Indian Journal of Chemistry

Vol. 51B, May 2012, pp. 739-745

Synthesis and antibacterial evaluation of benzazoles

tethered dihydro[1,3]oxazines
Davinder Prasada, Rajesh Kumar Rohillab, Nilanjan Royb, Mahendra Nath*a
a
Department of Chemistry, University of Delhi, Delhi 110 007, India
b
Department of Biotechnology, National Institute of Pharmaceutical Education and Research,
SAS Nagar, Punjab 160 062, India
E-mail: mnath@chemistry.du.ac.in
Received 29 August 2011; accepted (revised) 17 February 2012

Synthesis of various benzazoles tethered 1,3-oxazines such as 1H-benzo[d]azolophenyl-3,4-dihydro-2H-benzo-

[e][1,3]oxazines 2a-j, 2-(3-(1H-benzo[d]oxazol-2-yl)phenyl)-2,3-dihydro-1H-naphtho[1,2-e][1,3]oxazine 3, 2-(3-(1H-
benzo-[d]imidazol-2-yl)phenyl)-2,3-dihydro-1H-naphtho[1,2-e][1,3]oxazine 4 and 3-(3-(benzo[d]oxazol-2-yl)phenyl)-7-
(benzo[d]thiazol-2-yl)-3,4-dihydro-2H-benzo[e][1,3]oxazine 5 has been accomplished through three-component one-pot
Mannich type condensation-cyclization reaction of substituted phenols or β-naphthol with formaldehyde and benzazolo-
phenyl amines at 80-90°C. All the newly synthesized compounds have been characterized on the basis of elemental and
spectral analyses. Moreover, these compounds have been evaluated for their in vitro antibacterial efficacy against two Gram-
positive bacteria viz S. epidermidis and S. aureus and two Gram-negative bacteria viz E. coli and P. aeruginosa.
Interestingly, the compounds, 2f-i have displayed significant antibacterial efficacy with IC50 (0.208-1.106 µg/mL).

Keywords: One-pot methodology, antibacterial efficacy, benzazoles, dihydro[1,3]oxazines, polymerization

The 1,3-oxazine derivatives are an important class of Results and Discussion

heterocyclic compounds which occupy an unique
place in material and medicinal chemistry due to
their diverse physical and biological properties such
as monomer for polymer formation1, photochromic
agents2, antibacterial3,4, non-steroidal progesterone
receptor modulators5, antifungal4, antimalarial6,
antituberculor7, antitumor8,9 and anti-HIV agents10,11.
In addition, 1,3-oxazine nucleus is a part of many
biologically important natural products12,13 and other
synthetic bioactive molecules14-17 including
Efavirenz, a benzoxazinone derivative for the
treatment of HIV-1 infections18. Similarly,
benzazoles such as benzoxazoles, benzthiazoles and
benzimidazoles are known to display a wide range of
pharmacological properties19-28 and some of them are
common to 1,3-oxazines. Thus, it was contemplated
to synthesize benzazole tethered dihydro-1,3-oxazine
derivatives in order to obtain more potent
antibacterial agents. Hence, a series of novel
heteroarene tethered dihydro[1,3]oxazines has been
prepared by using one-pot methodology to evaluate
their antibacterial activity.
Chemistry
The synthetic routes of title compounds are
outlined in Scheme I. The starting compounds 2-(3-
aminophenyl)benzoxazole, 2-(3-aminophenyl)benz-
thiazole and 2-(3-aminophenyl)-1H-benzimidazole
were synthesized according to the literature
procedure29 via the condensation reaction of m-
aminobenzoic acid with corresponding o-substituted
aryl amines in polyphosphoric acid at 160°C, while 2-
(3-hydroxy-phenyl)benzthiazole was prepared30 by
the reaction of 2-aminothiophenol with 3-
hydroxybenzaldehyde in the presence of ceric
ammonium nitrate and 30% H2O2 in acetonitrile at
25°C. The physical and spectral data of these known
compounds are in agreement with those of reported
data31-34. The benzazolophenyl amines 1 on reaction
with corresponding phenols or β-naphthol and 37%
aqueous formaldehyde solution at 80-90°C afforded
1H-benzo[d]azolophenyl-3,4-dihydro-2H-benzo[e]-
[1,3]oxazines 2a-j, 2-(3-(1H-benzo[d]oxazol-2-yl)-
phenyl)-2,3-dihydro-1H-naphtho-[1,2-e][1,3]oxazine
740 INDIAN J. CHEM., SEC B, MAY 2012

