1. INTRODUCTION:

4-Thiazolidinone ring system contains sulphur and nitrogen heterogenous at position 1 and 3 respectively and keto-group at position 4. It encounter in many of Biosynthetic and Semisynthetic products(N. J. Delgado). Eg. Benzylpenicillin , Dicloxacillin , Cloxacillin . Azetidinones are the carbonyl derivatives of azetidines containing carbonyl group at the position-2. These are also none as 2-azetidinones or more commonly β-lactam chemistry is of great importance because of the use of β-lactam derivatives as antibacterial agents(L. Thomas). The molecular mode of action(L. Thomas) of the β-lactam antibiotic is selective and irreversible inhibition of enzymes processing the developing peptidoglycan layer. It is clear from the literature review that a number of Azetidinone and Thiazolidinone derivatives are known for the Anti-viral(G. Kucukguzel 2006), Cytotoxic(D. P. Maia 2009), Anti-HIV(J. Balzarini 2007), Analgesic(K. C. Asati 2006, B. M. Gurupadayya 2008), Anti-tubercular(R. Govindarajan 2003), antibacterial(R. Govindarajan 2003, S. Jubie 2009), antifungal(R. Govindarajan 2003), anticonvulsant(Archana 2002) and Antiinflammatory(M. G. Vigorita 2001) activities properties.

The present studies were performed with the objective of Synthesis of new series of 4-Thiazolidinone and 2-Azetidinone derivatives. In the present study Pyridine-3-Carbonyl Hydrazine was synthesized by refluxing Nicotinamide with hydrazine hydrate. Pyridine-3-Carbonyl Hydrazine was refluxed with aryl aldehyde to form Schiff base. Characterization of newly synthesized compounds by spectra methods viz infrared spectra (IR spectra), Nuclear magnetic resonance spectra (¹H NMR spectra) and Mass spectra. Screening of anti-TB, anti-convulsant anti-bacterial and anti-fungal activities of the newly synthesized compounds.

2. MATERIALS AND METHODS:

2.1 Materials:

All the reactions requiring anhydrous conditions were conducted in flame dried apparatus. Solvents and reagents used were of laboratory grade. The synthesis was carried out in room temperature. The synthesized compounds were purified by recrystallization and their melting points were determined by open capillary method and were uncorrected. The purity of the compounds was confirmed by TLC method. IR spectra was recorded on FTIR spectrometer using KBr pellets technique and expressed in cm-¹.¹H NMR spectra was recorded on BRUKER 300MHZ NMR spectrophotometer. Using (CDCl₃) as solvent and Chemical Shift value are reported as values in ppm, relative to TMS (δ=0) as an internal standard.

Mass spectra were recorded on JEOL GCmate Inlet Direct Probe. The mass spectra were recorded at room temperature.

2.2 synthetic methods:

I. Synthesis of Pyridine-3-Carbonyl Hydrazine: Nicotinamide (0.1 mol, 12.2 gm) was refluxed with (0.1 mol, 12 gm) of hydrazine hydrate in presence of methanol 40 ml for 6 h at about 100-110 ⁰C. The solid which is formed were separated on chilling and was recrystallized in ethanol.

II. Synthesis of Schiff’s Bases [II (a-e)] from Pyridine-3-Carbonyl Hydrazine (I):

Pyridine-3-Carbonyl Hydrazine (I) (0.1 mol, 13.7 gm) and aryl aldehyde (0.1 mol) were dissolved in methanol 30 ml. The contents were refluxed for a period of 3 h. The reaction mixture was concentrated under reduced pressure and cooled. The solid obtained was washed with cold water and recrystallized from ethanol.

III. Synthesis of 4-Thiazolidinone [III (a-e)] from Schiff’s Bases [II (a-e)]:

To a solution of Schiff’s Base [II (a-e)] (0.1 mol) and thioglycolic acid (0.15 mol, 13.8 ml) dissolved in acetone with vigorous stirring for 30 min. Added pinch amount of Zinc Chloride (ZnCl₂) at the start of reaction. The reaction mixture was refluxed for 3 h. Solid products were obtained after cooling to give adducts [III (a-e)]. This adducts was purified by recrystallization from ethanol.

