Department of Chemistry, Science Faculty, Ataturk University, Erzurum, Turkey.
J Enzyme Inhib Med Chem. 2010 Aug;25(4):476-9. doi: 10.3109/14756360903257900.
The inhibitory effects of some drugs on 6-phosphogluconate dehydrogenase from human erythrocytes have been investigated. For this purpose, initially, erythrocyte 6-phosphogluconate dehydrogenase was purified 3364 times in a yield of 58% by using ammonium sulfate precipitation and 2',5'-ADP Sepharose 4B affinity gel. A temperature of +4 degrees C was maintained during the purification process. Enzyme activity was determined with the Beutler method by using a spectrophotometer at 340 nm. This method was utilized for all kinetic studies. Many commonly used drugs were investigated in this study. Some drugs (ketotifen (K(i): 8.3 +/- 1.7 microM), dacarbazine (K(i): 10.1 +/- 0.7 microM), meloxicam (K(i): 50.9 +/- 13.2 microM), furosemide (K(i): 127 +/- 37.8 microM), methotrexate (K(i): 136.7 +/- 25.3 microM), metochloropramide hydrochloride (K(i): 2.1113 +/- 0.6979 mM), ritodrine hydrochloride (K(i): 6.0353 +/- 1.2783 mM), and gadopentetic acid (K(i): 73.4 +/- 21.9 mM)) inhibited enzyme activity in vitro. K(i) constants for the enzyme were found by means of Lineweaver-Burk graphs. All drugs showed non-competitive inhibition. In addition, IC(50) values of the drugs were determined by plotting activity percent vs [I].
已经研究了一些药物对人红细胞 6-磷酸葡萄糖酸脱氢酶的抑制作用。为此，最初通过使用硫酸铵沉淀和 2'，5'-ADP Sepharose 4B 亲和凝胶，在 +4°C 的温度下将红细胞 6-磷酸葡萄糖酸脱氢酶纯化 3364 倍，收率为 58%。酶活性通过 Beutler 法在分光光度计上在 340nm 处测定。该方法用于所有动力学研究。在这项研究中研究了许多常用药物。一些药物（酮替芬（K(i)：8.3 +/- 1.7 microM）、达卡巴嗪（K(i)：10.1 +/- 0.7 microM）、美洛昔康（K(i)：50.9 +/- 13.2 microM）、呋塞米（K(i)：127 +/- 37.8 microM）、甲氨蝶呤（K(i)：136.7 +/- 25.3 microM）、盐酸甲氧氯普胺（K(i)：2.1113 +/- 0.6979 mM）、盐酸利托君（K(i)：6.0353 +/- 1.2783 mM）和钆喷替酸（K(i)：73.4 +/- 21.9 mM））在体外抑制酶活性。通过 Lineweaver-Burk 图确定酶的 K(i)常数。所有药物均表现出非竞争性抑制。此外，通过绘制活性百分比与 [I] 的关系图来确定药物的 IC(50)值。
