Supporting Information
Efficacy of phenylquinolinephenol derivatives as COX-2 inhibitors;
an approach to emergent the small molecules as the antiinflammatory and analgesic therapeutics
Manikandan Aa, Ravichandran Sb, Sathiyanarayanan K.Ic, and Sivakumar Aa*
a
b
School of Bio-Science and Technology, VIT University, Vellore-632014, Tamil Nadu, India.
Center of Advanced Study in Marine Biology, Annamalai University, Parangipettai- 608 502, India.
*Corresponding
Author,
Email:
[email protected],
Mobile:+919047699029, Fax: 0416-2243092
Telephone:
0416-224309,
Ext:
2523,
Graphical abstract:
Table of Contents Pages
Chemistry Part
A) Scheme 1. General synthesis of phenyl quinoline phenols 4a-l
2
B) Scheme 2. Proposed mechanism of 4a-l
2
C) Scheme 3. Achieved structures of 4a-l
3
D) NMR Spectra 4a-l
4-19
Biological evaluations part
E) Table 1. In-vitro COX-2 inhibition activity results of 4a-l
20
F) Table 2. In-vitro anti-inflammatory activity (HRBC) results of 4a-l
20
G) Figure 1-24. Molecular docking (COX-2 & PQPDs interactions)
21-32
H) Figure 25. COX-2 PyMol view
33
1
Chemistry Part
General procedure for the preparation of compound 4a-l:
According to Scheme 1 depicted here, a mixture of the substituted salicylaldehyde 1 (10 mmol)
and substituted aromatic amine 2 (10 mmol) was stirred in nitromethane at room temperature for
10 minutes. After addition of 10 mol % anhydrous iodine and 20 mol % of Cu2O, phenyl
acetylene 3 (10 mmol) was added and the reaction mixture was refluxed for 24 h. The reaction
was monitored by TLC and the reaction mixture was cooled and diluted with saturated
NaHCO3 solution (80 mL). This was extracted with ethyl acetate (3 x 20 mL). The combined
organic extract was washed with brine and dried over (Na2SO4). The solvent was distilled off and
the resulting product was purified by column chromatography over silica gel (60-120 mesh)
using n-hexane-ethyl acetate mixture (9:1) as eluent to give the compounds (4a-l) and the
compound formed in two steps as mentioned in the proposed mechanism below here.
R1
OH
1
CHO
+
R2
Molecular I2,
Cu2O
R2
CH3NO2, Reflux
2
NH2
+
N
24 h
R1
HO
3
4a-l
Scheme 1. Synthesis of substituted 2-(4-phenylquinolin-2-yl) phenol (4a-4l)
Step 1 (imine formation)
NH2
OH
I2
OH
OH
O
OH
I2
H
N
HN
Step 2 (1, 2- Cycloaddition)
N
N
HO
HO
Scheme 2. Proposed mechanism for the formation of PQPDs (4a-l)
2
N
N
N
OH
N
OH
OH
4a Y = 88%
4b Y = 73%
4c Y = 65%
OH
4d Y = 78%
OMe
Cl
OEt
F
Br
N
Cl
N
N
OH
OH
OH
4e Y = 78%
4f Y = 71%
Br
Me
N
OH
4i Y = 66%
OH
4g Y = 90%
OMe
N
N
Me
N
OH
4j Y = 87%
4h Y = 88%
Me
N
OH
4k Y = 86%
OH
4l Y = 61%
Scheme 3. Proposed structures and yield details of PQPDs 4a-l
Note: Y-Yield
3
2-(4-phenylquinolin-2-yl)phenol (4a):
Yellow solid; Melting point: 153-155 0C: IR (KBr): 3263, 3088, 3057, 3030, 2917, 2876, 2331,
1737, 1585, 1467, 1433, 1398, 1274, 1213, 1120, 893, 752, 698, 675,663, 646 cm-1:
1
H NMR
(400 MHz, CDCl3) δ ppm: 6.95-6.91 (t, J = 8.0 Hz, 1H), 7.10-7.08 (t, J = 8.0 Hz, 1H), 7.37-7.33
(t, J = 8.0 Hz, 1H), 7.50-7.46 (t, J = 8.0 Hz, 1H), 7.58-7.51 (m, 5H), 7.75-7.71 (t, J = 8.0 Hz,
1H), 7.88-7.86 (d, J = 8.0 Hz, 1H), 7.95-7.94 (d, J = 4.0 Hz, 2H), 7.50-7.46 (t, J = 8.0 Hz, 1H),
8.09-8.07 (d, J = 8.0 Hz, 1H), 15.29 (s, 1H); 13C NMR (100 MHz, CDCl3) δ ppm: 117.5, 118.6,
118.7, 118.9, 121.2, 125.3, 125.8, 126.7, 126.9, 126.9, 127.9, 128.7, 128.7, 129.4, 129.4, 130.3,
132.0, 137.9, 145.2, 150.2, 157.4, 161.1; HRMS for C21H15NO Calculated [M+] m/z 297.1154,
Found 297.1150.
