Inhibiting Early-Stage Events in HIV-1 Replication by Small-Molecule Targeting of the HIV-1 Capsid
Inhibiting Early-Stage Events in HIV-1 Replication by Small-Molecule Targeting of the HIV-1 Capsid
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DOI：
10.1128/jvi.05006-11
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发表时间：
2012-08-01
影响因子：
5.4
通讯作者：
Smith, Amos B., III
中科院分区：
文献类型：
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作者：
Kortagere, Sandhya;Madani, Navid;Smith, Amos B., III
HIV-1衣壳蛋白（CA）在病毒复制的早期和晚期都起着至关重要的作用，并已成为一种新的药物靶点。我们报告了基于混合结构的虚拟筛选，以识别可能与HIV-1 CA的n端结构域（NTD）相互作用的小分子，并破坏HIV-1复制周期的早期预整合步骤。小分子4,4'-[二苯并[b，d]呋喃-2,8-二基双（5-苯基-1 -咪唑-4,2-二基）]二苯甲酸（CK026）在单轮和多轮感染中具有抗hiv -1活性，但不能抑制病毒在外周血单核细胞（PBMCs）中的复制。合成并评价了三种体积更小、类药物性能更好的CK026类似物。化合物I-XW-053（4-（4,5-二苯基- 1h -咪唑-2-基）苯甲酸）保留了亲本化合物的所有抗病毒活性，并抑制了多种原代HIV-1分离株在pbmc中的复制，同时没有明显的细胞毒性。这种抗病毒活性是针对HIV-1的，因为I-XW-053对SIV的复制或对一组非逆转录病毒没有作用。利用表面等离子体共振和等温滴定量热法定量了I-XW-053与野生型和突变型CA蛋白的直接相互作用。Ile37和Arg173是与化合物I-XW-053相互作用所必需的，它们的突变使病毒在整合前的早期阶段遭到破坏。通过定量PCR，我们证明了I-XW-053在多种细胞类型中抑制HIV-1逆转录，间接指出了脱膜过程中的功能障碍。总之，我们已经确定了一种cas特异性化合物，该化合物靶向并抑制NTD-NTD界面中的一个新区域，影响脱衣，并具有广谱抗hiv -1活性。
The HIV-1 capsid (CA) protein plays essential roles in both early and late stages of virl replication and has emerged as a novel drug target. We report hybrid structure-based virtual screening to identify small molecules with the potential to interact with the N-terminal domain (NTD) of HIV-1 CA and disrupt early, preintegration steps of the HIV-1 replication cycle. The small molecule 4,4'-[dibenzo[b,d]furan-2,8-diylbis(5-phenyl-1H-imidazole-4,2-diyl)]dibenzoic acid (CK026), which had anti-HIV-1 activity in single- and multiple-round infections but failed to inhibit viral replication in peripheral blood mononuclear cells (PBMCs), was identified. Three analogues of CK026 with reduced size and better drug-like properties were synthesized and assessed. Compound I-XW-053 (4-(4,5-diphenyl-1H-imidazol-2-yl)benzoic acid) retained all of the antiviral activity of the parental compound and inhibited the replication of a diverse panel of primary HIV-1 isolates in PBMCs, while displaying no appreciable cytotoxicity. This antiviral activity was specific to HIV-1, as I-XW-053 displayed no effect on the replication of SIV or against a panel of nonretroviruses. Direct interaction of I-XW-053 was quantified with wild-type and mutant CA protein using surface plasmon resonance and isothermal titration calorimetry. Mutation of Ile37 and Arg173, which are required for interaction with compound I-XW-053, crippled the virus at an early, preintegration step. Using quantitative PCR, we demonstrated that treatment with I-XW-053 inhibited HIV-1 reverse transcription in multiple cell types, indirectly pointing to dysfunction in the uncoating process. In summary, we have identified a CAs-pecific compound that targets and inhibits a novel region in the NTD-NTD interface, affects uncoating, and possesses broad-spectrum anti-HIV-1 activity.