Development of a Novel Medication for Alcohol Use Disorder with an Active IND Dual Inhibitor of T-Type Calcium Channel and Soluble Epoxide Hydrolase

7.总结 酒精使用障碍（AUD）是世界范围内最普遍的物质使用障碍，通常与 慢性疼痛、焦虑、抑郁和阿片依赖。不幸的是，目前可用的药物 AUD的疗效有限，有不良副作用。为了解决这一未得到满足的医疗需求，我们提出了SBIR I期U43项目响应PAR-22-102“研究性新药（IND）-使能和早期阶段 治疗酒精使用障碍和酒精相关器官损伤的药物开发。“我们的项目 将专注于我们的活性IND（157314）AFA-281的概念验证研究，AFA-281是一种新型T型双重抑制剂 钙通道（Cav 3）和可溶性环氧化物水解酶（sEH），用于AUD治疗。这些概念验证 研究将使用完善的啮齿动物AUD模型和测试范例。 AfaSci从Cav 3通道和sEH的一系列专利双调节剂中发现了AFA-281， 阿片类镇痛药我们通过合理的药物设计和迭代筛选， 膜片钳记录和酶测定。进行了先导物优化，包括选择性筛选 对hERG和关键心脏离子通道的影响，以及对81个停药靶点的CEREP验证。这些放映在一起 体外使用人心肌细胞的研究证明AFA-281没有心脏安全性问题。 在药代动力学和药效学（PK/PD）研究中，AFA-281显示出优异的口服生物利用度， 可接受的t1/2、CNS渗透性、对神经性和炎性疼痛的广泛镇痛作用和良好的安全性 利润率最近，AFA-281的IND申请被FDA接受，用于神经性疼痛的主要适应症。 我们对AUD的初步研究表明，AFA-281能够减少酒精摄入量， 饮酒和偏好，以及抑制啮齿动物的酒精寻求行为。 在这个拟议的SBIR I期U43项目中，我们将进行临床前概念验证研究，以调查 AFA-281对AUD及其相关慢性疼痛和负性情感障碍（如焦虑和抑郁）的影响 通过三个具体目标对大鼠进行抑郁症治疗。目标1将评估AFA-281在减少饮酒方面的PK/PD 并通过自我管理寻求行为。目的2研究AFA-281对痛觉过敏的影响， 焦虑和抑郁的大鼠从慢性酒精消费中撤出。目标3将探讨机制 通过评估AFA-281对酒精改变的神经细胞的调节， 通过c-Fos表达和脑神经炎症指示的活性，以及Cav 3通道活性的评价，以及 外侧缰核的细胞兴奋性，这是一个与AUD有关的大脑区域。 在我们的第一阶段项目的成功将证明，AFA-281可以减少酒精消费和寻求 行为，提供证据基础，为在U44中过渡到临床概念验证研究做准备 SBIR项目。鉴于AUD的重大公共卫生负担，开发更有效和安全的药物 可获得的治疗和预防AUD将对公共卫生产生相当大的积极影响。

7. SUMMARY Alcohol use disorder (AUD) is the most prevalent substance use disorder worldwide and often co-occurs with chronic pain, anxiety, depression, and opioid dependence. Unfortunately, currently available medications for AUD have limited efficacy with unwanted side effects. To address this unmet medical need, we propose an SBIR Phase I U43 project in response to PAR-22-102 "Investigational New Drug (IND)-enabling and Early-Stage Development of Medications to Treat Alcohol Use Disorder and Alcohol-Associated Organ Damage." Our project will focus on proof-of-concept studies of our active IND (157314), AFA-281, a novel dual inhibitor of T-type calcium channels (Cav3) and soluble epoxide hydrolase (sEH), for AUD treatment. These proof-of-concept studies would use well-established rodent AUD models and testing paradigms. AfaSci discovered AFA-281 from a series of patented dual modulators of Cav3 channels and sEH as a non- opioid analgesic. We completed lead identification through rational drug design and iterative screenings using patch-clamp recordings and enzymatic assays. Lead optimization was conducted, including selectivity screening on hERG and key cardiac ion channels, and CEREP proofing of 81 off-drug targets. These screenings together with studies using human cardiomyocytes ex vitro demonstrated that AFA-281 has no cardiac safety concerns. In pharmacokinetic and pharmacodynamic (PK/PD) studies, AFA-281 has shown excellent oral bioavailability, acceptable t1/2, CNS penetration, broad analgesic effects on neuropathic and inflammatory pain, and good safety margins. Recently, AFA-281's IND application was accepted by the FDA for a lead indication of neuropathic pain. Our preliminary studies on AUD have demonstrated that AFA-281 is capable of decreasing alcohol intake, binge drinking and preference, and suppressing alcohol-seeking behavior in rodents. In this proposed SBIR Phase I U43 project, we will conduct preclinical proof-of-concept studies to investigate AFA-281's effects on AUD, and its associated chronic pain and negative affective disorders, such as anxiety and depression in rats via three Specific Aims. Aim 1 will evaluate PK/PD of AFA-281 in reducing alcohol consuming and seeking behavior via operant self-administration. Aim 2 will investigate AFA-281's effects on hyperalgesia, anxiety, and depression in rats withdrawn from chronic alcohol consumption. Aim 3 will explore the mechanism of action of AFA-281 underlying its efficacy in AUD via assessing AFA-281’s modulation on alcohol-altered neural activity indicated by c-Fos expression and brain neuroinflammation, and evaluation of Cav3 channel activity, and cell excitability in the lateral habenula, a brain area implicated in AUD. Success in our Phase I project would demonstrate that AFA-281 can reduce alcohol consumption and seeking behavior, provide an evidence base to prepare for the transition to clinical proof-of-concept studies in the U44 SBIR project. Given the significant public health burden of AUD, developing a more effective and safe medication accessible for treatment and prevention of AUD would have a considerable positive impact on public health.