INTRODUCTION:

4-Thiozolidinone derivatives have been reported for local anesthetics¹, antibacterial^2-7,antifungal^8-10,insecticidal¹¹, anticonvulsant¹², antitubercular^13,14, antiinflammatory^15,16, CNS and CVS activitvities¹⁷, antithyroid¹⁸, antidiabetic^19,20, Anti HIV and YFV activities²¹. It was found that when one biodynamic heterocyclic system was coupled with another heterocyclic system enhanced biological activity was produced.

The present investigation was aimed at synthesizing substituted 4-thiazolidinone derivative compounds. Various reports describing the synthesis and activities of 4-thiazolidinones coupled with azetidinones, coumarin and other heterocyclic systems at C-2 position have been reported.

A survey of existing literature revealed that there wee no reports describing the synthesis and activity of heterocyclic system in which substituted Schiff’s base moiety has been linked with thioglycollic acid and produce substituted 4-thiazolinone nucleus at C-4 position. Hence it is thought worthwhile to synthesize and explore the activity of these compounds.

MATERIAL AND METHODS:

The melting points were determined in open capillaries and are uncorrected. IR spectra (KBr in cm^-1) were recorded on PerkinElmerFTIR 8400 series, ¹H NMR were recorded on Brucker –Ultra Sheild (300MHZ) spectrometer using DMSO-d₆ as solvent and TMS as internal standard (chemical shifts in δ ppm). The purity of the compound was monitored by TLC using precoated 0.1 mm silica gel plates.

GENERAL METHODS OF SYNTHESIS:

STEP I: Synthesis of substituted benzoyl chloride:

Weigh about (0.12 mol) of substituted benzoic acids and transfer it into round bottom flask and added equivalent moles of thionyl chloride to it. 5-7 ml of benzene was added to the above mixture (as a solvent). Refluxed for 3 hrs continously in a water bath.Cool the mixture the solid residue obtained was substituted benzoyl chloride

Synthesis of substituted benzohydrazides:

STEP II

Take the substituted benzoyl chloride in round bottom flask and added aquivalent moles of hydrazide (0.12 mol)and 10 ml of methanol as a solvent. Reflux the mixture for 6 hrs, the product obtained was substituted Benzohydrazide.

Synthesis of Various substituted Schiff’s bases:

STEP III

Take 0.01mol of substituted benzohydrazides and to this added(.o.12mol) of various substituted aromatic aldehydes in 10ml of ethanol.The reaction mixture was refuxed for 5 hrs, then the resulting solid was collected.Recrystallized from ethanol to give various substituted schiff’s bases.

Synthesis of Various Substituted 4-thiazolidinones:

STEP IV

Take substituted Schiff’s bases in a round bottom flask and add (0.01mol) of thioglycollic acid in 30ml of DMF and add a pinch of anhydrous zinc chloride. Refluxed for 6 hrs, the product obtained was substituted 4-thiazolidinones.

Spectral analysis of synthesized compounds:-^22,
23

Compound Name: N-(2-(4-Flourophehyl)-4-oxothiazolidin-3-yl) benzamide

(KBr)cm^-1:3447(secondary NH stretching),2926(C-H Stretching),1702 (C=O),1633(C=O,Str,thiosolidinone),1602(NHstreching),619(C-S-C):¹HNMR (CDCl₃):7.2(N-CH proton),7.5 (aromatic proton),4.2(S-CH2-CO-proton),8(-CONH protons)

Compound Name: N-(2-(4-hydroxyphehyl)-4-oxothiazolidin-3-yl) benzamide

(KBr)cm^-1:3447(secondary NH stretching),2926(C-H Stretching),1741 (C=O),1654(C=O,Str,thiosolidinone),1602(NHstreching),619(C-S-C):

¹HNMR (CDCl₃):7.2(N-CH proton),7.5 (aromatic proton),3.9(S-CH2-CO-proton),8(-CONH protons)

Compound Name: N-(2-(4-Methoxyphenyl)-4-oxothiazolidin-3-yl)benzamide

(KBr)cm^-1:3449(secondary NH stretching),2926(C-H Stretching),1750 (C=O),1654(C=O,Str,thiosolidinone),619(C-S-C):¹HNMR(CDCl₃):7.2(N-CH proton),7.5 (aromatic proton),4.0(S-CH2-CO-proton),8.6(-CONH protons)

IIa

Compound Name: 4-ChloroN-4-oxo-(2-(4-Trimethoxyphehyl)-thiazolidin-3-yl)benzamide

