target compounds were shown in Fig. 4 (Xiong et al. 2007;
Abe et al. 2000).
The result analysis of cytotoxicities was shown in Fig. 5,
among the eight compound, six compound (1h, 1n, 1o, 1p,
1q, 1v) had higher cell survival rate than picotamide at the
dose of 10 μM/L. And the cell survival rate of three com-
pound (1h, 1p, 1q) was higher than that of picotamide at the
dose of 100 μM/L. Among them, compound 1h and 1q had
the lowest cytotoxicities at the dose of 10 μM/L and
100 μM/L.
100
10
100
10
100
10
100
10
100
10
100
10
100
Preliminary structure activity relationship (SAR)
exploration
1. The effect of anti-platelet aggregation activity when the
substitutions were saturated hydrocarbons
The order of inhibition of platelet was: in o-position of the
phenyl rings: 1l#a>1d > 1g > 1aa. in m-position: 1h > 1e >
1ba. in p-position: 1i > 1k > 1j. That meant the activity
could be enhanced when the saturated hydrocarbons with
bigger steric hindrance were introduced in the phenyl rings.
2. The effect of anti-platelet aggregation activity when the
substitution were unsaturated hydrocarbons
The order of inhibition of platelet was: 1o > 1n > aspirin >
picotamide > 1m. the result showed that cyano- substituted
compounds were more potent than the alkynyl-substituted
analogs. Therefore, the relatively strong electron-
withdrawing substituent were introduced into the phenyl
rings maybe could serve to promote the anti-platelet
aggregation activity.
Fig. 5 Cell Survival Rate of Compounds
activity increased significantly. When
a
electron-
withdrawing group (Cl) was introduced into the o-position
at the same time, Anti-platelet aggregation activities are
significantly improved (the inhibition rate: 1s > 1r).
3. The effect of anti-platelet aggregation activity when the
substitutions were trifluoromethyl
4. The effect of anti-platelet aggregation activity when two
substitutions were introduced into the phenyl rings
The order of inhibition of platelet was: 1q > 1p > 1s >
aspirin > picotamide > 1r. The result indicated that when the
trifluoromethyl was introduced into the o-position and m-
position of phenyl rings, the anti-platelet aggregation
Two methyl groups were introduced into the phenyl rings,
compounds 1t and 1u were obtained. But compounds 1t
and 1u almost had no anti-platelet aggregation activity.

Medicinal Chemistry Research (2018) 27:1971–1983
1983
Methyl and chloro groups were simultaneously intro-
duced into the phenyl rings, compounds 1v and 1w were
obtained. The order of inhibition of platelet was: 1v > 1w >
aspirin > picotamide. Therefore, methyl and electron-
withdrawing groups were simultaneously introduced into
the phenyl rings might help to promote the anti-platelet
aggregation activity.
To sum up, the anti-platelet aggregation activity will
increase when the saturated hydrocarbons with bigger steric
hindrance were introduced in the side chain phenyl rings.
The introduction of cyano and trifluoromethyl could
enhance the activity on anti-platelet aggregation. Simulta-
neous introduction of two methyl groups was unfavorable to
the activity, while the introduction of methyl and chloro
groups could significantly enhance the activity on anti-
platelet aggregation.
controlled study. The ADEP Group. Circulation 87
(5):1563–1569
Berrettini M, Cunto MD, Parise P (1990) “In vitro” and “ex vivo”
effects of picotamide, a combined thromboxane A 2 -synthase
inhibitor and -receptor antagonist, on human platelets. Eur J Clin
Pharmacol 39(5):495–500
Born GV (1962) Aggregation of platelets by ADP and its reversal.
Nature 194(4832):927–928
Brito FCF, Kummerle AE, Lugnier C, Fraga CAM, Barreiro EJ,
Miranda ALP (2010) Novel thienylacylhydrazone derivatives
inhibit platelet aggregation through cyclic nucleotides modulation
and thromboxane A2 synthesis inhibition. Eur J Pharmacol 638
(1-3):5–12
Celestini A, Violi F (2007) A review of picotamide in the reduction of
cardiovascular events in diabetic patients. Vasc Health Risk
Manag 3(1):93–98
Eskandariyan Z, Esfahani ZM, Haj METK, Mashayekhi V, Kobarfard
F (2014) Synthesis of thioether derivatives of quinazoline-4-one-
2-thione and evaluation of their antiplatelet aggregation activity.
