Synthesis of Some New Substituted Quinazolin-4-3H-Ones as potent anticonvulsant agents Neha Garg#, Trilok Chandra, S. Lata, K.K Saxena and Ashok Kumar* Medicinal Chemistry Division Department of Pharmacology Medical College, Meerut-250004 (U.P.) India Abstract: All the synthesized compounds (1-32) were screened for their anticonvulsant activity at a dose of 30 mg/kg.Some of the compounds were found to exhibit good anticonvulsant activity. All the synthesized compounds have been characterized by elemental (C,H,N) and spectral (IR,1H-NMR) analysis.The Compound 31 was found to be most potent compound of this series showed 90% protection against MES. Keywords: Benzylidenoquinazolinones,thiazolyquinazolnones,thiazolidinylthiazolylquinazolidinones,anticonvulsant activity, acute toxicity studies. Introduction: Quinazolinone nucleus have been gaining prominence due to the fact that it’s derivative have been found to possess wide spectrum of activities like anticonvulsantactivity[1,2,3,4],anti-infammotaryactivity[5],have patented quinazolinone[6] ,derivatives as potent anticonvulsantactivity. Moreover thiadiazole[7], ,thiazolidinones[8,9], are other important pharmacodynamic heterocyclic nuclei, which incorporated in different heterocyclic templates have been reported to possess potent anticonvulsant activity.In the light of the above observation was throught worthwhile to *For Correspondence : Ashok Kumar, Professer Medicinal Chemistry Division, Department of Pharmacology, L.L.R.M. Medical College, Meerut (U.P.) -250004 , India.Tel:+91-09917053074 ,Fax:+91(0121)760888, Email.-rajputak@gmail.com # Part of the thesis work synthesized some new derivatives of quinazilinone by incorporate both thiazolyl and thaiazolydinoyl moieties at second position of quinazolinone nucleus with the hope to posses better anticonvulsant activity. Chemistry: Bromination of 3-substitutedphenyl -6,8-dibromo-2-methylquinazolin-4(3H)-ones i.e compounds 3-4 yielded 3-substitutedphenyl -2 bromomethyl-6,8-dibromoquinazolin4(3H)-ones i.e compounds 5-6.These brominated products on treatment with 99% hydrazine hydrate afforded dibromoquinazolin-4(3H)-ones chloroacetylchloride gave 7-8. 3-substitutedphenyl-2–hydrazinomethyl-6,8Compounds 7-8 on reaction with 3-substitutedphenyl-2–chloroacetylhydrazinomethyl-6,8- dibromoquinazolin-4(3H)-ones 9-10. further compounds 9-10 were converted in to thiazol congeners i.e. 3-substitutedphenyl-2–(2'-aminothiazol-4'-yl)hydrazinomethyl -6,8dibromoquinazolin-4(3H)-ones 11-12 by the reaction of thiourea.further compounds 1112 reacted with different aromatic aldehydes to give 3-substitutedphenyl-2–(2'benzylidenylimino-thiazol-4'-yl)hydrazinomethyl-6,8-dibromoquinazolin-4(3H)-ones 1322.substituted benzylidene congeners 13-22 were cyclized on reacting with thioglycolic acid and anhydrous ZnCl2 to yield 3-substitutedphenyl-2–[2'- (2''-phenylthizolidine-4''on-3''-yl]hydrazinomethyl-6,8-dibromoquinazolin-4(3H)-ones-23-32. Meterials and Methods: The melting points of compounds were determined in open capillaries and are uncorrected. Homogeneity of the synthesized compounds was routinely checked by thin layer chromatography on Silica Gel-G plates. The eluent was a mixture of different polar and nonpolar solvent in different proportion and spots were located by iodine. The IR spectra were recorded on Bruker IFS-66 V FTIR( max in cm–1). The 1H NMR spectra were recorded by Brucker DRX-400 FTNMR instrument using CDCl3 and DMSO-d6 as solvent and tetramethyl silane (TMS) as internal reference standard.All Chemical shift() values were recorded in ppm..Elemental analysis (C,H,N)of these newly synthesized compounds were performed on a Carlo Erba-1108 elemental analyzer. Synthesis of 