IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 Synthesis o f 1, 2, 4- Triazole Derivatives And Their Biological Activity Study E. M. Hussain Departme nt of Chemistry, College of Education, I bn- Al-Haitham Unive rsity of Baghdad Abstract This study includes the sy nthesis of new derivatives of 1, 2, 4- Triazole which are contain Schiff bases derived from 1, 4, 5, 6- tetrahy dropy rimidine. The st ructures of these derivatives were characterized fro m their melting points, infrared sp ectroscopy and elemental analysis. T hese derivatives were tested for inhibition of E-coli and were all found to be active. Introductions The sy nthesis of 1. 2, 4- triazole derivatives has att racted widesp read att ention due to their diverse biolo gical activities, in cluding antimicrobial, anti- inflammatory , analgesic, and antitumoral [1-7]. Nurhan et al [8] h ave p repared some new 1, 2, 4- Triazole derivatives with antimicrobial activity [Figure 1]. H3C NN N CH3 NHC O H3C H3C C O O NN N CH3 NH2 N NN N C H3 NH C O C H3 N NN N C H3 HN C H2 R R= C O OCH3 HO OC H OH, , , , , [Figure 1] Neslihan [9] has sy nthesized comp ounds incorp orating both 1, 2, 4- triazole and 1, 3, 4-thiadiazole due to t heir p ossible diverse pharmacological prop erties [Figure 2]. IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 R NN N CH2 NN S S O H R= - CH3 , -CH2 Ph , -Ph H2C O NN SH [Figure 2] Abeer [10] has sy nthesized different derivatives of bis- 1, 2, 4-triazole of p ossible biolo gical activity [Figure 3] Am C CH2 S NN N H NN N S CH2 C C CH2 Am H Am C CH2 S NN N H NN N S CH2 C C CH2 Am H (CH2)4 N NH NAm=( ), CH2 C CH2 C [Figure 3] Biocidal activity of Schiff bases have also been well established. These have been att ributed to the toxop horic C=N linkage in them [11]. Amina [12] h as sy nthesized different heterocyclic comp ounds derived fro m D- galactose incorp oratoring both 1, 2, 4- trizole and Schiff bases [Fi gure 4]. These derivatives were test ed for inhibition of E-coli and Staphy lococcus and were all found to be active. O NN N O O O O H3C CH3 CH3 CH3 CH2 O CH2 SH HN CH R R= (P-MeC6H4,P-Me2NC6H4, m-O 2NC6 H4) [Figure 4] In this study new derivatives of 1, 2, 4- triazole which are contain schiff bases were sy nthesized and study their biological activity . IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 Experime ntal Materials All chemical used were sup p lied from Ried el- De Ha en AG, BDH chemicals, Acros Organics, Janssen chemical, M erk chemicals Fluska AG and Hop kin and Wiliams. E lemental analyzer were carried out by using C arlo Erba / M od 1106, Infrared sp ectra were record ed using Shimadzu 408 (KBr disc) and meltin g p oints were recorded usin g Electrot hermal meltin g p oint app aratus. T he biometrials were obtained from Bio merieux Ltd. S ynthbesis of 1, 4, 5, 6- tetrahydropyrimi dine -2- thi ol. 1 Compound 1 was used as st arting material, this derivative was p repared by dissolving1,3- diaminoprop ane (1 ml) in ethanol (10 ml) and carbon disulphide (2 ml) was added. The mixture was stirred for (15) minutes then refluxed for (3) hrs, the mixture was cooled to room