2-(4-phenylbenzo[h]quinolin-2-yl)phenol (4b):
Yellow solid; Melting point: 180-182 0C: IR (KBr): 3256, 3092, 3057, 3013, 2976, 2337, 2328,
2297, 1944, 1815, 1705, 1583, 1492, 1365, 1296, 1280, 1215, 1195, 1068, 876, 829, 704, 628
cm-1:
1
H NMR (400 MHz, CDCl3) δ ppm: 6.96-6.92 (t, J = 8.0 Hz, 1H), 7.15-7.13 (d, J = 8.0
Hz, 1H), 7.39-7.35 (t, J = 8.0 Hz, 1H), 7.55-7.53 (m, 5H), 7.78-7.68 (m, 4H), 7.89-7.87 (d, J =
8.0 Hz, 1H), 7.97-7.95 (d, J = 8.0 Hz, 1H), 8.05 (s, 1H), 8.98-8.96 (d, J = 8.0 Hz, 1H), 15.54 (s,
1H) ;
13
C NMR (100 MHz, CDCl3) δ ppm: 118.3, 118.5, 118.9, 119.2, 122.7, 123.2, 124.1,
126.9, 127.7, 128.2, 128.7, 128.7, 128.7, 129.6, 129.7, 131.9, 133.8, 138.2, 143.7, 150.3, 155.8,
160.6; HRMS for C25H17NO Calculated [M+] m/z 347.1310, Found 347.1312.
2-methoxy-6-(4-phenylquinolin-2-yl)phenol (4c):
Yellow solid; Melting point: 164-166 0C: IR (KBr): 3250, 3087, 3053, 2985, 2945, 2825, 2336,
1741, 1591, 1550, 1431, 1363, 1240, 1049, 902, 758, 705, 601 cm-1:
1
H NMR (400 MHz,
CDCl3) δ ppm: 3.97 (s, 3H), 6.90-6.86 (t, J = 8.0 Hz, 1H), 6.99-6.97 (d, J = 8.0 Hz, 1H), 7.527.48 (t, J = 8.0 Hz, 1H), 7.59-7.56 (m, 6H), 7.77-7.73 (t, J = 8.0 Hz, 1H), 7.89-7.87 (d, J = 8.0
Hz, 1H), 7.96 (s, 1H), 8.09-8.07 (d, J = 8.0 Hz, 1H), 15.88 (s, 1H); 13C NMR (100 MHz, CDCl3)
δ ppm: 56.1, 113.3, 117.8, 118.6, 118.8, 125.3, 125.8, 126.7, 127.8, 128.7, 128.7, 129.4, 130.4,
137.9, 145.0, 149.5, 150.2, 151.6, 157.60; HRMS for C22H17NO2 Calculated [M+] m/z 327.1259,
Found 327.1260.