3, 5-dibromoanthralic acid (1): It was prepared according to the method of wheeler and oates[10].Bromine (0.8mol) in acetic acid (20ml) was added dropwise to the solution of anthranilic acid (0.4 mol) in absolute ethanol (60ml).The solid product thus crystallized out washed with hot water and dried .It was recrytalilized from methanol.compound 1: m.p 219C(Reported m.p 218C). Synthesis of 2-methhylbenzoxazinone-6,8-dibromo(2): It was prepared according to the method of Bogert and Seil [11] .A mixture of 3,5- dibromoanthralic acid (0.01 mol) and acetic anhydride (0.02 mol) were refluxed for 2-3 hours with constant stirring. The excess of acetic anhydride was distilled off on cooling a solid separated out which was filtered, washed with petroleum ether (60-80 ºC) and dried in vacuum compound 2: m.p 182C (Reported m.p 184C). Synthesis of 3-(p-hydroxyphenyl)-6,8-dibromo-2-methylquinazolin-4(3H)-one(3): A mixture of compound 2 (0.2 mol) and substituted aniline (0.2 mol) was heated on a free flame for 10-20 minutes in conical flask.After the disappearance of water droplets in conical flask, it was kept at room temperature.On cooling a jelly like mass obtained which was dissolved in methanol,refluxed and poured into water .The solid thus obtained was filtered, dried and finally recrystallized with ethanol. compound 3: m.p 202C; yield 62%;molecular formula C15H10N2O2Br2. IR (KBr) max in Cm-1 610 (C-Br),1635 (C=N), 1550 (C C of aromatic ring),1720(C=O of quinazolin ring),1734 (OH): 1H-NMR (CDCl3) in ppm 2.30 (s, 3H, CH3),7.25-7.90(m,6H,Ar-H),9.30(s,1H,,OH exchangable). The compound 4 have been prepared by employing the aforementioned method and their physical and analytical data are shown in Table-I Synthesis of 3-(p-hydroxyphenyl)-2-bromomethyl-6,8-dibromoquinazolin-4(3H)- one(5): Bromine (0.4 mol) in acetic acid (20ml) was added dropwise to the solution of compound 3 (0.2 mol) in acetic acid (50ml).The reaction mixture was poured onto crushed ice then left overnight at room temperature The precipitate thus obtained was filtered ,washed with water ,dried and recrystallized with ethanol Compound 5: m.p 214C; yield 70%;molecular formula C15H9N2O2Br3. IR (KBr) max in Cm-1 608 (C-Br), 1550 (C C of aromatic ring), 1632 (C=N), 1715 ( C=O of quinazolin ring), 3425 (OH):1HNMR(CDCl3)inppm2.75(s,2H,CH2),7.207.85(m,6H,ArH),9.86(s,1H,,OHexch angable). The compound 6 have been prepared by employing the aforementioned method and their physical and analytical data are shown in Table-I Synthesis of 3-(p-hydroxyphenyl)2–hydrazinomethyl-6,8-dibromoquinazolin-4(3H)one(7): A mixture of compound 5 (0.1 mol) and hydrazine hydrate (99%) (0.2 mol) in methanol was refluxed for 10 hours The excess of solvent was distilled off and the reaction mixture was poured onto ice. The solid thus obtained was filtered, washed with water,dried and recrystallized from methanol compound 7.m.p.222ºC;yield64%; molecular formula C15H12N4O2Br2. IR (KBr) max in Cm-1 610 (C-Br),1270(N-N),1300 (C-N), 1550 (C C of aromatic ring),1720(C=O of quinazolin ring) ,1620(C=N), 3425(OH) ,3300(NH,NH2) : 1 H- NMR (CDCl3) in ppm 2.64(s,2H,CH2),7.22-7.82(m,6H,Ar- H),9.85(s,1H,,OHexchangable). The compound 8 have been prepared by employing the aforementioned method and their physical and analytical data are shown in Table-I Synthesis of 3-(p-hydroxyphenyl)-2-chloroacetyhydrazinomethyl-6,8–dibromo- quinazolin-4 (3H)-one (9) : To the solution of compound 7 (0.01 mol) in dry benzene (60) chloroacetylchloride (0.02 mol) was added gradually with stirring under cool condition. The reaction mixture was further stirred for another 2 hours at room temperature and then refluxed for 4 hours .Benzene was removed by distillation ,to yield