(KBr)cm^-1:3449(secondary NH stretching),2927(C-H Stretching),1702 (C=O),1654(C=O,Str, thiosolidinone),620(C-S-C):¹HNMR(CDCl₃): 7.5 (aromatic proton),4.1(S-CH₂-CO-proton),3.9(OCH₃ protons)

IIb

Compound Name :4-ChloroN-4-oxo-(2-(4-Hydroxyphehyl)-thiazolidin-3-yl)benzamide

(KBr)cm^-1:3483(secondary NH stretching),2927(C-H Stretching),3296(OH stretching),1736 (C=O),1684(C=O,Str,thiosolidinone),1513(C=N str)762(C-S-C str),827(C-Cl str):¹HNMR(CDCl₃): 7.1 (aromatic proton),7.9(N-CH proton),3.9(S-CH₂-CO-proton),8.9(-CONHproton).

IIc

Compound name: 4-ChloroN- (2-(4-Methoxyphenyl)-4-Oxothiazolidin-3-yl)benzamide

IR(KBr)cm^-1:3483(secondary NH stretching), 1736 (C=O),1684(C=O,Str,thiosolidinone), 762(C-S-C str),824(C-Cl str):¹HNMR(CDCl₃): 7.2 (aromatic proton),3.6(OCH₃proton) 4.0(S-CH₂-CO-proton),8.6(-CONHproton).

IIIa

Compound Name: 4-MethoxyN- (2-(4-methoxyphehyl)-4-Oxothiazolidin-3-yl)benzamide

IR(KBr)cm^-1:3449(secondary NH stretching),2927(C-H Stretching), 1702 (C=O),1684(C=O,Str,thiosolidinone),1623(C=N str)630(C-S-C str): ¹HNMR(CDCl₃): 7.5 (aromatic proton),7.9(N-CH proton),3.0 (OCH₃ proton)3.9(S-CH₂-CO-proton),8.7(-CONHproton).

IIIb

CompoundName:4-MethoxyN-(4-Oxo-(2-(4-Trimethoxyphehyl)thiazolidin-3-yl)benzamide

IR(KBr)cm^-1:3430(secondary NH stretching),2930(C-H Stretching), 1655 (C=O),1623(C=O,Str,thiosolidinone),1580(C=N str),618(C-S-C str): ¹HNMR(CDCl₃): 7.5 (aromatic proton),7.9(N-CH proton),3.0 (OCH₃ proton)3.8(S-CH₂-CO-proton).

IIIc

CompoundName:N-(2-(4-Chlorophehyl)-4-oxothiazolidin-3-yl)4-methoxybenzamide

IR(KBr)cm^-1:3433(secondary NH stretching),2927(C-H Stretching), 1685 (C=O),1684(C=O,Str,thiosolidinone),1623(C=N str),702(C-S-C str): ¹HNMR(CDCl₃): 7.5 (aromatic proton),7.9(N-CH proton),3.9 (OCH₃ proton)4.2(S-CH₂-CO-proton).

IVa

Compound Name:N-(2-(4-Flourophehyl)-4-oxothiazolidin-3-yl)4-methylbenzamide

IR(KBr)cm^-1:3448(secondary NH stretching),2924(C-H Stretching), 1741 (C=O),1654(C=O,Str,thiosolidinone),1602(C=N str)813(C-S-C str),1167(C-FStr): ¹HNMR(CDCl₃): 7.2 (aromatic proton),8.6(CONH proton),3.8(S-CH₂-CO-proton)1.5(-CH₃ proton)

IVb

Compound Name:N-(2-(4-Methoxyphehyl)-4-oxothiazolidin-3-yl)4-methylbenzamide

IR(KBr)cm^-1:3570(secondary NH stretching),2928(C-H Stretching), 1686 (C=O),1655(C=O,Str,thiosolidinone), 815(C-S-C str): ¹HNMR(CDCl₃): 7.1 (aromatic proton),8.9(CONH proton),3.8(S-CH₂-CO-proton),3.9(OCH₃),1.5(-CH₃ proton)

IVc

Compound Name: N-(2-(4-Dimethylaminophehyl)-4-oxothiazolidin-3-yl)4-methylbenzamide

IR(KBr)cm^-1:3432(secondary NH stretching),2939(C-H Stretching), 1723 (C=O),1659(C=O,Str,thiosolidinone), 619(C-S-C str): ¹HNMR(CDCl₃): 7.2 (aromatic proton), 4.2(S-CH₂-CO-proton), 1.5(-CH₃ proton)