IV. Synthesis of 2-Azetidinones [IV (a-e)] from Schiff’s Bases [II (a-e):

To a solution of Schiff’s Base [II (a-e)] (0.01 mol) in acetone, triethylamine (0.005 mol, 0.795 ml) was added. To this, a solution of Chloroacetyl Chloride (0.01 mol, 1.13 ml) was added drop wise with stirring. The mixture was refluxed up to 3 h. The triehtylamine hydrochloride formed was filtered and washed several times with acetone. The filtrate and the washings were mixed and concentrated under reduced pressure. The residue obtained was washed with petroleum ether (40-60 ⁰C) to remove the unreacted Schiff’s base and the solid obtained was recrystallized from ethanol. It is shown in scheme 1. The different aromatic aldehydes are shown in Table 1.

2.3 ANTI-TUBERCULAR ACTIVITY:

2.3.1 REMA plate method:

Test compound concentrations prepared directly in the medium were 1.25, 2.5, 3.75, 5.0, 6.25, 7.5, 8.75 and 10.0 mg/L. A standard bacterial suspension equivalent in turbidity to that of a no. 1 McFarland standard was prepared and diluted 1:20 in 7H9 broth; a 100 mL inoculum was used to inoculate each well of the plate (M. D. Kakwani 2011). A growth control containing no test compound and a sterile control without inoculum were also included e. Plates were sealed and incubated at 37 ⁰C for 1 week. Twenty-five microlitres (25μL of 0.02% resazurin (Sigma Chem.Co.) solution was added to each well; plates were re-incubated for an additional 2 days. A change in colour from blue to pink indicated the growth of bacteria, and the MIC was read as the minimum test compound concentration that prevented the colour change in resazurin solution and is shown in tables 2-5.

2.4 Anti-convulsant activity:

Procedure:

Six mice in each group of either sex with a weight of 18 to 22 gm are taken (F. A. Ragab 1997). Group I control animals received 30% aqueous PEG 400 only, group II received standard (diazepam 10 mg/kg) i.p. Group III-VII are treated with the test compound (4a-4e) by oral administration. The synthesized compounds (4a-4e) were suspended in 30% aqueous solution of PEG 400 and administered orally in a standard volume of 0.5 ml/20 g body weight at 30 mg kg^-1 doses. 30 min after i.p. or 60 min after p.o. treatment the animals are injected with a subcutaneous dose of 300 mg/kg isoniazid (isonicotinic acid hydrazide).The occurrence of clonic seizures, tonic seizures and death is recorded. Anti-convulsant activity38 was expressed as percentage of tonus and clonus mortality and shown in tables 6 and 7 and figures 1 and 2.

TABLE - 1 Synthesis of 4-Thiazolidinone and 2-Azetidinones derivatives

2.5 SCREENING OF ANTI-BACTERIAL AND ANTI-FUNGAL ACTIVITY:

Gram-positive- Bacillus subtilis (ATCC 9372), Staphylococcus aureus (NCTC 8325)

Gram-negative - Proteus mirabilis (NCIM 826), Salmonella typhi (ATCC NCIM 2479)

Fungal strains - Aspergillus niger (NCIM 1207), Candida albicans (NCIM 3484), Auricularia polytricha (NCIM 1303) and Monilinia fruticola (NCIM 1011).

A well was prepared in the plates with the help of a cork- borer (6 mm) four holes per plates were made into agar medium. A total of 0.2 ml test solution of synthesized compounds was poured into the wells using a dropping pipette under aseptic condition.

