ISSN: 0970-020 X

ORIENTAL JOURNAL OF CHEMISTRY CODEN: OJCHEG

An International Open Free Access, Peer Reviewed Research Journal
2017, Vol. 33, No. (2):
Pg. 821-828
www.orientjchem.org

Synthesis of Benzo[g]quinoxaline-5,10-dione Based

Pyridine Derivatives and their Antimycobacterial Activity
SHIV KUMAR1*, NITIN KUMAR2 and SUSHMA DRABU3

1
Faculty of Pharmacy, Uttrakhand Technical University, Dehradun-248007, India.
2
Oxford College of Pharmacy, Masoori Industrial Area, U.P.S.I.D.C., M.G. Road,
Ghaziabad-201001, India.
3
Maharaja Surajmal Institute of Pharmacy, Janak Puri, New Delhi-110058, India.
*Corresponding author E-mail: [email protected]

http://dx.doi.org/10.13005/ojc/330230

(Received: January 11, 2017; Accepted: March 25, 2017)

ABSTRACT

Thirteen new compounds belonging to series 2-amino-6-(5,10-dioxo-2,3-diphenyl-

5,10-dihydrobenzo[g]quinoxalin-7-yl)-4-(substituted)phenylpyridine-3-carbonitrile (6a-m) were
synthesized by multistep synthetic scheme. The newly synthesized compounds were screened for
their antimycobacterial activity by L.J. Slope (Conventional) Method. Compound 6h with 4-N(CH3)2
group at phenyl ring of above mentioned position has been reported as most active antimycobacterial
compound and compound 6k with 4-CH3 substitution at phenyl above mentioned position has been
reported as the least active antimycobacterial compound.

Keywords: Quinoxaline, Benzo[g]quinoxaline-5,10-dione, Pyridine, Antimycobacterial.

INTRODUCTION estimated 9.6 million new TB cases were reported

and 1.5 million deaths were caused due to TB
Quinoxaline and pyridine derivatives worldwide16. Quinoxaline ring is also a an important
constitute the important part of various antibiotics part of antileprotic drug clofazimine (CZM) which
viz. hinomycin, levomycin and actinoleutin that is also widely indicated for treatment of multidrug
inhibit the growth various bacteria and are found resistant tuberculosis. In Mycobacterium tuberculosis,
to be cytotoxic against various transplantable CZM is reduced by NADH-dehydrogenase (NDH-2)
tumours1-7. Quinoxaline derivatives also exhibited a to release reactive oxygen17 and CZM also act as
wide range of biological activities viz. antifungal8-10, competitor with menaquinone (MK-4), which acts
antibacterial11-12, antitubercular8-9,12-15. as a key factor for its reduction by NDH-218. First
line antitubercular drug isoniazid (INH) and second
Tuberculosis (TB) is a major health problem line antitubercular drug ethionamide (Etm) are
worldwide now-a-days. In year 2014, there were an derivatives of six membered nitrogen containing
822 KUMAR et al., Orient. J. Chem., Vol. 33(2), 821-828 (2017)