4
2-ethoxy-6-(4-phenylquinolin-2-yl)phenol (4d):
Yellow solid; Melting point: 157-159 0C: IR (KBr): 3258, 3099, 3053, 2981, 2904, 2872, 2337,
1717, 1593, 1548, 1462, 1440, 1357, 1238, 1049, 1031, 902, 875, 858, 769, 702, 574 cm-1: 1H
NMR (400 MHz, CDCl3) δ ppm: 1.57-1.53 (t, J = 8.0 Hz, 3H), 4.20-4.14 (q, J = 8.0 Hz, 2H),
3H), 6.87-6.83 (t, J = 8.0 Hz, 1H), 6.98-6.96 (d, J = 8.0 Hz, 1H), 7.50-7.46 (t, J = 8.0 Hz, 1H),
7.56-7.55 (m, 6H), 7.75-7.71 (t, J = 8.0 Hz, 1H), 7.88-7.86 (d, J = 8.0 Hz, 1H), 7.95 (s, 1H),
8.07-8.05 (d, J = 8.0 Hz, 1H), 15.94 (s, 1H);
13
C NMR (100 MHz, CDCl3) δ ppm: 15.0, 64.4,
114.6, 117.8, 118.6, 118.8, 125.3, 125.8, 126.7, 127.7, 128.7, 128.7, 129.4, 130.4, 137.9, 145.0,
148.8, 150.1, 151.7, 157.6; HRMS for C23H19NO2 Calculated [M+] m/z 341.1416, Found
341.1416.
4-bromo-2-(4-phenylquinolin-2-yl)phenol (4e):
Yellow solid; Melting point: 184-186 0C: IR (KBr): 3267, 3091, 3041, 3011, 2987, 2335, 1737,
1579, 1489, 1467, 1392, 1304, 1255, 1207, 1182, 1085, 975, 883, 812, 705, 680, 628, 597 cm-1:
1
H NMR (400 MHz, CDCl3) δ ppm: 6.99-6.97 (d, J = 8.0 Hz, 1H), 7.43-7.40 (dd, J = 16.0, 4.0
Hz, 1H), 7.60-7.49 (m, 6H), 7.77-7.73 (t, J = 8.0 Hz, 1H), 7.90-7.88 (d, J = 8.0 Hz, 2H), 8.04 (s,
1H), 8.09-8.07 (d, J = 8.0 Hz, 1H), 15.34 (s, 1H);
C NMR (100 MHz, CDCl3) δ ppm: 110.4,
13
117.3, 120.5, 125.5, 125.9, 127.0, 127.9, 128.8, 128.9, 129.4, 129.4, 130.5, 134.5, 137.6, 145.1,
150.6, 156.0, 160.2; HRMS for C21H14BrNO Calculated [M+] m/z 375.0259, Found 375.0264.
2-(7-chloro-4-phenylquinolin-2-yl)phenol (4f):
Yellow solid; Melting point: 140-142 0C: IR (KBr): 3270, 3096, 3057, 2939, 2871, 2331, 1726,
1573, 1544, 1487, 1460, 1415, 1371, 1296, 1278, 1199, 1156, 1153, 1076, 981, 877, 821, 737,
626 cm-1: 1H NMR (400 MHz, CDCl3) δ ppm: 6.97-6.93 (q, J = 4.0 Hz, 1H), 7.13-7.11 (d, J =
8.0 Hz, 1H), 7.67-7.28 (m, 8H), 7.84-7.82 (d, J = 8.0 Hz, 1H), 7.96-7.91 (m, 2H), 8.10-8.04 (m,
2H), 14.82 (s, 1H);
13
C NMR (100 MHz, CDCl3) δ ppm: 117.7, 118.3, 118.9, 121.2, 123.0,
123.7, 126.9, 127.5, 127.7, 127.8, 128.0, 128.8, 129.0, 129.9, 131.2, 132.4, 132.5, 136.3, 137.4,
140.7, 145.8, 146.9, 149.8, 150.1, 157.1, 158.5, 161.1; HRMS for C21H14ClNO Calculated [M+]
m/z 331.0764, Found 337.0767.
5
2-(6-chloro-4-phenylquinolin-2-yl)phenol (4g):
Yellow solid; Melting point: 156-158 0C: IR (KBr): 3263, 3091, 3059, 3026, 2920, 2848, 2397,
2343, 1764, 1743, 1587, 1485, 1417, 1352, 1298, 1274, 1124, 1072, 869, 819, 738, 700, 684, 617
cm-1:
1
H NMR (400 MHz, CDCl3) δ ppm: 6.98-6.94 (t, J = 8.0 Hz, 1H), 7.13-7.11 (d, J = 8.0
Hz, 1H), 7.42-7.38 (t, J = 8.0 Hz, 1H), 7.64-7.55 (m, 5H), 7.71-7.69 (d, J = 8.0 Hz, 1H), 7.877.86 (d, J = 8.0 Hz, 1H), 7.95-7.94 (d, J = 4.0 Hz, 1H), 7.99 (s, 1H), 8.05-8.03 (d, J = 8.0 Hz,
1H), 14.90 (s, 1H);
13
C NMR (100 MHz, CDCl3) δ ppm: 118.4, 118.7, 118.9, 119.2, 122.4,
124.7, 126.0, 127.0, 128.9, 129.0, 129.3, 129.5, 131.2, 132.3, 132.6, 133.4, 137.2, 143.7, 149.4,
157.7, 161.0, 162.9; HRMS for C21H14ClNO Calculated [M+] m/z 331.0764, Found 337.0763.
2-(6-fluoro-4-phenylquinolin-2-yl)phenol (4h):
Yellow solid; Melting point: 167-169 0C: IR (KBr): 3270, 3091, 3059, 2991, 2971, 2351, 2330,
1726, 1624, 1587, 1550, 1508, 1494, 1425, 1382, 1355, 1251, 1195, 1109, 925, 873, 746, 702,
655 cm-1: 1H NMR (400 MHz, CDCl3) δ ppm: 6.95-6.91 (t, J = 8.0 Hz, 1H), 7.09-7.07 (d, J = 8.0
Hz, 1H), 7.37-7.33 (t, J = 8.0 Hz, 1H), 7.56-7.48 (m, 7H), 7.93-7.91 (d, J = 8.0 Hz, 1H), 7.96 (s,
1H), 8.07- (s, 1H), 14.88 (s, 1H);
13
C NMR (100 MHz, CDCl3) δ ppm: 109.4, 109.7, 118.2,
118.6, 118.8, 118.8, 120.2, 120.4, 126.1, 126.2, 126.9, 128.9, 129.0, 129.2, 130.3, 130.4, 132.1,
137.4, 142.3, 149.6, 149.7, 156.9, 156.9, 159.4, 160.8, 161.9; HRMS for C21H14FNO Calculated
[M+] m/z 315.1059, Found 315.1061.
2-(6-bromo-4-phenylquinolin-2-yl)phenol (4i):
Yellow solid; Melting point: 166-168 0C: IR (KBr): 3280, 3092, 3057, 3028, 2669, 2397, 2330,
1957, 1851, 1737, 1610, 1583, 1543, 1500, 1485, 1369, 1350, 1294, 1244, 1120, 1039, 871, 823,
746, 700, 669, 630 cm-1: 1H NMR (400 MHz, CDCl3) δ ppm: 6.99-6.95 (t, J = 8.0 Hz, 1H), 7.137.11 (d, J = 8.0 Hz, 1H), 7.42-7.38 (t, J = 8.0 Hz, 1H), 7.65-7.55 (m, 5H), 7.85-7.83 (dd, J =
16.0, 4.0 Hz, 1H), 7.98-7.96 (d, J = 4.0 Hz, 2H), 8.00 (s, 1H), 8.04-8.03 (d, J = 4.0 Hz, 1H),
14.91 (s, 1H);
C NMR (100 MHz, CDCl3) δ ppm: 117.3, 118.4, 118.7, 118.9, 120.7, 122.8,
13
126.5, 127.0, 128.0, 128.9, 129.0, 129.3, 129.6, 132.3, 132.5, 133.7, 137.2, 144.0, 149.3, 157.8,
161.0, 161.1, 163.0; HRMS for C21H14BrNO Calculated [M+] m/z 375.0259, Found 375.0259.
6
2-(6-methoxy-4-phenylquinolin-2-yl)phenol (4j):
Yellow solid; Melting point: 144-146 0C: IR (KBr): 3266, 3088, 3043, 2999, 2943, 2331, 1778,
1620, 1585, 1543, 1506, 1494, 1386, 1267, 1215, 1120, 1029, 906, 887, 754, 698, 624, 590 cm-1:
1
H NMR (400 MHz, CDCl3) δ ppm: 3.80 (s, 3H), 6.94-6.90 (t, J = 8.0 Hz, 1H), 7.09-7.07 (d, J =
8.0 Hz, 1H), 7.17-7.16 (d, J = 4.0 Hz, 1H), 7.35-7.31 (t, J = 8.0 Hz, 1H), 7.42-7.38 (dd, J = 16.0,
4.0 Hz, 1H), 7.57-7.53 (m, 5H), 7.91 (s, 1H), 7.93-7.92 (d, J = 4.0 Hz, 1H), 8.01-7.99 (d, J = 8.0
Hz, 1H), 15.13 (s, 1H);
C NMR (100 MHz, CDCl3) δ ppm: 55.5, 104.2, 117.9, 118.5, 118.7,
13
119.1, 122.4, 126.3, 126.6, 128.7, 128.8, 129.2, 129.4, 131.5, 138.2, 141.1, 148.8, 155.1, 158.1,
160.7; HRMS for C22H17NO2 Calculated [M+] m/z 327.1259, Found 327.1259.
