رفة والتطبیقیة المجلد 2009) 3( 22مجلة ابن الهیثم للعلوم الص -بیوتان-4,1یة تحضیر و تقییم الفعالیة المضادة للبكتریا لبعض مشتقات ثنائ اوكسادایزول-4,3,1 حنان ابراهیم عمر الدین ، جامعة بغداد ، كلیة الصیدلة قسم علوم المختبرات السریریة الخالصة من حمض االدبیك ثنائي الهیدازید [IIIa-j]اوكسادایزول -4,3,1- بیوتان-4,1تم تحضیر سلسلة من مشتقات ثنائیة مع بعض الحوامض االروماتیة المختلفة بوجود كلورید الفسفوریل، و قد تم تشخیص هذه السلسلة بأستخدام تقنیة االشعة تحت .الحمراء و تحلیل العناصر و طیف الكتلة م دراسة تأثیر هذه المركبات على الفعالیة المضادة للبكتریا من نوع و قد ت (Gram + و Gram)- و اظهرت الدراسة ان قسم .من هذه المركبات لها فعالیة بایولوجیة ضد انواع من البكتریا IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (3) 2009 Synthesis and Antimicrobial Evaluation o f Some Bis-1, 3, 4-Butane-1-3, 4-Oxadiazole Derivatives H.I. Omar–El deen Departme nt of Clinical Laboratory Sciences, College of Pharmacy, Unive rsity of Baghdad Abstract A series of new Bis-1,4-Butane -1,3,4 – Oxadizole der ivatives [III a-j] were sy nthesized from adip ic acid dihydrazide and different aromatic acids in the presence of phosp hours oxy chloide. There comp ounds were characterized by their IR, microanalysis, and mass sp ectral data. In vitro antimicrobial were sy nthesized. In vitro antimicrobial activity of these comp ounds against (Gram negative) and (Gram p ositive) were rep orted, some of these comp ounds p rep ared derivatives exh ibited antimicrobial activity . Introduction M any comp ounds contain 1,3,4-oxadiazole ring sy stem which p ossesses p ossible biological activity (1-2-3). 3-substituted aminomethy l-5-substitued-1,3-4-oxadiazole-2-thione are tuberculostatic (4) and fungicidal, similarly 3,5-disubst ituted – 1,3,4- oxadiazole-2-thione have st rong p esticidal (5) and hy p -oglycemic (6) activity . Some are bis-(5-mer cap to-1, 3, 4- oxadiazole-2-y l) alk anes and they revealed the antifun gal activity (7); they also showed that a slight increase in activity takes p lace as the nu mber of methy lene group s increases (8). In this work ethy l adip ate [I] readily reacted with hy drazine hy drate to give adip ic dihydrazide [II], p hosp hours oxy chloride was emp loyed in the p rep aration of the bis-1,4-butane -1,3,4- oxadizole derivatives [III a-j] from adip ic dihydrazide and differ ent aromatic acids (scheme 1). All comp ounds were isolated and purified in satisfactory y ield and their p hy sical data element analysis, melting p oint mass-sp ectrum as shown in Table (1), and IR sp ectra are list ed in the exp erimental section. Ant imicrobial activity of some sy nthesized comp ounds was carried out against three typ es bacteria: Staphylococcus aurevs ATCC 25923, Escherichia coli AT CC 29522, and Pseudomonas aeruginosa AT CC 27853. Some of the sy nthesized comp ounds [IIIa-j], showed antibacterial activity as shown in Table (1). Experime nt The purity of the resultant comp ounds was checked by the melting p oints which are uncorrected and were taken on a“Electrot hermal“melting p oints apparatus (M ettle), micro analytical samples were analyzed by Iraqi Petroleum Comp any, mass sp ectra were recorded on shimadzo Qp 1000, Gas mass sp ectrometer (Gc-M s), by using a d irect insertion sy st em from the range of m/z 10-1000 and ionization ener gy (EI), of 20ev or 70ev. IR sp ectra were measured by using a perkin –Elmer 1310 infrared sp ectrop