Investigation of the In vitro Effects of Sultamycillin, Cefdinir, Vancomycin Hydrochloride and Colistimethate Sodium Agents Used as Antibiotics on Enzyme Activity of Human Serum Paraoxonase-I

Yıl 2019, , 349 - 357, 24.03.2019

Esra Dilek , Merve Berivan Özçelik Serhat Yıldırım

https://doi.org/10.18185/erzifbed.467627

Öz

Paraoxonase (PON1; E.C.3.1.8.1) is synthesized in the liver and hydrolyzes its specific substrate, paraoxone. It is an enzyme from the group of A-esterases bound to high density lipoprotein (HDL). The aim of this study was to investigate the in vitro effects of sultamicillin, cefdinir, vancomycin hydrochloride and colistimethate sodium active substances on the PON1 enzyme activity in human serum. The PON1 enzyme from the human serum was purified using ammonium sulfate precipitation (in the range of 60-80%), ion exchange chromatography, and gel filtration chromatography techniques approximately 320,7 fold with 44,6% yield. The paraoxonase activity of the enzyme at 412 nm was investigated to investigate the inhibition effects of the active substances. Paraoxone was used as the substrate. Sultamycillin and vancomycin hydrochloride showed noncompetetive inhibition for the PON1 enzyme, whereas cefdinir showed a uncompetitive inhibition and colistimethate sodium showed a competitive inhibition effect. Ki values are determined for sultamycillin as 38.3 mM; for vancomycin hydrochloride as 5.08 mM; for cefdinir as 0.86 mM; and for colistimethate sodium as 17.01 µM. According to the results, it was determined that the most effective inhibitor colistimethate sodium (IC50: 13.59 µM) was found on the PON1 enzyme with 50% reduction in activity at very low concentration

Anahtar Kelimeler

Paraoxonase1, sultamycillin, cefdinir, vancomycin hydrochloride, colistimethate sodium

Antibiyotik Olarak Kullanılan Sultamisilin, Sefdinir, Vankomisin Hidroklorür ve Kolistin Etken Maddelerinin İnsan Serum Paraoksonaz1 Enzim Aktivitesi Üzerine in vitro Etkilerinin İncelenmesi

Öz

Paraoksonaz (PON1;
E.C.3.1.8.1), karaciğerde sentezlenir ve spesifik substratı olan paraoksonu
hidroliz eder. Yüksek yoğunluklu lipoprotein (HDL)’ne bağlı arilesterazlar
grubundan bir enzimdir. Bu çalışmanın amacı, sıklıkla kullanılan antibiyotik
ilaçların sultamisilin, sefdinir, vankomisin hidroklorür ve kolistimetat sodyum
etken maddelerinin insan serumunda bulunan PON1 enzim aktivitesi üzerine in vitro etkilerini araştırmaktır. İnsan
serumundan PON1 enzimi amonyum sülfat çöktürmesi (%60-80 aralığında), iyon
değişim kromatografisi ve jel filtrasyon kromatografisi teknikleri kullanılarak
%44,6 verimle
yaklaşık 320,7 kat saflaştırıldı. Etken maddelerin inhibisyon etkilerini incelemek üzere enzimin 412
nm’deki paraoksonaz aktivitesine bakıldı. Substrat olarak paraokson kullanıldı.
PON1 enzimi için sultamisilin ve vankomisin hidroklorür yarışmasız inhibisyon
gösterirken, sefdinir yarı yarışmalı inhibisyon, kolistimetat sodyum ise
yarışmalı inhibisyon etkisi gösterdi. Ki değerleri ise sultamisilin için 38.3
mM; vankomisin hidroklorür için 5.08 mM; sefdinir için 0.86 mM ve kolistimetat
sodyum için 17.01 µM olarak hesaplandı. Bu sonuçlara göre enzim aktivitesi
üzerine etki gösteren ilaç etken maddelerinin içinde, enzim aktivitesini düşük
konsantrasyonda %50 azaltması ile PON1 enzimi üzerinde de en kuvvetli inhibitör
kolistimetat sodyum (IC₅₀: 13.59 µM) olarak tespit edildi. 