Anti convulsant activity:^24-26

Healthy albino wistar rats weighing from 200-250g were selected. They were kept in separate cages, fed with balanced diet water ad libitum. Then the animals were divided into 14 groups each group containing six animals.The first group of animals were served as control, which received 0.5 ml DMSO. Second groups served as std which received phenytoin (100 mg/kg). Third group treated with compound Ia (100mg /kg). Fourth group treated with compound Ib (100 mg/kg). Fifth group treated with compound Ic (100mg/kg). Sixth group treated with compound IIa (100mg/kg). Seventh group treated with compound IIb (100mg/kg). Eigth group treated with compound II c(100 mg/kg). Ninenth group treated with compound IIIa (100mg/kg). Tenth group treated with compound IIIb (100mg/kg). Eleventh group treated with compound IIIc (100mg/kg). Twelth group treated with compound Iva (100 mg/kg). Thirteenth group treated with compound IV b (100mg/kg). Fourteenth group treated with compound IVc (100 mg/kg). All the test compound were dissolved in DMSO and administrative through intra peritoneal route. The evaluation was started 30 mins after administration of test compounds Pinna electrodes with the intensity of 150 mA current were used to deliver the stimuli. Inhibition of seizure relative to the control was calculated and the data shown on the table No.4.

ANTI HYPERGLYCEMIC ACTIVITY:^27-31

Male albino rats weighing 200-250 gm were used in the present study. All rats were kept at room temperature in the animal house. They were maintained on food pellets and water ad libitum. 45 rats included for the study, were divided into 15 groups, each consists of 3 animals out of 15 groups 14 were made diabetic with a single dose of alloxan (150mg/kg) by intraperitoneal route. Diabetes was confirmed by the determination of fasting blood glucose concentration (> 250mg/kg) on the fifth day of post administration of alloxan. Rats were divided into the following groups. Group-I Control group received 0.5 ml of DMSO/day/daily. Group-II Diabetic control received Alloxan with saline.(150 mg/kg/once a day/daily). Group-III Received standard anti-diabetic drug glicazide.(25 mg/kg/orally). Group-IV - XV Received various synthetic drugs.(100 mg/kg/i.p)

SUB ACUTE TREATMENT:

In sub acute treatment, the administration of drugs were continued for 10 days once daily.Blood samples were collected from the top of the tail just prior to end on days 1,3,7 and 10 of the drug administration. The blood glucose levels were determined for all the samples by using a cutrend kit (Glucose strip). Effect of sub acute treatment of various synthetic compounds on blood glucose level in alloxan induced diabetic rats. (10 days).

TABLE 1: Physical Data Of Synthesized Compounds

S.NO	CODE	MOLECULAR FORMULA	MOLECULAR WEIGHT	MELTING POINT	%YIELD	R_f VALUE
1.	Ia	C₁₆H₁₃N₂O₂SF	316.21	195ºC	85.59%	0.75
2	Ib	C₁₆H₁₃N₂O₂SCl	336.67	245ºC	72.52%	0.71
3	Ic	C₁₇H₁₆N₂O₃S	328.22	170ºC	68.08%	0.75
4	IIa	C₁₉H₁₆N₂O₅SCl	4190.83	200ºC	66.36%	0.61
5	IIb	C₁₆H₁₃N₂O₃SCl	348.80	173ºC	53.14%	0.76
6	IIc	C₁₇H₁₅O₃N₂Cl	362.81	205ºC	69.28%	0.77
7	IIIa	C₁₈H₁₈N₂O₄S	358.22	195ºC	57.62%	0.76
8	IIIb	C₂₀H₂₂N₂O₆S	418.28	210ºC	72.66%	0.64
9	IIIc	C₁₇H₁₅N₂O₃SCl	362.69	215ºC	84.16%	0.70
10	IVa	C₁₇H₁₅N₂O₂SF	330.22	185ºC	53.92%	0.66
11	IVb	C₁₇H₁₆N₂O₃S	328.22	215ºC	56.41%	0.84
12	IVc	C₁₆H₁₇N₂O₃S	317.47	230ºC	72.22%	0.64

