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

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

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