Pharmacology of HIV Viral DNA and Retroviral Integrases

HIV 病毒 DNA 和逆转录病毒整合酶的药理学

• 批准号: 10014290
• 负责人: YVES POMMIER
• 金额: $ 30.29万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10014290
• 关键词: Active Sites; Animal Testing; Antiviral Agents; Binding; Biochemical; Biology; CCR; Catalytic Domain; Chelating Agents; Chemicals; Clinical; Collaborations; Collection; Complex; Crystallization; DNA; Dose; Drug Kinetics; Drug resistance; Enzymes; FDA approved; Family; Formulation; Goals; Grant; HIV; HIV Integrase Inhibitors; Healthcare; Institutes; Integrase; Integrase Inhibitors; Ions; Journals; Laboratories; Lead; Legal patent; London; Malignant Neoplasms; Mediating; Metals; Modification; Molecular; Mutation; Nature; Patients; Peptides; Pharmaceutical Preparations; Pharmacology; Proteins; Proviruses; Publications; Publishing; Reaction; Recombinant Proteins; Recombinants; Research; Resistance; Resistance development; Series; Site; Specificity; Spumavirus; Structural Models; Structure; Testing; Therapeutic Index; Work; clinical development; clinically relevant; compliance behavior; cytotoxicity; data modeling; design; divalent metal; drug development; drug discovery; drug sensitivity; experimental study; flexibility; inhibitor/antagonist; interfacial; molecular modeling; mutant; novel; off-patent; pill; pol genes; preclinical development; prototype; therapeutic development; viral DNA

Integrase (IN) is encoded by the Pol gene from the HIV provirus. Our laboratory can efficiently express IN as an active recombinant protein, and has pioneered the integrase inhibitors research field (our landmark publication: PNAS 1993), discovered several families of lead inhibitors, demonstrated that IN inhibitors act as interfacial inhibitors (Nature Rev Drug Discovery 2012), and been granted several patents for IN inhibitors for therapeutic development. Our current studies are focused on the optimization of our novel chemotype integrase inhibitors to overcome resistance to raltegravir, elvitegravir and dolutegravir and target novel sites of IN. We have published and patented novel synthetic chemotypes as IN strand transfer inhibitors (INSTIs) including phtalimide and quinolinonyl derivatives in collaborations with Dr. Terrence Burke, Chemical Biology Laboratory (CCR, NCI). We have developed a panel of recombinant IN proteins bearing the mutations observed in patients that develop resistance to raltegravir, elvitegravir and dolutegravir. Using our resistant IN mutants, we have characterized the molecular pharmacology of elvitegravir, dolutegravir and our novel inhibitors, comparing them to raltegravir. We have shown that raltegravir, elvitegravir, dolutegravir and our novel series are highly selective for the strand transfer reaction, while being more than 100-fold less potent against the 3'-processing reaction, and almost inactive against the disintegration reaction mediated by integrase. The selective activity against strand transfer (one of the 3 reactions mediated by integrase) demonstrates the very high specificity of the clinically developed IN strand transfer inhibitors (INSTIs). It is consistent with our pharmacological hypothesis (Nature Drug Discovery 2012) that the strand transfer inhibitors trap the IN-viral DNA complex by chelating the divalent metals in the enzyme catalytic site following 3'-processing of the viral DNA and with our co-crystal structure and molecular modeling data. We have characterized the biochemical enzymatic activities and drug sensitivities of the IN mutants that confer clinical drug resistance. We have expanded these studies to double-mutants in the integrase flexible loop that commonly arise in raltegravir-resistant patients. Our results support the value of dolutegravir to overcome resistance to raltegravir and elvitegravir and facilitate patient compliance. We have determined additional crystal structures of wild-type and mutant prototype foamy virus (PFV) intasomes bound to our new series of inhibitors in collaboration with Dr. Peter Cherepanov at the Crick Institute, Cancer UK Center in London. The ability to structurally adapt to the structural changes associated with drug resistance is now achievable to rationally develop our new INSTIs. This year, we have also performed biochemical experiments demonstrating the feasibility of designing peptide inhibitors of IN. Because our XZ compounds possess high therapeutic index without cytotoxicity up to 250 microM (the highest dose tested), 3 of them, XZ419, XZ434 and XZ446 have been selected for preclinical development and animal testing by the NCI-CCR Drug Development Collaborative group (DDC). It is likely that further chemical modifications will be necessary to optimize formulation and pharmacokinetics. IATAP support will allow us to continue our collaborative work with Terrence Burke and XueZhi Zhao and with Stephen Hughes to achieve these goals.

