P2' benzene carboxylic acid moiety is associated with decrease in cellular uptake: evaluation of novel nonpeptidic HIV-1 protease inhibitors containing P2 bis-tetrahydrofuran moiety.
P2' benzene carboxylic acid moiety is associated with decrease in cellular uptake: evaluation of novel nonpeptidic HIV-1 protease inhibitors containing P2 bis-tetrahydrofuran moiety.
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P2 苯甲酸部分与细胞摄取减少相关：含有 P2 双四氢呋喃部分的新型非肽 HIV-1 蛋白酶抑制剂的评估。
DOI：
10.1128/aac.00868-13
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发表时间：
2013
影响因子：
4.9
通讯作者：
Mitsuya,Hiroaki
中科院分区：
文献类型：
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作者：
Yedidi,RavikiranS;Maeda,Kenji;Fyvie,WSean;Steffey,Melinda;Davis,DavidA;Palmer,Ira;Aoki,Manabu;Kaufman,JoshuaD;Stahl,StephenJ;Garimella,Harisha;Das,Debananda;Wingfield,PaulT;Ghosh,ArunK;Mitsuya,Hiroaki
GRL 007和GRL 008，两种结构相关的非肽类人类免疫缺陷病毒1型（HIV-1）蛋白酶抑制剂（PI），分别含有3（R），3a（S），6a（R）-双-四氢呋喃基氨基甲酸酯（bis-THF）作为P2部分和由苯羧酸和苯甲酰胺组成的磺酰胺等排体作为P2 ′部分，评价了它们的抗病毒活性和与野生型蛋白酶（PRWT）的相互作用。GRL 007（对PRWT的Ki为12.7pM）和GRL 008（对PRWT的Ki为8.9pM）在体外均能高效抑制PRWT。对PRWT Tin与GRL 007或GRL 008配合物的X-射线晶体学分析表明，两种化合物的双-THF部分与PRWT的Asp29和Asp30的主链酰胺氮原子有三个直接的极性接触。两种化合物的P2 ′部分均与Asp30 ′的主链直接接触，并通过水分子与Gly48 ′桥接极性接触。基于细胞的抗病毒试验表明，GRL 007对野生型HIV-1无活性（50%有效浓度[EC50]> 1 μ M），而GRL 008对野生型HIV-1具有高度活性（EC50为0.04 μ M）。基于高效液相色谱（HPLC）/质谱的细胞摄取测定显示，在人MT-2和MT-4细胞提取物中，GRL 008的细胞内浓度分别比GRL 007高8.1倍和84倍。因此，尽管GRL 007具有良好的酶抑制活性和蛋白酶结合特性，但在基于细胞的测定中表现出缺乏抗病毒活性，这很可能是由于其与P2 ′苯羧酸部分相关的细胞摄取受损。GRL 008的抗HIV-1效力、有利的毒性和结合特征表明，进一步优化P2 ′部分可能会改善其抗逆转录病毒特性。
GRL007 and GRL008, two structurally related nonpeptidic human immunodeficiency virus type 1 (HIV-1) protease inhibitors (PIs) containing 3(R),3a(S),6a(R)-bis-tetrahydrofuranylurethane (bis-THF) as the P2 moiety and a sulfonamide isostere consisting of benzene carboxylic acid and benzene carboxamide as the P2′ moiety, respectively, were evaluated for their antiviral activity and interactions with wild-type protease (PRWT). Both GRL007 (Kiof 12.7 pM with PRWT) and GRL008 (Kiof 8.9 pM) inhibited PRWTwith high potencyin vitro. X-ray crystallographic analysis of PRWTin complex with GRL007 or GRL008 showed that thebis-THF moiety of both compounds has three direct polar contacts with the backbone amide nitrogen atoms of Asp29 and Asp30 of PRWT. The P2′ moiety of both compounds showed one direct contact with the backbone of Asp30′ and a bridging polar contact with Gly48′ through a water molecule. Cell-based antiviral assays showed that GRL007 was inactive (50% effective concentration [EC50] of >1 μM) while GRL008 was highly active (EC50of 0.04 μM) against wild-type HIV-1. High-performance liquid chromatography (HPLC)/mass spectrometry-based cellular uptake assays showed 8.1- and 84-fold higher intracellular concentrations of GRL008 than GRL007 in human MT-2 and MT-4 cell extracts, respectively. Thus, GRL007, in spite of its favorable enzyme-inhibitory activity and protease binding profile, exhibited a lack of antiviral activity in cell-based assays, most likely due to its compromised cellular uptake associated with its P2′ benzene carboxylic acid moiety. The anti-HIV-1 potency, favorable toxicity, and binding profile of GRL008 suggest that further optimization of the P2′ moiety may improve its antiretroviral features.