pyridine ring. Isoniazid and Ethionamide, after 756 (ortho-disubstituted phenyl ring), 856 (meta-
bioactivation by mycobacterial catalase-peroxidase disubstituted phenyl ring), 1160 (aryl C-Cl), 1384
enzyme complex, inhibit synthesis of mycolic acids19- (aromatic C=C); 1H-NMR (CDCl3 300 MHz) d : 8.16-
23
, that are important components of mycobacterial 8.20 (q, J=5-6 Hz, A2B2 pattern, 2H, Ar-H), 7.74-7.79
cell wall. (m, J=4.5-6 Hz, 2H, Ar-H), 7.50-7.53 (m, J=1-4 Hz,
2H, Ar-H)
EXPERIMENTAL
Synthesis of 2,3-Diaminonaphthalene (2)
Chemistry Compound 1 (2 gm) was dissolved in
Melting points were measured in open absolute ethanol. Ammonium chloride (1.5 gm),
capillary tubes and are uncorrected. All the Fourier- dil. HCl (in catalytic amount) and excess of strong
Transform Infra Red (FT-IR) spectra were recorded ammonium solution were added to it and refluxed
on Shimdazu FT-8400 Spectrophotometer using KBr for 6 h on water bath till appearing of permanent dark
pallets.The 1H- NMR spectra were recorded on Bruker- greenish colour. After completion of reaction, solution
Spectrospin DCX NMR spectrometer using DMSO-d6 was cooled and kept in deep freezer for overnight.
and CDCl3 as solvents and tetramethylsilane (TMS)
as an internal standard (Chemical shifts expressed in Greenish crystals were filtered, dried and
d/ppm The purity of the compounds was checked by recrystallized with methanol. Dark greenish crystals;
thin layer chromatography (TLC) on Merck Silica Gel Yield 75%; Rf 0.89, Rf 0.82; mp 150oC; IR (nmax,
60F254 precoated sheets using Toluene : Ethylacetate KBr, cm-1): 758, 854, 1473 (aromatic =C-N), 1500
: Formic acid (5:4:1) solution as mobile phase. (aromatic C=C), 1681 (N-H), 3049 (N-H); 1H-NMR
(CDCl3 500 MHz): 7.45-7.46 (dd, J=3.5-4 Hz, 2H,
Synthesis of 2,3-Dichloronaphthalene (1) Ar-H), 7.12-7.14 (dd, J=3-3.5 Hz, 2H, Ar-H), 7.09 (s,
2,3-dihydroxynaphthalene (5 gm) was Ar-H, 2H), 7.05 (s, Ar-NH2,4H).
dissolved in phosphorusoxychloride (90 mL) in parts
at 0o-5oC with occasionally stirring and refluxed for 2,3-Diphenylbenzo[g]quinoxaline (3)
about 4 h till the appearance of clear dark pinkish Equimolar quantities of compound 2
solution. This solution was cooled and poured (0.001 mol) and benzil (0.001 mol) were dissolved in
into ice-cold water. A dark pinkish solid appeared ethanol (30 mL) and refluxed for 2 h. After completion
and was filtered, dried and recrystallized with of reaction, reaction mixture was cooled, kept for
dimethylformaide (DMF). Pinkish crystalline solid; overnight, yellow crystalline solid was filtered, dried
Yield 85%; Rf 0.72; mp 185oC. IR (nmax, KBr, cm-1): and recrystallized with ethanol : methanol (1:1)

NH2 N
OH Cl
(i) (ii) (iii)
N
OH Cl NH2

(2) (3)
(1)
(iv)
Compound R O
O
6a H N
N
6b 4-Cl
(v)
H3C
6c 4-OH N
N

6d 4-NO2 O O
O
(4)
(5)
6e 4-OCH3

6f 3-OH-4-OCH3 (vi) O

6g 2-Cl N

6h 4-N(CH3)2
N
6i 4-CF3
N
O
6j 3,4-(OCH3)2 R
N
C NH2
6k 4-CH3
(6a...)
6l 4-NH2

6m 2-OH

Reagents & Conditions : (i) POCl3, Reflux for 4 h (ii) NH4Cl, NH3 in excess, EtOH, Reflux at water-bath for 6 h
(iii) Benzil, EtOH, Reflux for 2 h (iv) CrO3/AcOH, Reflux for 2 h (v) CH3COCl, CCl4, AlCl3, Reflux for 6 h (vi)
Malononitrile, R-C6H4-CHO, CH3OONH4, NH3, EtOH, dil. HCl, Reflux for 48-50 h

