Development of NLRP3 inhibitors for HIV-associated neuroinflammation

摘要 有效的联合抗逆转录病毒疗法(CART)的出现延长了HIV-1的预期寿命 然而，这些患者仍然容易合并疾病，例如与hiv相关的神经认知。 疾病(手)，影响高达50%的艾滋病毒感染者。持续的炎症是由于 先天性免疫系统的过度激活是手部疾病的主要潜在原因之一。核苷酸结合 富含亮氨酸重复序列结构域含有蛋白-3(NLRP3)的炎症体是一种 用于治疗HIV-1相关神经病理的可用药靶点。NLRP3炎症体是 被显示为对一系列病原体和危险相关的分子模式做出反应而被激活 (分别为Pamps和Dumps)。激活过程中的一个关键步骤是 NLRP3中的吡咯结构域和一个适配蛋白，即含有一个卡片的凋亡相关斑点样蛋白 (ASC)。NLRP3激活导致促炎细胞因子的释放，如白细胞介素1(IL-1)和 IL-18，导致神经元性下垂和死亡。通过小分子干扰NLRP3信号，如 据报道，MCC950在转基因小鼠模型中显示出有益的效果。我们最近的研究发现 一种名为AMS-17的小分子在体外和体内都能抑制N9小胶质细胞中NLRP3的激活。 随后的机制分析表明，AMS-17的NLRP3抑制活性归因于其能力 与NLRP3吡林区结合，从而阻止NLRP3与ASC之间的相互作用。这项建议是 重点开发生物活性提高、毒性低、类药物活性高的AMS-17类似物。 本提案中描述的目标1侧重于计算机辅助设计、合成和化学 AMS-17类似物的表征。目标2将涉及在人性化小鼠中测试领先候选者 手型。拟议的研究具有重要意义，因为它们将提供新的治疗方案 将艾滋病毒相关的神经认知障碍降至最低。该提案包含了以下领域的专门知识 合成药物化学(Kulkarni博士)、生物筛选(Bukrinsky博士)和计算机辅助药物 设计(阿德朱贝博士)。这与本RFA的目标完全一致，并有望定义NLRP3抑制 化合物通过一种不同于任何其他目前使用的药物的新机制发挥作用。

ABSTRACT The advent of effective combination antiretroviral therapies (cART) has increased the life expectancy for HIV-1 patients, however, these patients are still prone to comorbidities, such as HIV-associatiated neurocognitive disorders (HAND), which affect up to 50% of HIV-infected individuals. Persistent inflammation due to the overactivation of the innate immune system is one of the main underlying causes of HAND. Nucleotide binding domain, leucine rich repeat pyrin domain containing protein-3 (NLRP3) inflammasome has emerged as a druggable target for the management of HIV-1-associated neuropathologies. The NLRP3 inflammasome is shown to be activated in response to a wide array of pathogen- and danger-associated molecular patterns (PAMPs and DAMPs, respectively). A key step in the activation process is a homotypic interaction between the pyrin domains in NLRP3 and an adapter protein, apoptosis-associated speck-like protein containing a CARD (ASC). NLRP3 activation leads to the release of pro-inflammatory cytokines, such as, interleukin-1 (IL-1) and IL-18, causing neuronal pyroptosis and death. Disruption of NLRP3 signaling via small molecules, such as MCC950, is reported to display beneficial effects in the transgenic mouse models. Our recent studies identified a small molecule, AMS-17, that thwarted the NLRP3 activation in N9 microglia both in vitro and in vivo. Subsequent mechanistic analysis revealed that the NLRP3 inhibitory activity of AMS-17 is attributed to its ability to bind to NLRP3 pyrin domain, thus preventing the interaction between NLRP3 and ASC. This proposal is focused on developing AMS-17 analogues with improved biological activity, low toxicity, and high drug-likeness. Aim 1 described in this proposal is focused on the computer-assisted design, synthesis and chemical characterization of AMS-17 analogues. Aim 2 will involve testing of the lead candidates in the humanized mouse model of HAND. The proposed studies are highly significant since they will provide new therapeutic options to minimize HIV-associated neurocognitive dysfunction. The proposal incorporates expertise in the area of synthetic medicinal chemistry (Dr. Kulkarni), biological screening (Dr. Bukrinsky), and computer-assisted drug design (Dr. Adzhubei). It is fully consistent with the goals of this RFA and is expected to define NLRP3 inhibitory compounds working through a novel mechanism different from that of any other currently used drug.