Bazı Antibiyotiklerin Ksantin Oksidaz Enzimi Üzerine In Vitro Afinite Etkileri

Ksantin oksidaz KO , pürin katabolizmasının en son enzimidir. KO, amonyum sülfat çöktürmesi ve afinite kromatografisi ile saflaştırıldıktan sonra tıpta yaygın olarak kullanılan antibiyotiklerin etkileri incelendi. Etkisi araştırılan antibiyotikler; klaritromisin, gentamisin sülfat, sodyum ampisilin, sodyum sefazolin, klindamisin fosfat, rifamisin SV, kanamisin sülfattır. Antibiyotiklerin inhibisyon etkileri IC50 değerleri ile verilirken, ksantin oksidazın enzim aktivitesi, substrat olarak ksantin bileşiğini kullanılarak ölçüldü. İnhibisyon konsantrasyonuna karşı % enzim aktivitesi grafiği çizildi ve grafikten yararlanılarak IC50 değerleri hesaplandı. Ksantin oksidaz enzimi üzerine makrolid grubu antibiyotiklerinden gentamisin sülfat ve kanamisin sülfatın etkisi incelendi, gentamisin sülfat ksantin oksidazın enzim aktivitesini artırırken kanamisin sülfat enzim aktivitesini azaltmıştır. Ayrıca sodyum ampisilin ve rifamisin SV de enzim aktivitesini artırmıştır. Sefazolin sodyum, klaritromisin ve klindamisin fosfat enzim aktivitesini inhibe etmişlerdir. Özellikle, sefazolin sodyum, 5.4 x 10–4 mg/mL IC50değeri ile çalışılan antibiyotikler arasında en etkili inhibitördür. Sefazolin sodyum için bulunan IC50 değeri, ksantin oksidazın bilinen diğer inhibitörleri ile kıyaslandığında oldukça yakın bir değer olduğu görülmektedir

Affinity Effects of Some Antibiotics on Xanthine Oxidase Enzyme Activities In Vitro

Xanthine oxidase XO is the last enzyme of purine catabolism. XO was separately purified by ammonium sulphate precipitation and affinity chromatography. The effect of some antibiotics which is commonly used in clinical on purified xanthine oxidase was determined in vitro. The name of antibiotics was gentamycin sulfate, sodium ampicillin, cefazolin sodium, chlarithromycin, rifamycin SV, clindamycin phosphate and kanamycin sulfate. XO was determined using xanthine as a substrate and IC50 values of these antibiotics exhibiting inhibition effects were found from graphs of activity % by plotting concentration of the antibiotics. Macrolid group of antibiotics, the effects of gentamycin sulfate and kanamycin sulfate were determinated on xanthine oxidase. Gentamycin sulfate increased xanthine oxidase enzyme activity but kanamycin sulfate caused an inhibitory effect on xanthine oxidase enzyme activity. In addition, sodium ampicillin and rifamycin SV caused activation on enzyme activity. Cefazolin sodium, chlarithromycin and clindamicin phosphate indicated inhibitory effect on xanthine oxidase enzyme activity. Especially, cefazolin sodium is the most effective inhibitor in studied antibiotics with the value of 5.4 x 10–4 mg/mL. This value is close to the other values found for XO’s classical inhibitors.

___

  • 1. R. Hille, Structure and Function of Xanthine Oxidoreductase, Eur. J. Inorg. Chem., 2006 (2006) 1913.
  • 2. T. Nishino, K. Okamoto, B.T. Eger, E.F. Pai, T. Nishino, Mammalian xanthing oxidoreductase-mechanism of transition from xanthing dehydrogenase to xanthine oxidase, FEBS Journal, 275 (2008) 3278.
  • 3. W.R. Waud, K.V. Rajagopalan, Preparation of Bovine Milk Xanthine Oxidase as a Dehydrogenase Form , Arch. Biochem.Biophys., 172 (1976) 365.
  • 4. T.A. Krenitsky, J.V. Tuttle, Xanthine oxidase activities: Evidence for two catalytically different types, Arch. Biochem. Biophys, 185 (1978) 370.