Fig. 1: Reaction Scheme

 KUMAR et al., Orient. J. Chem., Vol. 33(2), 821-828 (2017) 823

mixture. Yellow crystalline solid; Yield 72%; Rf 0.83; Equimolar quantities of compound 5
mp 170oC; IR (nmax,KBr, cm-1) : 725, 785, 820, 875, (0.01 mol), appropriate benzaldehyde (0.01 mol),
1475, 1510 (aromatic C=C), 1660 (quinoxaline ammonium acetate (2 gm) and malononitrile (0.01
C=N); 1H-NMR (DMSO-d6 500 MHz) d: 8.73-8.77 (m, mol) and were dissolved in ethanol (30 mL) and
J=2.5-4 Hz, 10H, Ar-H), 8.28 (s, 2H, Ar-H), 8.03-8.07 was refluxed under anhydrous conditions for 48-50
(m, 4H, J=2.5-4 Hz, Ar-H). h till the permanent appearance of crystalline solid.
Solid was filtered, dried and recrystallized with
Synthesis of 2,3-Diphenylbenzo[g]quinoxaline- chloroform.
5,10-dione (4)
Compound 3 (1.2 gm) was dissolved in 2-Amino-6-(5,10-dioxo-2,3-diphenyl-5,10-
glacial acetic acid (30 mL) and chromium trioxide dihydrobenzo[g]quinoxalin-7-yl)-4-phenyl
(1.1 gm) in 12 mL of glacial acetic acid : water (1:1) pyridine-3-carbonitrile (6a)
solution was added to it. This solution was refluxed Yellow crystalline solid; Yield 74%; Rf 0.84;
at 80oC for 2 h and then poured into ice-chilled mp 230oC; IR (nmax, KBr, cm-1): 719, 756, 820, 875,
water (1500 mL). Resulting solid was filtered, dried, 1296 (C=N, pyridine), 1384 (C-N), 1608 (>C=O,
recrystallized with methanol. White solid; Yield 65%; cyclic, conjugated), 1642 (N-H, primary amine,
Rf 0.78; mp 162oC; IR (nmax, KBr, cm-1): 718, 795, bend), 1697 (C=N, quinoxaline), 2358 (C-N, nitrile),
875, 1450, 1594 (>C=O, cyclic, conjugated), 1676; 3417 (N-H, primary amine, str.); 1H-NMR (CDCl3 500
1
H-NMR (CDCl3 500 MHz) d: 8.13-8.15 (d, 2H, J=8.5 MHz) d: 12.47 (s, 1H, Ar-H), 11.91 (s, 2H, Ar-H),
Hz, Ar-H), 7.85-7.88 (t, J=2.5-10.5 Hz, 4H, Ar-H), 11.73 (bs, 2H, Ar-NH2), 9.03-9.04 (d, J=5 Hz, 1H,
7.81-7.82 (d, J=8 Hz, 2H, Ar-H), 7.50-7.53 (t, J=7.5 Ar-H), 8.01-8.02 (d, 2H, J=8 Hz, Ar-H), 7.75-7.79
Hz, 6H, Ar-H); Elemental Anal. Found (Calcd.) for (t, J=8-12 Hz, 3H, Ar-H), 7.59-7.63 (t, J=8.5-9.5 Hz,
C24H14N2O2: Found: C, 79.42 (79.55); H 3.91 (3.89); 4H, Ar-H), 7.12-7.30 (m, J=7.5-14.5 Hz, 6H, Ar-H);
N 7.68 (7.73); O 8.95 (8.83). MS-ESI: 555.05 (m/z, M+); Elemental Anal. Found
(Calcd.)for C36H21N5O2 : C, 77.80 (77.83); H, 3.82
Synthesis of 7-Acetyl-2,3-diphenylbenzo[g] (3.81); N, 12.60 (12.61); O, 5.76 (5.76).
quinoxaline-5,10-dione (5)
Anhydrous aluminium trichloride (1 gm) 2-Amino-(4-chlorophenyl)-6-{5,10-dioxo-2,3-
was dissolved in carbon tetrachloride (30 mL) and diphenyl-5H,10H-benzo[g]quinoxalin-7-yl}
acetyl chloride (2 mL) was added to it under cold pyridine-3-carbonitrile (6b)
conditions (0o-5oC). Compound 4 (120 mg) was Yellow crystalline solid; Yield 80%; Rf 0.80;
added and stirred at room temperature for 6 h. Yellow mp 232oC; IR (nmax, KBr, cm-1): 723, 760, 819, 875,
crystalline solid was filtered, dried and recrystallized 1162 (aryl C-Cl), 1284, 1383, 1592, 1645, 1697,
with chloroform. 2363, 3404; 1H-NMR (DMSO-d6 500 MHz) d: 8.73-
8.77 (m, J=2.5-4 Hz, 10H, Ar-H), 8.03-8.07 (m,
Yellow crystalline solid; Yield 48%; Rf 0.74; J=3.0-3.5 Hz, 4H, Ar-H), 7.70-7.78 (q, J=7.5-15 Hz,
mp 225oC; IR (nmax, KBr, cm-1): 725, 775, 810, 875, 1H, Ar-H), 7.36-7.38 (d, J=8.5 Hz, 1H, Ar-H), 7.28-
1450, 1500, 1593 (>C=O, cyclic, conjugated), 1672 7.30 (d, J=7.5 Hz, 1H, Ar-H), 7.16-7.19 (t, J=8-7.5
(C=N, quinoxaline), 1974 (>C=O, aliphatic), 3063 Hz, 1H, Ar-H), 6.94-6.96 (d, J=8 Hz, 2H, Ar-NH2);
(C-H, sp3, CH3); 1H-NMR (CDCl3 500 MHz) d: 7.97- MS-ESI: 589.08 (m/z, M+); Elemental Anal. Found
7.98 (d, J=8 Hz, 4H, Ar-H), 7.65-7.68 (t, J=7.5 Hz, (Calcd.) for C36H20ClN5O2 : C, 73.27 (73.28); H, 3.43
3H, Ar-H), 7.50-7.53 (t, 6H, J=7.5-8 Hz, Ar-H), 7.26 (3.42); Cl, 5.98 (6.01); N, 11.89 (11.87); O, 5.42
(s, 3H, CH3CO); MS-ESI: 404.14 (m/z, M+); Elemental (5.42).
Anal. Found (Calcd.) for C26H16N2O3 : C 77.23 (77.22);
H 3.98 (3.99); N 6.97 (6.93); O 11.82 (11.87). 2-Amino-6-{5,10-dioxo-2,3-diphenyl-5H,10H-
benzo[g]quinoxalin-7-yl}-4-(4-hydroxyphenyl)
Synthesis of 2-amino-6-(5,10-dioxo-2,3- pyridine-3-carbonitrile (6c)
diphenyl-5,10-dihydrobenzo[g]quinoxalin-7-yl)- Yellow crystalline solid; Yield 72%; Rf 0.78;
4-(substituted) phenylpyridine-3-carbonitrile mp 250oC; IR (nmax, KBr, cm-1) : 718, 765, 827, 895,
(6a-m) 1256 (phenolic C-O phenolic), 1351, 1402, 1596,
824 KUMAR et al., Orient. J. Chem., Vol. 33(2), 821-828 (2017)