temp erature, then the white precipitate was formed filtered and recry st allized from ethanol to give 1. S ynthesis of 3, 4, 5, 6- Tetrahydropyrimi dine -2- thioacetic acid. 2 Compound 1 (3g, 25.86 mmo l) and sodium hy droxide (1.9, 27.5 mmol) wer e dissolved in ethanol (25 ml) and reflu xed for (2h). A solution of chloro acetic acid (2.44 g, 25.82 mmol) in ethanol (15 ml) was added. The reaction mixture was st irred for (24 hrs). T he white precipitate was filtered and recry st allized from ethanol to give 2. S ynthesis of 3, 4, 5, 6- Tetrahydropyrimi dine - 2- methylthioacetate. 3 Compound 2 (5g, 28.73 mmol) was dissolved in acetone (100ml) and anhydrous sodium carbonate (3 g, 28.3 mmo l) was added. The mixture was reflu xed for (1h). Dimethy lsulp hate (3.5 ml) was added and the mixture was further refluxed for (24 hrs). The solvent was removed under p ressure and the residue diluted with water and extracted with ethy lacetate (2x30 ml). The Organic lay er was dried and removed to give 3 as solid. Compound 3 was recrystallized from ethanol: water (8:2). S ynthesis of 3, 4, 5, 6,-Tetrahydropyrimi de- 2- thi oacetichydrazi de. 4 Compound 3 (3g, 15.95 mmol) and hy drazine hy drate (15 ml) were dissolved in ethanol (25 ml). The reaction mixture was reflu xed for (24 hrs). The p recipitate which sep arated on cooling was fi ltered and recryst allized from ethanol to give 4. Sy nthesis of 3, 4, 5, 6- Tetrahy dropy rimdine- 2- thiomethy lpotassiumxanthate. 5 IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 To a mixture of potassium hydroxide (0.9 g, 16.07 mmo l) and comp ound 4 (3 g, 15.95 mmol) in absolute ethanol (25 m l), carbon disulfide (1.5 ml) was added. The reaction mixture was diluted with ethanol (25 ml) and st irred over night at room temp erature. It was them diluted with dry ether (30 ml) y ieldin g a p ale y ellow p recipitate which was filtered, washed with ether and dried at room temp erature to give the p otassium salt 5 in quantitative y ield. The product was utilized in the next st ep without further p urification. S ynthesis of 2- [5- Mercapto- 4. Amino-1, 2, 4-triaz o-3-yl] thiomethyl-3, 4, 5, 6- tetrahydropyrimdine. 6 The p otassium salt 5 (2g, 6.62 mmol) was suspended directly in hy drazine hy drate (98%, 5 ml) and heated under reflu x for (3hrs). The mixture was cooling, diluted with water (75 ml) and f iltered. The filterate was neutralized with (10 %) HCl The p recipitate formed was filtered, dried and r ecry st allized from ethanol to give 6 as solid. Sy nthesis of 2- [M ercapto-4-ary lidineimino-1, 2, 4-triazol-3-yl]- thiomethy l-3, 4, 5, 6- tetrahy dropy rimdine. (7-11) General procedure: A hot ethanolic solution of comp ound 6 (3g, 12.29 mmol) was mixed with a solution of the selected aldehy de (12.29 mmol) in (25 ml) ethanol. The resulting mixture was then reflu xed for (3 hrs). The p roduct was filtered and recry st allized from ethanol. Results and Discussion The Strategy used for the Sy nthesis new derivatives of 1, 2, 4-triazole was started with derivatve 1 in a ser ies of reactions. [Scheme 1]. Compound 1 was synthesized from the reaction of 