Design and Synthesis of HIV Integrase as Potential Anti-AIDS Drugs

fda批准的HIV-1 IN抑制剂属于一类被称为“整合酶链转移抑制剂”（insts）的药物，因为与酶的3'-加工（3'-P）反应相比，它们能够优先阻断酶的链转移（ST）反应。不幸的是，突变形式的内隐素的出现导致临床对这些内隐素的耐药性。这进一步推动了开发下一代药物的持续需求，这些药物能够对新出现的insi耐药病毒株保持高抗病毒效力。利用我实验室的设计和合成能力，我们与药理学家（Yves Pommier博士，NCI），病毒学家（Hughes博士，NCI）和结构生物学家（Robert Craigie博士，NIDDK， Dmitry Lyumkis博士，Salk研究所和Cherepanov博士，弗朗西斯克里克研究所，英国）合作，更好地了解iniss与内溶体（多聚整合酶与DNA底物和金属辅助因子）的相互作用以及突变在下调这些相互作用中所起的作用。Cryo-EM在这些努力中发挥了关键作用。我们最好的与内泌体结合的insis的冷冻电镜结构揭示了一个复杂的、动态的水分子网络围绕着结合的insis，其中许多水似乎是保守的，并且在未配体和insi结合的结构中占据相似的位置。然而，由于INSTI的结合，一些水被置换或转移；其他的只有在结合时才发现，这表明由结合引起的构象变化稳定了它们的位置。我们的结论是，在“底物包膜”（由宿主和病毒DNA结合定义的区域）中，催化口袋的几何形状、它们的总体体积、水合作用的附近模式以及其他特征的差异，都对理解INSTI相互作用很重要。我们还与伦敦帝国理工学院Goedele Maertens博士的实验室合作，研究我们针对人类t细胞白血病病毒1型（HTLV-1）的多种in抑制剂，我们的许多合成构建物显示出针对该靶标的显着效力。我们最好的inst在单轮复制试验中表现出比目前fda批准的药物更好的抗病毒特性，该试验采用了100多种耐药突变形式的整合酶。为了在我们最好的insit上生成关键的药代动力学（PK）数据，我们与NCI发明开发计划（IDP）合作获取早期PK数据，包括在啮齿动物身上的研究。

FDA-approved inhibitors of HIV-1 IN belong to a class of drugs called " integrase strand transfer inhibitors" (INSTIs), due to their ability to preferentially block the enzyme's strand transfer (ST) reaction as compared to the enzymes 3'-processing (3'-P) reaction. Unfortunately, mutant forms of IN arise that lead to clinical resistance against these INSTIs. This adds impetus to a continuing need to develop next-generation agents that have the ability to retain high antiviral efficacy against emerging strains of INSTI-resistant virus. Utilizing my laboratory's design and synthetic capabilities, we have teamed with pharmacologists (Dr. Yves Pommier, NCI), virologists (Dr. Hughes, NCI) and structural biologists (Dr. Robert Craigie, NIDDK, Dr. Dmitry Lyumkis, the Salk Institute and Dr. Cherepanov, the Francis Crick Institute, UK) to better understand the interactions of INSTIs with intasomes (multimeric integrase with DNA substrate and metal cofactor) and the roles that mutations play in down-regulating these interactions. Cryo-EM has played a key role in these efforts. Cryo-EM structures of our best INSTIs bound to intasomes revealed a complex and dynamic network of water molecules surrounding bound INSTIs, with many of these waters appearing to be conserved and occupying similar positions in the unliganded and INSTI-bound structures. However, some waters are displaced or shifted as a consequence of binding of the INSTI; others are found only when INSTIs are bound, suggesting that the conformational changes induced by the binding stabilize their position. We concluded that within the "substrate envelope" (the region defined by the binding of host and viral DNA), differences in geometry of the catalytic pockets, their overall volume, the nearby patterns of hydration, among other features, all matter for understanding INSTI interactions. We also have a collaboration in place with the Imperial College London laboratory of Dr. Goedele Maertens to examine a diverse selection of our IN inhibitors against human T-cell leukemia virus type 1 (HTLV-1) and a number of our synthetic constructs show significant potency against this target. Our best INSTs exhibit better antiviral profiles than current FDA-approved agents when assayed in single round replication assays that employ a panel of more than 100 drug-resistant mutant forms of integrase. In order to generate key pharmacokinetic (PK) data on our best INSITs, we have partnered with the NCI Invention Development Program (IDP) to obtain early-stage PK data, including studies in rodents.