1679, 1683, 2358, 3146, 3367 (O-H phenolic); 1H- (C-O, phenolic), 1342, 1424 (C-N), 1611, 1660, 1673,
NMR (DMSO-d6 500 MHz) d: 9.12 (s, 1H, Ar-H), 2360, 3326, 3381 (O-H, phenolic); 1H-NMR (CDCl3
8.72-8.76 (m, J=2.5-4 Hz, 10H, Ar-H), 8.56-8.58 (d, 500 MHz) d: 7.80-7.81 (d, J=7 Hz, 10H, Ar-H), 7.96-
J=9 Hz, 1H, Ar-H), 8.48-8.49 (d, J=9 Hz, 1H, Ar-H), 7.97 (d, J=7.5 Hz, 1H, Ar-OH), 7.09 (s, 2H, Ar-NH2),
7.99-8.11 (t, J=3-5 Hz, 1H, Ar-H), 7.68-7.72 (dd, J=9 7.33-7.36 (t, J=7.5-8 Hz, 3H, Ar-H), 7.48-7.51 (t,
Hz, 2H, Ar-H), 7.44-7.49 (dd, J=9 Hz, 2H, Ar-H), 6.98 J=2.5-7.5 Hz, 2H, Ar-H), 7.63-7.66 (t, J=7-8 Hz, 1H,
(s, 2H, Ar-NH2), 6.86-6.88 (t, J=3-4 Hz, 1H, Ar-OH); Ar-H), 7.16-7.20 (t, J=7.5-8.5 Hz, 1H, Ar-H), 3.31 (s,
MS-ESI: 571.11 (m/z, M+); Elemental Anal. Found 3H, Ar-OCH3); MS-ESI: 601.16 (m/z, M+); Elemental
(Calcd.) for C36H21N5O3 : C, 75.62 (75.65); H, 3.69 Anal. Found (Calcd.) for C37H23N5O4 : C, 73.85 (73.87);
(3.70); N, 12.27 (12.25); O, 8.39 (8.40). H, 3.86 (3.85); N, 11.65 (11.64); O, 10.63 (10.64).