Arch Pharm Res 37(3):332–339
Liu XJ, Shi XX, Zhong YL, Liu N, Liu K (2012) Design, synthesis
and in vitro activities on anti-platelet aggregation of 4-methox-
ybenzene-1,3-isophthalamides. Bioorg Med Chem Lett 22
(21):6591–6595
Conclusion
Liu XJ, Wang SQ, Zhang J, Zhang FX, Li GZ, Wang BJ, Shao YL,
Zhang LG, Fang L, Cheng MS (2006) Design, synthesis, and
activities of novel derivatives of isophthalamide and benzene-1,3-
disulfonamide. Chem Res Chin U 22(3):356–359
In summary, 19 novel 4-methoxyisophthalamide analogs
were synthesized and evaluated for anti-platelet activities, the
compounds with higher activities were selected to continue
research on cytotoxicities. The result exhibited that com-
pounds 1h, 1p, 1q, 1v showed stronger anti-platelet activity
and lower cytotoxicities. And, more remarkably, compound
1q, that was a N1,N3-bis (3-trifluoromethylphenyl)-4-meth-
oxyisophthalamides, had significant anti-platelet activity
when the inducers were ADP and collagen and it had lower
cytotoxicity at the dose of 100 μM/L. Thus, 1q is potential to
be studied and may help in developing new anti-platelet drug
with elevated activity and lower toxicity.
Liu XJ, He X, Cheng LS, Meng J, Shao YL, Si HQ, Hu T (2011)
Synthesis and in vitro activities on anti-platelet aggregation of N,
N’-di(2-substituted-phenyl)-4-methoxyisophthalamides and ben-
zene-1,3-disulfonamides. Chin Chem Lett 22(10):1139–1142
Liu XJ, Wang CQ, Meng J, Shi XX, Yan YN, Liu XG (2017a) Design,
synthesis and biological evaluation of 4-methoxy diaryl iso-
phthalates as antiplatelet agents. Med Chem Res 1:1–9
Liu XJ, Yan YN, Wang SQ, Li X, Liu XG (2017b) New synthesis of
4-methoxyisophthalic acid. Russ J Org Chem 53(3):459–461
Neri Serneri GG, Coccheri S, Marubini E, Violi F (2005) Picotamide,
a combined inhibitor of thromboxane A2 synthesis and receptor,
reduces 2-year mortality in diabetics with peripheral arterial
disease: the DAVID study. J Cheminform 24(52):1845–1852
Sartori E, Camy F, Teulon JM, Caussade F, Virone-Oddos A (1993)
Synthesis and activities of new arylsulfonamido thromboxane A
2, receptor antagonists. J Cheminform 24(52):625–632
Shaikh MH, Subhedar DD, Khedkar VM, Jha PC, Khan FAK,
Sangshetti JN, Shingate BB (2016) 1,2,3-Triazole tethered acet-
ophenones: synthesis, bioevaluation and molecular docking
study. Chin Chem Lett 27(07):1058–1063
Violi F, Ghiselli A, Iuliano L, Iuliano L, Praticò D (1988) Inhibition by
picotamide of thromboxane production in vitro and ex vivo. Eur J
Clin Pharmacol 33(6):599–602
Xiong JW, Xiao H, Zhang ZX (2007) An experimental research on
different detection conditions between MTT and CCK-8. Acta
Laser Biol Sin 16(5):559–562
Xu SY, Bian RL, Chen X (1991) Pharmacological experimental
methods, 2nd edn. People’s medical Publishing House, Beijing,
pp 1438–1442
Yip S, Benavente O (2011) Antiplatelet agents for stroke prevention.
Neurotherapeutics 8(3):475–487
Acknowledgements The authors are grateful to the National Science
Foundation of China (11341014) & the Committee of Science and
Technology of Tianjin of China (15JCZDJC33100) for the financial
supports and Shenyang Pharmaceutical University of China for run-
ning platelet aggregation assays.
Compliance with ethical standards
Conflict of interest The authors declare that they have no conflict of
interest.
References
Abe S, Ochi H, Takahashi Y, Ishijima SA, Osumi M, Yamaguchi H
(2000) Characteristic biological effects of itraconazole on L929
fibroblasts and their cell membrane. J Infect Chemother 6
(1):35–40
Balsano F, Violi F (1993) Effect of picotamide on the clinical pro-
gression of peripheral vascular disease. A double-blind placebo-