Table – 2 MIC determination of 4-Thiazolidinone derivatives Anti-TB Activity against Mycobacterium tuberculosis

Comd.	Conc. of Test Sample/ Minimum Inhibitory Concentration (MIC)Colour Change
Comd.	1.25 µg/ml	2.5 µg/ml	3.75 µg/ml	5.0 µg/ml	6.25 µg/ml	7.5 µg/ml	8.75 µg/ml	10.0 µg/ml
IIIa	-----	Blue Colour to Pink	-----	-----	-----	-----	-----	-----
IIIb	-----	-----	-----	Blue Colour to Pink		-----	-----	-----
IIIc	-----	-----	Blue Colour to Pink	-----	-----	-----	-----	-----
IIId	-----	Blue Colour to Pink	-----	-----	-----	-----	-----	-----
IIIe	-----	-----	-----	-----	Blue Colour to Pink	-----	-----	-----

Compound IIIa and ID shown good Anti-TB activity

Table - 3 MIC determination of 2-Azetidinone derivatives Anti-TB Activity against Mycobacterium tuberculosis

Comd.	Conc. of Test Sample/ Minimum Inhibitory Concentration (MIC)Colour Change
Comd.	1.25 µg/ml	2.5 µg/ml	3.75 µg/ml	5.0 µg/ml	6.25 µg/ml	7.5 µg/ml	8.75 µg/ml	10.0 µg/ml
Iva	-----	Blue Colour to Pink	-----	-----	-----	-----	-----	-----
IVb	-----	-----	Blue Colour to Pink	-----	-----	-----	-----	-----
IVc		-----	-----	Blue Colour to Pink	-----	-----	-----	-----
IVd	-----	Blue Colour to Pink	-----	-----	-----	-----	-----	-----
IVe	-----	-----	-----	-----	Blue Colour to Pink	-----	-----	-----

Compound IVa and IVd shown good Anti-TB activity.

Table - 4 Anti tubercular activity of synthesized 4-Thiazolidinone derivatives against Mycobacterium tuberculosis

Compound	MIC Concentration (µg/ml)
IIIa	2.5
IIIb	5.0
IIIc	3.75
IIId	2.5
IIIe	6.25

Table - 5 Anti tubercular activity of synthesized 2-Azetidinone derivatives against Mycobacterium tuberculosis

IVa	2.5
IVb	3.75
IVc	5.0
IVd	2.5
IVe	6.25

So compound IIIa, IIId, IVa and IVd shows good activity against Mycobacterium tuberculosis.

Table - 6 Anticonvulsant activity of compounds (IIIa-IIIe)

Groups	Latency of clonus (min.)	% of clonus	%of tonus and clonus mortlity.
Control	3.20 ± 0.1797	100	100
Standard	8.57 ± 0.2144**	100	0
IIIa	6.85 ± 0.0038^**	100	14.67
IIIb	4.52 ± 0.1366	100	83.34
IIIc	4.03 ± 0.1806^*	100	16.57
IIId	8.15 ± 0.0551^**	100	66.67
IIIe	7.34 ± 17.58^*	100	55.00

N=6; dunnets t test; * P<0.05; ** P<0.01; ***P<0.001 when compared with control.

Table - 7 Anticonvulsant activity of compounds (IVa-IVe)

Groups	Latency of clonus (min.)	% of clonus	% of tonus and clonus mortlity.
Control	3.20 ± 0.4365	100	100
Standard	8.15± 0.0230**	100	0
Iva	7.53 ± 0.7438^**	100	16.78
IVb	4.52 ± 0.0743	100	84.34
IVc	4.03 ± 0.0345^*	100	65.67
IVd	8.15 ± 0.0551^**	100	16.67
IVe	6.40 ± 17.84^*	100	52.20

Figure-1 Anti-convulsant effect of synthesized compounds (% latency of clonus)

Figure - 2 Anti-convulsant effect of synthesized compounds

(% tonus and clonus mortality)

Table - 8 Anti –Bacterial Activity of 4-Thiazolidinone derivatives

Comp.	Conc.	Zone of Inhibition (mm)
Comp.	Conc.	Bacillus subtilis (ATCC 9372)	Staphylococcus aureus (NCTC 8325)	Proteus mirabilis (NCIM 8268)	Salmonella typhi (ATCC NCIM 2479)
IIIa	1000 µg/ml	21	18	19	18
	500 µg/ml	18	16	17	15
	250 µg/ml	15	15	15	13
	Std. 1000 µg/ml	28	28	28	28
IIIb	1000 µg/ml	19	18	15	16
	500 µg/ml	16	15	13	13
	250 µg/ml	14	14	12	11
	Std. 1000 µg/ml	28	28	28	28
IIIc	1000 µg/ml	24	25	19	20
	500 µg/ml	21	23	17	18
	250 µg/ml	17	18	14	15
	Std. 1000 µg/ml	28	28	28	28
IIId	1000 µg/ml	18	19	16	18
	500 µg/ml	16	17	14	15
	250 µg/ml	14	15	11	13
	Std. 1000 µg/ml	28	28	28	28
IIIe	1000 µg/ml	19	17	16	18
	500 µg/ml	18	16	17	16
	250 µg/ml	15	14	15	15
	Std. 1000 µg/ml	28	28	28	28