2-Amino-6-{5,10-dioxo-2,3-diphenyl-5H,10H- 2-Amino-4-(2-chlorophenyl)-6-{5,10-dioxo-
benzo[g]quinoxalin-7-yl}-4-(4-nitrophenyl) 2,3-diphenyl-5H,10H-benzo[g]quinoxalin-7-yl}
pyridine-3-carbonitrile (6d) pyridine-3-carbonitrile (6g)
Yellow crystalline solid; Yield 68%; mp White crystalline solid; Yield 66%; Rf 0.72;
210oC; Rf 0.80; IR (nmax, KBr, cm-1): 720, 758, 811, mp 180oC; IR (nmax, KBr, cm-1): 736, 761, 815, 892,
854, 1284, 1383 (-NO2, sym.), 1410, 1545 (-NO2, 1164 (aryl C-Cl), 1311, 1363, 1591, 1662, 2356,
asym.), 1594, 1669, 1684, 2359, 3335; 1H-NMR 3305; 1H-NMR (DMSO-d6 500 MHz) d: 8.74-8.76
(DMSO-d6 500 MHz) d: 9.23-9.27 (dd, J=9.5 Hz, 2H, (q, J=2.5-3.5 Hz, 4H, Ar-H), 8.04-8.07 (m, J=3.5-5
Ar-H), 9.11-9.11 (d, J=3 Hz, 1H, Ar-H), 8.87-8.92 (dd, Hz, 10H, Ar-H), 7.96 (s, 1H, Ar-H), 7.91-7.93 (t,
J=9 Hz, 2H, Ar-H), 8.82-8.86 (m, J=2.5-5 Hz, 10H, J=2.5–8.5 Hz, 1H, Ar-H), 7.68-7.71 (d, 2H, Ar-NH2),
Ar-H), 8.76-8.78 (d, J=11 Hz, 1H, Ar-H), 8.68-8.70 7.51-7.53 (d, J=8.5 Hz, 1H, Ar-H), 7.37-7.39 (d, J=8
(d, J=11 Hz, 1H, Ar-H), 8.59-8.60 (t, J=3-4.5 Hz, 1H, Hz, 1H, Ar-H); MS-ESI: 589.14 (m/z, M+); Elemental
Ar-H), 7.01 (s, 2H, Ar-NH2); MS-ESI: 600.14 (m/z, Anal. Found (Calcd.) for C36H20ClN5O2 : C, 73.28
M+); Elemental Anal. Found (Calcd.) for C36H20N6O4 (73.28); H, 3.41 (3.42); Cl, 6.00 (6.01); N, 11.88
: C, 71.98 (71.99); H, 3.34 (3.36); N, 14.02 (13.99); (11.87); O, 5.42 (5.42).
O, 10.65 (10.66).
2-Amino-4-[4-(dimethylamino)phenyl]-6-{5,10-
2-Amino-6-{5,10-dioxo-2,3-diphenyl-5H,10H- dioxo-2,3-diphenyl-5H,10H-benzo[g] quinoxalin-
benzo[g]quinoxalin-7-yl}-4-(4-methoxyphenyl) 7-yl}pyridine-3-carbonitrile (6h)
pyridine-3-carbonitrile (6e) Lemon crystalline solid, yield 76%; R f
White crystalline solid; Yield 74%; Rf 0.78; 0.80; mp 185oC; IR (nmax, KBr, cm-1): 723, 759, 819,
mp 200oC; IR (nmax, KBr, cm-1): 719, 758, 826, 853, 837, 1325, 1411, 1593, 1647, 2360, 2958 (C-H, sp3,
1130 (C-O-C, sym.), 1253 (C-O-C, asym.), 1340, CH3), 3282; 1H-NMR (DMSO-d6 500 MHz) d: 8.73-
1420, 1595, 1645, 1677, 2360, 3315; 1H-NMR 8.77 (m, J=3-4 Hz, 10H, Ar-H), 8.03-8.07 (m, J=3-4
(DMSO-d6 500 MHz) d: 8.73-8.76 (m, J=2.5-4 Hz, Hz, 4H, Ar-H), 8.28 (s, 1H, Ar-H), 7.93 (s, 1H, Ar-H),
10H, Ar-H), 8.03-8.07 (m, J=3-4 Hz, 4H, Ar-H), 7.74 7.65 (s, 1H, Ar-H), 7.28 (s, 1H, Ar-H), 7.14 (s, 2H,
(s, 1H, Ar-H), 7.67 (s, 1H, Ar-H), 7.44-7.46 (d, J=7.5 Ar-NH2), 3.49 (s, 6H, N-CH3); MS-ESI: 598.31 (m/z,
Hz, 1H, Ar-H), 7.28-7.30 (d, J=7.5 Hz, 1H, Ar-H), M+); Elemental Anal. Found (Calcd.) for C38H26N6O2
7.13-7.17 (t, 2H, Ar-NH2), 3.91 (s, 3H, Ar-OCH3); : C, 76.22 (76.24); H, 4.37 (4.38); N, 14.05 (14.04);
MS-ESI: 585.13 (m/z, M+); Elemental Anal. Found O, 5.34 (5.35).
(Calcd.) for C37H23N5O3 : C, 75.90 (75.89); H, 3.95
(3.96); N, 11.97 (11.96); O, (8.18) 8.20. 2-Amino-6-{5,10-dioxo-2,3-diphenyl-5H,10H-
benzo[g]quinoxalin-7-yl}-4-[4-(trifluoromethyl)
2-Amino-6-{5,10-dioxo-2,3-diphenyl-5H,10H- phenyl]pyridine-3-carbonitrile (6i)
benzo[g]quinoxalin-7-yl}-4-(3-hydroxy-4- Brown crystalline solid; Yield 78%; Rf 0.80;
methoxyphenyl)pyridine-3-carbonitrile (6f) mp 178oC; IR (nmax, KBr, cm-1): 723, 741, 843, 863,
Brown crystalline solid; Yield 72%; Rf 0.81; 1285 (C-F), 1320, 1420, 1603, 1645, 1675, 2365,
mp 210oC; IR (nmax, KBr, cm-1): 720, 757, 838, 865, 3236; 1H-NMR (DMSO-d6 500 MHz) d: 8.73-8.77 (m,
1124 (C-O-C, sym.), 1252 (C-O-C, asym.), 1295 J=3-4 Hz, 4H, Ar-H), 8.28 (s, 1H, Ar-H), 8.03-8.07
 KUMAR et al., Orient. J. Chem., Vol. 33(2), 821-828 (2017) 825

