Small molecule inhibitor of CD40 signaling for the control of inflammatory bowel disease

项目摘要 尽管炎症性肠病（IBD）的治疗取得了进展，但许多患者对炎症性肠病（IBD）的治疗没有反应。 疗法因此，需要找到针对IBD的新的治疗方法。CD 40-CD 154通路是一种 已知靶点是IBD和其它炎性疾病。临床试验表明，CD 40阻断剂 抗CD 154抗体减少炎症。然而，抗CD 154抗体引起血栓形成， （与CD 40的抑制无关）。此外，还预测了其他导致CD 40全面抑制的途径 增加机会性感染的风险。鉴定抑制CD 40诱导的炎症的策略 不诱导血栓形成或机会性感染的药物可能是IBD治疗的主要进展。 我们发现，阻断CD 40和细胞内衔接蛋白之间的相互作用， 在某些实施方案中，该方法包括抑制由CD 40诱导的炎症反应，同时保持对机会性病原体的保护完好。 我们发现了一种小分子，它可以结合衔接蛋白，阻断CD 40信号传导，减少前 在IBD的小鼠模型中，本发明的组合物在体外抑制炎症反应并减少肠道炎症。的化合物 并不削弱对机会致病菌的抵抗力。 该化合物具有次优的溶解度和微粒体稳定性。虽然迄今为止设计的一些类似物 显示溶解度或微粒体稳定性有所改善，需要进一步优化。客观 本申请的目的是开发一种优化的抑制剂，该抑制剂将在小鼠和人IBD系统中进行测试。 中心假设是具有改善的溶解度和微粒体稳定性的有效类似物将最佳地 阻断CD 40信号传导，并显著抑制IBD小鼠模型中的炎症以及CD 40驱动的 IBD患者肠细胞的炎症反应。为了验证这一假设，我们将设计和 产生化合物的类似物，测试它们的性质，在报道细胞中进行信号传导研究， 并在IBD动物模型中测试铅抑制剂。在第一个具体目标中，我们将使用结构 活性关系，以设计和生成化合物的类似物， 以改善溶解度和微粒体稳定性。我们将研究它们抑制CD 40信号传导的能力， 对衔接蛋白的亲和力。在第二个目标中，我们将测试最有效的类似物，以确定它们是否 在体内抑制CD 40信号传导。在第三个目标中，我们将确定铅类似物是否能减少肠道 在IBD小鼠模型中的炎症和抑制CD 40诱导的炎症分子的表达， IBD患者的肠细胞。这项工作可能会导致一种新的治疗IBD的策略， 抑制CD 40信号传导的方法。

PROJECT SUMMARY Despite advances in the treatment of inflammatory bowel disease (IBD), many patients fail to respond to therapy. Thus, there is a need to find new therapeutic approaches against IBD. The CD40-CD154 pathway is a known target against IBD and other inflammatory disorders. Clinical trials indicated that CD40 blockade with anti-CD154 antibodies reduced inflammation. However, the anti-CD154 antibodies caused thrombosis (unrelated to inhibition of CD40). Moreover, other approaches to cause global inhibition of CD40 are predicted to increase the risk of opportunistic infections. Identification of a strategy to inhibit CD40-induced inflammation that does not induce thrombosis or opportunistic infections can be a major advance in the treatment of IBD. We uncovered that blocking the interaction between CD40 and an intracellular adaptor protein inhibits pro- inflammatory responses induced by CD40 while leaving protection against an opportunistic pathogen intact. We identified a small molecule that binds the adaptor protein, blocks CD40 signaling, reduces pro- inflammatory responses in vitro and diminishes intestinal inflammation in mouse models of IBD. The compound did not impair resistance against an opportunistic pathogen. The compound has suboptimal solubility and microsomal stability. While some analogs designed to date showed some improvement in solubility or microsomal stability, further optimization is necessary. The objective of this application is to develop an optimized inhibitor that will be tested in mouse and human IBD systems. The central hypothesis is that a potent analog with improved solubility and microsomal stability will optimally block CD40 signaling, and markedly suppress inflammation in mouse models of IBD as well as CD40-driven inflammatory responses in intestinal cells from patients with IBD. To test this hypothesis, we will design and generate analogs of the compound, test their properties, perform signaling studies in reporter cells and intestinal cells and test the lead inhibitor in animal models of IBD. In the first specific aim we will use structure activity relationships with the aid of a docking model to design and generate analogs of the compound in order to improve solubility and microsomal stability. We will examine their ability to inhibit CD40 signaling and their affinity for the adaptor protein. In the second aim, we will test the most potent analogs to determine if they inhibit CD40 signaling in vivo. In the third aim, we will determine if the lead analog reduces intestinal inflammation in mouse models of IBD and inhibits CD40-induced expression of inflammatory molecules in intestinal cells from IBD patients. The proposed work may lead to a new strategy to treat IBD based on a novel approach to inhibit CD40 signaling.