Anahtar Kelimeler

Paraoksonaz1, sultamisilin, sefdinir, vankomisin hidroklorür, kolistin

Kaynakça

• Dilek E. İnsan Serumundan Paraoksonaz-1 ve İnsan Trombositlerinden Siklooksigenaz-1 Enziminin Saflaştırılması, Bazı İlaçların Bu Enzimlerin Aktiviteleri Üzerine Etkilerinin İncelenmesi. Doktora Tezi, Atatürk Üniversitesi Fen Bilimleri Enstitüsü, 2012, Erzurum.
• Durrington P. N., Mackness B., Mackness, M.J. Paraoxonase and atherosclerosis. Arteriosclerosis, thrombosis, and vascular biology. 2001;21(4), 473-480.
• Ng C. J., Shih D. M., Hama S. Y., Villa N., Navab M., Reddy S. T. The paraoxonase gene family and atherosclerosis, Free Radical Biology and Medicine, 2005;38(2), 153-163.
• Mackness B., Durrington P.N., Mackness M.I. Human serum paraoxonase. General Pharmacology: The Vascular System, 1998;31(3), 329-336.
• Canales A. Sanchez-Muniz F.J. Paraoxonase, something more than an enzyme? Medicina clinica, 2003;121(14), 537-548.
• Blatter M.C., James R.W., Messmer S., Barja F. Pometta D. Identification of a distinct human high‐density lipoprotein subspecies defined by a lipoprotein‐associated protein, K‐45: Identity of K‐45 with paraoxonase. European journal of biochemistry, 1993;211(3), 871-879.
• Gan K N., Smolen A., Eckerson H.W., La Du B.N. Purification of human serum paraoxonase / arylesterase. Evidence for one esterase catalyzing both activities. Drug Metabolism and Disposition, 1991;19(1), 100-106.
• Aviram M., Rosenblat M., Bisgaier C.L., Newton R.S., Primo-Parmo, S.L., La Du B.N. Paraoxonase inhibits high-density lipoprotein oxidation and preserves its functions. A possible peroxidative role for paraoxonase. The Journal of clinical investigation, 1998;101(8), 1581-1590.
• Lee J., Prohaska J.R. Thiele D.J. Essential role for mammalian copper transporter Ctr1 in copper homeostasis and embryonic development. Proceedings of the National Academy of Sciences, 2001;98(12), 6842-6847.
• Mackness M., Mackness, B. Paraoxonase 1 and atherosclerosis: is the gene or the protein more important? Free Radical Biology and Medicine, 2004;37(9), 1317-1323.
• Seres I., Paragh G., Deschene E., Fulop Jr T., Khalil A. Study of factors influencing the decreased HDL associated PON1 activity with aging. Experimental gerontology, 2004;39(1), 59-66.
• Kudchodkar B.J., Lacko A.G., Dory L., Fungwe T.V. Dietary fat modulates serum paraoxonase 1 activity in rats. The Journal of nutrition, 2000;130(10), 2427-2433.
• Kleemola P., Freese R., Jauhiainen M., Pahlman R., Alfthan G., Mutanen M. Dietary determinants of serum paraoxonase activity in healthy humans. Atherosclerosis, 2002;160(2), 425-432.
• Nelson D.L., Cox M.M. Lehninger principles of biochemistry. WH Freeman & Co, New York, 2005,4, 1100.
• Bayram E, Şentürk M, Küfrevioğlu Ö.İ, Supuran C.S. Invitro inhibition of salicylic acid derivatives on human cytosolic carbonic anhydrase isozymes I and II. Bioorganic and Medicinal Chemistry. 2008; 16: 9101-9105.
• Sarikaya S.B, Sisecioglu M, Cankaya M, Gulcin I, Ozdemir H. Inhibition profile of a series of phenolicacids on bovine lactoperoxidase enzyme. J. Enzyme Inhib. Med. Chem. 2014; 8: 1-5.
• Dilek E.;Caglar S. Effects of mono and dinuclear copper (II) complexes derived from non-steroidal anti-inflammatory drug naproxen on human serum paraoxanase1 (PON1) activity. Intl. J. Pharm. Chem. 2015; 5(6), 189-195.
• Dilek E. Polat M.F. In Vitro Inhibition Of Three Different Drugs Used In Rheumatoid Arthritis Treatment On Human Serum Paraoxanase 1 Enzyme Activity. Protein andPeptideLetters. 2016; 23, 3-8
• Ciftçi M, Küfrevioglu OI, Gündogdu M, Ozmen I. Effects of some antibiotics on enzymeactivity of glucose-6-phosphate dehydrogenase from human erythrocytes. Pharmacol Res. 2000; 41:109-13.
• Renault F., Chabrière E., Andrieu J. P., Dublet B., Masson P. Rochu D. Tandem purification of two HDL-associated partner proteins in human plasma, paraoxonase (PON1) and phosphate binding protein (HPBP) using hydroxyapatite chromatography. Journal of Chromatography B, 2006;836(1-2), 15-21.
• Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry, 1976;72(1-2), 248-254.
• Dilek E.B., Küfrevioğlu Ö.İ. Beydemir Ş. Impacts of some antibiotics on human serum paraoxonase 1 activity. Journal of enzyme inhibition and medicinal chemistry, 2013;28(4), 758-764.
• Dilek E., Caglar S., Erdogan K., Caglar B. Sahin O. Synthesis and characterization of four novel palladium (II) and platinum (II) complexes with 1‐(2‐aminoethyl) pyrrolidine, diclofenac and mefenamic acid: In vitro effect of these complexes on human serum paraoxanase1 activity. Journal of biochemical and molecular toxicology, 2018;32(4), e22043.
• Segel I. H. Enzyme Kinetics, John Wiley and Sons, 1975; New York.
• Telefoncu A. Temel ve Uygulamalı Enzimoloji. Ege üniversitesi, Fen Edebiyat Fakültesi Yayını (Der), 1986;59 s, İzmir.
• Ekinci D. Beydemir S. Effect of some analgesic on purified paraoxonase-1 from human serum”, Journal of Enzyme Inhibition and Medicinal Chemistry, 2009; 24(4), 1034-1039.
• Türkeş C. İnsan Serumundan Paraoksonaz-1 Enziminin Saflaştırılması ve Bazı İlaçların Enzim Aktivitesi Üzerine Etkilerinin İncelenmesi. Yüksek Lisans Tezi, Atatürk Üniversitesi Fen Bilimleri Enstitüsü, 2010;Erzurum.