N
O

N
3

b X= O
H
N X

N X N
b a
N N
O X = NH N O
1 NH2
R3 R1

c X=O R2
4 2a-j
O
N
O
N S

N
5
Reagents and conditions: a) Substituted phenol, HCHO, 80-90ºC, 4 hr; b) β-Naphthol, HCHO, 80-90ºC, 4 hr; c) 2-(3-
Hydroxyphenyl)benzothiazole, HCHO, 80-90ºC, 5 hr.
Scheme I

3 and 2-(3-(1H-benzo-[d]imidazol-2-yl)phenyl)-2,3-
dihydro-1H-naphtho[1,2-e][1,3]oxazine 4 in moderate
to good yields (Scheme 1). Under similar reaction
conditions, the scope of the reaction was further
explored by reacting 2-(3-hydroxyphenyl)benz-
thiazole with 37% aqueous formaldehyde solution and
2-(3-aminophenyl)benzoxazole to obtain 3-(3-
(benzo[d]oxazol-2-yl)phenyl)-7-(benzo[d]-thiazol-2-
yl)-3,4-dihydro-2H-benzo[e][1,3]-oxazine 5 in 25%
yield. All the newly prepared compounds were
characterized on the basis of spectral and elemental
analyses. The IR spectrum of compound 2c has shown
two characteristic absorption peaks at 1217 cm-1 due
to asymmetric stretching of C-O-C bond and at 1048
cm-1 which corresponds to the symmetric stretching of
C-O-C bond. In the proton NMR of 2c, two chara-
cteristic peaks of benzoxazine ring were observed at δ
5.44 corresponding to N-CH2-O protons and at δ 4.73
corresponding to N-CH2-Ar protons. Similarly, two
characteristic carbon signals at δ 79.1 and 50.3 in the
13
C NMR of 2c were assigned to N-CH2-O and N-
CH2-Ar groups, respectively. The mass spectral
analysis gave further evidence for the formation of 3-
(3-(benzo[d]thiazol-2-yl)phenyl)-6-chloro-3,4-dihyd-
ro-2H-benzo[e][1,3]oxazine 2c by showing [M]+ ion
peak at m/z 378.0157 for the molecular formula,
C21H15ClN2OS. In addition, the thermally-activated
cure behavior of synthesized dihydro-1,3-oxazines
was studied by differential scanning calorimetry
(DSC). The DSC thermogram of compound 2c has
shown an endothermic peak at 134.84°C which was
attributed to its melting point and the characteristic
exothermic peak with onset at 265.92°C and maxi-
mum at 270.90°C was observed due to ring opening
polymerization of the oxazine ring system35. The
amount of heat of polymerization was found to be
175.8 J/g. The typical synthetic methods, characteri-
zation and DSC data of compounds 2a-j, 3, 4 and 5
are presented in the experimental section.
 PRASAD et al.: SYNTHESIS OF BENZAZOLES TETHERED DIHYDRO[1,3]OXAZINES 741

Table I — In vitro antibacterial activity of compounds 2a-j, 3, 4 and 5

Compd X R1 R2 R3 Yield IC50 (µg/mL)

(%) Se Sa Ec Pa
2a O H Cl H 80 * * * *
2b O H Br H 72 * * * *
2c S H Cl H 84 * * * *
2d S H Br H 84 * * * *
2e NH H F H 58 * * * *
2f NH H Cl H 85 0.852 0.208 1.106 0.957
2g NH H Br H 82 0.402 0.208 1.106 0.947
2h NH H I H 77 * 0.419 * 0.957
2i NH Cl Cl H 86 * * * 0.957
2j NH H CH3 H 85 * * * *
3 - - - - 86 * * * *
4 - - - - 85 * * * *
5 - - - - 25 * * * *
Tetracycline - - - - - 0.080 0.120 0.090 0.060
Se, S. epidermidis; Sa, S. aureus; Ec, E. coli; Pa, P. aeruginosa
*Activity: >500 µg/mL.