Scheme of work

TABLE2: Synthesizied Compounds

S. No	Code	Chemical Name
1.	Ia	N-(2-(4-flourophenyl)-4-oxathiazolidin-3-yl)benzamide
2.	Ib	N-(2-(4-Hydroxyphenyl)-4-oxathiazolidin-3-yl)benzamide
3.	Ic	N-(2-(4-methoxyphenyl)-4-oxathiazolidin-3-yl)benzamide
4.	IIa	4-chloro-N-(4-oxo-2-(3,4,5-trimethoxyphenyl) thiazolidin-3-yl)benzamide
5.	IIb	4-chloroN-(2-(4-Hydroxy phenyl)-4-oxathiazolidin-3-yl)benzamide
6.	IIc	4-chloro N-(2-(4-methoxy phenyl)-4-oxathiazolidin-3-yl)benzamide
7.	IIIa	4-methoxy N-(2-(4-methoxy phenyl)-4-oxathiazolidin-3-yl)benzamide
8.	IIIb	4-methoxy-N-(4-oxo-2-(3,4,5-trimethoxyphenyl) thiazolidin-3-yl)benzamide
9.	IIIc	N-(2-(4-chlorophenyl)-4-oxathiazolidin-3-yl)-4-methoxybenzamide
10.	IVa	N-(2-(4-flourophenyl)-4-oxathiazolidin-3-yl)-4-methylbenzamide
11.	IVb	N-(2-(4-methoxyphenyl)-4-oxathiazolidin-3-yl)-4-methylbenzamide
12.	IVc	N-(2-(4-dimethylamino)phenyl)-4-oxathiazolidin-3-yl)-4-methylbenzamide

TABLE 3: Anti Convulsant Activity

Group	Body Weight(Grams)	Drug	Dose	Duration of Extension Phase in seconds	%Inhibition of Extension Phase
I	200-250 g	DMSO	0.5 ml	13.480.35	-
II	200-250 g	Std (Phenytoin)	25mg/kg	2.900.44	78.48
III	200-250 g	Ia	100 mg/kg	11.450.48	15.05
IV	200-250 g	Ib	100 mg/kg	4.100.96	69.5**
V	200-250 g	Ic	100 mg/kg	5.61.02	58.4*
VI	200-250 g	IIa	100mg/kg	8.48 0.92	37.09
VII	200-250 g	IIb	100 mg/kg	4.20.88	68.84**
VIII	200-250 g	IIc	100 mg/kg	9.621.04	28.63
IX	200-250 g	IIIa	100 mg/kg	3.460.22	74.33**
X	200-250 g	IIIb	100 mg/kg	9.620.46	28.63
XI	200-250 g	III c	100 mg/kg	4.020.48	70.17**
XII	200-250 g	Iva	100 mg/kg	11.051.06	18.02
XIII	200-250 g	IVb	100 mg/kg	4.80.26	64.39**
XIV	200-250 g	IVc	100 mg/kg	8.760.48	35.0

Statically tool:

1. Values are expressed as mean SEM. 2.Values are findout BY using one wayANOVA followed by Newman Keul’s Multiple range test. 3.*values are significantly different from control p (<0.05). 4. ** values are significantly different from control (P<0.01).

TABLE 4:Anti Hyperglycemic Activity

Group	Treatment of Dose	Blood Glucose Level(mg/dt)
Group	Treatment of Dose	Basal Value	Day 1	Day 3	Day 7	Day 10
I	Control Normal0.5 ml Of DMSO	76.00±1.26	76.00±1.26	81.00±2.48	88.00±2.56	85.50±1.76
II	Diabeticcontrol Alloxan150mg/kg	298.5±8.96	296.00±7.28	301.5±4.38	290.00±3.68	288.00±2.46
III	Standard Glicazide 25mg/kg	302.5±2.76	286.5±1.97	248.5±2.96	196.5±3.86	148.5±4.83
IV	Ia	296.5±6.98	278.5±5.36	256.00±4.38	226.5±4.96	215.0±5.38
V	Ib	292.5±2.86	268.00±1.46	242.00±1.46	202.5±4.96	**186.5±5.32
VI	Ic	301.5±4.82	272.52. ±32	238.5±4.12	208.5±5.62	**180.5±5.62
VII	IIa	294.00±6.96	280.5±4.96	260.3±.62	242.0±2.98	230.5±4.08
VIII	IIb	292.00±1.96	270.00±4.16	240.5±1.86	212.5±4.10	**186.5±3.06
IX	IIc	296.00±2.02	264.5±2.32	248.5±2.68	216.5±3.06	**176.5±4.08
X	IIIa	290.5±1.98	276.53.08	258.5±1.46	232.5±3.16	**202.5±3.86
XI	IIIb	296.5±2.06	280.5±4.02	260.5±2.06	230.00±2.96	**215.5±2.80
XII	IIIc	301.5±1.06	276.001.98	256.5±4.32	222.5±3.86	**188.5±2.36
XIII	IVa	290.00±7.36	278.00±3.60	258.5±2.82	240.5±1.98	238.5±2.06
XIV	IVb	292.5±2.46	270.5±1.90	250.00±4.02	226.5±3.06	**202.5±1.06
XV	IVc	288.5±2.10	272.00±3.68	258.00±3.96	238.5±1.90	230.5±3.06