(m, J=3-4.5 Hz, 10H, Ar-H), 7.73-7.84 (d, J=8.5 6.92-6.93 (t, J=2.5-3 Hz, 4H, Ar-H), 6.89-6.91 (d,
zHzHHHHz, 1H, Ar-H), 7.35-7.37 (d, J=8 Hz, 1H, J=10 Hz, 2H, Ar-H), 6.85 (s, 1H, Ar-H), 6.06 (bs, 4H,
Ar-H), 7.76 (s, 1H, Ar-H), 7.02-7.03 (d, J=4.5 Hz, 2H, Ar-NH2); MS-ESI: 570.18 (m/z, M+); Elemental Anal.
Ar-NH2); MS-ESI: 623.14 (m/z, M+); Elemental Anal. Found (Calcd.) for C36H22N6O2 : C, 75.79 (75.78); H,
Found (Calcd.) for C37H20FN5O2 : C, 71.26 (71.27); H, 3.90 (3.89); N, 14.73 (14.73); O, 5.58 (5.61).
3.24 (3.23); F, 9.12 (9.14); N, 11.24 (11.23); O, 5.13
(5.13). 2-Amino-6-{5,10-dioxo-2,3-diphenyl-5H,10H-
benzo[g]quinoxalin-7-yl}-4-(2-hydroxyphenyl)
2-Amino-4-(3,4-dimethoxyphenyl)-6-{5,10-dioxo- pyridine-3-carbonitrile (6m)
2,3-diphenyl-5H,10H-benzo[g]quinoxalin-7-yl} Yellow crystalline solid; Yield 72%; Rf 0.86;
pyridine-3-carbonitrile (6j) mp 220oC; IR (nmax, KBr, cm-1): 725, 760, 820, 870,
White crystalline solid; Yield 66%; Rf 0.75; 1280, 1311 (C-O, phenolic), 1382, 1595, 1650, 2358,
mp 144oC; IR (nmax, KBr, cm-1): 720, 752, 821, 854, 3380 (O-H, phenolic), 3406; 1H-NMR (DMSO-d6
1125 (C-O-C, sym.), 1246 (C-O-C, asym.), 1337, 500 MHz) d: 8.78 (bs, 1H, Ar-H), 8.64-8.66 (d, J=10
1418, 1608, 1645, 1682, 2357, 3246; 1H-NMR Hz, 1H, Ar-H), 8.56-8.59 (t, J=4-10 Hz, 1H, Ar-H),
(DMSO-d6 500 MHz) d: 8.98 (s, 1H, Ar-H), 8.83- 8.48-8.50 (t, J=4-5 Hz, 1H, Ar-H), 8.22-8.31 (m,
8.94 (m, J=3.5-12 Hz, 10H, Ar-H), 8.11-8.12 (d, J=2.5-8.5 Hz, 10H, Ar-H), 7.12-7.18 (m, J=2.5-10
J=8 Hz, 1H, Ar-H), 8.01-8.03 (d, J=8 Hz, 1H, Ar-H), Hz, 4H, Ar-H), 6.94 (bs, 2H, Ar-NH2), 6.86-6.87
7.92-7.94 (d, J=8.5 Hz, 1H, Ar-H), 7.83 (bs, 2H, Ar- (d, J=3 Hz, 1H, Ar-OH); MS-ESI: 571.14 (m/z, M+);
NH2), 7.24-7.26 (d, J=10 Hz, 1H, Ar-H), 7.21-7.31 Elemental Anal. Found (Calcd.) for C36H21N5O3 : C,
(d, J=9.5 Hz, 1H, Ar-H), 6.91-6.93(d, J=10 Hz, 1H,
O
Ar-H), 3.38 (s, 6H, Ar-OCH3); MS-ESI: 615.19 (m/z,
M+); Elemental Anal. Found (Calcd.) for C38H25N5O4 N
: C, 74.16 (74.14); H, 4.07 (4.09); N, 11.38 (11.38);
O, 10.39 (10.40).
N
N
2-Amino-6-{5,10-dioxo-2,3-diphenyl-5H,10H- O
benzo[g]quinoxalin-7-yl}-4-(4-methylphenyl) R
NC NH2
pyridine-3-carbonitrile (6k)
Yellow crystalline solid; Yield 70%; Rf 0.70; Table 1: Antimycobacterial activity of
mp 140oC; IR (nmax, KBr, cm-1): 723, 750, 817, 850, synthesized compounds against M.
1328, 1406, 1606, 1650, 2358, 2839 (C-H, sp3, CH3), tuberculosis H37Rv by L. J. Slope Method
3209; 1H-NMR (DMSO-d6 500 MHz) d: 8.88 (s, 1H,
Ar-H), 8.56-8.58 (d, J=10 Hz, 1H, Ar-H), 8.48-8.51 Compound R MIC (mg/mL)
(d, J=10.5 Hz, 1H, Ar-H), 8.42-8.43 (t, J=3.5-4 Hz,
1H, Ar-H), 8.11-8.18 (m, J=2.5-7.5 Hz, 10H, Ar-H), 6a H 125
7.82-7.99 (m, 4H, Ar-H), 6.88 (s, 2H, Ar-NH2), 3.58- 6b 4-Cl 62.5
3.59 (t, J=1.5-5 Hz, 3H, CH3); MS-ESI: 569.17 (m/z, 6c 4-OH 100
M+); Elemental Anal. Found (Calcd.) for C37H23N5O2 6d 4-NO2 500
: C, 78.03 (78.02); H, 4.06 (4.07); N, 12.30 (12.29); 6e 4-OCH3 —-
O, 5.60 (5.62). 6f 3-OH-4-OCH3 500
6g 2-Cl 62.5
2-Amino-4-(4-aminophenyl)-6-{5,10-dioxo- 6h 4-N(CH3)2 50
2,3-diphenyl-5H,10H-benzo[g]quinoxalin-7-yl} 6i 4-CF3 100
pyridine-3-carbonitrile (6l) 6j 3,4-(OCH3)2 250
Yellow crystalline solid; Yield 74%; Rf 0.84; 6k 4-CH3 500
mp 180oC; IR (nmax, KBr, cm-1): 725, 750, 820, 860, 6l 4-NH2 —-
1325, 1415, 1650 (N-H, primary amine, bend), 1658, 6m 2-OH —-
2358, 3240; 1H-NMR (DMSO-d6 500 MHz) d: 7.24- Rifampicin —- 0.25
7.26 (q, J=2.5-3 Hz, 10H, Ar-H), 7.01 (s, 1H, Ar-H), Isoniazid —- 0.20
826 KUMAR et al., Orient. J. Chem., Vol. 33(2), 821-828 (2017)