In vitro antibacterial efficacy donating methyl groups, respectively at position 6

The in-vitro antibacterial activity of compounds
2a-j, 3, 4 and 5 and standard drug tetracycline was
carried out against two Gram-positive bacteria viz. S.
epidermidis and S. aureus and two Gram-negative
bacteria viz. E. coli and P. aeruginosa. The activity
data are presented in Table I. Though, target
compounds were not as active as standard drug,
tetracycline but compounds 2f-i were found signi-
ficantly potent with low IC50 values in the range of
0.208-1.106 µg/mL. The most active compound 2g
possessing bromo substituent at position 6 in the
fused aryl ring of dihydro-1,3-benzoxazine moiety has
displayed better antibacterial effect against S.
epidermidis, S. aureus, E. coli and P. aeruginosa with
IC50 values 0.402, 0.208, 1.106 and 0.947 µg/mL,
respectively. The other compound 2f with chloro
substituent at position 6 was found equipotent to 2g
except it showed slightly higher IC50 values against S.
epidermidis (0.852 µg/mL) and P. aeruginosa (0.957
µg/mL). An exchange of bromo substituent at position
6 in 2g by iodo group as in 2h led to exhibit
significant antibacterial effect only against S. aureus
(IC50 0.419 µg/mL) and P. aeruginosa (IC50 0.957
µg/mL). Introduction of two chlorine atoms at
positions 6 and 8 in 2i did not improve the activity but
was selectively effective against P. aeruginosa (IC50
0.957 µg/mL). In addition, the compounds 2e and 2j
containing electron withdrawing fluoro and electron
were found inactive against all the tested bacterial
strains. These results imply that instead of the nature,
the size of substituent at position 6 of benzoxazine
ring is crucial for the antibacterial activity. Further, an
introduction of O or S in place of NH in the
benzimidazole nucleus as in compounds 2a-d led to
complete loss of antibacterial activity against tested
bacterial strains. In contrast, their benzazolophenyl-
2,3-dihydro-1H-naphtho[1,2-e][1,3]-oxazine counter-
parts 3 and 4 and 3-(3-(benzo[d]oxazol-2-yl)phenyl)-
7-(benzo[d]thiazol-2-yl)-3,4-dihydro-2H-benzo[e]-
[1,3]oxazine 5 were presenting antibacterial activity at
more than 500 µg/mL concentration.
Conclusions
In summary, various benzazole tethered di-
hydro[1,3]oxazines have been successfully prepared
by using simple one-pot methodology and evaluated
for their in vitro antibacterial efficacy. Among all,
compounds 2f and 2g have displayed significant
activity with IC50 values in the range of 0.208-1.106
µg/mL against all the tested bacterial strains. Further,
it was noticed that the size of the substituent present
at position 6 in the fused aryl ring of dihydro-1,3-
benzoxazine moiety is crucial for the activity in
addition to 1H-benzimidazole nucleus. Within the
series, the compound 2i possessing chloro substituent
in the 6,8 positions of dihydro-1,3-benzoxazine ring,
742 INDIAN J. CHEM., SEC B, MAY 2012
displayed significant and selective efficacy against P.
