Hydrolase Activity of the Genetic Variants of Human Alpha-1-Acid Glycoprotein
Hydrolase Activity of the Genetic Variants of Human Alpha-1-Acid Glycoprotein
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DOI：
10.1021/acs.molpharmaceut.1c00614
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发表时间：
2022-03-07
影响因子：
4.9
通讯作者：
Maruyama，Toru
中科院分区：
文献类型：
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作者：
Kono，Kenta;Fujimura，Rui;Maruyama，Toru
在人血浆中，左旋多巴的酯型前药（命名为ONO-2160）的主要水解剂是α-1-酸性糖蛋白（AGP），它是摩尔比为3：1至2：1的F1*S和A变体的混合物。在这项研究中，通过评估F1*S和A变体对ONO-2160水解的贡献和鉴定AGP水解酶活性位点，研究了AGP酯酶样活性的机制。我们发现，虽然这两种变体水解ONO-2160，但它们的水解酶活性不同。F1*S变体的固有血浆清除率（0.441 mL/h/mg蛋白）约为A变体（0.0148 mL/h/mg蛋白）的30倍，表明F1*S变体对AGP酯酶样活性的贡献最大。为了确定水解酶活性位点的AGP，我们进行了抑制ONO-2160水解的研究，使用12个AGP结合药物与不同的配体结合常数和结合选择性的两个AGP变体。活性抑制与配体结合F1*S变体的常数呈正相关。此外，对F1*S变体具有高亲和力的化合物对ONO-2160水解的抑制作用最大。总之，我们的数据表明ONO-2160主要在其配体结合位点被F1*S变体水解。
In human plasma, the main agent of hydrolysis of the ester-type prodrug of levodopa, designated ONO-2160, is alpha-1-acid glycoprotein (AGP), which is a mixture of the F1*S and A variants at molar ratios of 3:1 to 2:1. In this study, the mechanism of AGP esterase-like activity was investigated by evaluating the contribution of the F1*S and A variants to ONO-2160 hydrolysis and identifying the AGP hydrolase active site. We found that although both variants hydrolyzed ONO-2160, their hydrolase activities were different. The intrinsic plasma clearance of the F1*S variant (0.441 mL/h/mg protein) was approximately 30 times higher than that of the A variant (0.0148 mL/h/mg protein), indicating that the F1*S variant contributed the most to AGP esterase-like activity. To identify the hydrolase active site of AGP, we performed inhibition studies of ONO-2160 hydrolysis using 12 AGP-binding drugs with various ligand-binding constants and binding selectivities to the two AGP variants. Inhibition of activity was positively correlated with the constant of ligand binding to the F1*S variant. In addition, compounds with high affinity to the F1*S variant inhibited ONO-2160 hydrolysis the most. Together, our data indicate that ONO-2160 is predominantly hydrolyzed by the F1*S variant at its ligand-binding site.