Synthesis and Antimicrobial Screening of Some 4-Substituted-3-Chloro-2-Oxo-Azetidine Derivatives
Rakesh Patel1* and Anil Bhandari2
1Charak Institute of Pharmacy, Mandleshwar (Khargone) M P
2Department of Pharmacy, Jodhpur National University, Jodhpur (Raj)
*Corresponding Author E-mail: patelcip@gmail.com
ABSTRACT:
A new series of 2-oxo-azetidine derivatives of isoniazid have been synthesized and evaluated for their Anti-bacterial, Antifungal and Anti-tubercular activity. Synthesized compounds show significant activity against bacterial, fungal and mycobacterium strains. Their structures were established on the basis of elemental analysis, IR, 1H NMR and Mass Spectral data.
KEYWORDS: INZ, Isoniazid, azetidine, antimycobacterial, TB.
Although deaths from bacterial and fungal infection have dropped currently, still those are the major cause of death in the world.1 Over the few past decades the bacterial resistance to antibiotics, anti-fungal and anti-tuberculotic drugs has become one of the most challenging problem in the infections treatments. Tuberculosis (TB) is the world’s oldest known infectious disease that kills three million deaths each year. The urgency to develop new and effective drugs is due to the resistance development by strains against current medications and grooving problem of co-infection in immunocompromised patients.2, 3
Several research has been done and currently in progress to develop new and better chemical entity against infections. Literature survey reveals that 2-oxo-azetidines have shown various biological activities along with antimicrobial activity. 4-11 In view of these findings some 2-oxo- azetidine derivatives of Isoniazid have been synthesized and evaluated for Anti-bacterial, Antifungal and Anti-tubercular activity.
MATERIAL AND METHOD:
All the chemicals used were purchased from E Merck, S D Fine and Loba Chem and were purified by established methods (whenever needed). Various Substituted Isoniazid (Schiff Base) derivatives were prepared according to the procedure outline in scheme-I. 2-oxo azetidine derivatives were synthesized by formation of imines (from Schiff base) and ketenes (from 2-chloro acetylchloride) followed by cycloaddition of ketenes to imines, in the presence of 1:4 Dioxane, in a single step reaction. Melting points were determined by open capillary tube method and are uncorrected. Purity of synthesized compounds was checked by TLC plates (Silica Gel G) and visualized by iodine vapor. The infra red absorption spectra of the synthesized compounds were recorded using KBr disc on FTIR 8010 Shimadzu model. 1H NMR spectra were recorded on Brucker Spectrospin DPX 300 spectrophotometer. Mass spectra were recorded on Jeol SR-102 FAB Mass spectrometer. CHN analyses of synthesized compounds were done on Perkin-Elmer-240 analyzer.
Scheme: I
General procedure for synthesis of substituted Isoniazid (Schiff Base), I (a-f):
A mixture of Isoniazid (0.01 mol), substituted aldehyde (0.01 mol) and ethanol (30-35ml) were reflux for 3 Hrs. The reaction mixture were cooled at room temperature and allowed to stand for 5 hrs. Solid products were separated out, filtered, washed with ice-cold water, dried and recrystallized from ethanol.
General procedure for synthesis of 2-oxo-azetidine derivatives of Isoniazid, IIa-f:
Substituted Isoniazid (0.01 mol) (Ia-f) were dissolved in 1:4 Dioxane (20ml) with constant stirring, triethylamine (0.01 mol) was added followed by drop wise addition of 2-chloro acetyl chloride (0.01 mol). Content was stirred vigorously for 15 minutes and refluxed for 5 hrs. Mixture was cooled at room temperature, filtered, washed with ice cooled water, dried and recrystallized from ethanol. Physical Characteristics were shown in Table No.: 1
IIa: N-(3-chloro-2-oxo-4-methylazetidine-1-yl) isonicotinamide:
IR (KBr, cm-1): 3390 (N-H Sec. Amide), 3060 (C-H Ar), 1710 (C=O Cyclic β-Lactam), 1660 (C=N, Pyridine), 1630 (C=O acyclic). 1H NMR (DMSO-d6 δ ppm) 7.8-8.4 (m, 4H, CH Pyridine), 8.6 (s, 1H, NH Amide), 4.1-4.3 (s, 1H, CH-Cl of β-Lactam), 2.5 (s, 1H, CH), 2.3 (t, 3H, Methyl), Elemental analysis % found (% calculated): C-58.72 (58.82), H-4.86 (4.90), N-20.48 (20.59).