  • 5. L. Stryer, Biochemistry, Fifth edition, Freeman, New York, 2002.
  • 6. C. E. Cross, Oxygen Radicals and Human Disease, Ann. Internal Med, 107 (1987) 526.
  • 7. P.A. Southorn, G. Powis, Free radicals in medicine. II Involvement in human disease, Mayo Clin. Proc., 63 (1988) 390.
  • 8. A. J. Hilliker, B. Duyf, D. Evans, J.P. Phillips, Uratenull rosy mutants of drosophila melanogaster are hypersensitive to oxygen stress, Proc. Natl. Acad. Sci. USA, 89 (1992) 4343.
  • 9. T.M. Millar, C.R. Stevens, N. Benjamin, R. Eisenthal, R. Harrison, D.R. Blake, Xanthine oxidoreductase catalyses the reduction of nitrates and nitrite to nitric oxide under hypoxic conditions, FEBS Letts, 427 (1998) 225.
  • 10. E. Edward, M.D. Morse, Toxic effects of drugs on erythrocytes. Ann. Clin. Sci., 18 (1988) 13.
  • 11. G. Jacobasch, S.M. Rappoport, Hemolytic anemias due to erythrocyte enzyme deficiencies. Mol. Asp. Med., 17 (1996) 143.
  • 12. M. Çiftçi, V. Türkoğlu, S. Aldemir, Effects of some antibiotics on glucose 6-phosphate dehydrogenase in sheep liver, Vet. Med., 47 (2002) 283.
  • 13. M. Ciftci, S. Beydemir, H. Yilmaz, et al., Effects of some drugs on rat erythrocyte 6-phosphogluconate dehydrogenase: An in vitro and in vivo study, Polish Journal Of Pharmacology, 54 (2002) 275.
  • 14. S. Umeki, Anti-inflammatory action of gentamycin through inhibitory effect on neutrophil NADPH oxidase activity, Comp. Biochem. Physiol. B Biochem. Mol. Biol., 110 (1995) 817.
  • 15. S. Beydemir, D. Kulacoglu, F. Ciftçi, O.I. Küfrevioglu, The effects of some antibiotics on sheep lens glucose 6-phosphate dehydrogenase in vitro, Eur. J. Ophthalmol, 13 (2003) 155.
  • 16. A.T. Çoban, B. Nalbantoglu, M.Y. Çil, H. Özdemir, O.I. Küfrevioğlu, Investigation of the inhibition effects of some antibiotics on human erythrocyte carbonic anhydrase isozymes, Tr. J. Med. Sci., 28 (1998) 407.
  • 17. A.D. Rodrigues, E.M. Roberts, D.J. Mulford, Y. Yao, D. Ouellet, Oxidative metabolism of clarithromycin in the presence of human liver microsomes. Major role for the cytochrome P4503A (CYP3A) subfamily, Drug Metab. Dispos., 25 (1997) 623.
  • 18. M.A. Bruce, S.D. Hall, B.D. Haehner-Daniels, J.C. Gorski, In vivo effect of clarithromycin on multiple cytochrome P450s, Drug Metab. Dispos., 29 (2001) 1023.
  • 19. C.M. Liu, Y.K. Chen, T.H. Yang, S.Y. Hsieh, M.H. Hung, E.T. Lin, High-performance liquid chromatographic determination of clindamycin in human plasma or serum: application to the bioequivalency study of clindamycin phosphate injections, J. Chromatogr. B, 696 (1997) 298.
  • 20. J.W. Munson, E.J. Kubiak, A high-performance liquid chromatographic assay for clindamycin phosphate and its principal degradation product in bulk drug and formulations, J. Pharm. Biomed. Anal. 3 (1985) 523.
  • 21. J. Schlessinger, A. Menter, M. Gold, et al., Clinical safety and efficacy studies of a novel formulation combining 1.2% clindamycin phosphate and 0.025% tretinoin for the treatment of acne vulgaris, J Drugs Dermatol , 6 (2007) 607.
  • 22. G. Mancuso, N. Masara, Contact urticaria and severe anaphylaxis from rifamycin SV., Contact Dermatitis, 27 (1992) 124.
