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

项目摘要 标题为“用于酒精用途的新型CRFBP-CRFR 2复合物调节剂的先导优化”的R 01申请 疾病”是对PAR-17-336“发现体内化学探针用于新的脑靶点（R 01）”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 主要可预防的死亡原因。仍然有一个关键的需求没有得到满足， 治疗AUD。压力是AUD发展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-肾上腺轴的激活中起着一定的作用 以及随后响应于应激而释放糖皮质激素。CRF通过与两个 受体（CRF 1和CRF 2）和37 kD CRF结合蛋白[CRFBP（37 kD）]。CRFBP通过以下方式发挥关键作用： CRF 2通过腹侧被盖区（VTA）的作用调节乙醇消耗。我们有 证明CRF通过增强N-甲基-D-天冬氨酸介导的兴奋性突触传入来调节突触输入。 突触后电流通过CRFBP/CRF 2相互作用在这个区域。最近，我们的数据表明， CRFBP的作用，其中CRFBP（27 kD）片段起终止CRF效应的作用，并且其中CRFBP（27 kD）片段起 （10 kD）片段具有潜在的兴奋功能。这些数据支持CRFBP具有以下特征的假设： 功能超越螯合CRF，它与CRF 2的相互作用可能代表了一种新的靶点， 治疗AUD。我们开发并进行了一种新的高通量筛选利用拴系受体 CRFBP（10 kD）和CRF 2之间的复合物，以及鉴定的新型小分子CRFBP-CRF 2阴性 对CRF起非竞争性作用的别构调节剂（NAM）。这些NAM不抑制CRF 2 在CRFBP（10 kD）不存在的情况下或抑制CRF 1。我们的构效关系研究导致了 开发可用于全面化学优化的先导和备用CRFBP-CRF 2 NAM 以提供适合于离体研究的化合物。因此，我们的总体目标是开发口服活性药物， CRFBP-CRF 2调节剂适用于治疗AUD的先进体内概念验证研究。我们 具体目的是：1）设计和合成在体内具有口服活性的优化的CRFBP-CRF 2 NAM; 2） 在相关体外试验中评估CRFBP-CRF 2 NAM的效力和选择性; 3）描述吸收， CRFBP-CRF 2 NAM的体外分布、代谢和排泄（ADME）特性和药代动力学 (PK)体内性质;和4）在AUD的离体啮齿动物模型中表征前导CRFBP-CRF 2 NAM探针。 生成的CRFBP-CRF 2 NAM将为测试CRFBP-CRF 2的作用提供强大的工具 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 将用于体内概念验证研究的具有优化PK特性的CRFBP-CRF 2 NAM。这 多学科研究计划具有重大的科学和医学影响的潜力， 发现治疗AUD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.