Farnet C M, Wang B, Lipford J R, Bushman F D
Infectious Disease Laboratory, Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9742-7. doi: 10.1073/pnas.93.18.9742.
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.
为了进行复制，HIV-1必须将病毒RNA基因组的cDNA拷贝整合到宿主的染色体中。整合系统是抗逆转录病毒药物的一个有前景的靶点，但迄今为止尚未鉴定出临床上有用的整合抑制剂。以前针对整合酶抑制剂的筛选检测了对从大肠杆菌表达系统纯化的HIV-1整合酶的反应抑制作用。在这里，我们比较了抑制剂在体外对纯化的整合酶以及从感染HIV-1的淋巴细胞中分离出的亚病毒预整合复合物（PIC）的作用。我们发现许多对纯化整合酶有活性的抑制剂对PIC无活性。使用PIC检测作为主要筛选方法，我们鉴定出了三种对PIC以及纯化整合酶都有活性的新型蒽醌抑制剂。我们提出，PIC检测是体外最接近体内整合的方法，因此特别适合于鉴定有前景的整合抑制剂。