aeruginosa with IC50 value 0.957 µg/mL. Henceforth,
these results are useful and could be guide for the
development of potent antibacterial agents in the
future.
Experimental Section
All the chemicals were purchased from Sigma-
Aldrich and used without any further purification.
Thin layer chromatography was performed on
precoated Merck silica gel 60 F254 plates and spots
were developed under UV light (254 nm) or in iodine
chamber. All the compounds were purified by column
chromatography using silica gel (60-120 mesh). The
1
H and 13C NMR spectra were recorded on Bruker
300 or 400 MHz spectrometer. The IR spectra were
obtained on a Perkin Elmer IR spectrometer and
peaks are given in reciprocal centimeter (cm-1). Mass
spectra were recorded on Waters Micromass LCT
mass spectrometer. Elemental analyses were
determined on Elementar Analysensysteme GmbH
VarioEL V3.00 and CHN values were found within
±0.4 of theoretical values for all the final compounds.
The melting points were obtained by Perkin-Elmer
Differential Scanning Calorimetry.
General procedure for the synthesis of 2a-j, 3 and 4
A mixture of amine 1 (1 mmol), substituted phenol
or β-naphthol (1 mmol) and formalin (37%, w/v, 2
mmol) was heated at 80-90°C for 2 hr. The additional
amount of formalin (2 mmol) was added to the
reaction mixture and heating was continued for
another 2 hr. The reaction mixture was diluted with
water (10 mL) and product was extracted with ethyl
acetate (20 mL × 3 times). The organic layers were
combined and washed with 10% NaOH solution (15
mL × 3 times) followed by water (15 mL × 3 times).
The organic layer was dried over anhydrous sodium
sulfate and evaporated under reduced pressure to
obtain the crude compound. The product was purified
by column chromatography on silica gel using ethyl
acetate/hexane as eluent.
3-(3-(Benzo[d]oxazol-2-yl)phenyl)-6-chloro-3,4-
dihydro-2H-benzo[e][1,3]oxazine, 2a: Rf: 0.85 (30%
EtOAc in hexane); m.p. 98.3°C; Tmax (DSC
exotherm): 280.3°C; IR (Nujol): 1603 (C=N), 1552,
1476, 1456, 1376, 1234, 1160, 998, 936, 874, 821,
743 cm-1; 1H NMR (300 MHz, CDCl3): δ 8.00 (s, 1H,
ArH), 7.84 (d, J = 7.6 Hz, 1H, ArH), 7.81-7.78 (m,
1H, ArH), 7.62-7.59 (m, 1H, ArH), 7.46-7.36 (m, 3H,
ArH), 7.28-7.26 (m, 1H, ArH), 7.11-7.06 (m, 2H,
ArH), 6.79 (d, J = 8.5 Hz, 1H, ArH), 5.45 (s, 2H, N-
CH2-O), 4.72 (s, 2H, N-CH2-Ar); 13C NMR (100
MHz, CDCl3): δ 162.83, 152.79, 150.65, 148.43,
141.93, 129.92, 128.15, 128.01, 126.43, 125.65,
125.14, 124.56, 121.89, 120.70, 120.67, 119.94,
118.39, 116.85, 110.57, 78.90, 50.20; HRMS (ESI):
m/z [M]+ calcd. for C21H15ClN2O2: 362.0822; found:
362.1564. Anal. Calcd. for C21H15ClN2O2.0.6H2O: C,
67.51; H, 4.37; N, 7.50. Found: C, 67.42; H, 4.45; N,
7.45%.
3-(3-(Benzo[d]oxazol-2-yl)phenyl)-6-bromo-3,4-
dihydro-2H-benzo[e][1,3]oxazine, 2b: Rf: 0.70 (20%
EtOAc in hexane); m.p. 153.9°C; Tmax (DSC
exotherm): 260.5°C; IR (KBr): 1605 (C=N), 1548,
1463, 1377, 1246, 1001, 972, 931, 863, 818, 793, 760,
744, 722, 688 cm-1; 1H NMR (300 MHz, CDCl3): δ
7.98 (s, 1H, ArH), 7.83 (d, J = 7.4 Hz, 1H, ArH),
7.79-7.76 (m, 1H, ArH), 7.60-7.57 (m, 1H, ArH),
7.44-7.34 (m, 3H, ArH), 7.25-7.18 (m, 3H, ArH), 6.73
(d, J = 8.6 Hz, 1H, ArH), 5.43 (s, 2H, N-CH2-O), 4.71
(s, 2H, N-CH2-Ar); 13C NMR (100 MHz, CDCl3): δ
162.83, 153.32, 150.67, 148.43, 141.96, 130.92,
129.94, 129.38, 128.19, 125.14, 124.58, 122.47,
120.73, 119.97, 118.86, 116.92, 112.95, 110.59,
78.92, 50.16; HRMS (ESI): m/z [M]+ calcd. for
C21H15BrN2O2: 406.0317; found: 406.5157. Anal.
Calcd. for C21H15BrN2O2.0.4H2O: C, 60.86; H, 3.84;
N, 6.76. Found: C, 60.90; H, 3.89; N, 6.56%.
3-(3-(Benzo[d]thiazol-2-yl)phenyl)-6-chloro-3,4-
dihydro-2H-benzo[e][1,3]oxazine, 2c: Rf: 0.69 (20%
EtOAc in hexane); m.p. 134.8°C; Tmax (DSC
exotherm): 270.9°C; IR (Nujol): 1597, 1452, 1365,
1260, 1217, 1161, 1048, 986, 951, 880, 819, 759, 726,
682 cm-1; 1H NMR (400 MHz, CDCl3): δ 8.08 (d, J =
10 Hz, 1H, ArH), 7.91 (d, J = 8.8 Hz, 1H, ArH), 7.87
(s, 1H, ArH), 7.61 (d, J = 8.8 Hz, 1H, ArH), 7.52-7.47
(m, 1H, ArH), 7.41-7.36 (m, 2H, ArH), 7.21 (d, J =
7.6 Hz, 1H, ArH), 7.09-6.99 (m, 2H, ArH), 6.78 (d, J
= 10 Hz, 1H, ArH), 5.44 (s, 2H, N-CH2-O), 4.73 (s,
2H, N-CH2-Ar); 13C NMR (75 MHz, CDCl3): δ 167.9,
154.0, 152.9, 148.6, 135.1, 134.7, 130.0, 128.0, 126.5,
126.3, 125.7, 125.2, 123.2, 122.0, 121.6, 121.0, 120.2,
118.4, 116.8, 79.1, 50.3; HRMS (ESI): m/z [M]+
calcd. for C21H15ClN2OS: 378.0594; found: 378.0157.
Anal. Calcd. for C21H15ClN2OS.0.4H2O: C, 65.33; H,
4.12; N, 7.26. Found: C, 65.59; H, 3.97; N, 7.20%.
3-(3-(Benzo[d]thiazol-2-yl)phenyl)-6-bromo-3,4-
dihydro-2H-benzo[e][1,3]oxazine, 2d: Rf: 0.75 (20%
EtOAc in hexane); m.p. 159.2°C; Tmax (DSC
exotherm): 255.1°C; IR (Nujol): 1597, 1508, 1462,
 PRASAD et al.: SYNTHESIS OF BENZAZOLES TETHERED DIHYDRO[1,3]OXAZINES
1377, 1313, 1263, 1160, 1049, 951, 876, 819, 760,
726, 687 cm-1; 1H NMR (400 MHz, CDCl3): δ 8.08 (d,
J = 9.2 Hz, 1H, ArH), 7.90 (d, J = 9.2 Hz, 1H, ArH),
7.87 (s, 1H, ArH), 7.61 (d, J = 10.8 Hz, 1H, ArH),
7.52-7.48 (m, 1H, ArH), 7.41-7.36 (m, 2H, ArH),
7.26-7.19 (m, 3H, ArH), 6.68 (d, J = 10.4 Hz, 1H,
ArH), 5.46 (s, 2H, N-CH2-O), 4.71 (s, 2H, N-CH2-
Ar); 13C NMR (100 MHz, CDCl3): δ 167.90, 154.02,
153.35, 148.60, 135.05, 134.71, 130.92, 130.00,
129.39, 126.32, 125.22, 123.23, 122.53, 121.60,
121.04, 120.28, 118.86, 116.84, 112.94, 79.10, 50.22;
HRMS (ESI): m/z [M]+ calcd. for C21H15BrN2OS:
422.0088; found: 422.0244. Anal. Calcd. for
C21H15BrN2OS.0.5H2O: C, 58.34; H, 3.73; N, 6.48.
Found: C, 58.21; H, 3.63; N, 6.33%.
3-(3-(1H-Benzo[d]imidazol-2-yl)phenyl)-6-fluo-
ro-3,4-dihydro-2H-benzo[e][1,3]oxazine, 2e: Rf:
0.70 (50% EtOAc in hexane); m.p. 211.5°C; Tmax
(DSC exotherm): 216.1°C; IR (KBr): 3040 (NH),
1604 (C=N), 1497, 1476, 1449, 1410, 1364, 1245,
1225, 1198, 1167, 1132, 1011, 968, 929, 866, 815,
795, 766, 745, 688 cm-1; 1H NMR (300 MHz, DMSO-
d6): δ 12.96 (s, 1H, NH), 7.99 (s, 1H, ArH), 7.73-7.71
(m, 2H, ArH), 7.59 (m, 1H, ArH), 7.50-7.44 (m, 1H,
ArH), 7.32-7.27 (m, 3H, ArH), 7.13-7.02 (m, 1H,
ArH), 7.00-6.96 (m, 1H, ArH), 6.86-6.82 (m, 1H,
ArH), 5.58 (s, 2H, N-CH2-O), 4.83 (s, 2H, N-CH2-
Ar); 13C NMR (75 MHz, DMSO-d6): δ 157.61,
154.47, 151.28, 150.14, 150.11, 148.02, 131.04,
129.79, 122.55, 122.46, 118.66, 118.47, 117.56,
117.46, 114.85, 114.52, 114.21, 113.54, 113.23,
78.30, 48.75; HRMS (ESI): m/z [M]+ calcd. for
C21H16FN3O: 345.1277; found: 345.3566. Anal.
Calcd. for C21H16FN3O.0.5H2O: C, 71.17; H, 4.84; N,
11.86. Found: C, 70.94; H, 4.59; N, 11.60%.
3-(3-(1H-Benzo[d]imidazol-2-yl)phenyl)-6-chlo-
ro-3,4-dihydro-2H-benzo[e][1,3]oxazine, 2f: Rf:
0.75 (50% EtOAc in hexane); m.p. 208.9°C; Tmax
(DSC exotherm): 212.7°C; IR (KBr): 3047 (NH),
1604 (C=N), 1585, 1476, 1448, 1409, 1364, 1249,
1232, 1194, 1159, 1121, 1096, 1011, 967, 950, 881,
870, 817, 794, 765, 745, 688, 635 cm-1; 1H NMR
(300 MHz, DMSO-d6): δ 12.92 (s, 1H, NH), 8.00 (s,
1H, ArH), 7.74-7.71 (m, 2H, ArH), 7.57 (m, 1H,
ArH), 7.48-7.46 (m, 1H, ArH), 7.33-7.17 (m, 5H,
ArH), 6.84 (d, J = 8.7 Hz, 1H, ArH), 5.62 (s, 2H, N-
CH2-O), 4.84 (s, 2H, N-CH2-Ar); HRMS (ESI): m/z
[M]+ calcd. for C21H16ClN3O: 361.0982; found:
361.9693. Anal. Calcd. for C21H16ClN3O.0.5H2O: C,
68.02; H, 4.62; N, 11.33. Found: C, 68.30; H, 4.40;
N, 11.02%.
743
3-(3-(1H-Benzo[d]imidazol-2-yl)phenyl)-6-bro-
mo-3,4-dihydro-2H-benzo[e][1,3]oxazine, 2g: Rf:
0.72 (50% EtOAc in hexane); m.p. 201.8°C; Tmax
(DSC exotherm): 210.3°C; IR (KBr): 3053(NH), 1608
(C=N), 1476, 1448, 1409, 1365, 1275, 1228, 1157,
1120, 1096, 1009, 973, 943, 855, 746, 609 cm-1; 1H
NMR (300 MHz, DMSO-d6): δ 12.83 (s, 1H, NH),
7.92 (s, 1H, ArH), 7.64 (m, 2H, ArH), 7.51 (m, 1H,
ArH), 7.40-7.37 (m, 2H, ArH), 7.24-7.22 (m, 4H,
ArH), 6.74-6.70 (m, 1H, ArH), 5.55 (s, 2H, N-CH2-
O), 4.77 (s, 2H, N-CH2-Ar); HRMS (ESI): m/z [M]+
calcd. for C21H16BrN3O: 405.0477; found: 405.4429.
Anal. Calcd. for C21H16BrN3O.H2O: C, 59.45; H,
4.28; N, 9.90. Found: C, 59.52; H, 4.22; N, 9.51%.
3-(3-(1H-Benzo[d]imidazol-2-yl)phenyl)-3,4-di-
hydro-6-iodo-2H-benzo[e][1,3]oxazine, 2h: Rf: 0.75
(50% EtOAc in hexane); m.p. 215.3°C; Tmax (DSC
exotherm): 218.7°C; IR (KBr): 3047(NH), 1607
(C=N), 1585, 1534, 1474, 1447, 1403, 1364, 1319,
1276, 1228, 1189, 1157, 1122, 1009, 968, 940, 853,
821, 766, 747, 683, 598, 476 cm-1; 1H NMR (300
MHz, DMSO-d6): δ 12.90 (s, 1H, NH), 7.99 (s, 1H,
ArH), 7.73-7.70 (m, 2H, ArH), 7.57 (m, 2H, ArH),
7.48-7.43 (m, 2H, ArH), 7.30-7.23 (m, 3H, ArH), 6.65
(d, J = 8.7 Hz, 1H, ArH), 5.61 (s, 2H, N-CH2-O), 4.82
(s, 2H, N-CH2-Ar); HRMS (ESI): m/z [M]+ calcd. for
C21H16IN3O: 453.0338; found: 453.1175. Anal. Calcd.
for C21H16IN3O: C, 55.64; H, 3.56; N, 9.27. Found: C,
55.79; H, 3.87; N, 8.92%.
3-(3-(1H-Benzo[d]imidazol-2-yl)phenyl)-6,8-di-
chloro-3,4-dihydro-2H-benzo[e][1,3]oxazine, 2i: Rf:
0.74 (50% EtOAc in hexane); m.p. 238.4°C; Tmax
(DSC exotherm): 242.9°C; IR (KBr): 3081(NH), 1608
(C=N), 1590, 1541, 1482, 1465, 1433, 1399, 1363,
1316, 1240, 1187, 1147, 1118, 1010, 974, 931, 859,
831, 794, 764, 747, 710, 683 cm-1; 1H NMR (300
