C-5-Modified Tetrahydropyrano-Tetrahydofuran-Derived Protease Inhibitors (PIs) Exert Potent Inhibition of the Replication of HIV-1 Variants Highly Resistant to Various PIs, including Darunavir
C-5-Modified Tetrahydropyrano-Tetrahydofuran-Derived Protease Inhibitors (PIs) Exert Potent Inhibition of the Replication of HIV-1 Variants Highly Resistant to Various PIs, including Darunavir
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DOI：
10.1128/jvi.01829-15
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发表时间：
2016-03-01
影响因子：
5.4
通讯作者：
Mitsuya, Hiroaki
中科院分区：
文献类型：
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作者：
Aoki, Manabu;Hayashi, Hironori;Mitsuya, Hiroaki
我们鉴定了三种非肽类HIV-1蛋白酶抑制剂（PI），GRL-015、GRL-085和GRL-097，它们含有一个C-5羟基的四氢吡喃-四氢呋喃（TP-THF）。这三种化合物对野生型实验室HIV-1株（HIV-1（WT））有效，50%有效浓度（EC（50）s）为3.0至49 nM，并显示出最小的细胞毒性，GRL-015、-085和-097的50%细胞毒性浓度（CC 50）分别为80、>100和>100 μ M。所有三种化合物均有效抑制使用每种当前可用PI选择的高度PI耐药HIV-1变体和从携带多药耐药HIV-1变体（HIVMDR）的患者中获得的重组临床HIV-1分离株的复制。重要的是，地瑞那韦（DRV）对一种高度耐药的HIV-1变体（HIV-1（DRVRP 51））的活性降低了1,000倍以上;这三种化合物对HIV-1（DRVRP 51）仍然具有活性，仅降低了6.8至68倍。此外，与DRV的情况相比，对这三种化合物具有耐药性的HIV-1变异体的出现大大延迟。特别是，即使使用两种不同的高DRV抗性HIV-1变体作为起始群体，也没有出现对GRL-085和GRL-097具有抗性的HIV-1变体。在结构分析中，GRL-015、GRL-085和GRL-097的Tp-THF显示与HIV-1蛋白酶活性位点氨基酸残基（Asp 29和Asp 30）的骨架原子的强氢键相互作用。TP-THF的羟基部分和Gly 48的羰基氧原子之间形成强氢键是新发现的。目前的研究结果表明，这三种化合物值得进一步研究，作为可能的治疗药物，用于治疗携带野生型HIV和/或HIVMDR.IMPORTANCEDarunavir（DRV）的个体抑制大多数现有的多药耐药HIV-1株的复制，并具有较高的遗传屏障。然而，最近在体内和体外观察到高DRV抗性HIV-1株（HIVDRVR）的出现。在此，我们鉴定了三种新型HIV-1蛋白酶抑制剂（PI），其含有具有C-5羟基的四氢吡喃-四氢呋喃（Tp-THF）部分（GRL-015，-085和-097），可有效抑制HIV DRVR的复制。此外，与DRV的情况相比，对这三种化合物耐药的HIV-1菌株的出现大大延迟。在结构分析中，C-5羟基与蛋白酶Gly 48的羰基氧原子形成强的氢键相互作用。有趣的是，具有缺乏羟基部分的Tp-THF的化合物显著降低了针对HIV DRVR的活性。这三种新型化合物应进一步开发为治疗携带野生型和多PI耐药HIV变体以及HIVDRVR的个体的潜在药物。
We identified three nonpeptidic HIV-1 protease inhibitors (PIs), GRL-015, -085, and -097, containing tetrahydropyrano-tetrahydrofuran (Tp-THF) with a C-5 hydroxyl. The three compounds were potent against a wild-type laboratory HIV-1 strain (HIV-1(WT)), with 50% effective concentrations (EC(50)s) of 3.0 to 49 nM, and exhibited minimal cytotoxicity, with 50% cytotoxic concentrations (CC50) for GRL-015, -085, and -097 of 80,>100, and >100 mu M, respectively. All the three compounds potently inhibited the replication of highly PI-resistant HIV-1 variants selected with each of the currently available PIs and recombinant clinical HIV-1 isolates obtained from patients harboring multidrug-resistant HIV-1 variants (HIVMDR). Importantly, darunavir (DRV) was > 1,000 times less active against a highly DRV-resistant HIV-1 variant (HIV-1(DRVRP51)); the three compounds remained active against HIV-1(DRVRP51) with only a 6.8- to 68-fold reduction. Moreover, the emergence of HIV-1 variants resistant to the three compounds was considerably delayed compared to the case of DRV. In particular, HIV-1 variants resistant to GRL-085 and -097 did not emerge even when two different highly DRV-resistant HIV-1 variants were used as a starting population. In the structural analyses, Tp-THF of GRL-015, -085, and -097 showed strong hydrogen bond interactions with the backbone atoms of active-site amino acid residues (Asp29 and Asp30) of HIV-1 protease. A strong hydrogen bonding formation between the hydroxyl moiety of Tp-THF and a carbonyl oxygen atom of Gly48 was newly identified. The present findings indicate that the three compounds warrant further study as possible therapeutic agents for treating individuals harboring wild-type HIV and/or HIVMDR.IMPORTANCEDarunavir (DRV) inhibits the replication of most existing multidrug-resistant HIV-1 strains and has a high genetic barrier. However, the emergence of highly DRV-resistant HIV-1 strains (HIVDRVR) has recently been observed in vivo and in vitro. Here, we identified three novel HIV-1 protease inhibitors (PIs) containing a tetrahydropyrano-tetrahydrofuran (Tp-THF) moiety with a C-5 hydroxyl (GRL-015, -085, and -097) which potently suppress the replication of HIVDRVR. Moreover, the emergence of HIV-1 strains resistant to the three compounds was considerably delayed compared to the case of DRV. The C-5 hydroxyl formed a strong hydrogen bonding interaction with the carbonyl oxygen atom of Gly48 of protease as examined in the structural analyses. Interestingly, a compound with Tp-THF lacking the hydroxyl moiety substantially decreased activity against HIVDRVR. The three novel compounds should be further developed as potential drugs for treating individuals harboring wild-type and multi-PI-resistant HIV variants as well as HIVDRVR.