Leofloxacin was used as standard for bacterial strains (R. Cha 2003), while Amphotericin B for fungal strain (V. P. Patel 2011). The plates were maintained at room temperature for 3-5 h to allow diffusion of the solution into the medium and inoculated with 0.2 ml of different bacterial and fungal species mixed well with the petri dishes used for antibacterial screening were incubated 371 °C for 24 h. The diameter of zones of inhibition (mm) surrounding each of the well was recorded in tables 8-11.

3. RESULTS AND DISCUSSION:

3.1 Chemistry:

3.1.1 Compound IIIa:

Mol.wt- 313, m.p. 173-175⁰C, R_f value-0.78; IR (KBr, ν_max, cm^-1): 1734.46 (Ar- C=O str.), 1395.59 (Ar- C=N str.), 3648.90 (Ar- -OH str.), 3398.22 (N-H str.), 3100.14 (Ar- C-H str.), 1181.56 (C-S str. Thiazolidinone); ¹H NMR (δppm): 6.61-6.89 (m. 4H of 5-Benzene), 5.0 (s. H of Ar- OH), 7.63-9.17 (m. 4H Pyridine), 8.0 (s. NH- CONH), 4.77 (s. H CH-Thiazolidinone), 3.54 (d. H of CH₂Thiazolidinone); MS m/z: 312.24(M+), 250.59, 168.73, 119.02, 102.50; Elemental anal. (%), calcd. For C₁₅H₁₃N₃O₃S: C, 57.5; H, 4.15; N, 13.41;

3.1.2 Compound IIIb:

Mol.wt- 299, m.p. 192-195⁰C, R_f value-0.89; IR (KBr, ν_max, cm^-1): 1682.94 (Thiazolidinone, C=O str.), 1653.45 (Amide C=O str.), 1616.58 (C=N str.), 1716.12 (CONH-), 3523.48

(>N-H str.), 3058.06 (Ar- C-H str.), 1182.16 (C-S str. of Thiazolidinone), 1472.81 (C=C, Ar- str.), 1395.39

(Pyridine, C=N str.); ¹H NMR (δppm): 7.06-7.14 (m. 4H Benzene), 7.63-9.17 (m. 4H of 1-Pyridine), 8.0 (s. NH- CONH), 4.50 (s. H -CH Thiazolidinone), 3.67 (d. H -CH₂ Thiazolidinone ring); MS m/z: 298.24(M+), 243.39, 214.87, 114.43, 93.55; Elemental anal. (%),calcd. For C₁₃H₁₁N₃O₃S: C, 60.20; H, 4.34; N, 14.05;

3.1.3 Compound IIIc:

Mol.wt- 289, m.p. 183-185⁰C, R_f value-0.73; IR (KBr, ν_max, cm^-1): 1701.38 (Thiazolidinone, C=O str.), 1616.58 (CONH- str.), 3488.48 (>N-H str.), 3023.48 (Ar. C-H str.), 1244.23 (C-O str.), 1177.88 (C-S str. of Thiazolidinone), 1421.19 (C=C, Ar. str.), 1030.41 (C-N str.), 1303.22 (Pyridine, C=N str.); ¹H NMR (δppm): 6.06-7.28 (m.3H Furan) 7.63-9.17 (m. 4H-Pyridine), 8.0 (s. NH- CONH), 5.01 (s. H -CH Thiazolidinone), 3.77 (d. H CH₂ Thiazolidinone); MS m/z: 288.26(M+), 196.61, 108.89, 90.03; Elemental anal. (%),calcd. For C₁₃H₁₁N₃O₃S: C, 53.98; H, 3.80; N, 14.53;