the product ,which was fjnally recrystallized from methanol to afford compound 9.m.p.242 ºC ; yield 70%; molecular formula C17H12N4O3Br2Cl. IR (KBr) max in Cm-1 610(C-Br), 760(C-Cl), 1270(N-N), 1320 (C-N), 1550 (C C of aromatic ring), 1715 ( C=O of quinazolin ring), 1618 (C=N), 3480 (OH) ,3320(N-H), 2956(C-H aliphatic) :,3055 (C-H aromatic) ,3480 (OH) 1 H- NMR(CDCl3)in ppm 2.60 (s, 2H, CH2 ),(s,2H,-CHCl2) , 7.25-7.80(m,6H,ArH),9.85(s,1H,,OH exchangable). The compound 10 have been prepared by employing the aforementioned method and their physical and analytical data are shown in Table-I Synthesis of 3-(p-hydroxyphenyl)-2–(2'-aminothiazol-4'-yl)hydrazinomethyl-6,8- dibromoquinoazolin-4(3H)-one(11): A mixture of compound 9, (0.02 mol) thiourea (0.02 mol) and acetone (60 ml) was refluxed for 12 hours. The completion of the reaction was monitored by TLC it was then concerted and cooled, where upon the solid separated out was filtered and then recrystallized from methanol.The solid thus obtained was washed with 2% saturated sodium carbonate solution and water to librate the base completely dried and recrystallized from ethanol to give compound 11: m.p.250 ºC; yield 68%; molecular formula C18H14N6O2SBr2. IR (KBr) max in Cm-1 612(C-Br), 690(C-S-C), 1270(N-N), 1220 (C-N), 1580 (C C of aromatic ring), 1715 ( C=O of quinazolin ring), 1616 (C=N), 3485 (OH) ,3390(N-H).1H-NMR(CDCl3)in ppm 2.55 (s, 2H, CH2 NH), 6.40(s,2H,NH2 exchangable with D2O), 7.25-7.84(m,7H,Ar-H),7.89 (brs,2H,NH.NH2),10.00(s,1H,,OH exchangable). The compound 12 have been prepared by employing the aforementioned method and their physical and analytical data are shown in Table-I Synthesis of 3-(p-hydroxyphenyl)-2–(2'-benzylidenylimino-thiazol-4'-yl) hydrazinomethyl-6,8-dibromoquinoazolin-4(3H)-one(13): To the solution of compound 11 (0.01 mole) in methanol (80ml), benzaldehyde (0.01) with few drops of glacial acetic acid was added and then reaction mixture refluxed for 10 hours ,completion of the reaction was monitored by TLC. After distillation of excess of solvent the reaction mixture was cooled ,diluted with cold water and filtered. The solid thus collected was recrystalized from ethanol to furnish compound 13 :m.p.260 ºC ; yield 58%; molecular formula C25H18N6O2SBr2. . IR (KBr) max in Cm-1 608 (C-Br), 690(C-SC), 1270(N-N), 1219 (C-N), 1575 (C C of aromatic ring), 1717 ( C=O of quinazolin ring), 1620 (C=N), ,3382(N-H) 3485 (OH).1H-NMR(CDCl3)in ppm 2.56 (s, 2H, CH2NH),7.05-7.90(m,12H,Ar-H),7.85 (brs,2H,NH.NH exchangable with D2O),4.624(s,1H,=CH-Ar),10.04(s,1H,,OH exchangable). The compounds 14-22 have been prepared by employing the aforementioned method and their physical and analytical data are shown in Table-I Synthesis of 3-(p-hydroxyphenyl)-2-[2'-(2''-phenylthizolidine-4''on-3''yl] hydrazinomethyl-6,8-dibromoquinoazolin-4(3H)-one(23): A mixture of compound 13 (0.01), thioglycolic acid (0.01) and anhydrous ZnCl2 (2 gm)in absolute ethanol was refluxed for 10 hours.The progress and completion of reaction was checked by TLC. After refluxing , excess of solvent was distilled off and residue was poured in cold water , filtered, dried and finally the product was recrystalized from benzene to furnish compound 23 :m.p.268 ºC ; yield 60%; molecular formula C27H20N6O3S2Br2.. . IR (KBr) max in Cm-1 610 (C-Br), 682(C-S-C), 1217 (C-N), 1574 (C C of aromatic ring), 1740 ( C=O of quinazolin ring), 1622 (C=N), ,3370(N-H) 3515 (OH).1H-NMR(CDCl3)in ppm 2.45 (s, 2H, -CH2NH), 4.05(s,2H,-CH2 of thiazolidinone ring7 ),707-7.10(m,12H,Ar-H),7.94 (brs,2H,NH.NH exchangable with D2O),4.60(s,1H,=CH-Ar),10.08(s,1H,,OH exchangable). The compounds 24-32 have been prepared by employing the aforementioned method and their physical