TABLE 5: ANTIBACTERIAL ACTIVITY OF VARIOUS SYNTHESIZED COMPOUNDS

Name of the Compound	Organisms used Zone of inhibition in mm		Inference
Name of the Compound	*Staphylococcus aureus*	*Salmonella typhi*	Inference
Standardd (Amikacin)	16	22
Test compound	10 µg /ml	10 µg /ml
Ia	10	10	Active	Active
Ib	9	7	Active	Not Active
Ic	7	10	Not Active	Active
IIa	11	9	Active	Active
IIb	12	11	Active	Active
IIc	12	10	Active	Active
IIIa	12	11	Active	Active
IIIb	10	12	Active	Active
III c	13	13	Active	Active
Iva	10	12	Active	Active
IVb	13	11	Active	Active
IVc	8	8	Not active	Not active

Standard – Amikacin (10 µg g/disc), Control – DMSO.

ANTIBACTERIAL ACTIVITY:³²

Assay was carried out by diffusion plate method. The method followed was spread plate technique. The agar plates free from contamination were spread with 50l of 48h old culture of bacterial test organism using sterile buds. The std disc of Amikacin (sterile) of 5 mm diameter was in the Petri plates. Then the filter paper discs (sterile) of 5 mm were soaked in 1ml (1µg/ml) of the test solution and in solvent control DMSO. After evaporating the solvent in a sterile atmosphere the drug impregnated discs were placed in Petri plates. The plates were refrigerated for 1h to arrest the growth and for easier diffusion of test compounds. Then the plates were removed from refrigerator and incubated at 37⁰C over night is an inverted position. The clear zones of inhibition were measured using Hi media zone reader scale. The values are tabulated. The zones of test solutions were compared with standard Amikacin.

RESULTS AND DISCUSSION:

Anticonvulsant activity:

The table values shows that various synthetic drugs such as Ic, IIc, IIIa, IIIc, IVb posses significant anticonvulsant activity. Because it reduces the duration of extension phase after convulsion induced by MES method.

Anti hyperglycemic activity:

The table values shows that various synthetic compounds such as Ia, Ic, Ib, IIc and IIIc has shown statically significant (p<0.01) and compounds such as IIIa, IIIb, IVb significantly reduce the blood glucose level at(p<0.05) in alloxan reduced diabetic rats. But rest of the compounds like Ia, IIIc, IVa, IVc does not posses significant hypoglycemic effect. The subacute treatment with Ia, Ic, IIc, IIIa, IIIb, IIIc and IVc in alloxan induced diabetic rats produced consistent reduction in the blood glucose level as compared with diabetic control.

Antibacterial activity:

The antibacterial activity of the compounds was evaluated against gram positive organism Staphylococcus aureus and negative organism Salmonella typhi.The zone of inhibition was measured as the parameter of activity. Amikacin10μg/disc was used as standard compound. Gram positive organism staphylococcus aureus showed a zone of inhibition of 16mm, where as Ia, Ib, IIa, IIb, IIc, IIIa, IIIb, IIIc, IVa, IVb (10μg/disc mg/ml) showed activity near to amikacin(16mm) .And the gram negative organism salmonella typhi showed a zone of inhibition of 22mm,where as Ia, Ic, IIa, IIb, IIc, IIIa, IIIb, IIIc, IVa, IVb showed activity near to standard. Table 1.

CONCLUSION:

The present study was aimed at synthesis and characterization of some novel substituted schiff’s bases bearing 4-thiazolidinone nucleus compounds. The compounds were screened for anticonvulsant, anti hyperglycemic and anti bacterial activities and were found to possess considerable activity.

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Received on 10.07.2009 Modified on 14.09.2009

Asian J. Research Chem. 2(4):Oct.-Dec. 2009 page 529-535