75.64 (75.65); H, 3.69 (3.70); N, 12.27 (12.25); O, followed by dehydration to yield 2,3-diphenylbenzo[g]
8.39 (8.40). quinoxaline (3). Compound 3 undergoes aromatic
oxidation at C-5 and C-10 on refluxing with equimolar
In-vitro Antimycobacterial Screening mixture of chromium trioxide and glacial acetic acid
Ten compounds 6a, 6b, 6c, 6d, 6f, 6g, 6h, 6i, in water, till the permanent appearance of dark green
6j and 6k were submitted for their antimycobacterial colour, furnished 2,3-diphenylbenzo[g]quinoxaline-
activity against Mycobacterium tuberculosis H37Rv 5,10-dione (4). Compound 4 on acetylation reaction
(MTCC-200) by L.J. Slope (Conventional) Method24- with acetylchloride and anhydrous aluminium
25
. Each compound was diluted to 2000 ìg/mL chloride in carbon tetrachloride yields 7-acetyl-
concentration (as a stock solution). Inoculum Size 2,3-diphenylbenzo[g]quinoxaline-5,10-dione (5).
for Mycobacterium tuberculosis was adjusted to 1 Compound 5 on refluxing with equimolar quantity of
mg/mL. L.J. Medium was used as nutrient medium. substituted aromatic aldehydes, ammonium acetate,
Isoniazid (0.20 µg/mL) and rifampicin (0.25 µg/mL) malononitrile, ammonia and catalytic amount of
were used as standard drug. dil. HCl undergoes Heitzch-Pyridine synthesis to
yield 2-amino-6-(5,10-dioxo-2,3-diphenyl-5,10-
Primary screen dihydrobenzo[g]quinoxalin-7-yl)-4-(substituted)
In primary screening, the compounds phenylnicotinonitrile (6a-m). Structure of all newly
were diluted to 500 µg/mL, 250 µg/mL and 125 µg/ synthesized thirteen compounds were confirmed by
mL concentrations. Compounds found active in this FT-IR, 1H-NMR, MS-ESI spectral data interpretation
primary screening were further tested in a second and their elemental analysis.
set of dilution against Mycobacterium tuberculosis.
Biological Activity (In-vitro Antimycobacterial
Secondary screen Screening)
The compounds found active in primary The ten newly synthesized compounds
screening were further diluted to 100 µg/mL, 50 µg/ viz. 6a, 6b, 6c, 6d, 6f, 6g, 6h, 6i, 6j and 6k were
mL, 25 µg/mL, 12.5 µg/mL, 6.250 µg/mL, 3.125 µg/ screened for their antimycobacterial screening
mL and 1.5625 µg/mL concentrations. against Mycobacterium tuberculosis H37Rv by L.J.
Slope (Conventional) Method to observe the effect
Reading Result of substitution at phenyl ring attached to C-4 of
The highest dilution showing at least 99% pyridine ring of 2-amino-6-(5,10-dioxo-2,3-diphenyl-
inhibition was considered as Minimum Inhibitory 5,10-dihydrobenzo[g]quinoxalin-7-yl)-4-(substituted)
Concentration. The inoculum size of test should phenylpyridine-3-carbonitrile (Compound 6a-m).
contain 108 organism/mL. Isoniazid and Rifampicin were used as standard
drugs.
RESULT and DISCUSSIONs
Unsubstituted phenyl ring (6a) on the
Chemistry 4-position of pyridine ring in above mentioned
Thirteen new compounds belonging to nucleus exhibited moderate antimycobacterial
series 2-amino-6-(5,10-dioxo-2,3-diphenyl-5,10- activity. Substitution with electron donating
dihydrobenzo[g]quinoxalin-7-yl)-4-(substituted) para-dimethylamino group (6h) on the phenyl
phenylpyridine-3-carbonitrile (6a-m) were synthesized ring present at C-4 of pyridine ring increases
by multistep synthetic scheme (Figure 1). 2,3- antimycobacterial activity and produces the most
Dihydroxynaphthalene on refluxing with phosphorus active antimycobacterial compound of the above
oxychloride yielded 2,3-dichloronaphthalene (1). mentioned newly synthesized series. Substitution
Compound 1 on refluxing with ammonium chloride with electron withdrawing 4-Cl group (6b) and
in excess of ammonia, catalytic amount of dil. HCl 2-Cl (6g) at the phenyl ring also increases the
and ethanol undergoes amination (nucleophilic antimycobacterial activity but lesser than that of
substitution) & furnished 2,3-diaminonaphthalene substitution with para-dimethylamino group (6h).
(2). Compound 2 on refluxing with equimolar quantity Substitution with para-hydroxy (6c) and electron
of benzil in ethanol undergoes nucleophilic addition withdrawing para-trifluoromethyl (6i) at the pheny
 KUMAR et al., Orient. J. Chem., Vol. 33(2), 821-828 (2017) 827