1, 3- diaminop rop ane with carbon disulphide [13]. The IR sp ectrum of 1 showed stretching bands at 3220 cm -1 , 1050 cm -1 and 1625 cm -1 for (NH), (C=S) and (C=N) resp ectively. Tables (1) and (2) showed the characteristic IR absorp tion bands and p hysical prop erties for all new deriv atives. Treatment of comp ound 1 with chloroacetic acid in b asic medium under ref lux gave 2. The IR sp ectrum of compound 2 showed the disapperance of st retching band at 1050 c m -1 for thion group with app erance of st retching band at 1660 cm -1 for (C=O) group resp ectively. Compound 3 was synthesized by the reaction of compound 2 with dimethy lsulphate in aceton. The IR Sp ectrum of 3 showed the disapp erance of st retching broad band of (OH) group at (3200-2300) cm -1 with apperance of (C=O) group at 1720 cm -1 which is disp lacement to high frequency . Gatterman method [14] was used for the sy nthesis of IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 derivative 4. The IR Sp ectrum of 4 showed the disp lacement of (C=O) group to low frequency at 1675 cm -1 with apperance of stretching band at 3355 cm -1 for (NH2) group . The salt 5 was indicated by its solubility and infrared sp ectrum which showed multip le (NH) st retching bands at (3300-3100) cm -1 , a stretching band at 1640 c m -1 due to a mide group . The sp ectrum, also showed absorp tion at1055 cm -1 , 1215 cm -1 which is att ributed to (C=S) and (N-N) stretching vibrations resp ectively. The cyclization of 5 to 6 could be accomplished by the suggested st eps [15] in [Scheme 2]. The IR sp ectrum of compound 6 showed a sp lit broad band at 3360 and 3210 cm - 1 which was assign ed to the asy mmetric and sy mmetric st retching bands of (NH2 and NH) group s resp ectively. The IR sp ectrum also showed a bending b and at 1645 cm -1 for (NH) group , st retching band at 1595 cm -1 for (C=N) endocy lic, appearance band at 1496 cm -1 due to (-N-C=S) toutomeric [ Figur e 5]. The compounds 7-11 were sy nthesized from the reaction of comp ound 6 with different substituted benzaldehy des in p ara p osition. The IR sp ectra of these derivatives showed the disapp earance of st retching bands due to (NH2) of the triazole and (CO) group of different substituted benzaldehy des with app earance of st retching band in the range (1610- 1650) cm -1 att ribute to the imine group . Compound 7-11 exhibited a biolo gical activity against E-coli bacteria. Co mpound 11 exh ibited higher degree of activity than the others Table (3). IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 [Scheme 1] CH2 CH2 CH2 NH2 NH2 + CS2 C2H5OH N N C H SH N N C S H H (1) (1a) N N C H S CH2 C O OH a N C N H S CH2 (2) C O OCH3 b (3) N C N H S CH2 C O NH (4) NH2 c N C N H S CH2 C O NH (5) C S S-K+NH d N N C H S CH2 (6) N N N H NH2 S N N C H S CH2 N NN S NH2 N N C N N C NN N H S CH2 NN N S N CH R H H S CH2 S N CH (7-11) e R H Taut . T hion Thiol H Taut . Thion Thiol a: ClCH2COOH, NaOH, C2H5OH. b: Na2CO3, (CH3)2SO4, Acetone. c: NH2-NH2, C2H5OH. d: CS2, C2H5OH, KOH. e: NH2-NH2. H2O (98%). f: R CHO , C2H5OH Where R= H M e M eO CN Compound No. 7 8 9 10 11 N M e M e IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 [Scheme 2] R C O NHNH C S S K + NH2N H2 R C O NH NH C S S K - + NH H N H2 R C O NH N H C SH N HH 2N S K - R C O NH N H C S K N H2N H +-+ -H2 S R C O H N H N C NH2HN S K +- R NN N NH2 S K - + R H NH2 S K N NN H2 O +- H3O + - NN N N H2 R SH HCl+ KCl- R= ( N N C H S CH2 ) R C O NHNH2 KOH/CS2 C2H5OH +H + IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 Re ferences 1: Neslihan, D.; Ahmet, D.; Sengul, A.and Sancak, K. (2004), "Sy nthesis and antimicrobial activities of some n ew 1,2,4-triazolre deriv atives", Eur. J. M ed. Chem. 39:793-804. 2: Neslihan, D.; Ahmet, D.; Sengu l, A., and Elif. C. (2005) " Sy nthesis and antimicrobial activities of some new [1,2,4] triazolo [3,4-b] [1,3,4] thiadiazines" ARKIVOC (i). 75- 91. Ant imicrobial activities", ARKIVOC (i), 75- 91. 3: Neslihan, D.; Ahmet, D.and Sen gul, A. K. (2005), " Sy nthesis and biological activity of new 1,2,4-triazol-3-one derivatives", Russian J. Bioor g. Chem. 39 : 387- 97. 4: Sabir, H.; Gy oti, S.and M ohd, H. (2008), "Sy nthesis and antimicrobial activities of 1,2,4-triazole and 1,3,4-thiadiazole derivatives of 5- amino-2-hydroxy benzoic acid", E. J. Chem. 5 (4), 963- 968. 5: Aiy alu, R. and Kalasalingam, A. R. (2009), " Sy nthesis of some novel triazole derivatives as anti-nociceptive and anti-inflammatory agents", 59: 355- 364. 6: M udasir, R. B.and Abdul, R. (2009), "Subst ituted 1,2,4-triazoles and thiazolidinonce from fatty acids: Sp ectral Charterization and antimicrobial activity ", Indian J. Chem., 48 B, 97-102. 7: M alii, R. R.; So mani, R. R. ; Toraskar, M . P.; M ali, K. K.and Shirodkar, P. Y. (2009) "Sy nthesis of some antifun gal and Ant i-tubercular 1,2,4-triazole analoges", Int. J. Chem Tech Res., 1 (2). 168-173. 8: Nurhan, G.; M evlut, S.; Elif, C.; Ali, S.and Neslihan, D. (2007), "Sy nthesis and antimicrobial activity of some new 1,2,4-triazole deriv atives", Turk. J. Chem. 31: 335-348. 9: Neslihan, D. (2005) "Sy nthesis and characterization of new trihetrocy clic comp ounds consisting of 1,2,4-triazol-3-one, 1,3,4-thiadiazole and 1,3,4-oxadiazole rings", Turk. J. Chem. 29:125-133. 10: Abeer, J. H. (2002), "Sy nthesis and antimicrobial evolution of some new bis-1,3,4- oxadiazole, b is-1,2,4-triazole and bis-1,3,4-thiadiazole deriv atives", M . Sc. Thesis, College of Education/ Ibn Al-Haitham Baghdad University , 29-34. 11: Hodett, E. M. and Dunn, W. D. (1970), "Structure – antitumor activity correlation of some Schiff bases". J. M ed. Chem., 13(4):768- 771. 12: Amina, A. F. (2008). " Sy nthesis of new heterocyclic comp ounds derived fro m D- glactose and evolution of their biological activity of these derivatives" Ph. D. T hesis, College of Education/ Ibn Al-Haitham, Baghdad University ; 38-47 IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 13: Brown, D. J.and Evans, R. F. (1962), "hy dropy rimidines. Part I. 1,4,5,6- Tetrahy dropy rimidine and its derivatives", J. Chem. So c. 527- 533. 14: Gatterman, L.and Weiland, H. (1952), "Laboratory methods of organic chemistry ", M acillon Co. P. 135. 