Naresh K et al., PharmaScient, Volume 1, Issue 1, 2012; 1-4.

Evaluation of Anti-Bacterial and Anti-Fungal of Novel Thiophene Derivatives

Madhukar. A, Anjali shah M, Sandeep Kumar C, Swetha M, Bharath Kumar B, Naresh K* Department of Pharmaceutical Chemistry, St.Marys College of Pharmacy, Secunderabad. A.P. INDIA. *naresh4nani@gmail.com

ABSTRACT
Thiophene nucleus has been established as the potential entity in the largely growing chemical world of heterocyclic compounds possessing promising pharmacological characteristics. The similar compounds synthesized through different routes bear variable magnitudes of biological activities. In the same way compounds with azitidinone moiety possess a varied number of biological activities. In view of this it is worthwhile to prepare different thiophene derivatives that bear an azitidinone moiety at C-2. Different 2-amino thiophene-3-carboxylic acid ethyl esters were prepared by using Gewalds reaction. 2-amino thiophene 3-caboxy hydrazides were prepared by condensing Phenyl hydrazine with 2-amino thiophene-3-carboxylic acid ethyl esters. From 2-amino thiophene 3-caboxy hydrazides different Schiffs bases were prepared by reacting with various aromatic aldehydes (viz. Benzaldehyde, Cinnamaldehyde, Para amino Dimethyl Benzaldehyde, Indole-3caboxaldehyde). Finally the Schiffs bases thus obtained were treated with chloroacetylchloride in presence of triethyl amine to afford the title compounds. The title compounds were subjected to evaluate for their antibacterial and anti-fungal activities. Key words: Thiophene, azitidinone, anti-bacterial activity and anti-fungal activity.

INTRODUCTION Thiophene nucleus is a biologically important heterocyclic moiety and attracting attention in chemistry research due to its diverse range of biological activities. An intensive literature review on thiophene, azitidinone and their derivatives revealed that they were found to possesses different biological activities such as antibacterial1, mycolytic1 antimicrobial activity2,3, anti-mitotic4, antidepressant activity5. Azetidinone derivatives which contain -lactam ring in their structure show potent anti-cunvulsant6, atni-tubercular7, antiinflammatory and anti-tumor activities.8 In view of the biological importance and the past research on the thiophene, azitidinone and their derivatives; it is worthwhile to synthesize some novel azatidinone incorporated thiophene derivatives. The synthesis of title compounds was achieved by a systematic approach is outlined in the Scheme-I.

MATERIALS AND METHODS Melting points of all the compounds were determined in open capillaries using Toshniwal and Cintex melting point apparatus and are uncorrected. IR spectra of the compounds were recorded on SCHIMADZU FT-IR Spectrophotometer by using KBr discs. Progress of the each reaction in the present investigation was monitored by TLC using E-Merck 0.25mm silica gel plates.

EXPERIMENTAL METHODS Synthesis of 2-amino-4,5-sustituted thiophene-3carboxylate (Ib&Ic): An equimolar mixture of sulphur (0.1moles), ethyl cyano acetate (0.1mole) and ketone (cyclohexanone for I and buta-2-one for II), (0.1mole), were taken in a conical flask containing 10 to 15mL of ethanol. The mixture was stirred for 5minutes and morpholine (0.1mole) was slowly added with stirring for 15minutes. This was irradiated at 180W for four minutes and it was cooled to room temperature and kept in refrigerator

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Naresh K et al., PharmaScient, Volume 1, Issue 1, 2012; 1-4.

overnight. The crystals thus obtained were collected by filtration under reduced pressure. The collected crystals were washed with cold carbinol and recrystallised using ethanol. Synthesis of 4,5-substituted 2-amino thiophene-3carboxyphenyl hydrazides (IIb & IIc): An equimolar mixture of phenyl hydrazine and respective 2-amino-4, 5-sustituted thiophene-3carboxylate (Ib&Ic) were taken and dissolved in 20mL of ethanol and irradiated at 450W for 3minutes. The crystals were collected under reduced pressure and washed with cold carbinol and recrystallised using ethanol. Synthesis of 2[(aryl)methylene amino] 4,5substituted thiophene-3-carboxyhydrazides (IIIah): Equimolar quantities of compound IIb & IIc were taken separately and dissolved in required amount of ethanol and then acidified with glacial acetic acid. The reaction mixture further treated with various
O