ring exhibits moderate activity lesser than that of 7-yl)-4-(substituted)phenylpyridine-3-carbonitrile

compounds 6h, 6g and 6b and more than that of (6a-m), which were screened for antimycobacterial
compound with unsubstituted phenyl ring (6a). screening against Mycobacterium tuberculosis H37Rv
While, substitution with bulky electron withdrawing by L. J. Slope (Conventional) Method, compound 6h
group para-nitro (6d), electron donating para-methyl with 4-N(CH3)2 at phenyl ring attached at pyridine
(6k) and bulky electron donating groups 3-OH- group of 2-amino-6-(5,10-dioxo-2,3-diphenyl-5,10-
4-OCH 3 (6f) and 3,4-dimethoxy (6j) decreases dihydrobenzo[g]quinoxalin-7-yl)-4-(substituted)
the antimycobacterial activity to great extent and phenylpyridine-3-carbonitrile (6a-m) exhibited the
produces the least active compounds of the newly maximal potency, while compounds with 4-Cl (6b)
synthesized series. and 2-Cl (6g) substitutions at the above mentioned
position exhibited better antimycobacterial potency
From the above observations, Structure as compared to other compounds. Compounds
Activity Relationship (SAR) may be established as with 4-OH (6c), 4-CF3 (6i) and with no substitution
“substitution with smaller electron withdrawing and (6a) at phenyl ring of the above mentioned position
donating groups at the ortho and para positions exhibited moderate antimycobacterial activity,
of phenyl ring attached to C-4 of pyridine ring, while compounds with 3,4-(OCH3)2 (6j), 4-NO2 (6d),
present at 7-position of 2-amino-6-(5,10-dioxo-2,3- 3-OH-4-OCH3 (6f) and 4-CH3 (6k) showed minimal
diphenyl-5,10-dihydrobenzo[g]quinoxalin-7-yl)-4- antimycobacterial potency against Mycobacterium
(substituted)phenylpyridine-3-carbonitrile increases tuberculosis H37Rv strain under observation.
antimycobacterial activity, while substitution with
bulky electron donating groups at the phenyl ring ACKNOWLEDGEMENT
attached to above mentioned position decreases
antimycobacterial activity.” Authors are grateful to Microcare
Laboratory and TRC, Surat, Gujarat (India) for
CONCLUSION providing antimycobacterial activity data and
Advanced Instrumentation Research Facilities
Among the newly compounds synthesized (AIRF), Jawaharlal Nehru University, New Delhi,
compounds of series phenyl ring of 2-amino-6-(5,10- India for providing spectral data.
dioxo-2,3-diphenyl-5,10-dihydrobenzo[g]quinoxalin-

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