Neamati Nouri, Lin Zhaiwei, Karki Rajeshri G, Orr Ann, Cowansage Kiriana, Strumberg Dirk, Pais Godwin C G, Voigt Johannes H, Nicklaus Marc C, Winslow Heather E, Zhao He, Turpin Jim A, Yi Jizu, Skalka Anna Marie, Burke Terrence R, Pommier Yves
Department of Pharmaceutical Sciences, University of Southern California, School of Pharmacy, 1985 Zonal Avenue, Los Angeles, California 90089, USA.
J Med Chem. 2002 Dec 19;45(26):5661-70. doi: 10.1021/jm0201417.
Human immunodeficiency virus type 1 integrase (HIV-1 IN) is an essential enzyme for effective viral replication. Therefore, IN inhibitors are being sought for chemotherapy against AIDS. We had previously identified a series of salicylhydrazides as potent inhibitors of IN in vitro (Neamati, N.; et al. J. Med. Chem. 1998, 41, 3202-3209.). Herein, we report the design, synthesis, and antiviral activity of three novel mercaptosalicylhydrazide (MSH) derivatives. MSHs were effective against the IN catalytic core domain and inhibited IN binding to HIV LTR DNA. They also inhibited catalytic activities of IN in IN-DNA preassembled complexes. Site-directed mutagenesis and molecular modeling studies suggest that MSHs bind to cysteine 65 and chelate Mg(2+) at the active site of HIV-1 IN. Contrary to salicylhydrazides, the MSHs are 300-fold less cytotoxic and exhibit antiviral activity. They are also active in Mg(2+)-based assays, while IN inhibition by salicylhydrazides is strictly Mn(2+)-dependent. Additionally, in target and cell-based assays, the MSHs have no detectable effect on other retroviral targets, including reverse transcriptase, protease, and virus attachment, and exhibit no detectable activity against human topoisomerases I and II at concentrations that effectively inhibit IN. These data suggest that MSHs are selective inhibitors of HIV-1 IN and may serve as leads for antiviral therapeutics.
人类免疫缺陷病毒1型整合酶（HIV-1 IN）是病毒有效复制所必需的一种酶。因此，人们正在寻找IN抑制剂用于艾滋病的化疗。我们之前已鉴定出一系列水杨酰肼在体外是IN的有效抑制剂（Neamati, N.等人，《药物化学杂志》，1998年，第41卷，3202 - 3209页）。在此，我们报告三种新型巯基水杨酰肼（MSH）衍生物的设计、合成及抗病毒活性。MSH对IN催化核心结构域有效，并抑制IN与HIV长末端重复序列（LTR）DNA的结合。它们还抑制IN-DNA预组装复合物中IN的催化活性。定点诱变和分子模拟研究表明，MSH与半胱氨酸65结合，并在HIV-1 IN的活性位点螯合Mg(2+)。与水杨酰肼不同，MSH的细胞毒性低300倍且具有抗病毒活性。它们在基于Mg(2+)的检测中也有活性，而水杨酰肼对IN的抑制严格依赖于Mn(2+)。此外，在基于靶点和细胞的检测中，MSH对其他逆转录病毒靶点（包括逆转录酶、蛋白酶和病毒附着）没有可检测到的影响，并且在有效抑制IN的浓度下对人拓扑异构酶I和II没有可检测到的活性。这些数据表明，MSH是HIV-1 IN的选择性抑制剂，可能成为抗病毒治疗的先导药物。
