Differential inhibition of HIV-1 preintegration complexes and purified integrase protein by small molecules
Differential inhibition of HIV-1 preintegration complexes and purified integrase protein by small molecules
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DOI：
10.1073/pnas.93.18.9742
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发表时间：
1996-09-03
影响因子：
11.1
通讯作者：
Bushman, FD
中科院分区：
文献类型：
--
作者：
Farnet, CM;Wang, BB;Bushman, FD
为了复制，HIV-1必须将病毒RNA基因组的cDNA拷贝整合到宿主的染色体上，整合系统是抗逆转录病毒药物的一个有希望的靶标，但迄今为止还没有发现临床上有用的整合抑制剂。先前的整合酶抑制剂筛选已经检测了含有从大肠杆菌表达系统纯化的HIV-1整合酶的反应的抑制作用。在这里，我们比较了抑制剂在体外对纯化整合酶和从感染HIV-1的淋巴样细胞分离的亚病毒预整合复合物（PICs）的作用。我们发现许多对纯化整合酶有活性的抑制剂对PICs无活性，使用PIC测定作为主要筛选，我们已经鉴定出三种新的蒽醌抑制剂对PICs和纯化整合酶都有活性。我们认为PIC测定法是体外最接近体内整合的方法，因此，特别适合于鉴定有希望的整合抑制剂。
To replicate, HIV-1 must integrate a cDNA copy of the viral RNA genome into a chromosome of the host, The integration system is a promising target for antiretroviral agents, but to date no clinically useful integration inhibitors have been identified, Previous screens for integrase inhibitors have assayed inhibition of reactions containing HIV-1 integrase purified from an Escherichia coli expression system, Here we compare action of inhibitors in vitro on purified integrase and on subviral preintegration complexes (PICs) isolated from lymphoid cells infected with HIV-1. We find that many inhibitors active against purified integrase are inactive against PICs, Using PIC assays as a primary screen, we have identified three new anthraquinone inhibitors active against PICs and also against purified integrase. We propose that PIC assays are the closest in vitro match to integration in vivo and, as such, are particularly appropriate for identifying promising integration inhibitors.