Synthesis, Antimicrobial and Anticancer Evaluation of 2-Azetidinones Clubbed with Quinazolinone

Article abstract of DOI:10.1007/s11094-016-1392-3

A novel series of 2-azetidinones clubbed with quinazolinone was synthesized using anthranilic acid. All the synthesized compounds were evaluated for their antimicrobial activity against two Gram positive bacterial strains (Bacillus subtilis and Staphyloco

Full text of DOI:10.1007/s11094-016-1392-3

DOI 10.1007/s11094-016-1392-3
Pharmaceutical Chemistry Journal, Vol. 50, No. 1, April, 2016 (Russian Original Vol. 50, No. 1, January, 2016)
SYNTHESIS, ANTIMICROBIAL AND ANTICANCER
EVALUATION OF 2-AZETIDINONES CLUBBED
WITH QUINAZOLINONE
Aakash Deep,1 Pradeep Kumar,1 Balasubrmanian Narasimhan,1,*
Lim Siong Meng,2 Kalavathy Ramasamy,2 Rakesh Kumar Mishra,3
and Vasudevan Mani3
Original article submitted January 10, 2014.
A novel series of 2-azetidinones clubbed with quinazolinone was synthesized using anthranilic acid. All the
synthesized compounds were evaluated for their antimicrobial activity against two Gram positive bacterial
strains (Bacillus subtilis and Staphylococcus aureus), one Gram negative bacterial strain (Escherichia coli)
and two fungal strains (Candida albicans and Aspergillus niger). All the synthesized compounds were also
screened for their anticancer activity against human breast cancer cell line, MCF-7. Results of antimicrobial
and anticancer study indicate that compounds 12 and 5 (IC50 = 49.52 mM) are the most potent antimicrobial
and anticancer agents, respectively.
Keywords: antibacterial; antifungal; anticancer activity.
INTRODUCTION
has been laid on the development of new antimicrobial
agents [2]. The 2-azetidinone structural motifs have attracted
a great deal of interest due to their broad biological activity
profile including antimicrobial [3], anticancer [4],
antimycobacterial [5], anti-inflammatory, analgesic [6], and
antiviral [7] activities. In the light of facts mentioned above
and in continuation of our previous research effort in the
field of synthesis, antimicrobial and anticancer evaluation
[8], the present article reports on the synthesis, antimicrobial,
Cancer is a disease characterized by a shift in the con-
trolled mechanisms that govern cell proliferation and differ-
entiation. Malignancy is caused by abnormalities in cells,
which might be due to inherited genes or caused by outside
exposure of the body to chemicals, radiation, or even infec-
tious agents. Several techniques were adopted for the treat-
ment and eradication of cancerous cells. These techniques in-
clude surgery, radiation, immunotherapy, chemotherapy, and
chemoprevention. Ideal anticancer drugs would eradicate
cancer cells without harming normal tissues. Unfortunately,
no currently available agents meet this criterion and clinical
use of drugs involves weighing benefits against toxicity in
the search of favorable therapeutic index [1]. In recent de-
cades, problems of multidrug resistant microorganisms have
reached an alarming level around the world. These pose a se-
rious challenge to the scientific community; hence, emphasis
and anticancer evaluation of 2-azetidinone derivatives.
A series of 2-azetidinone derivatives (1 – 17) has been
synthesized using a synthetic procedure outlined in Fig. 1,
and their physicochemical properties are presented in Ta-
ble 1.
EXPERIMENTAL CHEMICAL PART
1
Reaction progress was observed by thin layer chroma-
tography (TLC) using commercial silica gel plates (Merck),
Faculty of Pharmaceutical Sciences, Maharshi Dayanand University,
Rohtak-124001, India.
2
Collaborative Drug Discovery Research Group, Faculty of Pharmacy,
1
Silica gel F254 on aluminum sheets. The H nuclear mag-
Campus Puncak Alam, Universiti Teknologi MARA (UiTM), 42300
Bandar Puncak Alam, Selangor, Malaysia.
netic resonance (1H NMR) spectra were measured using a
Bruker AV300 NMR spectrometer. The infrared (IR) spectra
were recorded on an Agilent Resolutions Pro FTIR spec-
trometer.
3
Brain Research Laboratory, Faculty of Pharmacy, Campus Puncak Alam,
Universiti Teknologi MARA (UiTM), 42300 Bandar Puncak Alam,
Selangor, Malaysia.
*
e-mail: naru2000us@yahoo.com.
24
0091-150X/16/5001-0024 © 2016 Springer Science+Business Media New York

Synthesis, Antimicrobial and Anticancer Evaluation
25
Fig. 1. Scheme of the synthesis of 2-azetidinone derivatives (1 – 17).
