Design, Synthesis, Characterization and Biological Evaluation of Certain New Azetidin-2-One

DESIGN, SYNTHESIS, CHARACTERIZATION AND BIOLOGICAL EVALUATION OF CERTAIN NEW AZETIDIN-2-ONE DERIVATIVES

M. Pharm. Dissertation Protocol

Submitted to the

Rajiv Gandhi University of Health Sciences, Karnataka.

Bangalore.

.

By

Mr. GIRISH K.

B. Pharm.

Under the guidance of

Prof. G. SUDHEENDRA

M Pharm. (Ph.D)

DEPARTMENT OF PHARMACEUTICAL CHEMISTRY

LUQMANCOLLEGE OF PHARMACY, GULBARGA

2011-12

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE

ANNEXURE II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

Name of the candidate And
address / Mr. GIRISH K.
s/o Dr. Pranesh K, H.No.2-2-4/20, VIJAYASHRI, H.R.B. LAY OUT,
OPP. MAHILA SAMAJ, JAIL ROAD,
RAICHUR -584101. KARNATAKA
Name of the institution / Luqman college of pharmacy,
BEHIND P&T QUARTERS,
old jewargi road,
gulbarga-585102
Course of study and subject / m.PHARM
(Pharmaceutical Chemistry)
Date of Admission of course / 10/06/2011
TITLE OF THE TOPIC / DESIGN, SYNTHESIS, CHARACTERIZATION AND BIOLOGICAL EVALUATION OF CERTAIN NEW AZETIDIN-2-ONE DERIVATIVES
6. / Brief Resume of the intended work:
6.1 Need for the study:
The chemistry of heterocyclic compoundsis a field of organic/pharmaceutical chemistry which is being continuously explored for the synthesis and biological evaluation of new chemical entities in search of better lead molecules.
AZETIDIN-2-ONES
Azetidin-2-ones, commonly known as β-lactams, are well-known heterocycliccompoundsamong the organic and medicinalchemists mainly because of their antimicrobial and diverse pharmacological activities.The β-lactam antibiotics are still the most prescribed antibiotics used in medicine. They areconsidered as an important contribution of science to humanity1,2. The most widely used antibiotics such as the Penicillins, Cephalosporins, Carumonam, Aztreonam, Thienamycine and the Nocardicins contain β-lactam (azetidin-2-one) rings3. The long-term use of β-lactam antibiotics exert selective pressure on bacteria and permit the proliferation of resistant organisms4. A comparative study of current antibiotics with those from previous decades shows an alarming increase in bacterial resistance to β-lactam antibiotics5,6. The development of several synthetic and semi-synthetic β-lactam antibiotics by the pharmaceutical industry was due to the growing resistance of bacteria towards the β-lactam antibiotics and the need for medicines with a more specific antibacterial activity7.
A large number of antibiotics contain amide linkage. Several derivatives of amides were prepared and found to possess antimicrobial activities. Literature survey reveals that various drugs e.g. penicillin8 (antibacterial), pyrazinamide9 (antitubercular), indinavir10, ritonavir11. (Protease inhibitors as anti-AIDS) etc contain their particular activities due to the amide linkage present in their structure.
2-Azetidinones are the monocyclic β-lactams, are well-known heterocyclic compounds among the organic and medicinal chemists12,13. A large number of 3-chloro monocyclic β-lactams possess powerful antibacterial, antimicrobial, anti-inflammatory, anticonvulsant and antitubercular activities14. They also function as enzyme inhibitors and are effective on the central nervous system15. β-Lactams also serve as synthon for various biologically important classes of organic compounds16.
QUINAZOLIN-4-ONES
Quinazolin-4-one derivatives are versatile nitrogen heterocyclic compounds which have long been known as a promising class of biologically active compounds possessing wide variety of biological and pharmacological activities like antibacterial17, anthelmintic18, neuroleptic19, antitubercular20,platelet anti-aggregating21,antifungal22,anticancer23,anti-inflammatory24,antiviral25,CNS depressant activity26, antiparkinson27,bronchodilator28 etc.It is observed during the literature survey that, Quinazolinone system possesses the variable sites like position 2 and 3 which can be suitably modified to yield potent chemotherapeutic and pharmacotherapeutic agents21.
The literature survey also reveals that, the better drug molecules would be obtained by bringing changes in its biological behavior of the lead compound either by structure variation and/or combination of two or more biologically active moieties into one molecular framework.
Prompted by the above observations, here in we propose for the design and synthesis of some new structural hybrids of azetidin-2-ones to which another important class of heterocycles, substituted quinazoline-4-one isbridged appropriatelythrough an amide linkage to yield the title compounds. And the various derivatives synthesized would be screened for their antimicrobial properties. This combination suggested is an attempt to investigate the influence of such hybridization and structure variation on the anticipated biological activities hoping the possibility that the target derivatives might be more efficacious as antimicrobial agents.
6.2 Objective of the study:
It is well established that various derivatives of azetidin-2-oneand quinazoline-4-one exhibit broad spectrumantimicrobial and diverse biological activities. Several derivatives of amides were prepared and found to possess antimicrobial activities; literature survey also reveals that, a large number of antibiotics contain amide linkage eg. penicillin(antibacterial), pyrazinamide (antitubercular), indinavir, ritonavir(Protease inhibitors as anti-AIDS) etc contain their particular activities due to the amide linkage present in their structure.
Prompted by all the above observations, we have planned to prepare some new 2-azetidinone derivatives bridged through an amide linkage to suitably substituted quinazoline-4-one nucleus and evaluate them for antimicrobial activities.
The present investigations includes the following:
Substituted 1,3,4 benzoxazinone prepared are reacted with active hydrogen atoms of aminegroup bearing moiety by conventional synthetic methods to form 2,3-substituted quinazolinone nucleus following known methods. The hydrazide of which is then reacted with different aryl aldehydes to yield Schiff bases following literature methods. These are further cyclised to prepare azetidin-2-one derivatives by the reaction with appropriate cyclising agent(s).
The chemical structure of the compounds synthesized could be established on the basis of elemental analysis and IR, 1HNMR and Mass spectral studies.
The compounds of the above type containing different heterocyclic moieties would be evaluated for their antimicrobial properties.
6.3 Review of Literature:
A) AZETIDIN-2-ONES: Abundant literature is available that reveals the use of azetidin-2-one nucleus as synthon for the preparation of various types of derivatives and evaluation for their microbiological and pharmacological profiles. Few of the important literature of azetidin-2-ones are here as under:

• Meshram J S et al (2011)29 performed an efficient synthesis of novel bioactive azetidinones and thiazolidinones of 1,5-dimethyl-2-phenyl-1H-pyrazole-3(2H)-one and screened their antibacterial activity using bacterial strains (E. coli, B. subtilis, Pseudomonas sp., Rhodococci, B. stearothermopelus) by measuring the zone of inhibition on agar plates.

• Taj T et al (2011)30 performed an expeditious green synthesis of Schiff bases and azetidinones derivatised with 1,2,4-triazoles and evaluated their antimicrobial and antitubercular activities. The antimicrobial activity was carried out by using the MIC (Minimum Inhibition Concentration) technique.

• Srivastava Y K et al (2011)31 performed microwave induced synthesis of some biologically active azetidinones. All the compounds were subjected to antibacterial activity against E. coli , P. vulgaris . K. pneumoneae and S. aureus.

• Jubie S et al (2009)32 synthesized some 2-azetidinone derivatives and evaluated their antimicrobial activity. The synthesized compounds were subjected to antimicrobial screening by cup plate method for zone of inhibition. The antibacterial activity was tested against various gram +ve bacteria (S. faecalis, S. aureus) and gram –ve bacteria (P. aeruginosa, E. coli) and antifungal activity was tested against fungi (C. albicans, A. niger).

• Toraskar M P et al (2009)33 synthesized some azetidinones and evaluated their antifungal activity. All the compounds were screened for invitro antifungal activity against C.albicans using cup-plate agar diffusion methodby measuring the zone of inhibition.

• Vijay Kumar M M J et al (2009)34 synthesized new novel anti-inflammatory agents as N-substituted-3-chloro-2-azetidinones.

• Bhat I K et al (2007)35 synthesized azetidinone derivatives with the para-anisidine moiety and evaluated their antimicrobial study. The bacterial strains employed were Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. The fungal strain used was Candida albicans. This activity was assayed using the cup-plate agar diffusion method by measuring the zone of inhibition.

• Pai N R et al (2007)36 synthesized N-substituted-3-chloro-2-azetidinones and evaluated their biological activity. The compounds were tested for their antibacterial activity against bacteria such as gm +ve (S. aureus, B. subtilis) and gm –ve (P.aeruginosa, E.coli) and for their antifungal activity against fungi such as C. tropicans, A. niger and F. heterosporium.