15: Hoggarth, E. (1952), "2-benzoly ldithiocarbazinic acid and related comp ounds" J. Chem. So c., 4811- 4817. Table (1): Characteristic IR a bsorption bands of the ne w derivatives Compound No. Infrared data (m axcm -1 ) (KBr disc) 1 (NH) 3220; (C=S) 1050; (C=N) 1625. 2 (NH) 3215; (OH) (3200-2300); (C=O) 1660; (C=N) 1620. 3 (NH) 3230; (C=O) 1720; (C=N) 1620. 4 (NH2) 3355; (NH) 3215; (C=O) 1675; (C=N) 1615. 5 M ultiple (NH) (3300- 3100); (C=O) amide 1640; (C=N) 1615; (C=S) 1055; (N-N) 1215. 6 (NH2) 3360; (NH) 3210; (NH) bending (1645); (C=N) 1595; (-N-C=S) 1496; (C=S) 1055. 7 (NH) 3225; (C=S) 1055; (C=N) 1640; (C=C) (1590); (-N-C=S) 1490; (C=C) bendin g 820. 8 (NH) 3230; (C=S) 1050; (C=N) 1630; (C=C) 1600; (-N-C=S) 1495. (C=C)bending 815. 9 (NH) 3220; (C=S) 1050; (C=N) 1650; (C=C) 1595; (-N-C=S) 1495; (C=C) bending 820. 10 (NH) 3225; (C=S) 1050; (C=N) 1650; (C=C) 1610; (-N-C=S) 1495; (C=C) bending 835. 11 (NH) 3230; (C=S) 1055; (C=N) 1625; (C=C) 1595: (-N-C=S) 1495; (C=C) bending 830. IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 Table (2): Physical propertie s for derivatives Compound No. Formula M elting p oint Cº Elemental analy sis calculated (Found) Yield % C% H% N% 1 C4H8N2S 210 41.37 (41.22) 6.89 (6.68) 24.13 (23.98) 78 2 C6H10N2O2S 172 41.37 (41.05) 5.74 (5.52) 16.09 (15.88) 82 3 C7H12N2O2S 188 44.68 (44.8) 6.38 (6.07) 14.59 (14.71) 77 4 C6H12N4OS 231 38.29 (38) 6.38 (6.11) 29.78 (29.56) 91 6 C7H12N6S2 240 34.42 (34.22) 4.91 (4.9) 34.42 (34.61) 65 7 C14H16N6S2 227 50.60 (50.92) 4.81 (4.53) 25.30 (24.98) 88 8 C15H18N6S2 235 54.21 (53.99) 5.20 (5.18) 24.27 (24.45) 95 9 C15H18N6OS2 196 49.72 (49.49) 4.97 (5.11) 23.02 (22.88) 78 10 C16H21N7S2 218 51.20 (51.00) 5.60 (5.82) 26.13 (26.33) 80 11 C15H15N7S2 226 50.42 (50.61) 4.20 (3.92) 27.45 (27.54) 96 IHJPAS IBN AL- HAITHAM J . FO R PURE & APPL. SC I VO L. 23 (3) 2010 Table (3): Effect of antimicrobial agents on Esche richia Coli Compound No. Effect of new derivatives on t he growt h of E-coli Bacteria 1 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0.0 9 Concentratio n gm/ml 7 - - - - - + 8 - - - - - - + 9 - - - - - - - - - + 10 - - - - - - - - + 11 - - - - - - - - - - + Blank + (-) No growt h (+) Growth IHJPAS 2010) 3( 23 مجلة ابن الھیثم للعلوم الصرفة والتطبیقیة المجلد فعالیة بایولوجیة محتملة وترایزول ذ -4،2،1تحضیر مشتقات جدیدة لـ محمد حسین إیمان جامعة بغداد ،ابن الهیثم -كلیة التربیة ،قسم الكیمیاء الخالصة ر . حتوي على قواعد شفترایزول ت -4، 2، 1یدة من مركبات مشتقات جد تحضر تم في هذا البحث حض مادة 1استخدم المشتق رقم . ثنائي امین بروبان مع ثنائي كبریتید الكاربون - 3، 1المركب ةمن مفاعل 1المشتق رقم مشتق ة علثم مفا. 6المشتق الىترایزول بعد مروره بسلسلة من التفاعالت وصوال - 4، 2، 1ابتدائیة لتحضیر مشتقات 1ال اعطى اذ ،الذي عومل بدوره مع كبریتات ثنائي المثیل 2اعطى المشتق ،اذوسط قاعدي فيمع حامض كلورید الخلیك ةحضر من مفاعل 5المشتق . امع الهیدرازین بوجود الكحول مذیب 3المشتق ة حضر من معامل 4المشتق. 3المشتق الذي عومل بدوره مع 6اعطى المشتق ،اذمع الهیدرازین 5یت ثعومل ملح الزان. مع ثنائي كبریتید الكاربون 4المشتق الفعالیة البایولوجیة تدرس. 11-7من االلدیهایدات المختارة لیتم الحصول على مشتقات لقواعد شف من ة یدعدانواع .11رها فاعلیة هو المشتق ضد بكتریا القولون وكان اكث 11-7للمشتقات من IHJPAS