  • 23. F. Erel, M. Karaayvaz, M. Deveci, N. Ozanguc, Severe anaphylaxis from rifamycin SV. Ann allergy asthma, Immunol, 81 (1998) 257.
  • 24. F. Garcia, J. Blanco, P. Carretero, et al., Anaphylactic reactions to topical rifamycin, Allergy, 54 (1999) 527.
  • 25. A. Magnan, L. Venemalm, F. Porri, D. Vervloet, Anaphylactic reaction to rifamycin SV: presence of specific IgE antibodies, J Allergy Clin Immunol, 103 (1999) 954.
  • 26. Y.Y. Lau, G.D. Hanson, et al., Determination of rifampin in human plasma by high-performance liquid chromatography with ultraviolet detection, J. Chromatogr. B, 676 (1996) 147.
  • 27. K.J. Swart, M. Papgis, Automated high-performance liquid chromatographic method for the determination of rifampicin in plasma, J. Chromatogr. A, 593 (1992) 21.
  • 28. K.C. Jindal, R.S. Chaudhary, S.S. Gangwal, A. Singla, K.S. Khanna, High-performance thin-layer chromatographic method for monitoring degradation products of rifampicin in drug excipient interaction studies, Chromatogr. A, 685 (1994) 195.
  • 29. M.I. Walash, F. Belal, M. E. Metwally, M.M. Hefnawy, Spectrophotometric determination of rifampin in presence of its metabolites and degradation products in pharmaceutical preparations, Anal. Lett., 26 (1993) 1905.
  • 30. A. Espinosa-Mansilla, M.I. Acedo Valenzuela, F. Salinas, Adsorption of mercury, cadmium and lead from aqueous solution on heat-treated and sulphurized activated carbon, Anal. Chim. Acta, 376 (1998) 365.
  • 31. A. Espinosa-Mansilla, M.I. Acedo Valenzuela, A. Munoz de la Pena, F. Salinas, Comparative study of partial least squares and a modification of hybrid linear analysis calibration in the simultaneous spectrophotometric determination of rifampicin, pyrazinamide and isoniazid, Anal. Chim. Acta, 427 (2001) 129.
  • 32. S.A. Benetton, E.R.M. Kedor-Hackmann, R.M. Santoro, V.M. Borges, Visible spectrophotometric and first-derivative UV spectrophotometric determination of rifampicin and isoniazid in pharmaceutical preparations, Talanta, 47 (1998) 639.
  • 33. S. Sinan, F. Kockar, N. Gencer, H. Yildirim, O. Arslan, Amphenicol and macrolide derived antibiotics inhibit paraoxonase enzyme activity in human serum and human hepatoma cells (HepG2) in vitro, Biochemistry (Moscow), 71 (2006) 46.
  • 34. S. Sinan, F. Kockar, N. Gencer, H. Yildirim, O. Arslan, Effects of some antibiotics on paraoxonase from human serum in vitro and from mouse serum and liver in vivo, Biol. Pharm. Bull., 29 (2006) 1559.
  • 35. M. Çiftçi, V. Türkoğlu, S. Aldemir, Effects of some antibiotics on glucose 6-phosphate dehydrogenase in sheep liver, Vet. Med. – Czech., 47 (2002) 283.
  • 36. T. Honjo, A. Watanabe, Clinical experience on sulfate/ cefoperazone in the pediatric field, Jpn J Antibiot, 37 (1984) 32.
  • 37. L.K. Pickering, D.M. O’Connor, D. Anderson, A.C. Bairan, R.D. Feigin, J.D. Cherry, Clinical and pharmacologic evaluation of cefazolin in children, J Infect Dis, 128 (1973) 407.
  • 38. M. Turck, R.A. Clark, H.N. Beaty, K.K. Holmes, W.W. Karney, L.B. Reller, Cefazolin in the treatment of bacterial pneumonia, J Infect Dis, 128 (1973) 382.
  • 39. S.M. Singhvi, A.F. Heald, H.H. Gadebusch, M.E. Resnick, L.T. Difazio, M.A. Leitz, Human serum protein binding of cephalosporin antibiotic in vitro, J Lab Clin Med., 89 (1977) 414.