and analytical data are shown in Table-I Biological Methods: The pharmacological activities of compounds were evaluated by the following standard procedures. All the newly synthesized compounds were evaluated for anticonvulsant activity and acute toxicity. Anticonvulsant activity was determined by supramaximal eletroshock seizures pattern test (SMES). Albino rats of either sex weighing between 90150 gm were divided into 10 groups of 10 animals in each group. Only mice were used for performing acute toxicity studies. The effect of unknown compounds were compared with the standard drug and propylene glycol treated group served as control. The statistical evaluation of observations was done by using appropriate statistical tests. All the compounds were screened for their anticonvulsant activity and approximate lethal dose (ALD50) Anticonvulsant activity1. Supramaximal electroshock seizure pattern test (SMES): This activity was performed by following the method of Toman et al[12] in albino rats. Rats of either sex weighing 90-150 gm were divided into groups of ten animal each.The test drugs and phenytoin sodium reference drug were administrated intraperitoneally in rats. After one hours they were subjected to a shock of 150 M.A for 0.2 seconds and the presence or absence of extensor response was noted. Animals in which extensor response was abolished were taken as protected rats. 2. Approximate lethal dose (ALD50): The LD50 was determined in mice weighing 25-30 gm of either sex by the method of smith[13]. The text compounds were administed orally in one group and the same volume of propylene glycol in another group of animals consisting six mice in graded doses. During the study the animals were allowed to take food and water adlibidum. After 24 hours of drug administration percent mortality in each group was observed. From the data obtained ALD50 was calculated. Result and Discussion: Newly synthesized compounds were tested in vivo in order to evaluate their anticonvulsant activity at a dose of 30mg/kg i.p. The anticonvulsant effects of all the compounds of this series reported in Table II.The characteristic feature of this series is the presence of a five membered thiazole ring at second position of 2-Methyl-6,8- dibromo-3-substitutedphenyl quinazolin -4 (3H)-onyl moieties which were further substituted with alkyl benzylidenyl groups at the second position of five membered thiadiazole ring .It was observed that compounds haveing 2-Methyl-6,8-dibromo-3-(4chiorophenyl) quinazolin-4(3H)-onyl moiety showed more degree of protection in comparison to 2-methyl-6,8-dibromo-3-phenylquinazolin-4(3H) onyl moieties. All the compounds 13-22 exhibiting promising anticonvulsant activity .It was observed that compounds having phenyl group (compounds13 and18) as subsituent showed least activity (50 and 60% respectively).while compounds (16 and 21 )substituted with 3mehoxy-4-hydroxy phenyl ring exhibited the maximum percent protection (70 and 80 %) respectively against seizures induced by MES. Compounds substituted with 4mehoxyphenyl group (15and20) exhibited 60 and 70% inhibition of seizures respectively. Compounds having 4-hydroxyphenyl and 4-N,N-dimethylamino phenyl also elicited remarkable anticonvulsant activity of 60%( compounds 14 and 17) and 70% (compounds 19 and 22 )respectively. Further the next step of the series was characterized by the presence of thiazolidinone (-thiolactum ring )in addition to thiadiazole ring .All compounds showed potent anticonvulsant activity compounds 26 and 31 substituted with 3-methoxy-4-hydroxyphenyl group have shown most potent activity of 80 and 90% respectively. However compound 26 showed equipotent activity while compound 31 have shown