MHz, DMSO-d6): δ 12.87 (s, 1H, NH), 7.93 (s, 1H,
ArH), 7.68 (d, J = 7.2 Hz, 1H, ArH), 7.58 (m, 2H,
ArH), 7.44-7.39 (m, 2H, ArH), 7.29 (s, 1H, ArH),
7.25-7.20 (m, 3H, ArH), 5.69 (s, 2H, N-CH2-O), 4.81
(s, 2H, N-CH2-Ar); HRMS (ESI): m/z [M]+ calcd. for
C21H15Cl2N3O: 395.0592; found: 395.4139. Anal.
Calcd. for C21H15Cl2N3O.0.4H2O: C, 62.51; H, 3.95;
N, 10.41. Found: C, 62.80; H, 4.20; N, 10.02%.
3-(3-(1H-Benzo[d]imidazol-2-yl)phenyl)-3,4-di-
hydro-6-methyl-2H-benzo[e][1,3]oxazine, 2j: Rf:
0.75 (50% EtOAc in hexane); m.p. 212.9°C; Tmax
(DSC exotherm): 218.4°C; IR (KBr): 3045 (NH),
1605 (C=N), 1582, 1501, 1493, 1459, 1444, 1409,
1367, 1313, 1292, 1276, 1259, 1227, 1174, 1138,
1120, 1009, 971, 952, 917, 812, 792, 764, 742, 682,
744 INDIAN J. CHEM., SEC B, MAY 2012
599, 467, 442 cm-1; 1H NMR (300 MHz, DMSO-d6):
δ 12.84 (s, 1H, NH), 7.91 (s, 1H, ArH), 7.65-7.63 (m,
2H, ArH), 7.51 (d, J = 7.2 Hz, 1H, ArH), 7.41-7.36
(m, 1H, ArH), 7.23-7.14 (m, 3H, ArH), 6.94 (s, 1H,
ArH), 6.88 (d, J = 8.1 Hz, 1H, ArH), 6.64 (d, J = 8.1
Hz, 1H, ArH), 5.49 (s, 2H, N-CH2-O), 4.71 (s, 2H, N-
CH2-Ar) 2.18 (s, 3H, CH3); HRMS (ESI): m/z [M]+
calcd. for C22H19N3O: 341.1528; found: 341.3740.
2-(3-(Benzo[d]oxazol-2-yl)phenyl)-2,3-dihydro-
1H-naphtho[1,2-e][1,3]oxazine, 3: Rf: 0.80 (20%
EtOAc in hexane); m.p. 156.4°C; Tmax (DSC
exotherm): 270.2°C; IR (KBr): 1598 (C=N), 1548,
1455, 1377, 1295, 1229, 1153, 1009, 942, 910, 821,
745, 682 cm-1; 1H NMR (300 MHz, CDCl3): δ 7.98 (s,
1H, ArH), 7.72-7.62 (m, 4H, ArH), 7.56 (d, J = 8.7
Hz, 1H, ArH), 7.47-7.41 (m, 2H, ArH), 7.30-7.15 (m,
5H, ArH), 6.98 (d, J = 8.7 Hz, 1H, ArH), 5.41 (s, 2H,
N-CH2-O), 4.95 (s, 2H, N-CH2-Ar); 13C NMR (100
MHz, CDCl3): δ 162.93, 152.19, 150.65, 149.04,
141.97, 131.08, 129.85, 128.99, 128.64, 128.40,
128.08, 126.73, 125.07, 124.52, 123.73, 120.90,
120.69, 120.42, 119.92, 118.74, 116.94, 112.32,
110.57, 78.63, 48.33; HRMS (ESI): m/z [M]+ calcd.
for C25H18N2O2: 378.1368; found: 378.0103. Anal.
Calcd. for C25H18N2O2: C, 77.50; H, 4.94; N, 7.23.
Found: C, 77.78; H, 5.25; N, 6.93%.
2-(3-(1H-Benzo[d]imidazol-2-yl)phenyl)-2,3-di-
hydro-1H-naphtho[1,2-e][1,3]oxazine, 4: Rf: 0.80
(50% EtOAc in hexane); m.p. 200°C; IR (KBr): 3047
(NH), 1605 (C=N), 1584, 1493, 1472, 1442, 1406,
1367, 1230, 1188, 1098, 1058, 1010, 952, 810, 742,
692 cm-1; 1H NMR (300 MHz, DMSO-d6): δ 12.84 (s,
1H, NH), 8.01 (s, 1H, ArH), 7.92 (d, J = 9 Hz, 1H,
ArH), 7.84 (d, J = 8.1 Hz, 1H, ArH), 7.74-7.63 (m,
3H, ArH), 7.56-7.51 (m, 2H, ArH), 7.42-7.33 (m, 3H,
ArH), 7.20-7.16 (m, 2H, ArH), 7.06 (d, J = 9 Hz, 1H,
ArH), 5.61 (s, 2H, N-CH2-O), 5.08 (s, 2H, N-CH2-
Ar); HRMS (ESI): m/z [M]+ calcd. for C25H19N3O:
377.1528; found: 377.4210. Anal. Calcd. for
C25H19N3O.H2O: C, 75.93; H, 5.35; N, 10.63. Found:
C, 76.05; H, 5.65; N, 10.42%.
Synthesis of 3-(3-(benzo[d]oxazol-2-yl)phenyl)-7-
(benzo[d]thiazol-2-yl)-3,4-dihydro-2H-benzo[e]-
[1,3]oxazine, 5
A mixture of 2-(3-aminophenyl)benzoxazole (100
mg, 0.475 mmol), 3-(benzo[d]thiazol-2-yl)phenol