  • 40. M. Ciftci, O.I. Kufrevioglu, M. Gundogdu, I. Ozmen, Effects of some antibiotics on enzyme activity of glucose 6-phosphate dehydrogenase from the human erythrocytes, Pharmacol Res, 41 (2000) 109.
  • 41. Ş. Beydemir, M. Çiftçi, İ. Özmen, M.E. Büyükokuroğlu, H. Özdemir, Ö. İ. Küfrevioğlu, Effects of some medical drugs on enzyme activities of carbonic anhydrase from human erythrocytes in vitro and from rat erythrocytes in vivo, Pharmacological Research, 42 (2000) 187.
  • 42. N. Özer, M. Müftüoğlu, D. Ataman, A. Ercan, I. H. Öğüş, Simple, High-Yield Purification Of Xanthine Oxidase From Bovine Milk, J. Biochem. Biophys. Methods, 39 (1999) 153.
  • 43. V. Massey, P.E. Brumby, H. Komai, Studies on milk xanthine oxidase: Some spectral and kinetic properties, J Biol Chem., 244 (1969) 1682.
  • 44. M.M. Bradford, A rapid and sensitive method for the quantition of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem. 72 (1976) 248.
  • 45. U.K. Laemmli, Cleavage of structual proteins during the assembly of the head of bacteriophage T4, Nature, 227 (1970) 680.
  • 46. C. R. Stevens, T. M. Millar, J. G. Clinch, J. M. Kanczler, T. Bodamyali, D.R. Blake, Antibacterial properties of xanthine oxidase in human milk, The Lancet, 356 (2000) 929.
  • 47. R.M. Hochster, M. Kates, J.H. Quastel, Metabolic inhibitors, Academic Pres, chap. 4, 1972. 66.
  • 48. E.O. Farombi, M. Ekor, Curcumin attenuates gentamicin-induced renal oxidative damage in rats, Food Chem. Toxicol., 44 (2006) 1443.
  • 49. A. Kuhad, N. Tirkey, S. Pilkhwal, K. Chopra, Effect of Spirulina, a blue green algae, on gentamicininduced oxidative stress and renal dysfunction in rats, Fundam. Clin. Pharmacol., 20 (2006) 121.
  • 50. B.H. Ali, Agents ameliorating or augmenting experimental gentamicin nephrotoxicity: some recent research, Food Chem Toxicol, 41 (2003) 1447.
  • 51. H.D. Humes, J.M. Weinberg, Toxic nephropathies. In: Brenner MB, Rector FE Jr, eds: Kidney, Philadelphia: Saunders, 1986, 453.
  • 52. J.M. Eisenberg, H. Koffer, H.A. Glick, M.L. Connell, L.E. Loss, G.H. Talbot, N.H. Shusterman, B.L. Strom, What is the cost of nephrotoxicity associated with aminoglycosides?, Ann Intern Med, 107 (1987) 900.
  • 53. A.A. Al-Majed, A.M. Mostafa, A.C. Al-Rikabi, O.A. Al-Shabanah, Protective effects of oral Arabic gum administration on gentamicin-induced nephrotoxicity in rats, Pharmacol Res, 46 (2002) 445.
  • 54. I. Karahan, A. Atezzahin, S. Yilmaz, A.O. Ceribazi, F. Sakin, Protective effect of lycopene on gentamicininduced oxidative stress and nephrotoxicity in rats, Toxicology, 215 (2005) 198.
  • 55. R. Dhanarajan, P. Abraham, B. Isaac, Protective effect of ebselen, a selenoorganic drug, against gentamicin-induced renal damage in rats, Basic Clin Pharmacol Toxicol, 99 (2006) 267.
  • 56. H. Parlakpinar, S. Tasdemir, A. Polat, A. Karabulut, N. Vardi, M. Ucar, M. Yanilmaz, A. Kavakli, A. Acet, Protective effect of chelerythrine on gentamicininduced nephrotoxicity, Cell Biochem Funct., 24 (2006) 41.