2-(6-methyl-4-phenylquinolin-2-yl)phenol (4k):
Yellow solid; Melting point: 168-170 0C: IR (KBr): 3250, 3089, 3047, 2926, 2858, 2330, 1741,
1583, 1546, 1490, 1355, 1246, 1124, 1076, 962, 871, 752, 744, 705, 594 cm-1: 1H NMR (400
MHz, CDCl3) δ ppm: 2.46 (s, 3H), 6.94-6.90 (t, J = 8.0 Hz, 1H), 7.09-7.07 (d, J = 8.0 Hz, 1H),
7.36-7.32 (t, J = 4.0 Hz, 1H), 7.57-7.53 (m, 6H), 7.61 (s, 1H), 7.91 (s, 1H), 7.94-7.92 (d, J = 4.0
Hz, 1H), 7.99-7.97 (d, J = 8.0 Hz, 1H), 15.30 (s, 1H); 13C NMR (100 MHz, CDCl3) δ ppm: 21.8,
117.6, 118.5, 118.6, 119.1, 121.0, 124.6, 125.3, 126.8, 127.7, 128.6, 128.7, 129.4, 130.0, 131.8,
132.4, 136.8, 138.1, 143.7, 149.5, 156.5, 161.0, 161.7; HRMS for C22H17NO Calculated [M+]
m/z 311.1310, Found 311.1316.
2-(5,7-dimethyl-4-phenylquinolin-2-yl)phenol (4l):
Yellow solid; Melting point: 149-151 0C: IR (KBr): 3290, 3142, 3057, 2978, 2861, 2328, 1735,
1597, 1494, 1411, 1381, 1363, 1300, 1222, 1153, 765, 756, 868, 659, 619, 588 cm-1: 1H NMR
(400 MHz, CDCl3) δ ppm: 2.00 (s, 3H), 2.54 (s, 3H), 6.94-6.90 (t, J = 8.0 Hz, 1H), 7.12-7.10 (d,
J = 8.0 Hz, 1H), 7.14 (s, 1H), 7.40-7.28 (m, 3H), 7.52-7.49 (m, 3H), 7.79-7.77 (d, J = 8.0 Hz,
2H), 7.92-7.90 (d, J = 8.0 Hz, 1H), 15.44 (s, 1H);
C NMR (100 MHz, CDCl3) δ ppm: 21.4,
13
24.1, 118.5, 118.5, 118.7, 118.7, 118.9, 122.8, 125.9, 126.8, 128.0, 128.0, 128.6, 131.8, 132.3,
135.3, 140.1, 142.4, 146.6, 150.4, 155.9, 161.2; HRMS for C23H19NO Calculated [M+] m/z
325.1467, Found 325.1469.
7
NMR spectra of PQPDs (4a-l)
1H-NMR
spectrum of 4a
13
C-NMR spectrum of 4a
8
1
H-NMR spectrum of 4b
13
C-NMR spectrum of 4b
9
1
H-NMR spectrum of 4c
13
C-NMR spectrum of 4c
10
1
H-NMR spectrum of 4d
13
C-NMR spectrum of 4d
11
1
H-NMR spectrum of 4e
13
C-NMR spectrum of 4e
12
1
H-NMR spectrum of 4f
13
C-NMR spectrum of 4f
13
1
H-NMR spectrum of 4g
13
C-NMR spectrum of 4g
14
1
H-NMR spectrum of 4h
13
C-NMR spectrum of 4h
15
1
H-NMR spectrum of 4i
13
C-NMR spectrum of 4i
16
1
H-NMR spectrum of 4j
13
C-NMR spectrum of 4j
17
1
H-NMR spectrum of 4k
13
C-NMR spectrum of 4k
18
1
H-NMR spectrum of 4l
13
C-NMR spectrum of 4l
19
Bioactivity part
Table S1. In-vitro COX-2 inhibition activity results of 4a-l*.