3.1.4 Compound IIId:

Mol.wt- 333, m.p. 186-189⁰C, R_f value-0.69; IR (KBr, ν_max, cm^-1): 1749.08 (Thiazolidinone, C=O str.), 1303.22 (Pyridine, C=N str.), 3481.62 (>N-H str.), 3059.90 (Ar- C-H str.), 1188.94 (C-S, Thiazolidinone), 669.12 (C-Cl str.), 1421.19 (Ar- C=C, str.); ¹H NMR (δppm): 7.01-7.14 (m. 4H Benzene), 7.63-9.17 (m. 4H of 1-Pyridine), 8.0 (s. NH- of CONH), 4.77 (s. H -CH Thiazolidinone), 3.67 (d. H CH_2-Thiazolidinone); MS m/z: 332.27(M⁺), 243.35, 185.79, 60.50; Elemental anal. (%),calcd. For C₁₅H₁₂N₃O₂SCl: C, 54.05; H, 3.60; N, 12.61;

3.1.5 Compound IIIe:

Mol.wt- 329, m.p. 159-162⁰C, R_f value-0.63; IR (KBr, ν_max, cm^-1): 1734.36 (Thiazolidinone, C=O str.), 1458.06 (Pyridine, C=N str.)., 3490.28 (N-H str.), 3082.83 (Ar- C-H str.), 2862.36 (-CH₃ Str.), 1161.62 (C-S str. of Thiazolidinone); ¹H NMR (δppm): 6.67-6.95 (m. 4H Benzene), 7.63-9.17 (m. 4H-Pyridine), 8.0 (s. NH- CONH), 4.77 (s. H -CH Thiazolidinone), 3.67 (d. H of CH₂-Thiazolidinone); MS m/z: 328.14 (M⁺) 275.88, 243.35, 108.81, 90.03; Elemental anal. (%),calcd. For C₁₆H₁₅N₃O₃S: C, 58.35; H, 4.56; N, 12.77;

3.1.6 Compound IVa:

Mol.wt- 334, m.p. 181-184⁰C, R_f value-0.59; IR (KBr, ν_max, cm^-1): 1734.36 (Ar- C=O str.), 1421.19 (Pyridine, C=N str.), 3457.89 (>N-H stretching), 3011.90 (Aromatic Ring C-H stretching), 3621.62 (Aromatic –OH Stretching); ¹H NMR (δppm): 6.68-6.95 (m. 4H Benzene), 7.63-9.17 (m. 4H Pyridine), 5.00 (s. H OH Benzene), 5.44 (d. H >CH Azitidinone), 5.00 (s. H CH Azitidinone); MS m/z: 317.18 (M+), 243.35, 221.18, 60.50; Elemental anal. (%),calcd. For C₁₅H₁₂N₃O₃Cl: C, 56.60; H, 3.77; N, 13.21;

3.1.7 Compound IVb:

Mol.wt- 318, m.p. 189-192⁰C, R_f value-0.79; IR (KBr, ν_max, cm^-1): 1734.66 (Ar- C=O str.), 1340.09 (Pyridine, C=N str.), 3358.90 (>N-H str.), 3100.14 (Ar- C-H str.); ¹H NMR (δppm): 6.68-6.95 (m. 4H Benzene), 7.63-9.17 (m. 4H Pyridine ring), 5.00 (s. H OH Benzene), 5.44 (d. H >CH₂-Azitidinone), 5.00 (s. H of CH Azitidinone); MS m/z: 317.18 (M+), 243.35, 221.18, 60.50; Elemental anal. (%),calcd. For C₁₅H₁₂N₃O₂Cl: C, 59.60; H, 3.97; N, 13.91;