IIb: N-(3-chloro-2-oxo-4-phenylazetidine-1-yl) isonicotinamide:
IR (KBr, cm-1): 3370 (N-H Sec. Amide), 3050 (C-H Ar), 1715 (C=O Cyclic β-Lactam), 1660 (C=N, Pyridine), 1630 (C=O acyclic). 1H NMR (DMSO-d6 δ ppm) 7.8-8.4 (m, 4H, CH, Pyridine), 8.6 (s, 1H, NH Amide), 4.1-4.3 (s, 1H CH-Cl of β-Lactam), 2.5 (s, 1H, CH), 7.1-7.2 (m, 5H phenyl), Elemental analysis % found (% calculated): C-67.72 (67.67), H-4.48 (4.51), N-15.78 (15.84).
IIc: N-[3-chloro-2-oxo-4-(p-methoxyphenyl) azetidine-1-yl] isonicotinamide:
IR (KBr, cm-1): 3260 (N-H Sec. Amide), 3010 (C-H Ar), 1780 (C=O Cyclic β-Lactam), 1660 (C=N, Pyridine), 1610 (C=O acyclic), 2820 (CH Methoxy). 1H NMR (DMSO-d6 δ ppm) 7.8-8.4 (m, 4H, CH, Pyridine), 8.6 (s, 1H, NH Amide), 7.3-7.4 (m, 4H phenyl) 4.1-4.3 (s, 1H CH-Cl of β-Lactam), 3.2 (t, 3H methoxy), 2.5 (s, 1H CH), Elemental analysis % found (% calculated): C-64.83 (64.86), H-4.74 (4.78), N-14.21(14.19).
IId: N-[3-chloro-2-oxo-4-(p-hydroxyphenyl) azetidine-1-yl] isonicotinamide:
IR (KBr, cm-1): 3260 (N-H Sec. Amide), 2990 (C-H Ar), 1770 (C=O Cyclic β-Lactam), 1660 (C=N, Pyridine), 1610 (C=O acyclic), 1280 (OH Ar). 1H NMR (DMSO-d6 δ ppm) 7.8-8.4 (m, 4H, CH, Pyridine), 8.6 (s, 1H, NH Amide), 7.3-7.4 (m, 4H phenyl), 5.1 (s, 1H OH), 4.1-4.3 (s, 1H CH-Cl of β-Lactam), 2.5 (s, 1H CH), Elemental analysis % found (% calculated): C-63.78 (63.83), H-4.29 (4.26), N-14.87 (14.94).
IIe: N-[3-chloro-2-oxo-4-(p-chlorophenyl) azetidine-1-yl] isonicotinamide:
IR (KBr, cm-1): 3260 (N-H Sec. Amide), 2990 (C-H Ar), 1770 (C=O Cyclic β-Lactam), 1660 (C=N, Pyridine), 1610 (C=O acyclic), 825 (C-Cl). 1H NMR (DMSO-d6 δ ppm) 7.8-8.4 (m, 4H, CH, Pyridine), 8.6 (s, 1H, NH Amide), 7.3-7.4 (m, 4H phenyl), 4.1-4.3 (s, 1H CH-Cl of β-Lactam), 2.5 (s, 1H CH Ar), Elemental analysis % found (% calculated): C-59.92 (59.90), H-3.57 (3.66), N-14.01 (13.98).
IIf: N-[3-chloro-2-oxo-4-(p-nitrophenyl) azetidine-1-yl] isonicotinamide:
IR (KBr, cm-1): 3260 (N-H Sec. Amide), 2990 (C-H Ar), 1770 (C=O Cyclic β-Lactam), 1660 (C=N, Pyridine), 1610 (C=O acyclic), 1340 (NO2). 1H NMR (DMSO-d6 δ ppm) 7.8-8.4 (m, 4H, CH, Pyridine), 8.6 (s, 1H, NH Amide), 7.3-7.4 (m, 4H phenyl), 4.1-4.3 (s, 1H CH-Cl of β-Lactam), 2.5 (s, 1H CH Ar), Elemental analysis % found (% calculated): C-57.84 (57.88), H-3.59 (3.54), N-18.12 (18.01).