Aromatic aldehydes (0.36gms) and irradiated at 350W for three minutes, it was allowed to cool to room temperature. The crystals were collected by filtration under reduced pressure and recrystallized using ethanol. Synthesis of 2[(3-chloro, 4-aryl azetidine-2-one)]4,5-substituted thiophene-3-carboxy hydrazide: The products of the previous step 2[(aryl)methylene amino] 4,5-substituted thiophene-3carboxyhydrazides (IIIa-h) were dissolved in benzene separately. An equimolar mixture of chloroacetylchloride and tri ethylamine was added to above solution by keeping the conical flask in an Ice bath. Later it was allowed to cool to room temperature and irradiated at 600W for 4minutes. The reaction mixture was poured on to 200gm of crushed ice. The crude compound thus obtained was collected by filtration and dried. All the final compounds obtained were purified by recrystallisation.

CN

S + H 2C

+
COOC 2H 5

R1

H2 C

R2

ETHANOL, MORPHOLINE

R1

O C OC 2H 5

180W, 4 MIN

O R1 C

H N

H N

E, ZIN RA Y D OL H Y L AN EN ET H IN PH 3M W, 30 0

R2

NH2

Ia & Ib

R2

NH2

IIa & IIb

Ar-CHO
Glacial Acetic acid

O R1 C

H N

H N
Chloroacetyl Chloride, TEA

O R1 C

H N

H N

R2

N S

H C

Ar R2 S N

H C

Ar

IIIa-h

IVa-h

Cl

Ia, IIa, IIIa-d, IVa-d Ib, IIb, IIIe-h, IVe-h

[R1 = R2 = CH3 ] [R1= R2 = -(CH2 )4 -]

Ar = Phenyl, Cinnamyl, N,N Dimethyl amino Benzyl Indole-3-yl

Scheme I
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Naresh K et al., PharmaScient, Volume 1, Issue 1, 2012; 1-4.

EXPERIMENTAL SECTION Antibacterial activity: All the synthesized compounds were screened for their possible antibacterial activity by disc diffusion technique9,10,11 against Escheresia coli, Pseudomonas aeruginosa bacterial strains. The test compounds and standard were dissolved in DMSO, solvent and growth controls were kept for comparison. The zone of inhibition (mm) were measured after incubating the petri plates for 24h at 370C and the results are given in (Table 1) Antifungal activity: The synthesized compounds were also screened for antifungal activity9, 10, 11 against fungal strains Sachharomyces cerevisiae, Candida albicans. The diameter of zone of inhibition was noted. Fluconazole was used as standard to compare the activity of title compounds. The results are given in (Table 2)

Table 1: Physical data table Compound Ic Ib IIb IIc IIIa IIIb IIIc IIId IIIe IIIf IIIg IIIh IVa IVb IVc IVd IVe IVf IVg IVh Molecular formula C11H15NO2S C9H13NO2S C13H15N3OS C15H7N3OS C22H21N3OS C24H26N4OS C24H23N3OS C24H22N4OS C20H19N3OS C22H24N4OS C22H21N3OS C22H20N4OS C24H22N3O2S C26H27ClN4O2S C26H27ClN3O2S C26H23ClN4O2S C22H20ClN3O2S C24H25ClN4O2S C24H22ClN3O2S C24H21ClN4O2S Melting point 108-1100C 92-960C 125-1300C 182-1850C 70-720C 75-760C 70-730C 90-960C 73-780C 76-800C 90-950C 110-1150C 90-950C 94-1000C 101-1030C 104-1100C 120-1280C 110-1130C 112-1160C 110-1150C Yield (%) 75.8 75 75 65 73 70 71 76 78 76 69 74 76 74 76 74 73 72 72 78 Molecular weight 225.31 199.27 261.34 287.38 375.49 418.55 401.52 414.52 349.45 392.52 375.49 388.49 451.97 495.04 478.01 491.00 425.93 469.00 451.97 464.97

Table 2: Antibacterial and Anti-fungal data of title compounds Compound (1000g/mL) IVa IVb IVd IVe IVf IVg IVh Standard(100 g/mL) E.Coli 12 12 16 12 17 11 15 22 Zone of inhibition(in mm) P.aeruginosa C. albicans 11 13 12 12 15 16 11 12 16 15 13 11 16 17 23 24 S. cerevisiae 10 11 17 11 16 12 16 21

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Naresh K et al., PharmaScient, Volume 1, Issue 1, 2012; 1-4.