  • 57. P.D. Walker, Y. Barri, S.V. Shah, Oxidant mechanisms in gentamicin nephrotoxicity, Renal Fail, 21 (1999) 433.
  • 58. J. Pedraza-Chaverri, D. Barrera, P.D. Maldonado, Y.I. Chirino, N.A. Macias-Ruvalcaba, O.N. MedinaCampos, L. Castro, M.I. Salcedo, R.S. HernandezPando, Allylmercaptocysteine scavenges hydroxyl radical and singlet oxygen in vitro and attenuates gentamicin-induced oxidative and nitrosative stress and renal damage in vivo, BMC Clin Pharmacol, 4 (2004) 5.
  • 59. M. Kadkhodaee, H. Khastar, M. Faghihi, R. Ghaznavi, M. Zahmatkesh, Effects of co-supplementation of vitamins E and C on gentamicin-induced nephrotoxicity in rat, Exp Physiol., 90 (2005) 571.
  • 60. S. Cuzzocrea, E. Mazzon, L. Dugo, I. Serraino, R. Di Paola, D. Britti, A. De Sarro, S. Pierpaoli, A. Caputi, E. Masini, D. Salvemini, A role for superoxide in gentamicin-mediated nephropathy in rats, Eur J Pharmacol, 450 (2002) 67.
  • 61. K. Aoyagi, K. Akiyama, S. Shahrzad, C. Tomida, A. Hirayama, S. Nagase, K. Takemura, A. Koyama, S. Ohba, M. Narita, Formation of guanidinosuccinic acid, a stable nitric oxide mimic, from argininosuccinic acid and nitric oxide-derived free radicals, Free Radic Res, 31 (1999) 59.
  • 62. V. Tugcu, E. Ozbek, A.I. Tasci, E. Kemahli, A. Somay, M. Bas, C. Karaca, T. Altug, M.B. Cekmen, H.K. Ozdogan, Selective nuclear factor kappa-B inhibitors, pyrolidium dithiocarbamate and sulfasalazine, prevent the nephrotoxicity induced by gentamicin, BJU Int, 98 (2006) 680.
  • 63. M. Kavutcu, O. Canbolat, S. Ozturk, E. Olcay, S. Ulutepe, C. Ekinci, I.H. Gokhun, I. Durak, Reduced enzymatic antioxidant defense mechanism in kidney tissues from gentamicin-treated guinea pigs: effects of vitamins E and C, Nephron, 72 (1996) 269.
  • 64. E.F. Gale, E. Cundliffe, P.E. Reynolds, et al, The molecular basis of antibiotic action, Second edition, John Wiley & Sons, New York, 1981.
  • 65. S. Douthwaite, Interaction of the antibiotics clindamycin and lincomycin with Escherichia coli 23S ribosomal RNA, Nucleic Acids Res., 20 (1992) 4717.
  • 66. A. Ianaro, A. Ialenti, P. Maffia, L. Sautebin, L. Rombola, R. Carnuccio, T. Iuvone, F. D’Acquisto, M. Di Rosa, Anti-inflammatory activity of macrolide antibiotics, J. Pharmacol. Exp. Ther., 292 (2000) 156.
  • 67. M.T. Labro, Anti-inflammatory activity of macrolides: a new therapeutic potential, J. Antimicrob. Chemother., 41 (1998) 37.
  • 68. H. Biri, H.S. Ozturk, M. Kacmaz, K. Karaca, H. Tokucoglu, I. Durak, Activities of DNA turnover and free radical metabolizing enzymes in cancerous human prostate tissue, Cancer Invest., 17 (1999) 314.
  • 69. R. Memişogullari, S. Taysi, E. Bakan, I. Capoglu, Antioxidant status and lipid peroxidation in type II diabetes mellitus, Cell Biochem. Funct., 21(2003) 291.
  • 70. M.F. Polat, S. Taysi, M. Gul, O. Cikman, I. Yilmaz, E. Bakan, F. Erdogan, Oxidant/antioxidant status in blood of patients with malignant breast tumour and benign breast disease, Cell Biochem. Funct., 20 (2002) 327.
