Lead optimization of novel CRFBP-CRFR2 complex modulators for alcohol use disorder

项目摘要 这份R01申请题为《酒精用新型CRFBP-CRFR2复合调制剂的先导优化 无序“是对PAR-17-336”发现新的大脑靶点的活体化学探针(R01)“的回应。 在美国，酒精使用障碍(AUD)影响着1510万18岁以上的成年人，是第四大 是可预防的主要死因。仍然有一个关键的需求没有得到满足，需要开发出更有效的产品 治疗澳门氏症。应激是AUD发生和维持的重要组成部分。 促肾上腺皮质激素释放因子(CRF)在下丘脑-垂体-肾上腺轴激活中起重要作用 以及随后糖皮质激素的释放，以应对压力。CRF通过与两个 受体(CRF1和CRF2)和37kD的CRF结合蛋白[CRFBP(37kD)]。CRFBP通过以下途径发挥关键作用 CRF2通过腹侧被盖区(VTA)的作用调节酒精消耗。我们有 CRF通过增强N-甲基-D-天冬氨酸介导的兴奋性调节突触输入 该区域通过CRFBP/CRF2相互作用的突触后电流。最近，我们的数据表明， CRFBP的作用，其中CRFBP(27kD)片段作用于终止CRF效应以及CRFBP (10kD)片段具有潜在的兴奋作用。这些数据支持CRFBP具有 CRF的功能不仅仅是隔离CRF，它与CRF2的相互作用可能代表着CRF的一个新靶点 AUD的治疗。我们开发并进行了一种利用系留受体的新型高通量筛查 CRFBP(10kD)与CRF2的结合及鉴定的新型小分子CRFBP-CRF2阴性 变构调节剂(NAM)对CRF起非竞争性作用。这些NAM不抑制CRF2 在没有CRFBP(10kD)或抑制CRF1的情况下。我们的结构-活性-关系研究导致了 准备进行全面化学优化的先导和后备CRFBP-CRF2 NAMS的开发 以提供适合于体外研究的化合物。因此，我们的总体目标是发展口腔活动 CRFBP-CRF2调节剂适用于治疗AUD的先进体内概念验证研究。我们的 具体目标是：1)设计和合成具有体内口服活性的优化的CRFBP-CRF2 NAMS； 在相关的体外检测中评估CRFBP-CRF2 NAMS的效力和选择性；3)描述吸收， CRFBP-CRF2-NAMS的体外分布、代谢和排泄特性及药代动力学 (PK)体内性质；以及4)体外AUD啮齿动物模型中铅CRFBP-CRF2 NAM探针的特征。 生成的CRFBP-CRF2 NAM将为测试CRFBP-CRF2的作用提供强大的工具 活体内相互作用。更重要的是，我们处于有利地位，可以开发有效和选择性的小分子。 CRFBP-CRF2 NAMS具有优化的PK属性，将用于体内概念验证研究。这 多学科研究计划有可能对科学和医学产生重大影响 发现了治疗澳门氏症的新药。

Project Summary This R01 application entitled “Lead Optimization of Novel CRFBP-CRFR2 Complex Modulators for Alcohol Use Disorder” is in response to PAR-17-336 “Discovery of in vivo Chemical Probes for Novel Brain Targets (R01)”. In the United States, alcohol use disorder (AUD) affects 15.1 million adults over the age of 18 and is the 4th leading preventable cause of death. There remains a critical unmet need to develop more effective therapeutics to treat AUD. Stress is a significant component in the development and maintenance of AUD. Corticotropin releasing factor (CRF) plays an obligatory role in hypothalamic-pituitary-adrenal axis activation and subsequent release of glucocorticoids in response to stress. CRF exerts its effects by binding to two receptors (CRF1 and CRF2) and a 37 kD CRF binding protein [CRFBP (37kD)]. CRFBP plays a key role via CRF2 in the modulation of ethanol consumption through actions in the ventral tegmental area (VTA). We have demonstrated that CRF modulates synaptic input by potentiating N-methyl-D-aspartate-mediated excitatory postsynaptic currents through CRFBP/CRF2 interactions in this region. More recently, our data suggest a dual role for CRFBP where the CRFBP (27kD) fragment acts to terminate CRF effects and where the CRFBP (10kD) fragment has a potential excitatory function. These data support the hypothesis that CRFBP has functions beyond sequestering CRF and that its interaction with CRF2 may represent a novel target for the treatment of AUD. We developed and performed a novel high-throughput screen utilizing a tethered receptor complex between CRFBP (10kD) and CRF2 and identified novel, small molecule, CRFBP-CRF2 negative allosteric modulators (NAMs) that act noncompetitively with respect to CRF. These NAMs do not inhibit CRF2 in the absence of CRFBP (10kD) or inhibit CRF1. Our structure-activity-relationship studies led to the development of both lead and back-up CRFBP-CRF2 NAMs that are ready for full-scale chemistry optimization to provide compounds suitable for ex vivo studies. Thus, our overall objective is to develop orally active CRFBP-CRF2 modulators suitable for advanced in vivo proof-of-concept studies for the treatment of AUD. Our Specific Aims are: 1) Design and synthesize optimized CRFBP-CRF2 NAMs that are orally active in vivo; 2) Assess the potency and selectivity of CRFBP-CRF2 NAMs in relevant in vitro assays; 3) Profile the absorption, distribution, metabolism, and excretion (ADME) properties of CRFBP-CRF2 NAMs in vitro and pharmacokinetic (PK) properties in vivo; and 4) Characterize lead CRFBP-CRF2 NAM probes in ex vivo rodent models of AUD. The CRFBP-CRF2 NAMs generated will provide powerful tools for testing the role of the CRFBP-CRF2 interaction in vivo. More importantly, we are well-positioned to develop potent and selective small molecule CRFBP-CRF2 NAMs with optimized PK properties that will be utilized for in vivo proof-of-concept studies. This multidisciplinary research program has the potential for significant scientific and medical impact by contributing to the discovery of new medications for AUD.