整合酶 (IN) 由 HIV 原病毒的 Pol 基因编码。我们的实验室可以有效地将IN表达为活性重组蛋白，并开创了整合酶抑制剂研究领域（我们的里程碑出版物：PNAS 1993），发现了几个先导抑制剂家族，证明了IN抑制剂可作为界面抑制剂（Nature Rev Drug Discovery 2012） ），并获得多项 IN 抑制剂治疗开发专利。我们目前的研究重点是优化我们的新型化学型整合酶抑制剂，以克服对拉替拉韦、埃替拉韦和多替拉韦的耐药性，并靶向 IN 的新位点。我们与化学生物学实验室（CCR、NCI）的 Terrence Burke 博士合作，发表了新型合成化学型作为 IN 链转移抑制剂 (INSTI) 并获得了专利，其中包括邻苯二甲酰亚胺和喹啉衍生物。我们开发了一组重组 IN 蛋白，这些蛋白带有在对拉替拉韦、埃替拉韦和多替拉韦产生耐药性的患者中观察到的突变。使用我们的耐药IN突变体，我们表征了埃替拉韦、多替拉韦和我们的新型抑制剂的分子药理学，并将它们与拉替拉韦进行比较。我们已经证明，拉替拉韦、埃替拉韦、多替拉韦和我们的新系列对链转移反应具有高度选择性，而对 3' 加工反应的效力要低 100 倍以上，并且对整合酶介导的崩解反应几乎没有活性。针对链转移（整合酶介导的 3 个反应之一）的选择性活性证明了临床开发的 IN 链转移抑制剂 (INSTI) 具有非常高的特异性。这与我们的药理学假设（Nature Drug Discovery 2012）一致，即链转移抑制剂通过在病毒 DNA 3' 处理后与我们的共晶螯合酶催化位点中的二价金属来捕获 IN-病毒 DNA 复合物。结构和分子建模数据。我们已经表征了赋予临床耐药性的 IN 突变体的生化酶活性和药物敏感性。我们已将这些研究扩展到整合酶柔性环中的双突变体，这种突变体通常出现在拉替拉韦耐药患者中。我们的结果支持多替拉韦克服拉替拉韦和艾维拉韦耐药性并促进患者依从性的价值。我们与伦敦克里克研究所、英国癌症中心的 Peter Cherepanov 博士合作，确定了与我们的新系列抑制剂结合的野生型和突变型原型泡沫病毒 (PFV) 嵌体的其他晶体结构。现在可以实现结构上适应与耐药性相关的结构变化的能力，以合理开发我们的新 INSTI。今年，我们还进行了生化实验，论证了设计IN肽抑制剂的可行性。由于我们的 XZ 化合物具有高治疗指数且无细胞毒性，最高可达 250 µM（测试的最高剂量），因此其中 3 个 XZ419、XZ434 和 XZ446 已被 NCI-CCR 药物开发协作组 (DDC) 选择进行临床前开发和动物测试。 ）。可能需要进一步的化学修饰来优化配方和药代动力学。 IATAP 的支持将使我们能够继续与 Terrence Burke、XueZhi Zhu 以及 Stephen Hughes 合作，以实现这些目标。

项目成果

Peptidic HIV integrase inhibitors derived from HIV gene products: structure-activity relationship studies.

来自 HIV 基因产物的肽 HIV 整合酶抑制剂：结构-活性关系研究。

• 发表时间: 2010-09-15
• 影响因子: 3.5
• 作者: Suzuki, Shintaro;Maddali, Kasthuraiah;Hashimoto, Chie;Urano, Emiko;Ohashi, Nami;Tanaka, Tomohiro;Ozaki, Taro;Arai, Hiroshi;Tsutsumi, Hiroshi;Narumi, Tetsuo;Nomura, Wataru;Yamamoto, Naoki;Pommier, Yves;Komano, Jun A;Tamamura, Hirokazu
• 通讯作者: Tamamura, Hirokazu

3-Hydroxypyrimidine-2,4-diones as an inhibitor scaffold of HIV integrase.

3-Hydroxypyrimidine-2,4-diones 作为 HIV 整合酶的抑制剂支架。

• 发表时间: 2011-04-14
• 影响因子: 7.3
• 作者: Tang, Jing;Maddali, Kasthuraiah;Metifiot, Mathieu;Sham, Yuk Y;Vince, Robert;Pommier, Yves;Wang, Zhengqiang
• 通讯作者: Wang, Zhengqiang

Peptide HIV-1 integrase inhibitors from HIV-1 gene products.

来自 HIV-1 基因产物的肽 HIV-1 整合酶抑制剂。

• 发表时间: 2010-07-22
• 影响因子: 7.3
• 作者: Suzuki, Shintaro;Urano, Emiko;Hashimoto, Chie;Tsutsumi, Hiroshi;Nakahara, Toru;Tanaka, Tomohiro;Nakanishi, Yuta;Maddali, Kasthuraiah;Han, Yan;Hamatake, Makiko;Miyauchi, Kosuke;Pommier, Yves;Beutler, John A;Sugiura, Wataru;Fuji, Hideyoshi;Ho
• 通讯作者: Ho

Comparison of raltegravir and elvitegravir on HIV-1 integrase catalytic reactions and on a series of drug-resistant integrase mutants.

拉替拉韦和埃替拉韦对 HIV-1 整合酶催化反应和一系列耐药整合酶突变体的比较。

• 发表时间: 2008-09-09
• 影响因子: 2.9
• 作者: Marinello, Jessica;Marchand, Christophe;Mott, Bryan T;Bain, Anjali;Thomas, Craig J;Pommier, Yves
• 通讯作者: Pommier, Yves

Cell-permeable stapled peptides based on HIV-1 integrase inhibitors derived from HIV-1 gene products.

基于源自 HIV-1 基因产物的 HIV-1 整合酶抑制剂的细胞渗透性钉合肽。

• 发表时间: 2013-10-18
• 影响因子: 4
• 作者: Nomura, Wataru;Aikawa, Haruo;Ohashi, Nami;Urano, Emiko;Métifiot, Mathieu;Fujino, Masayuki;Maddali, Kasthuraiah;Ozaki, Taro;Nozue, Ami;Narumi, Tetsuo;Hashimoto, Chie;Tanaka, Tomohiro;Pommier, Yves;Yamamoto, Naoki;Komano, Jun A;Murakami, Tsut
• 通讯作者: Murakami, Tsut