3.1.8 Compound IVc:

Mol. wt- 292, m.p. 146-149⁰C, R_f value-0.75; IR (KBr, ν_max, cm^-1): 1716.12 (Ar- C=O str.), 1384.33 (Pyridine, C=N str.), 3407.23 (>N-H str.), 3086.88 (Ar- C-H stretching), 1244.23 (C-O str.); ¹H NMR (δppm): 6.06-7.28 (m. 3H Furan), 7.63-9.17 (m. 4H-Pyridine), 5.44 (s. H >CH-Azitidinone), 5.20 (d. H of CH₂ of 2-Azitidinone); MS m/z: 291.65 (M+), 269.88, 132.77, 108.89, 90.03; Elemental anal. (%),calcd. For C₁₃H₁₀N₃O₃Cl: C, 52.74; H, 3.43; N, 14.38;

3.1.9 Compound IVd:

Mol. Wt- 337, m.p. 162-165⁰C, R_f value- 0.82; IR (KBr, ν_max, cm^-1): 1682.94 (Ar- C=O str.), 1340.09(Pyridine, C=N str.), 3448.90 (>N-H str.), 3000.14 (Ar- C-H str.); ¹H NMR (δppm): 7.02-7.22 (m. 4H-Benzene), 7.63-9.17 (m. 4H-Pyridine), 5.44 (s. H >CH Azitidinone), 5.00 (d. H CH2 -Azitidinone); MS m/z: 336.14 (M+), 243.35, 185.79, 74.46, 60.50; Elemental anal. (%),calcd. For C₁₅H₁₁N₃O₂Cl₂: C, 53.41; H, 3.26; N, 12.46;

3.1.10 Compound IVe:

Mol. Wt- 332, m.p. 166-169⁰C, , R_f value- 0.84; IR (KBr, ν_max, cm^-1): 1734.86 (Ar- C=O str.), 1395.39 (Pyridine, C=N str.) , 3496.90 (>N-H str.), 3100.14 (Ar- C-H str.), 1229.48 (-O- Str.), 2856.48 (-CH₃ Str.); ¹H NMR (δppm): 6.72-7.01 (m. 4H-Benzene), 3.71 (s. H -OCH₃), 7.63-9.17 (m. 4H Pyridine), 5.00 (s. H OH- Benzene), 5.44 (s. H >CH Azitidinone), 5.00 (d. H CH₂ Azitidinone); MS m/z: 331.14 (M+), 243.35, 185.79, 128.25, 60.50; Elemental anal. (%), calcd. For C₁₆H₁₄N₃O₃Cl: C, 57.83; H, 4.22; N, 12.65;

Table - 9 Antibacterial Activity of 2-Azetidinone derivatives:-

Comp.	Conc.	Zone of Inhibition (mm)
Comp.	Conc.	Bacillus subtilis (ATCC 9372)	Staphylococcus aureus (NCTC 8325)	Proteus mirabilis (NCIM 8268)	Salmonella typhi (ATCC NCIM 2479)
Iva	1000 µg/ml	19	19	18	17
	500 µg/ml	16	17	16	14
	250 µg/ml	14	14	13	11
	Std. 1000 µg/ml	28	28	28	28
IVb	1000 µg/ml	20	19	17	18
	500 µg/ml	17	16	15	16
	250 µg/ml	15	14	13	14
	Std. 1000 µg/ml	28	28	28	28
IVc	1000 µg/ml	24	25	20	21
	500 µg/ml	21	23	17	18
	250 µg/ml	17	18	14	15
	Std. 1000 µg/ml	28	28	28	28
IVd	1000 µg/ml	21	18	19	18
	500 µg/ml	18	16	17	15
	250 µg/ml	15	14	15	13
	Std. 1000 µg/ml	28	28	28	28
IVe	1000 µg/ml	19	18	17	16
	500 µg/ml	18	16	15	14
	250 µg/ml	15	14	12	12
	Std. 1000 µg/ml	28	28	28	28