ho-meter on KBr disc. IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (3) 2009 Preparation of adipic dihydrazide [II] General Procedure (9) A mixture of diethy l adip ate [1] (0.01 mole) and excess hy drazine hy drate (0.02mole) were reflu xed for 30 min. The sep arated p reap ilate were filtered and washed with absolute ethanol and used without further p urification , yield 100% , m.p . = 182°c lit, 182°c (10 ). IR (KBr ) v max of these hy drazide show st retching bands (3300,3160,3060cm -1 ) NH2 and N-H group s, (1603 cm -1 ) C=0 amide I , (1540 cm -1 ) C=0 amid e II. Bis-1,4-(5-5-aryl -1,3,4-oxadizole-2y l ) butane [III a-j]. A mixture of acid d ihydrazide [II] (0.01mole) aromatic acid (0.02mole) and p hosp hours oxy chloride (5 ml) was reflu xed gently at (80-90) °C for 3 hours. Aft er cooling, the mixture was p oured into ice water and made basi c by adding sodium bicarbonate solution. The resulting solid was filtered, dried and recry-st allized from a prop er solvent to give the desired oxadiazole der ivatives. IR(KBr) v m axof these compounds show bands (2900-2825 cm -1 ) C-H aromatic ;(1600-1570 cm -1 ) C ═ N and C ═ C ; (1250-1200 cm -1 ) C-O-O, another p hy sical data are shown in Table (1). Bacterial Strains and Culture Media For antibacterial activity we used the following microor ganism : Staphyloco ccus aureus AT CC 25923, Escherich ia coli AT CC 25922 and Pseudomonas aeruginosa ATTC 27853, these were cultivated in Typticase Soy a agar (Difco), typt icase soy broth (Difco). Anti microbial activity The compounds of T able (1) were screened for their inhibitory effects against S.aureus, E. coli and Ps. aeruginosa by agar diffusion technique (11). The chemical comp ounds were dissolved in DM SO to give a final concentration of 1mg/ml. A 20ml of the sterilize tryp ticase Soy a agar med ia was p oured in a glass p lates of 9 cm in diameter and after solid ification aloop ful of overnight culture of each test org. was streaked on the surface of the p redried agar p lates, wells of 6mm were maele in the agar media by cork borer after the removal of the agar p illets, 100mg from each test comp ound were p laced in each well in a dup licate. The plates then were incubated at 37°C for 24 hours t o show the inhibition zone. Streptomy cin sulfate in a concentration of (0.1 mg/ml) were used as a standard growt h inhibitor for the bacteria. The inhibitory effect of DM SO was also examined which shows no inhibitory effect a gainst the test organisms. Results and Discussion S ynthesis of the compounds The sy nthesis of the bis-1,3 (5, 5aryl -1,3,4-oxadiazole-2-yl ) butane derivatives [IIIa-j] were accomp lished in accordance with t he sequence of r eactions dep icted in Scheme 1 Ethy l diester [I] were r eflu xed with 98% hy drazine hydrate to give after 30 min. the exp ected hy drazide [II], which was identified by melting p oint 182°C.lit, 182°C and by infrared sp ectroscopy . IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (3) 2009 The 2,5- disubst ituted – 1,3,4-oxadiazole were p repared by a route in which the acid dihydrazide [II] was condensed with the ap p rop riate aromatic acid in the p resence of p hosp hours oxy chloride . The st ructures of 1,3,4-oxadiazole derivatives [IIIa-j] were confirmed by infrared, C,H,N- analysis and mass sp ectroscopy . The IR sp ectra of [IIIa-j] were devoided