• Mehta A G et al (2006)37 synthesized azetidinone and thiazolidinone derivatives of 2-amino-6-(2-naphthalenyl)thiazolo[3,2-d]thiadiazole and evaluated their antifungal activity. The synthesized compounds were screened for their antifungal activity against various fungi such as Panicilium expansum, Botrydepladia thiobromine, Nigrospora sp. and Trichothesium sp.

• Mehta A G et al (2006)38 synthesized novel azetidinone and thiazoloidinones derivatives and evaluated their antimicrobial activity. Antimicrobial activity of all the compounds were studied against gram +ve bacteria (Bacillus Subtillies and staphycoccus aureus) and gram -ve bacteria (E.Coli and salmonella typhi).

• Guner V A et al (2005)39 performed antimicrobial activity of 4-substituted-styryl-2-azetidinones. Compounds were subjected to an antimicrobial screening procedure against Gram(+) and Gram(-) strains of Staphylococcus aureus ATCC 25923; Bacillus subtilis ATCC 6633; Escherichia coli ATCC 35218; Pseudomonas aeruginosa ATCC 10145; Candida albicans ATCC 90028, Candida glabrata ATCC 90030.

B) QUINAZOLIN-4-ONES:The extensive literature survey revealed that the compounds containing Quinazolinone derivatives are reported to possess the wide range of biological activities. There are few important literaturefor Quinazolinonesas under.

• Abbas S Y et al (2011)40 synthesized some biologically active 4(3H)-quinazolinones derived from 2,3-pyridine dicarboxylic anhydride. The synthesized compounds were evaluated for their antifungal activity against fungi such as Aspergillus ochraceus wilhelmand Penicillium chrysogenum Thom.

• Revanasiddappa H D et al (2010)41 synthesized new Schiff bases containing 4(3H)-quinazolinone ring system and evaluated their biological activity. All the synthesized compounds were tested against fungi such as Aspergillus Niger, Aspergillus flavus and Alternaria solani by disc diffusion method.

• Reddy P S N et al (2010)42 evaluated antibacterial, antifungal and antifeedant activity of quinazolinonyl-b-lactams/quinazolinones and bis (quinazolinonyl-b-lactams). The synthesized compounds were tested for antifungal activity against fungi such as Fusarium oxisporium and Macrophomina sorgina.

• Rajasekaran S et al (2010)43synthesized of some 2-phenyl-3-substituted quinazolin-4(3H)-ones and evaluated their antituberculor, antibacterial and antioxidant activities. The compounds were evaluated their antituberculor activity against mycobacterium tuberculosis by agar dilution method.

• Kaur P et al (2009)44developed new approachof quinazolinone peptides as potent medicinal agents. The synthesized compounds were evaluated their antifungal activity against Microsporam audouinii, Trichophyton mentagrophtes, Candida albicans and Aspergillus Niger.

• Khairy A M et al (2009)45prepared novel 4-(3H)-quinazolinone containing biologically active thiazole,pyrazole, 1,3-diathiazole, pyridine, chromene, pyrazolopyrimidine and pyranochromene of expected biological activity. All the synthesized compounds were evaluated for their antifungal activity against Aspergillus ochraceus Wilhelmand Fusarium oxysporium fungi.

• Al-Deeb A O et al (2008)46synthesized of some new 3H-quinazoli-4-one derivatives as potential antitubercular agents. All the compounds were evaluated against Mycobacterium tuberculosis.

• Dahiya R et al (2008)47synthesized some peptide derivatives of iodoquinazolinones and nitroimidazoles and evaluated their antimicrobial and anthelmintic activities. They were evaluated their antimicrobial activity such as Bacillus subtilis (NCIM 2063), Staphylococcus aureus (NCIM 2079), Pseudomonas aeruginosa (NCIM 2034) and Klebsiella pneumoniae (NCIM 2011) and fungal strainsMicrosporum audouinii (MUCC 545), Trichophyton mentagrophytes (MUCC 665), Candida albicans (MUCC 29) and Aspergillus Niger(MUCC 177).

• Desai A R et al (2005)48performed Niementowski reaction that is microwave induced and conventionalsynthesis of quinazolinones and 3-methyl-1H-5-pyrazolones and their antimicrobial activity. All compounds were screened for their antifungal against Candida albicansandCandida kruseiand antibacterial against B.subtilis, S.aureus as gram positive and E.coli,P.aeruginosa as gram negative bacteria.