Synthesis
flux for 3 h and then allowed to cool. The solid product was
collected and recrystallized from acetic acid to yield methyl
2-benzamidobenzoate, 0.2 M of which was refluxed with ex-
cess of hydrazine hydrate (0.30 M) in ethanol (250 mL) for
about 3 h and cooled. The solid was separated by filtration
and recrystallized from ethanol to afford 3-amino-2-phenyl-
quinazolin-4(3H)-one. A mixture of the product (0.025 M)
and appropriate aldehyde (0.025 M) was refluxed in metha-
A mixture of 0.25 M anthranillic acid and excess of
methanol (250 mL) with 1 mL of sulfuric acid was refluxed
for 3 – 4 h in round-bottom flask and then cooled. The pre-
cipitated methyl 2-aminobenzoate was separated by filtration
and recrystallized from methanol. A mixture of the product
(0.2 M) and phthalic anhydride (0.2 M) was heated under re-

26
Aakash Deep et al.
nol (50 mL) in the presence of a catalytic amount of glacial
acetic acid for about 2 h. The mixture was cooled and the
precipitated solid was separated by filtration and recrystalli-
zed from methanol to give the corresponding hydrazone,
0.02 M of which was refluxed with triethylamine (0.02 M)
and chloroacetyl chloride (0.02 M) in dimethylformamide
(40 mL) for 3 h on water bath to yield 2-azetidinone deriva-
tive. After cooling, the solution was poured on crushed ice to
precipitate the crude product, which was recrystallized from
rectified ethanol.
(C=N), 1280 (C-N), 1246 (N-N), 768 (C-Cl), 1177 (C-O-C
str., -OC H ), 3298 (OH-Ar); 1H NMR (DMSO-d
2
5
6,
400 MHz; d, ppm): 7.23 – 8.54 (m, 13H, ArH), 6.87 (d, 1H,
N-CH), 6.89 (d, IH, CH-Cl), 1.24 (t, 3H, CH of -OC H ),
3
2
5
3.33 (m, 2H, CH of -OC H ), 4.10 (s,1H, OH).
2
2
5
3-(3-Chloro-2-oxo-4-phenylazetidin-1-yl)-2-phenylqu
inazolin-4(3H)-one(5): m.p. (°C), 145 – 147; yield, 86.20%;
IR (KBr pellets; n, cm-1): 3050 (C-H aromatic),1622 (C=O),
1
1571 (C=N), 1210 (N-N), 750 (C-Cl); H NMR (DMSO-d
6,
400 MHz; d, ppm): 7.88 – 8.72 (m, 14H, ArH), 7.51 (d, 1H,
N-CH), 7.53 (d, IH, CH-Cl).
3-(3-Chloro-2-(2-methoxyphenyl)-4-oxoazetidin-1-yl)-
2-phenylquinazolin-4(3H)-one (1): m.p. (°C), 111 – 113;
yield, 67.04%; IR (KBr pellets; n, cm-1): 3001 (C-H aro-
matic), 1615 (C=C aromatic), 1665 (C=O), 1520 (C=N),
1291 (C-N), 1248 (N-N), 751 (C-Cl), 1156 (C-O-C str.,
-OCH ); 1H NMR (DMSO-d 400 MHz; d, ppm):
3-(3-Chloro-2-(2-chlorophenyl)-4-oxoazetidin-1-yl)-2-
phenylquinazolin-4(3H)-one (6): m.p. (°C), 188 – 190;
yield, 61.41%; IR (KBr pellets; n, cm-1): 3005 (C-H aro-
matic), 1615 (C=C aromatic), 1700 (C=O), 1563 (C=N),
1
1272 (C-N), 1209 (N-N), 681 (C-Cl); H NMR (DMSO-d
3
6,
6,
7.15 – 7.99 (m, 13H, ArH), 7.05 (d, 1H, N-CH), 7.04 (d, IH,
400 MHz; d, ppm): 7.51 – 8.16 (m, 13H, ArH), 7.49 (d, 1H,
N-CH), 7.48 (d, IH, CH-Cl).
CH-Cl) 3.89 (s, 3H, -OCH ).
