An Innovative Approach to Identify Correctors of Metabolic Complications in HIV

自采用联合抗逆转录病毒疗法(CART)以来，西方世界艾滋病毒携带者的存活率和生活质量不断提高。然而，HIV感染和CART与糖脂代谢紊乱、肥胖、代谢综合征患病率增加和心血管疾病的发生率较高有关。事实上，心血管疾病正在成为PLWH患者人群发病率和死亡率的主要原因。代谢副作用与使用目前最关键的CART疗法(例如，广泛使用的固定剂量组合Atripla和Kaletra，以及新的整合酶抑制剂和蛋白酶抑制剂)有关，尽管它们通常比早期抗逆转录病毒药物的此类效果麻烦得多。在这里，我们提出了一种创新的方法来识别小分子，通过靶向HIV和慢性炎症中调节失调的途径来调节和正常化代谢控制。为此，我们将在1536孔板格式中开展高通量筛选活动，并结合分层方法来筛选&gt；665,000化合物斯克里普斯药物发现文库(SDDL)，并确认~500个类药物分子的效力。将进行HIT-验证，以识别小分子调节剂并消除非特异性效应器，使用平行和正交分析以及使用多个对应屏的脱靶评估。为了确定热门脚手架系列的优先顺序，我们将从现有的复合库和其他商业来源中选择确认热门的类似产品。HIT支架将进行分类，以去除难处理的分子。我们将从最有希望的HITS中选择3-5个分子系列，并对其进行分析，以验证它们的选择性、有效性和无细胞毒性。利用申请实验室(加利福尼亚州斯克里普斯和佛罗里达州斯克里普斯)的精心安排的努力，2-4系列中的引线将被制定并重新测试效力/选择性，目的是推进能够在上述检测中引起适当的体外反应并可以评估对细胞表达水平的适当表型反应的引线。随后将进行体外和体内药代动力学(PK)研究，以确定1-2个顶级支架以供进一步研究。最后，最有希望的2-3个化合物具有良好的药物代谢和药代动力学(DMPK)特性，包括高口服生物利用度，将被选择用于在啮齿动物疗效模型中进行体内测试，例如HIV转基因大鼠。总之，我们提出了一种新的战略，以建立新的和更有效的治疗方法，以改善艾滋病毒相关的代谢并发症，这是一个重要的未得到满足的临床需求，也是NIDDK任务范围内高度优先的艾滋病毒/艾滋病研究领域。

Since the introduction of combination antiretroviral therapy (cART) the survival and quality of life of people living with HIV (PLWH) in the Western world has continued to improve. However, HIV infection and cART are associated with glucose and lipid metabolism dysregulation, obesity, an increased prevalence of metabolic syndrome, and a higher incidence of cardiovascular disease. In fact, cardiovascular disease is becoming a leading cause of morbidity and mortality in the PLWH patient population. The metabolic side effects are variably associated with the use of most current key cART therapeutics (e.g., the widely used fixed- dose combinations Atripla and Kaletra, and the new integrase inhibitors and protease inhibitors), although they are generally less troublesome than such effects from earlier antiretrovirals. Here, we propose an innovative approach to identify small molecules to modulate and normalize metabolic control by targeting a pathway dysregulated in HIV and chronic inflammation. To this end, we will carry out a high-throughput screening campaign in a 1536-well plate format in conjunction with a tiered approach to screen the >665,000-compound Scripps Drug Discovery Library (SDDL) and confirm the potency of ~500 drug-like molecules. Hit-validation will be performed to identify small-molecule regulators and eliminate nonspecific effectors, using parallel and orthogonal assays as well as off-target assessments using multiple counterscreens. To prioritize hit scaffold series, we will select analogs of confirmed hits from both the existing compound libraries and other commercial sources. Hit scaffolds will be triaged to remove intractable molecules. We will select 3-5 molecular series from the most promising hits, which will be profiled to verify their selectivity, potency, and lack of cytotoxicity. Using an orchestrated effort from the applicant laboratories (Scripps California and Scripps Florida), leads in 2-4 series will be formulated and retested for potency/selectivity with the aim of advancing leads that can elicit the appropriate in vitro response in the aforementioned assays and can be evaluated for appropriate phenotypic responses to cell-expression levels. This will be followed by in vitro and in vivo pharmacokinetics (PK) studies to identify 1-2 top scaffolds for further investigation. Finally, the most promising 2-3 compounds with favorable drug metabolism and pharmacokinetics (DMPK) properties including high oral bioavailability, will be selected for in vivo testing in rodent efficacy models such as HIV transgenic rats. Altogether, we propose a novel strategy to establish new and more effective therapies to ameliorate HIV-associated metabolic complications, which is an important unmet clinical need and an area of high priority HIV/AIDS research within the mission of the NIDDK.