7. / Materials and Methods:
7.1 Method of collection of data:
A. Synthesis of Target Molecules:
The synthesis of title compounds will be carried out following the given scheme for which the collection of data is as below:

1. The homogeneity of the compounds is monitored by TLC technique and Rfvalues are recorded.
2. Percentage of yield, physical constant, solubility and elemental analytical data for each compound will be determined and recorded.
3. Spectroscopic data of new compounds i.e. I.R., NMR, Mass spectral data will be recorded for structural confirmation of few synthesized compounds. IR spectra in KBr (cm-1) would be recorded on a Schimadzu FTIR-8000 series spectrophotometer and 1H NMR spectra (CDCl3/DMSO-d6) on EM 390 MHz spectrometer using TMS as internal standard (Chemical shifts are expressed in δ ppm). Mass spectra would be recorded on a Jeol JMSD-300 Mass Spectrometer operating at 70 eV.

B. Biological activity:

• Antimicrobial screening49,50:

Antibacterial activity of the compounds would be carried out against gram positive and gram negative bacteria and antifungal activityagainst fungal strains by disc diffusion or cup and plate method to determine zone of inhibition (in mm). During the study, appropriate standard antibacterial and antifungal drugs, respectively, will be used to compare the activities.
7.2 Synthetic strategy:
All the compounds in the present study would be synthesized by following given scheme. The starting material 1,3,4-benzoxazinone will be synthesized from anthranilic acid by known method. It is then reacted with active hydrogen atoms of amino group bearing moiety by conventional synthetic methods to form 2,3 disubstituted Quinazolinone nucleus. The hydrazide of which is then treated with different aryl aldehydes to yield Schiff bases which are further cyclized to prepare different azetidin-2-ones by the reaction with appropriate cyclising agent(s).
SCHEME

7.3Biological Activity:
Antimicrobial screening of the compounds could be carried out by cup and plate or disc diffusion method to determine zone of inhibition (in mm) and the activity will be compared with that of standard.
7.4 Source of data:

1. From available literature.
2. From library based books
3. Web sites

-
-
-
-
-
7.4-1 Assessment of toxic effect: ------Not applicable------
7.4-2 Screening of Statistical analysis:------
7.4-3 Does the study require any investigations or interventions to be conducted on patients or humans or animals? If so, please describe briefly. ------No------
7.4-4 Has ethical clearance been obtained from your institution in case of 7.4?
------Not applicable------
List of References:

1. Southgate R. The synthesis of natural β-lactam antibiotics.Contemp. Org. Synth.1994, 1, 417-431.
2. Morin R B, Gorman M.Chemistry and Biology of β-Lactam Antibiotics. Academic Press, New York, 1982.
3. Mata E G, Fraga M A, Delpiccolo C M L. An efficient stereo selective solid-phase synthesis of β-lactams using Mukaiyama’s Salt for the Staudinger Reaction.J Comb Chem. 2003, 5, 208-210.
4. Page E I.The Chemistry of β-Lactams. Blackie Academic and Professional. New York, 1992.
5. Niccolai D, Trasi L, Thomas R J. The renewed challenge of antibacterial chemotherapy.Chem Commun.1997, 2333-2342.
6. Chu D T W, Plattner J I, Katz L. New Directions in Antibacterial Research.J Med Chem. 1996, 39, 3853-3874.
7. Van der Steen F H, Van Koten G. Synthesis of 3-amino-2-azetidinones: A literature survey.Tetrahedron,1991, 47, 7503-7524.