3
3-(3-Chloro-2-(3-methoxyphenyl)-4-oxoazetidin-1-yl)-
2-phenylquinazolin-4(3H)-one (2): m.p. (°C), 137 – 139;
yield, 68.10%; IR (KBr pellets; n, cm-1): 3010 (C-H aro-
matic), 1601 (C=C aromatic), 1656 (C=O), 1556 (C=N),
1261 (C-N), 1220 (N-N), 785 (C-Cl), 1163 (C-O-C str.,
-OCH ); 1H NMR (DMSO-d 400 MHz; d, ppm):
3-(3-Chloro-2-oxo-4-styrylazetidin-1-yl)-2-phenylqui
nazolin-4(3H)-one (7): m.p. (°C), 179 – 181; yield, 59.29%;
IR (KBr pellets; n, cm-1): 2972 (C-H aromatic), 2876 (C-H
str.,-CH=CH-), 1629 (C=C aromatic), 1513 (C=N), 1279
(C-N), 769 (C-Cl); 1H NMR (DMSO-d 400 MHz; d, ppm):
6,
3
6,
7.25 – 8.54 (m, 14H, ArH), 7.23 (d, 1H, N-CH), 6.89 (d, IH,
CH-Cl), 6.87 (d, 1H, CH).
7.45 – 8.69 (m, 13H, ArH), 7.43 (d, 1H, N-CH), 7.44 (d, IH,
CH-Cl) 3.40 (s, 3H, -OCH ).
3
3-(3-Chloro-2-(4-methoxyphenyl)-4-oxoazetidin-1-yl)-
3-(3-Chloro-2-(3-chlorophenyl)-4-oxoazetidin-1-yl)-2-
phenylquinazolin-4(3H)-one (3): m.p. (°C); 155 – 157;
yield, 77.52%; IR (KBr pellets; n, cm-1): 3003 (C-H aro-
matic), 1626 (C=O), 1565 (C=N), 1263 (C-N), 1210 (N-N),
2-phenylquinazolin-4(3H)-one (8): m.p. (°C), 130 – 132;
yield, 79.77%; IR (KBr pellets; n, cm-1): 2967 (C-H aro-
matic), 1601 (C=C aromatic), 1509 (C=N), 1250 (C-N),
1216 (N-N), 780 (C-Cl), 1166 (C-O-C str., -OCH ); 1H NMR
780 (C-Cl); 1H NMR (DMSO-d 400 MHz; d, ppm):
3
6,
(DMSO-d 400 MHz; d, ppm): 7.80 – 8.63 (m, 13H, ArH),
7.07 (d, 1H, N-CH), 7.05 (d, IH, CH-Cl) 3.83 (s, 3H,
7.57 – 7.93 (m, 13H, ArH), 7.54 (d, 1H, N-CH), 7.56 (d, IH,
CH-Cl).
6,
3-(3-Chloro-2-(3-ethoxy-4-hydroxyphenyl)-4-oxoazeti
din-1-yl)-2-phenylquinazolin-4(3H)-one (4): m.p. (°C),
172 – 174; yield, 57.29%; IR (KBr pellets; n, cm-1): 2970
(C-H aromatic), 1597 (C=C aromatic), 1629 (C=O), 1514
-OCH ).
3
3-(3-Chloro-2-oxo-4-p-tolylazetidin-1-yl)-2-phenylqu-
inazolin-4(3H)-one (9): m.p. (°C), 135 – 137; yield,
66.86%; IR (KBr pellets; n, cm-1): 3031 (C-H aromatic),
TABLE 1. Physicochemical Characteristics and Anticancer Activity of 2-Azetidinone derivatives 1 – 17
*Rf
*Rf
Comp.
Formula
MW
Comp.
Formula
MW
IC50 (mM)
122.72
IC50 (mM)
>203.29
>208.01
>216.50
>232.63
>224.76
>239.33
>229.21
>252.63
6.00
1
2
3
4
5
6
7
8
9
C24H18ClN3O3
C24H18ClN3O3
C23H15Cl2N3O2
C25H20ClN3O4
C23H16ClN3O2
C23H15Cl2N3O2
C25H18ClN3O2
C24H18ClN3O3
C24H18ClN3O2
431.87
431.87
436.29
461.90
401.85
436.29
427.88
431.87
415.87
0.54
0.61
0.71
0.69
0.58
0.72
0.50
0.67
0.58
10
11
C26H22ClN3O5
C23H15BrClN3O2
C25H20ClN3O4
C24H16ClN3O3
C25H21ClN4O2
C23H16ClN3O3
C23H15Cl2N3O2
C21H18ClN3O3
491.92
480.74
461.90
429.86
444.91
417.84
436.29
395.84
0.61
0.52
0.67
0.75
0.70
0.56
0.73
0.57
>231.55
>229.21
>216.50
49.52
12
13
14
>229.21
>233.71
>231.55
>240.46
15
16
17
5-FU
* TLC mobile phase: benzene.

