Alım Zuhal, Kılınç Namık, İşgör Mehmet M, Şengül Bülent, Beydemir Şükrü
Department of Chemistry, Faculty of Sciences, Atatürk University, 25240, Erzurum, Turkey.
Department of Biochemistry, Faculty of Veterinary Sciences, Mustafa Kemal University, 31000, Hatay, Turkey.
Chem Biol Drug Des. 2015 Oct;86(4):857-63. doi: 10.1111/cbdd.12561. Epub 2015 Apr 7.
Carbonic anhydrases (CAs) are known as a drug-target enzymes. The inhibitors of the enzyme are important compounds for discovering new therapeutic agents and understanding in detail protein-drug interactions at the molecular level. For this purpose, the in vitro effects of some anti-inflammatory agents such as tenoxicam, fluorometholone acetate, and dexamethasone were investigated on esterase activity of human erythrocyte CA-I and CA-II in this study. hCA-I and hCA-II were purified by affinity chromatography with a yield of 47.25% and 87%, and a specific activity of 642.8 EU/mg proteins and 5576.9 EU/mg proteins, respectively. SDS-PAGE was performed to determine the purity of the enzymes. Inhibitory effects of the drugs on hCA-I and hCA-II were determined by spectrophotometric method. IC50 values for hCA-I and hCA-II were 0.198, 2.18, 11.7, 0.11, 17.5 and 14 μm using tenoxicam, fluorometholone acetate, and dexamethasone, respectively. For fluorometholone acetate and dexamethasone, Ki values from Lineweaver-Burk plots were obtained as 1.044 and 21.2 μm (noncompetitive) for hCA-I and 9.98 and 8.66 μm (non-competitive) for hCA-II. In conclusion, tenoxicam, fluorometholone acetate, and dexamethasone showed potent inhibitory effects on esterase activity of hCA-I and hCA-II isozymes under in vitro conditions.
碳酸酐酶(CAs)是一类已知的药物靶点酶。该酶的抑制剂是发现新治疗药物以及在分子水平详细了解蛋白质-药物相互作用的重要化合物。为此,本研究考察了替诺昔康、醋酸氟米龙和地塞米松等一些抗炎药对人红细胞CA-I和CA-II酯酶活性的体外作用。通过亲和层析纯化hCA-I和hCA-II,产率分别为47.25%和87%,比活性分别为642.8 EU/mg蛋白质和5576.9 EU/mg蛋白质。采用SDS-PAGE测定酶的纯度。通过分光光度法测定药物对hCA-I和hCA-II的抑制作用。替诺昔康、醋酸氟米龙和地塞米松对hCA-I和hCA-II的IC50值分别为0.198、2.18、11.7、0.11、17.5和14 μ m。对于醋酸氟米龙和地塞米松,根据Lineweaver-Burk图得到hCA-I的Ki值分别为1.044和21.2 μ m(非竞争性),hCA-II的Ki值分别为9.98和8.66 μ m(非竞争性)。总之,替诺昔康、醋酸氟米龙和地塞米松在体外条件下对hCA-I和hCA-II同工酶的酯酶活性显示出较强的抑制作用。