Compounds
% Protection±SD
IC50 µM**
4a
No inhibition
-
4b
49.43±1.98
4.674
4c
No inhibition
-
4d
No inhibition
-
4e
50.35±1.43
3.754
4f
78.22±2.75
0.181
4g
70.98±2.91
1.092
4h
84.24±1.04
0.088
4i
60.41±2.17
1.112
4j
82.01±1.45
0.102
4k
48.24±2.01
6.153
4l
68.32±1.31
1.162
Diclofenac
79.12±1.01
0.092
Control#
No inhibition
--
# Distilled Water, *S.D. = Standard deviation (Average of three determinations), ** All values are the mean of duplicate or triplicate
measurements
Table S2. In-vitro anti-inflammatory activity (HRBC) results of 4a-l*.
Compounds
Mean absorbance±SD*
% Protection
IC50 µM**
4a
0.2812±0.012
48.98
6.382
4b
0.3088±0.009
54.91
1.031
4c
0.1696±0.049
22.24
12.38
4d
0.1601±0.046
20.88
11.42
4e
0.3106±0.016
55.35
0.983
4f
0.3844±0.008
78.22
0.064
4g
0.3028±0.014
50.98
1.157
4h
0.3982±0.006
82.72
0.021
4i
0.3641±0.011
62.65
0.484
4j
0.3781±0.007
58.96
0.629
4k
0.1511±0.009
53.01
0.702
4l
0.3278±0.008
58.38
0.652
Diclofenac
0.3812±0.006
80.03
0.090
Control#
0.1886±0.028
No inhibition
--
# Distilled Water, *S.D. = Standard deviation (Average of three determinations), ** All values are the mean of duplicate or triplicate
measurements
20
Figure 1-24. Molecular docking (COX-2 & 4a-l interactions)
Fig. 1. 4a interaction with COX-2 (PDB ID: 1CX2)
Fig. 2. 4a in the binding pocket of COX-2 (PDB ID: 1CX2)
21
Fig. 3. 4b interaction with COX-2 (PDB ID: 1CX2)
Fig. 4. 4b in the binding pocket of COX-2 (PDB ID: 1CX2)
22
Fig. 5. 4c interaction with COX-2 (PDB ID: 1CX2)
Fig. 6. 4c in the binding pocket of COX-2 (PDB ID: 1CX2)
23
Fig. 7. 4d interaction with COX-2 (PDB ID: 1CX2)
Fig. 8. 4d in the binding pocket of COX-2 (PDB ID: 1CX2)
24
Fig. 9. 4e interaction with COX-2 (PDB ID: 1CX2)
Fig. 10. 4e in the binding pocket of COX-2 (PDB ID: 1CX2)
25
Fig. 11. 4f interaction with COX-2 (PDB ID: 1CX2)
Fig. 12. 4f in the binding pocket of COX-2 (PDB ID: 1CX2)
26
Fig. 13. 4g interaction with COX-2 (PDB ID: 1CX2)
Fig. 14. 4g in the binding pocket of COX-2 (PDB ID: 1CX2)
27
Fig. 15. 4h interaction with COX-2 (PDB ID: 1CX2)
Fig. 16. 4h in the binding pocket of COX-2 (PDB ID: 1CX2)
28
Fig. 17. 4i interaction with COX-2 (PDB ID: 1CX2)
Fig. 18. 4i in the binding pocket of COX-2 (PDB ID: 1CX2)
29
Fig. 19. 4j interaction with COX-2 (PDB ID: 1CX2)
Fig. 20. 4j in the binding pocket of COX-2 (PDB ID: 1CX2)
30
Fig. 21. 4k interaction with COX-2 (PDB ID: 1CX2)
Fig. 22. 4k in the binding pocket of COX-2 (PDB ID: 1CX2)
31
Fig. 23. 4l interaction with COX-2 (PDB ID: 1CX2)
Fig. 24. 4l in the binding pocket of COX-2 (PDB ID: 1CX2)
32
Fig. 25. COX-2 (PDB ID: 1CX2) PyMol view
33