(108 mg, 0.475 mmol) and formalin (37% w/v, 82 µL,
0.950 mmol) was heated at 80-90°C. A second lot of
formalin (37% w/v, 82 µL, 0.950 mmol) was added to
the reaction mixture after 2.5 hr and the heating was
continued for additional 2.5 hr. After completion of
the reaction, the mixture was cooled to RT and diluted
with water (10 mL). The product was extracted with
ethyl acetate (10 mL×3 times). The organic layers
were combined and washed with 10% aqueous NaOH
solution (10 mL× 3 times) followed by water (10
mL×3 times). The organic layer was dried over
anhydrous sodium sulfate and evaporated under
reduced pressure. The crude product was purified
over silica gel column by using 15% ethyl acetate in
hexane as eluent.
Rf: 0.70 (30% EtOAc in hexane); m.p. 196.5°C;
Tmax (DSC exotherm): 264.7°C; IR (CHCl3): 1602
(C=N), 1550, 1484, 1452, 1380, 1244, 1191, 1114,
1000, 935, 860, 791, 758, 684 cm-1; 1H NMR (300
MHz, CDCl3): δ 8.06 (s, 1H, ArH), 8.03 (s, 1H, ArH),
7.88 (d, J = 7.8 Hz, 1H, ArH), 7.83 (d, J = 7.5 Hz,
1H, ArH), 7.79-7.76 (m, 1H, ArH), 7.67-7.57 (m, 3H,
ArH), 7.50-7.25 (m, 6H, ArH), 7.18 (d, J = 7.8 Hz,
1H, ArH), 5.51 (s, 2H, N-CH2-O), 4.80 (s, 2H, N-
CH2-Ar); HRMS (ESI): m/z [MH]+ calcd. for
C28H20N3O2S: 462.1276; found: 462.0231.
In vitro antibacterial assay
The in-vitro antibacterial activity of all the prepared
compounds was evaluated against Gram-positive
bacteria viz. S. epidermidis and S. aureus and Gram-
negative bacteria viz. E. coli and P. aeruginosa and
results are shown in Table I. The antimicrobial
susceptibility testing was carried out by using National
Committee for Clinical Laboratory Standards
(NCCLS) micro-dilution assay. Briefly, the bacterial
strains were grown in standard media until exponential
growth was achieved. Tests were performed in a 96-
well microtitre plate in a final volume of 100 µL. Test
compounds were dissolved in 5% DMSO at an initial
concentration of 500 µg and serially diluted in plate.
Each well was then inoculated with ~2-5 × 105
bacterial cells and incubated at 37°C for 24 hr with
shaking at 200 rpm. One well containing cells and 5%
DMSO without any test compound (growth control),
and one well containing only growth medium (sterility
control) were used as controls. Growth of bacteria was
determined using Power wave200 microplate scanning
spectrophotometer (Bio-Tech Instruments, Winooski,
VT, USA). Percent survival was calculated using
growth without any compound as 100% survival. The
IC50 values are calculated using Grafit 4.0 software
(Erithacus Software Ltd., Horley, Surrey, UK).
 PRASAD et al.: SYNTHESIS OF BENZAZOLES TETHERED DIHYDRO[1,3]OXAZINES
Acknowledgements
The authors thank University of Delhi, India for
providing a research grant under the scheme to
strengthen R & D Doctoral Research programme. One
of the authors (DP) is grateful to Council of Scientific
and Industrial Research (CSIR), New Delhi, India, for
Senior Research Fellowship.
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