of the amid e bands in the sp ectrum of acid dihydrazide [II] at (3300,3160,3050 cm -1 ) and (1630 cm -1 ) but showed (C═ N) st retching vibration band (12) ,in the range(1570-1600 cm -1 ) a band in the range (1200-1250 cm -1 ) of C-O-C st retching vibration combined with N-N band of 1,3,4-o xadiazole moiety (13,14). Evidence of the presence of aromatic ring which is the p resence of C ═ C aromatic ring which is the p resence of C ═ aromatic stretching band (1400-1570 cm -1) and out of p lane bend ing substitut ed aromatic sy stems in the range (700-850 cm -1) . Furt her structural proof for the oxadiazole derivatives was obtained from mass sp ectra, the fragmentation of 2,5disubst itut ed- 1,3,4- oxadiazole is sp ecific and indicative for the st ructure (15). The most informative fragments that were observed in the mass sp ectra of 1,3,4- oxadiazole deriv atives are list ed in Table (1) which gives a st rong evidence for the presence of oxadiazole ring. Anti microbial activity Table (1) shows that oxadiazole derivatives containing aryl substituent 64 (a, c, d, f, g, h, I, j) showed antimicrobial activity towards all kinds of bacteria used, althou gh their effect is less than that of t he standard used, the resist ance of some of these bacteria may be due t o the permeability of these compounds through the cell wall (16). Re ferences 1. Goswami, B.N.; Kataky J.C.S. and Baruah J.N., (1984) J. Heterocylic. Chem. 21: 1255. 2. Goswam, B.N. I.; Kataky J.C.S. and Baruah J.N., (1984) Indian J. Chem. 2313: 796. 3. M astat, A.O.; Abussaud M .; Tashtoush H. and Al-Talib M ., (2002) Pol. J. Pharamacol., 54: 55-59. 4. Gladwell, H.C.; Shewald R.J. and Burkhalter J.H., (1958) J. Am. Pharm. Assoc. Sci. Edn, 47: 799. 5. Kubo, H.; Hamura I.; Osuga S. and Sato R., (1969) Z assokenky u, 8: 42, Chem. Abst., 73: 108594 t (1970). 6. Kurihara, T.; H. Ito; Takeda, H. and Sagawa, K., (1971) Tohoku Yakka Daigaku Kenky 4 Nempol, 7: 43, Chem. Abst . 75: 10246 r (1971). 7. M ishra, V.K. and Bahel S.C. (1983) J. Indi an Chem. Soc., Lx: 867. 8. M ahmoud, M .J.; M ust afa I.F. and Omar–Eldeen H.I., (1999) Iraqi J. of Che m. 25(2): 157. 9. M ahmoud, M .J.; M ustafa I.F. and At to A.T., (1996) J. for Research and Studies, 11(5): 155. 10. Vo gel, I.A. (1974) “A text book of p ractical organic chemistry ”3 rd Ed., Longman, London, 439: 58. 11. Barry, A.L., (1976):“The Ant imicrobial Susceptibility Test: Practical and Practices ”. (Illus Loc and Febr iger, Philadelp hia):180. 12. lancelot, J.C. ; M aume, D. and Robba M . , (1980) J. Heterocy lic Chem. , 17: 625. 13. Dutta M .M .; Goswani B.N. and Kataky J.C.S., (1986) J. Heterocy lic Chem., 23: 793. 14. I. Felamin g and D.H. Williams (1966) “Sp ectroscop ic M ethods in Organic Ch emistry ”M C Graw – Hill Publishing Company Ltd. London. 15. Peet N. (1981) J. Barbuch, Org. M ass Sp ectrum., 18: 1601. 16. Sarkis G. Y.; Skenderian N.Y. and Abdul Ghani Z.G. ,(1989) The Iraqi Chem. So c. 14(1): 50 IBN AL- HAITHAM J. FO R PURE & APPL. SC I VO L.22 (3) 2009 S cheme 1 Table (1): The diameter of growth i nhibiti on z one *=Growth (no inhibition) Com p. S . aureus E. coli Ps . ae rugi nosa III a __ _* 11.0 10.0 III c 13.0 12.0 12.0 III d 13.0 10.0 10.0 III e __ _ __ _ __ _ III f 13.0 __ _ 12.0 III g 12.0 13.0 9.0 III h 12.0 __ _ __ _ III i 13.0 10.0 10.0 III j 11.0 8.0 __ _ St rept omycin Sulph ate 21.0 23.0 21.0