Synthesis, Antimicrobial and Anticancer Evaluation
27
Fig. 2. Structural requirements for the antimicrobial and anticancer activity of 2-azetidinones.
(C-H str., Ar CH ); 1H NMR (DMSO-d 400 MHz; d, ppm):
7.70 – 8.50 (m, 13H, ArH), 6.78 (d, 1H, N-CH), 6.76 (d, IH,
1620 (C=C aromatic), 1511 (C=N), 1301 (C-N), 1212 (N-N),
1
779 (C-Cl), 2965(C-H str., Ar CH ); H NMR (DMSO-d
3
6,
3
6,
400 MHz; d, ppm): 7.76 – 8.66 (m, 13H, ArH), 7.33 (d, 1H,
N-CH), 7.31(d, IH, CH-Cl), 2.38 (s, 3H, CH ).
CH-Cl), 2.51 (s, 6H, N (CH ) ).
3 2
3-(3-Chloro-2-(4-hydroxyphenyl)-4-oxoazetidin-1-yl)-
2-phenylquinazolin-4(3H)-one (15): m.p. (°C), 188 – 190;
yield, 66.30%; IR (KBr pellets; n, cm-1): 3594(OH), 3049
(C-H aromatic), 1624 (C=C aromatic), 1592 (C=N), 1293
3
3-(3-Chloro-2-oxo-4-(3,4,5-trimethoxyphenyl)azetidin-
1-yl)-2-phenylquinazolin-4(3H)-one (10): m.p. (°C), 151 –
153; yield, 53.49%; IR (KBr pellets; n, cm-1): 2946 (C-H ar-
omatic), 1622 (C=C aromatic), 1502 (C=N), 1295 (C-N),
(C-N), 1210 (N-N), 703 (C-Cl); 1H NMR (DMSO-d
6,
1231 (N-N), 763 (C-Cl), 1184 (C-O-C str., -OCH ); 1H NMR
400 MHz; d, ppm): 7.90 – 8.72 (m, 13H, ArH), 7.60 (d, 1H,
N-CH), 7.58 (d, IH, CH-Cl), 3.51 (s, 1H, -OH).
3
(DMSO-d 400 MHz; d, ppm): 8.66 – 9.90 (m, 11H, ArH),
7.27 (d, 1H, N-CH), 7.23 (d, IH, CH-Cl) 3.99 (s, 9H,
6,
3-(3-Chloro-2-(4-chlorophenyl)-4-oxoazetidin-1-yl)-2-
-OCH ).
phenylquinazolin-4(3H)-one (16): m.p. (°C), 193 – 195;
yield, 62.66%; IR (KBr pellets; n, cm-1): 3008 (C-H aro-
matic), 1600 (C=C aromatic), 1655 (C=O), 1554 (C=N),
3
3-(2-(4-Bromophenyl)-3-chloro-4-oxoazetidin-1-yl)-2-
phenylquinazolin-4(3H)-one (11): m.p. (°C), 141 – 143;
yield, 68.01%; IR (KBr pellets; n, cm-1): 2995 (C-H aro-
matic), 1626 (C=C aromatic), 1585 (C=N), 1295 (C-N), 701
1
1261 (C-N), 1220 (N-N), 783 (C-Cl); H NMR (DMSO-d
6,
400 MHz; d, ppm): 7.90 – 8.08 (m, 13H, ArH), 7.89 (d, 1H,
N-CH), 7.88(d, IH, CH-Cl).
(C-Cl), 597 (C-Br); 1H NMR (DMSO-d 400 MHz; d, ppm):
6,
7.82 – 8.70 (m, 13H, ArH), 7.74 (d, 1H, N-CH), 7.72 (d, IH,
CH-Cl).
4-(3-Chloro-4-oxo-1-(4-oxo-2-phenylquinazolin-3(4H)-
yl)azetidin-2-yl)butanal (17): m.p. (°C), 148 – 150; yield,
3-(3-Chloro-2-(3,4-dimethoxyphenyl)-4-oxoazetidin-1-
80.12%; IR (KBr pellets; n, cm-1): 3013 (C-H aromatic),
1600 (C=C aromatic), 1656 (C=O), 1557 (C=N), 1260
(C-N), 1221 (N-N), 785 (C-Cl), 2895(C-H str.,-CH -CH -);
yl)-2-phenylquinazolin-4(3H)-one (12): m.p. (°C), 161 –
163; yield, 62.60%; IR (KBr pellets; n, cm-1): 3004 (C-H ar-
omatic), 1622 (C=C aromatic), 1509 (C=N), 1259 (C-N),
2
2
1H NMR (DMSO-d 400 MHz; d, ppm): 7.89 – 8.07 (m, 9H,
ArH), 7.88 (d, 1H, N-CH), 7.87 (d, IH, CH-Cl), 1.24 – 2.64
1238 (N-N), 754 (C-Cl), 1141 (C-O-C str., -OCH ); 1H NMR
6,
3
(DMSO-d 400 MHz; d, ppm): 7.37 – 8.64 (m, 12 H, ArH),
7.09 (d, 1H, N-CH), 7.07 (d, IH, CH-Cl) 3.83 (s, 3H,
6,
(m, 6H (CH ) ).