ANTIMICROBIAL ACTIVITY:
All the synthesized compounds were evaluated for their invitro antimicrobial activity against gram positive bacteria staphylococcus aureus (ATCC-24392), the gram negative bacteria Echerichia coli (ATCC-24391) in nutrient agar media, 12 fungi C Albicans (ATCC-436) in sabouraud dextrose medium 13 and mycobacterium tuberculosis (ATTC-27286) in tween-albumin medium 14. The zone of inhibition values were determined and compared with well known (standard) antibacterial (Ofloxacin), antifungal (Ketoconazole) and antituberculotic (Isoniazid) drugs. Table: 2 shows data obtained from the biological screening of synthesized compounds and reference drugs.
Table: 1-Physical characteristics of synthesized compounds
|
Compound |
R |
Molecular Formula |
Yield (%) |
m. p. (°C) |
Molecular Weight |
Rf |
|
IIa |
CH3 |
C10H10N3O2 |
58 |
235 |
204.20 |
0.6 |
|
IIb |
C6H5 |
C15H12N3O2 |
62 |
242 |
266.27 |
0.4 |
|
IIc |
C6H4OCH3 |
C16H14N3O3 |
71 |
295 |
296.30 |
0.7 |
|
IId |
p-OHC6H4 |
C15H12N3O3 |
74 |
286 |
282.27 |
0.7 |
|
IIe |
p-ClC6H4 |
C15H11N3O2Cl |
73 |
172 |
300.72 |
0.6 |
|
IIf |
p-NO2C6H4 |
C15H11N4O4 |
69 |
216 |
311.27 |
0.8 |
Table:2-Antimicrobial screening data of synthesized compounds.
|
Compounds |
Zone of Inhibition (in mm) at concentration of 20 µg/mL) |
|||
|
S. aureus |
E. Coli |
C. Albicans |
M. tuberculosis |
|
|
IIa |
09 |
13 |
22 |
24 |
|
IIb |
12 |
15 |
24 |
23 |
|
IIc |
11 |
14 |
26 |
27 |
|
IId |
18 |
22 |
28 |
31 |
|
IIe |
16 |
18 |
29 |
28 |
|
IIf |
23 |
29 |
32 |
33 |
|
Ofloxacin |
12 |
16 |
----- |
----- |
|
Ketoconazole |
----- |
----- |
26 |
----- |
|
Isoniazid |
----- |
----- |
----- |
29 |
RESULTS AND DISCUSSION:
Chemistry:
Yield of synthesized compounds were found to be satisfactory. The purity of synthesized compounds and completion of reactions were checked by TLC on silica Gel G plates in the solvent system methyl chloride: methanol (8:2 v/v) and visualized spots in iodine vapor. Proposed structures were confirmed by Spectral and microanalysis data. 1H NMR (δ ppm) spectra shows signals at 4.1-4.3 (CH-Cl of β-Lactam), 8.6 (N-H Secondary Amide) and 7.8-8.4 (C-H Pyridine). Presences of various functional groups were supported by the IR and Mass spectral data. Further elemental analysis data were also found in agreement with calculated values from proposed structures.
Antimicrobial Activity:
Antibacterial, antifungal and anti tuberculotic screening data of synthesized compounds showed good to moderate activity as compared to reference drug. Compound IIa and IIc showed moderate activity against all strains. Compound IIb showed good activity against S. aureus, E Coli and C. Albicans while showed moderate activity against mycobacterium tuberculosis. Compound IId, IIe and IIf showed good antibacterial, antifungal and anti tuberculotic activity against tested strains. The antimicrobial activity is due the presence of pharmacological active β-Lactam ring and increased by the addition p-substituted phenyl moiety at 4 position of β-Lactam ring. Amongst these IIf showed highest activity as compare to other synthesized compounds, is due to the presence the NO2 group at p- position of phenyl moiety.
CONCLUSION:
On the basis of above research work; the results and discussion showed that the synthesized compounds showed good antimicrobial activity as compared to reference antimicrobial drugs. These results concluded the need of development of such type of compounds in future for the progress of drug synthesis area.
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Received on 29.01.2014 Modified on 20.03.2014
Accepted on 05.04.2014 © AJRC All right reserved
Asian J. Research Chem. 7(11): November, 2014; Page 950-953