An Innovative Approach to Identify Correctors of Metabolic Complications in HIV

自从采用联合抗逆转录病毒疗法（cART）以来，西方世界艾滋病毒感染者（PLWH）的生存率和生活质量不断改善。然而，艾滋病毒感染和cART与葡萄糖和脂质代谢失调、肥胖、代谢综合征患病率增加以及心血管疾病发病率较高有关。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。代谢副作用与大多数当前关键的cART疗法的使用（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），尽管它们通常比早期抗逆转录病毒药物的这种作用更少麻烦。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孔板格式中进行高通量筛选活动，并结合分层方法筛选> 665，000种化合物Scripps药物发现库（SDDL），并确认约500种药物样分子的效力。将使用平行和正交试验以及使用多个反筛选的脱靶评估进行命中验证，以鉴定小分子调节剂并消除非特异性效应物。为了优先考虑命中支架系列，我们将从现有化合物文库和其他商业来源中选择确认命中的类似物。击中的支架将被分类以去除难处理的分子。我们将从最有希望的命中中选择3-5个分子系列，将对其进行分析以验证其选择性、效力和无细胞毒性。使用申请人实验室（Scripps加州和Scripps佛罗里达）的协调工作，将配制2-4系列中的先导化合物，并重新检测效价/选择性，目的是推进可在上述试验中引起适当体外反应的先导化合物，并可评价对细胞表达水平的适当表型反应。随后将进行体外和体内药代动力学（PK）研究，以确定1-2种顶级支架用于进一步研究。最后，将选择具有良好药物代谢和药代动力学（DMPK）特性（包括高口服生物利用度）的最有希望的2-3种化合物用于啮齿动物功效模型（例如HIV转基因大鼠）中的体内测试。总之，我们提出了一种新的策略，以建立新的和更有效的治疗，以改善艾滋病毒相关的代谢并发症，这是一个重要的未满足的临床需求和高优先级的艾滋病毒/艾滋病研究领域内的NIDDK的使命。

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.