more potent activity than standard drug phenytoin sodium cycloaddition of thioglycolic acid on schiffs bases (13-22)forms their corresponding thiazolidinone derivatives (23-32) (Table-II).which in general increases the anticonvulsant activity except in compounds 25 and 30 which showed the same degree of protection i.e. 70% against MES test.compounds 23 and 28 having phenyl ring at second position of thiazolidinone ring showed 60 and 70% protection respectively. Where as compounds 25 and 30 having 4-mehoxyphenyl group exhibited 70% protection each compound with 4hydroxyphenyl group i.e. compounds 24 and 29 have also shown remarkable percentage protection of 70 and 80 % respectively.Eighty percent protection was also exhibited by the compounds substituted with 4-N,N-dimethylaminophenyl group against seizures induced by MES. There fore considering the compounds of this step it may be concluded that compounds 21 and 31 have shown most potent response. These compounds were also tested at three graded doses i.e. 7.5,15 and 30 mg/kg i.p. Interestingly at all the three doses compound 21 exhibited activity equipotent to standard drug phenytoin sodium Moreover compound 31 was found to exhibit more potent activity than phenytoin sodium at all the three dose levels. 1. The compounds having 2-methyl-3-(4-chorophenyl)-6,8-dibromoquinazolin4(3H) onyl moiety showed more protection than the compounds having 2-methyl3-phenyl-6,8-dibromoquinazolin-4(3H)onyl moiety. 2. Thiazolidinones showed more potent anticonvulsant activity in comparison to their corresponding thiazoles. 3. Substitution with 3-methoxy-4-hydroxyphenyl group was found to be more benificial for the anticonvulsant activity. Ackowledgement: We are thankful to SAIF Punjab University Chandigarh for elemental and spectral analysis. Refererences: 1. Kumar A et al ,2006,Thiazolidinoyl and azetidinyl quinazolinones as potential anticonvulsant agents.,Oriental.J.Chem.,22:219-230. 2. Guan et al ,2007.,Synthesis of some quinazolin -2-(1H)-one and 1,2,4triazolo [4,3-a] quinazolin derivatives as potent anticonvulsants.J.Pharm.Pharmaceut. Sci 10(3):254-262. 3. Zappala.M et al 2003.,1-Aryl-6,7-methylenedioxy-3H-quinazolin -4-ones as anticonvulsant agents.,Bioorg and Medic.Chem.Laters 13: 4427-4430. 4. El-Helby AG.and Wahab M.H.A.,2003,Design and synthesis of some new derivatives of 3H-quinazolin-4-one with promising anticonvulsant activity .,Acta Pharm.,53:127-138. 5. Alagarsamy V et al 2006.Pharmacological evaluation of 2-substituted (1,3,4) thiadiazolo quinazolines.Indian J of Pharmaceu .Sci.,68:108-111. 6. Bajaj K.et al.,2003,Newer substituted benzoxaepinyl quinazolinones as potent antipsycotropic and anticonvulsant agents.,Arzeim-forsch/Drug research.,53: 480-485. 7. Mohan RR et al ,1985,Synthesis of some newer Quinazolinyl ,Oxadiazoles, Tiosemicarbazides,Thiadiazoles as pharmacologically active agents ,Indian J of Chem,24B:78-82. 8. Archana et al ,2002, Synthesis of newer thiadiazolyl and thiazolidinonyl quinazolin-4(3H)-ones as potent as anticonvulsant agents.,Eur.J Med Chem..,37:873-882. 9. Archana et al ,2003, Synthesis of newer indolyl thiadiazoles and their thiazodinones and formazanes as potential anticonvulsant agents.,Indian J Pharmaceu.sci.,65:358-362. 10. Wheeler A.S and oats W M ,1910.The bromination of anthranilic acid. J.Am.Chem.Soc,32:,770-773. 11. Bogert M T and Seli H A,1907.Resarches on quinazolines on dialkyl -4quinazolones and the products obtained by alkylating -2-alkyl-4quinazolones(2-alkyl-4-hydroxy quinazolones). J.Am.Chem.Soc,29 :517536. 12. Toman J.E.P,et al.,1946,J .Neuro Physo 9: 231-240. 13. SmithQ.E,1960, Phamacological screening tests progress in medicinal chemistry, Butterworths London .,1: