Carbonic anhydrase inhibitors: Cloning, characterization, and inhibition studies of the cytosolic isozyme III with sulfonamides
Carbonic anhydrase inhibitors: Cloning, characterization, and inhibition studies of the cytosolic isozyme III with sulfonamides
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DOI：
10.1016/j.bmc.2007.08.037
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发表时间：
2007-12-01
影响因子：
3.5
通讯作者：
Supuran, Claudiu T.
中科院分区：
文献类型：
--
作者：
Nishimori, Isao;Minakuchi, Tomoko;Supuran, Claudiu T.
用GST-融合蛋白法克隆并纯化了人胞质碳酸酐酶（hCA，EC 4.2.1.1）同工酶Ⅲ（hCA Ⅲ）。重组纯hCA III在20 ℃和pH 7.5下具有以下CO2水合反应的动力学参数：k（cat）为1.3 x 10（4）s（-1），k（cat）/ K-M为2.5 x 10（5）M-1 s（-1），与遗传相关的胞质异构体hCA I和II相比，其是生理反应的较慢催化剂。一个抑制研究与库的磺胺类药物和一个氨基磺酸盐，其中一些是临床使用的化合物，报告。与hCA I和II相比，hCA III不太容易被这些化合物抑制，对于hCA I和II，早期检测到许多低纳摩尔抑制剂。最好的hCA III抑制剂是prontosil、舒必利、indisulam、benzolamide、aminobenzolamide和4-amino-6-chloro-benzene-1，3-disulfonamide，其K（I）在2.3-18.1 μ M范围内。临床使用的化合物如乙酰唑胺、醋甲唑胺、乙氧唑胺、多佐胺、布林佐胺、托吡酯、唑尼沙胺、塞来昔布和伐地昔布是较不有效的hCA III抑制剂，其亲和力在154-2200 μ M的范围内。这是第一个研究中，低微摩尔hCA III抑制剂的报告。(C)2007爱思唯尔有限公司版权所有。
The cytosolic human carbonic anhydrase (hCA, EC 4.2.1.1) isozyme III (hCA III) has been cloned and purified by the GST-fusion protein method. Recombinant pure hCA III had the following kinetic parameters for the CO2 hydration reaction at 20 degrees C and pH 7.5: k(cat) of 1.3 x 10(4) s(-1) and k(cat)/ K-M of 2.5 x 10(5) M-1 s(-1), being a slower catalyst for the physiological reaction as compared to the genetically related cytosolic isoforms hCA I and II. An inhibition study with a library of sulfonamides and one sulfamate, some which are clinically used compounds, is reported. hCA III is less prone to be inhibited by these compounds as compared to hCA I and II for which many low nanomolar inhibitors were detected earlier. The best hCA III inhibitors were prontosil, sulpiride, indisulam, benzolamide, aminobenzolamide, and 4-amino-6-chloro-benzene-1,3-disulfonamide which showed K(I)s in the range of 2.3-18.1 mu M. Clinically used compounds such as acetazolamide, methazolamide, ethoxzolamide, dorzolamide, brinzolamide, topiramate, zonisamide, celecoxib, and valdecoxib were less effective hCA III inhibitors, with affinities in the range of 154-2200 mu M. This is the first study in which low micromolar hCA III inhibitors are reported. (C) 2007 Elsevier Ltd. All rights reserved.