  • 71. S. Taysi, M. Gul, R.A. Sari, F. Akcay, N. Bakan, Oxidant/ antioxidant status in serum of patients with systemic lupus erythematosus, Clin. Chem. Lab. Med., 40 (2002) 684.
  • 72. S. Taysi, I. Koçer, R. Memişoğullari, A. Kiziltunç, Serum oxidant/antioxidant status in serum of patients with Behçet’s disease, Ann. Clin. Lab. Sci., 32 (2002) 377.
  • 73. S. Taysi, F. Polat, M. Gul, R.A. Sari, E. Bakan, Lipid peroxidation, some extracellular antioxidants and antioxidant enzymes in serum of patients with rheumatoid arthritis, Rheumatol. Int., 21 (2002) 200.
  • 74. Yagi, K., Increased lipid peroxides initiates atherogenesis, Bioessays, 1, 58–60, 1984.
  • 75. C.E. Cross, B. Halliwell, E.T. Borish, W.A. Pryor, B.N. Ames, R.L. Saul, et al., Oxygen radicals and human disease, Ann. Intern. Med., 107 (1987) 526.
  • 76. F. Döner, N. Delibaş, H. Doğru, I. Sari, B. Yorgancigil, Malondialdehyde levels and superoxide dismutase activity in experimental maxillary sinusitis, Auris Nasus Larynx, 26, (1999) 287.
  • 77. C. Uslu, S. Taysi, N. Bakan, Lipid peroxidation and some antioxidant enzyme activities in experimental maxillary sinusitis, Ann. Clin. Lab. Sci., 33 (2003) 18.
  • 78. O. Akyol, H. Herken, E. Uz, E. Fadillioglu, S. Unal, S. Sogut, H. Ozyurt, H.A. Savas, The indices of endogenous oxidative and antioxidative processes in plasma from schizophrenic patients. The possible role of oxidant/antioxidant imbalance, Prog. NeuroPsych. Biol. Psych., 26 (2002) 995.
  • 79. K. Kiyomiya, N. Matsushita, S. Matsuo, M. Kurebe, Cephaloridine-induced inhibition of cytochrome c oxidase activity in the mitochondria of cultured renal epithelial cells (LLC-PK1) as a possible mechanism of its nephrotoxicity, Toxicology and Applied Pharmacology, 167 (2000) 151.
  • 80. D.Z. Hsu, C.T. Liu, Y.H. Li, P.Y. Chu, M.Y. Liu, Protective effect of daily sesame oil supplement on gentamicininduced renal injury in rats, Shock, 33 (2010) 88.
  • 81. M. Alirezai, B. Gerlach, A. Horvath, et al, Results of a randomized, multicentre study comparing a new water-based gel of clindamycin 1% versus clindamycin 1% topical solution in the treatment of acne vulgaris, Eur J Dermatol, 15 (2005) 274.
  • 82. W.J. Cunliffe, C. Fernandez, R. Bojar, et al., An observed-blind parallel-group, randomized, multicentre clinical and microbiological study of a topical clindamycin/zinc gel and a topical clindamycin lotion in patients with mild/moderate acne, J. Dermatol Treat., 16 (2005) 213.
  • 83. D. Chassard, R. Kanis, F. Namour, et al., A single centre, open-label, cross-over study of pharmacokinetics comparing topical zinc/clindamycin gel (Zindaclin) and topical clindamycin lotion (Dalacin T) in subjects with mild to moderate acne, J. Dermatol. Treat., 17 (2006) 154.
  • 84. B. Aktan, S. Taysi, K. Gümüştekin, H. Üçüncü, R. Memişoğullari, K. Save, N. Bakan, Effect of macrolide antıbiotics on nitric oxide synthase and xanthine oxidase activities, and malondialdehyde level in erythrocyte of the guinea pigs with experimental otitis media with effusion, Pol. J. Pharmacol., 55 (2003) 1105.
Hacettepe Journal of Biology and Chemistry-Cover
  • ISSN: 2687-475X
  • Yayın Aralığı: Yılda 4 Sayı
  • Başlangıç: 1972
  • Yayıncı: Hacettepe Üniversitesi, Fen Fakültesi