Table - 10 Antifungal Activity of 4-Thiazolidinone derivatives

Compound Name	Conc. of Compound	Zone of Inhibition (mm)
Compound Name	Conc. of Compound	Aspergillus niger (NCIM 1207)	Candida albicans (NCIM 3484)	Auricularia polytricha (NCIM 1303)	Monilinia fruticola (NCIM 1011)
IIIa	1000 µg/ml	18	20	16	19
	500 µg/ml	12	14	10	13
	250 µg/ml	0	9	0	5
	A*	22	25	20	24
IIIb	1000 µg/ml	17	18	15	16
	500 µg/ml	10	12	11	09
	250 µg/ml	4	4	0	0
	A*	22	25	20	24
IIIc	1000 µg/ml	19	18	16	18
	500 µg/ml	11	12	13	11
	250 µg/ml	9	5	9	5
	A*	22	25	20	24
IIId	1000 µg/ml	19	20	16	18
	500 µg/ml	13	14	10	12
	250 µg/ml	9	0	0	5
	A*	22	25	20	24
IIIe	1000 µg/ml	15	16	15	17
	500 µg/ml	12	13	11	14
	250 µg/ml	0	4	0	3
	A*	22	25	20	24

A* Indicates the Standard used= Amphotericin

Table - 11 Antifungal Activity of 2-Azetidinone derivatives

Compound Name	Conc. of Compound	Zone of Inhibition (mm)
Compound Name	Conc. of Compound	Aspergillus niger (NCIM 1207)	Candida albicans (NCIM 3484)	Auricularia polytricha (NCIM 1303)	Monilinia fruticola (NCIM 1011)
IVa	1000 µg/ml	18	19	16	17
	500 µg/ml	12	14	10	12
	250 µg/ml	0	9	0	5
	A*	22	25	20	24
IVb	1000 µg/ml	17	18	15	16
	500 µg/ml	10	12	11	09
	250 µg/ml	4	4	0	0
	A*	22	25	20	24
IVc	1000 µg/ml	18	17	19	18
	500 µg/ml	12	13	14	14
	250 µg/ml	5	8	7	9
	A*	22	25	20	24
IVd	1000 µg/ml	18	19	16	20
	500 µg/ml	13	14	10	12
	250 µg/ml	9	0	0	5
	A*	22	25	20	24
IVe	1000 µg/ml	15	16	15	17
	500 µg/ml	12	13	11	14
	250 µg/ml	0	4	0	3
	A*	22	25	20	24

A* Indicates the Standard used= Amphotericin B.

In pharmacological evaluation the newly synthesized compounds were tested in vivo in order to evaluate their anti-bacterial, anti-fungal, anti-tubercular and anti-convulsant activity. It was observed that compound IIIb and IVb exhibited lower activity; whether as the compounds IIIa, IIIc, IIId, IIIe, IVa, IVc, IVd and IVe substituted with 4-Hydroxyphenyl, Furyl, 2-Chlorophenyl and 4-Methoxyphenyl respectively in each compounds shown good activities.

Compounds IIIc and IVc shown best anti-fungal and anti-bacterial activity. Particularly the compounds IIIa , IIId, IVa and IVd have been shown found to be the most potent in the series.

On of the most important point regarding anti-tubercular Activity is that the all synthesized compounds have been showed the anti-tubercular Activity against Mycobacterium tuberculosis at concentration <6.25 μg/ml, which is less than the standard drug a first line drug of Anti-TB Pyrazinamide (12.0 μg/ml).

4. CONCLUSION:

In the present study, we have depicted the synthesis of substituted 4-Thiazolidinone and 2-Azetidinone derivatives, also anti-tubercular, anti-convulsant, anti-fungal and anti-bacterial activity of selective compounds. The synthesized compounds were characterized by TLC, melting point, elemental analysis, IR, NMR and Mass spectroscopy. These derivatives evaluated, in-vivo anti-tubercular activity by REPA plate method, anti-convulsant activity in rats and anti-microbial activity using plate hole diffusion method. All the synthesized compounds shown better anti-TB as well as anti-convulsant activity, whether as the several standard anti-TB drugs have the convulsion as a side effect.

5. ACKNOWLEDGMENTS:

The authors are grateful to the Chairman V. Shanmugan and S. Nandha Kumar Pradeep, Secretary of Nandha College of Pharmacy and Research Institute, Erode, Tamilnadu, India for their assistance.

6. REFERENCES:

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Received on 05.04.2012 Accepted on 08.05.2012

Asian J. Pharm. Res. 2(2): April-June 2012; Page 63-70