8.Abraham E P and Florey H W. Lancent II, 1941,177.
9.Emmerson A M, David. Green Wood’s Antimicrobial Chemotherapy, 1995,3,306.
10.Clereq D. Clin Microbial Rev.1997, 10, 674.
11.Alterman M and Samuelsson B. J Med Chem, 1998,41, 3782.
12.Alcaide B, Almendros P, Aragoncillo C. β-Lactams: Versatile building blocks for the stereoselective synthesis of non-β-lactam products. Chem Rev. 107, 11, 4437-4492 2007.
13.Bhalla A, Madan S, Venugopalan P, Bari S S. C-3 β-lactam carbocation equivalents: versatile synthons for C-3 substituted β-lactams. Tetrahedron 62 (21): 5054-5063 (2006).
14.Chavan A A, Pai N R. Synthesis and Biological Activity of N-Substituted-3-chloro-2-azetidinones. Molecules 12(11): 2467-2477 (2007).
15.Jocoby G A. Extended-spectrum β-lactamases and other enzymes providing resistance to oxyimino-β-lactams. Infect Dis Clin North Am. 11 (4): 875-887 (1997).
16.Conte L J.Fluorine Chem.1995, 70, 175.
17.Singh VKS, Gulati A and Shankar K. Antiparkinsonian agents from quinazolinyl thiazolidinones and azetidinones. Indian Journal of Chemistry. 1987;26:652-656.
18.Gupta DP, Ahmad S, Kumar A and Shankar K. Newer quinazolinone derivatives as anthelmintic agents. Indian Journal of Chemistry. 1988; 27(B):1060-1062.
19.Mukherji DD, Nautiyal SR, Prasad CR and Dhawan BN. CNS-depressant activity of some newly synthesized 4(3H)-quinazolones. Indian Journal of Medical Research. 1980;71:480-482.
20.Joshi V, Chaudhari RP. Synthesis, characterization and antibacterial activity of some new 2,3,6-trisubstituted quinazolin-4(3H)-ones. Indian Journal Chemistry.1987; 26(B):602.
21.Desai AR and Desai K.Niementowski reaction: microwave induced and conventionalsynthesis of quinazolinones and 3-methyl-1H-5-pyrazolonesand their antimicrobial activity.ARKIVOC. 2005 (xiii) 98-108
22.Chaurasia MR, Sharma SK. The synthesis of 6,8-disubstituted-2-phenyl-3-(substituted benzothiozol-2-yl)-4-(3H)-quinazolinones and their antifungal activity. Journal Indian Chemical Soc. 1972; 49:370.
23.Pandey VK and Lohani HC. The anti-tumour activity of 2-aryl/alkyl-3(2-amino ethyl-1, 3, 4-thiadiazol-5-yl) quinazolin-4(3H) ones. Journal Indian Chemical Soc. 1979; 56:415.
24.Ravi S, Devender RA, Malla RV and Sattur PB. The synthesis of new N4-(N-(6,8-dibromo-2-methyl-3-quinazolin-4(3H)-one)acetamido)-N1-substituted sulfanilamides. Current Science. 1984; 53:1069.
25.Mishra VS and Sunita D. The synthesis of 2-phenyl-3-benzimidazolyl-alkyl/aryl-6-bromoquinazoline-4(3H)-ones. Journal IndianChemicalSociety. 1978; 55:172.
26.Kumar R, Gupta TK and Surendra S. The synthesis of quinazolone azomethines for CNS depressant activity. Indian Journal Pharm. 1970;33:108.
27.Tiwari SS and Pandey VK.The synthesis of some new benzoxinones and their corresponding quinazolinones. Journal Indian Chemical Society. 1975;52:736.
28.Raghu RA and Rajesh HB. The synthesis and bronchodilator activity of benzimidazo (1, 2,-c) quinazolines .Indian Journal Chemistry. 1999; 38(B):434.
29.Meshram JS, Chopde HN, Pagadala R, Jetti V. An efficient synthesis of novel bioactive azetidinones and thiazolidinones. International Journal of Pharma and Bio Sciences. 2011;02:6299.
30.Taj T, Kamble RR, Gireesh T, Badami BV.An expeditious green synthesis of Schiff bases and azetidinones derivatised with 1,2,4-triazoles. J. Chem. Sci. 2011;123:657.
31.Srivastava YK, Malhotra G, Gothwal P.Microwave induced synthesis of some biologically active azetidinones. Der Chemica Sinica. 2011;2(3):47.
32.Jubie S, Gowrwmma B, Muthal NK, Kalirajan R, Gomathi S, Elango K. Synthesis and antimicrobial evaluation of some 2-azetidinone derivatives. International journal of Chem Tech Research. 2009;1(2):153-157.
33.Toraskar MP, Kadam VJ, Kulkarni VM. Synthesis and antifungal activity of some azetidinones. International Journal of Chem Tech Research. 2009;1(4):1194-1199.
34.Vijay Kumar MMJ, Nagaraja TS, Shameer H, Jayachandra E, Sreenivasa GM. N-substituted-3-chloro-2-azetidinones: Synthesis and characterization of new novel anti-inflammatory agents. Journal of Pharmaceutical Sciences and Research. 2009;1(2):83-92.
35.Bhat IK, Chaithanya SK, Satyanarayana PD, Kalluraya B. The synthesis and antimicrobial study of some azetidinone derivatives with the para-anisidine moiety. J. Serb. Chem. Soc. 2007;72(5):437-442.
36.Chavan AA, Pai NR. Synthesis and biological activity of N-substituted-3-chloro-2-azetidinones. Molecules. 2007;12:2467-2477.