2 3
-OCH ).
3
EXPERIMENTAL BIOLOGICAL PART
4-(3-Chloro-4-oxo-1-(4-oxo-2-phenylquinazolin-3(4H
)-yl)azetidin-2-yl)benzaldehyde (13): m.p. (°C), 185 – 187;
The antimicrobial activity of synthesized compounds
was tested against bacterial strains of S. aureus, B. subtilis,
and E. coli (37 ± 1°C for 24 h) and fungal strains of C.
albicans (37 ± 1°C for 48 h) and A. niger (25 ± 1°C for 7 d)
using the tube dilution method. All the compounds were dis-
solved in dimethyl sulfoxide (DMSO) to give a concentra-
tion of 10 mg mL-1. The stock solution was serially diluted to
give concentrations of 50 – 1.56 mg mL-1 [9].
yield, 54.77%; IR (KBr pellets; n, cm-1): 2993 (C-H aro-
matic), 1618 (C=C aromatic), 1689 (C=O), 1504 (C=N),
1298 (C-N), 1208 (N-N), 692 (C-Cl), 2816 (C-H str., CHO).
3-(3-Chloro-2-(4-(dimethylamino)phenyl)-4-oxoazeti
din-1-yl)-2-phenylquinazolin-4(3H)-one (14): m.p. (°C),
165 – 167; yield, 88.04%; IR (KBr pellets; n, cm-1): 2984
(C-H aromatic), 1599 (C=C aromatic), 1520 (C=N), 1228
(N-N), 743 (C-Cl), 1364 (C-N str., aryl tertiary amine), 2806

28
Aakash Deep et al.
Gram negative bacterium E. coli, compounds 6 and 14
TABLE 2. Antimicrobial Activity (pMIC in mM/mL) of Synthe-
sized 2-Azetidinone Derivatives 1 – 17
(pMIC = 2.15 mM/mL) were found to be most potent
ec
antimicrobial agents. Compound 7 (pMIC = 2.21 mM/mL)
No.
1
pMICbs
1.84
1.54
1.54
1.87
1.51
1.54
1.61
1.84
0.92
1.59
1.89
2.17
1.84
1.55
1.52
1.24
1.50
2.61*
pMICec
1.84
1.84
1.54
1.57
1.51
2.15
1.91
1.84
1.52
1.90
1.59
1.57
1.84
2.15
1.83
1.54
1.50
2.61*
pMICsa
0.94
0.94
1.54
1.57
1.21
1.54
1.61
1.24
1.82
1.90
1.89
2.17
2.14
1.55
1.52
1.84
1.50
2.61*
pMICan
1.54
1.84
1.24
1.57
1.81
1.24
2.21
1.24
1.52
1.29
1.59
1.27
1.54
0.95
1.22
1.54
1.50
2.64**
pMICca
1.24
1.54
0.94
1.27
1.81
1.24
1.91
1.24
0.92
1.29
1.59
2.17
1.84
1.25
1.22
1.54
1.80
2.64**
an
was found to be most potent antifungal agent against A.
niger. In general, compound 12 was found to be most potent
antimicrobial agent among the synthesized derivatives but
less active as compared to the standard drug.
2
3
Anticancer activity results (Table 1) indicated that the
synthesized compounds exhibited weak anticancer activity
against breast cancer cell line (MCF-7) as compared to stan-
4
5
6
dard drug. Compound 5 (IC = 49.52 mM) was found to be
50
7
most potent anticancer agent. Data on the structure – activity
relationships for antimicrobial and anticancer activity of syn-
thesized compounds are presented in Figure 2.
8
9
10
11
12
13
14
15
16
17
Std.
CONCLUSION
In the present study, a series of 2-azetidinone derivatives
(1 – 17) was synthesized and evaluated for their in vitro
antimicrobial and anticancer potentials. The synthesized
compounds exhibit more potent antimicrobial activity as
compared to their anticancer activity. Compounds 12 and 5
(IC 49.52 mM) were found to be most potent
antimicrobial and anticancer agents, respectively.
50 =
Standard drug: * norfloxacin; ** fluconazole.
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