RESULTS AND DISCUSSION Synthesis of seven different 2[(3-chloro, 4-aryl azetidine-2-one)]-4, 5-substituted thiophene-3carboxy hydrazide was carried out by employing microwave irradiation method by following a systematic approach, initially the starting material 2-amino thiophene- 3-carboxylate ethyl esters were (Ia & I b) obtained by the reaction of ethylactoacetate with a ketone and elemental sulphur in presence of morpholine. In the following step compounds (Ia & Ib) were further condensed with phenyl hydrazine to afford respective2- amino thiophene-3-carboxyhydrazides (IIa & IIb). The 2- amino group of resulted compounds was condensed with a variety of aromatic aldehyde to achieve respective schiffs bases. Formation of the schffs base was confirmed by the disappearance of -NH2 peak in the IR spectra that was observed in the 2-amino thiophene carboxy hydrazides. Finally all the schiffs base were again condensed with chloroacetyl chloride in TEA afford the respective 2[(3-chloro, 4-aryl azetidine-2-one)]-4, 5substituted thiophene-3-carboxy hydrazide. The results of in-vitro antibacterial activity revealed that among the all tested compounds, compounds IVd, IVf and IVh showed moderate antibacterial activity. The antifungal data of title compounds revealed that no compound is as good as standard drug fluconazole against tested fungal strains. However compounds IVd, IVf and IVh showed moderate antifungal activity. CONCLUSION Seven thiophene derivatives have successfully been synthesized by employing microwave irradiation from thiophene -3- carboxylate. Azitidinone moiety was incorporated into the thiophene moiety at second position. At the third carbon carbonyl group was hydrazinated. All the final compounds and compound IIIa-h (data not shown) were screened for their antimicrobial activity on selected strains of bacteria and fungi. The results demonstrated the importance of azitidinone moiety and its effect of incorporation on its antimicrobial activity. Anti fungal and antibacterial activity has been increased in all the tested and compounds IVd, IVf and IVh showed good activity among the tested compounds. www.pharmascient.com Hence this series with a substitution on azitidinone could be considered as a lead molecule for the design of future antimicrobial agents. REFERENCES
1. Sajal Srivastava and Barnali Das. Synthesis and evaluation of some novel thiophenes as potential antibacterial and mycolytic agents, IJPBS, 2011. 3(6): 103-111 2. Anand Sivadas & Subbaraya. Synthesis, Characterization and Antimicrobial Activity of Novel Thiophene Acrylate Derivatives, IJPSR, 2011. 2(4): 1007-1014 3. K.H.Patel and A.G.Mehta. Synthesis of Novel Azetidinone and Thiazolidinones Derivatives and Evaluation of Their Antimicrobial Efficacy, e-Journal of Chemistry, 2006, 3(11): 103-109. 4. Romeo Romagnoli, Bioorg. Synthesis of novel antimitotic agents based on 2-amino-3-aroyl-5(hetero)arylethynyl thiophene derivatives. Med. Chem. Lett. 21 (2011) 27462751. 5. Luis Berrade et al., ARKIVOC 2008 (xiii) 198-206. 1993. 6. N.Ramalakshmi, R.Vijayakumar, Ilango S, Arunkumar and A.Puratchikody. Synthesis and characterization of certain novel azetidinone derivatives as antibacterial and antifungal agents. Research journal of pharmaceutical, biological and chemical sciences, 2010, 2(1): 107. 7. Parul D Mehtha, N P S Senger and A K Pathek. 2azetidinone a new profile of various pharmacological activities. European Journal of Medicinal Chemistry, 2010, 45: 5541-5560. 8. Duguid J.P, Cruikshank R, Marmion B.P and Swain R.H.A, Medicinal Microbiology, 12th edition, 1975. 9. Arthrington-Skaggs B.A, Motley M, Warnock D.W and Morrison C.J. J Clin Microbiology, 2000, 38, 2254. 10. Collins A.H, Microbiological Methods, 2nd edition, Butterworth, 1976.