Development of an AlphaScreen-Based HIV-1 Integrase Dimerization Assay for Discovery of Novel Allosteric Inhibitors
Development of an AlphaScreen-Based HIV-1 Integrase Dimerization Assay for Discovery of Novel Allosteric Inhibitors
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DOI：
10.1177/1087057111436343
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发表时间：
2012-06-01
影响因子：
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通讯作者：
Christ, Frauke
中科院分区：
文献类型：
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作者：
Demeulemeester, Jonas;Tintori, Cristina;Christ, Frauke
近年来，HIV-1整合酶(IN)已成为抗逆转录病毒药物开发领域的既定靶点。然而，它唯一的临床批准的抑制剂，整合酶链转移抑制剂(INSTI)雷替格列韦，具有令人惊讶的低遗传屏障的耐药性。此外，目前仅有的两种处于高级临床试验中的整合酶抑制剂，elvitegravir和多洛替格韦，具有相同的作用机制和某些耐药途径。为了维持一系列的治疗选择，药物发现努力现在转向变构IN抑制剂，它应该不会与异烟肼交叉耐药。由于IN的活性需要几个低聚物种之间精确而动态的平衡，这种平衡的调节呈现出一个有趣的变构靶标。我们报告了一种基于AlphaScreen的高通量筛选HIV-1 IN二聚体调节子的方法的开发、特征和验证。在本实验中被确认为HITS的化合物被证明是变构IN抑制剂。此外，该分析还为研究IN二聚化提供了一个灵活的平台。
In recent years, HIV-1 integrase (IN) has become an established target in the field of antiretroviral drug discovery. However, its sole clinically approved inhibitor, the integrase strand transfer inhibitor (INSTI) raltegravir, has a surprisingly low genetic barrier for resistance. Furthermore, the only two other integrase inhibitors currently in advanced clinical trials, elvitegravir and dolutegravir, share its mechanism of action and certain resistance pathways. To maintain a range of treatment options, drug discovery efforts are now turning toward allosteric IN inhibitors, which should be devoid of cross-resistance with INSTIs. As IN requires a precise and dynamic equilibrium between several oligomeric species for its activities, the modulation of this equilibrium presents an interesting allosteric target. We report on the development, characterization, and validation of an AlphaScreen-based assay for high-throughput screening for modulators of HIV-1 IN dimerization. Compounds identified as hits in this assay proved to act as allosteric IN inhibitors. Additionally, the assay offers a flexible platform to study IN dimerization.