Advancing mGlu1 positive allosteric modulators as therapeutics to facilitate abstinence in cocaine use disorder

推进 mGlu1 正变构调节剂作为促进可卡因使用障碍戒断的治疗方法

• 批准号: 10577196
• 负责人: Marina Elizabeth Wolf
• 金额: $ 32.54万
• 依托单位: ELEUTHERIA PHARMACEUTICALS LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2024-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10577196
• 关键词: Abstinence; Adverse effects; Benchmarking; Biological Assay; Brain region; Businesses; Cell Line; Chemicals; Clinical; Cocaine; Cocaine Users; Cocaine use disorder; Collection; Couples; Cues; Development; Disease; Dose; Drug Screening; Epidemic; Exhibits; FDA approved; Family; Funding; Future; GRM1 gene; GRM5 gene; Glutamate Receptor; Glutamates; Goals; Grant; Human; In Vitro; Incubated; Injections; Laboratories; Lead; Libraries; Link; Long-Term Depression; Mediating; Metabotropic Glutamate Receptors; Methamphetamine; Modeling; National Institute of Drug Abuse; Nucleus Accumbens; Opioid; Oregon; Overdose; Patients; Permeability; Persons; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Positioning Attribute; Prefrontal Cortex; Property; Proteins; Rattus; Relapse; Reproducibility; Rewards; Sampling; Self Administration; Signal Pathway; Signal Transduction; Small Business Technology Transfer Research; Source; Substance Use Disorder; Synapses; Synaptic plasticity; Testing; Therapeutic; Therapeutic Agents; Translations; Universities; Ventral Tegmental Area; Wolves; Work; assay development; base; craving; cytotoxicity; design; drug craving; drug of abuse; high throughput screening; in vivo evaluation; member; metabotropic glutamate receptor type 1; novel; novel strategies; positive allosteric modulator; pre-clinical; scaffold; screening; screening services; small molecule; success; therapeutic target; tool

Project Summary A major challenge in treating cocaine use disorder is that vulnerability to cue-induced craving and relapse persist even after long periods of abstinence. There is presently no FDA-approved medication to lessen cocaine craving. The goal of Eleutheria Pharmaceuticals LLC is to develop metabotropic glutamate receptor 1 (mGlu1) positive allosteric modulators (PAMs) as therapeutic agents to reduce craving and thereby facilitate abstinence in persons with cocaine use disorder. The target, mGlu1, has been thoroughly validated by the lab of the PI and Company Founder, and by other work. Our studies have used the ‘incubation of drug craving’ model, in which cue-induced craving in rats progressively intensifies (‘incubates’) during abstinence and then remains high for months. Incubation of craving also occurs in humans. We showed that incubation depends on strengthening of glutamate synapses in a key brain region for drug craving (nucleus accumbens) via incorporation of atypical high conductance AMPA-type glutamate receptors (CP-AMPARs). mGlu1 PAMs reverse this plasticity by eliciting a form of long-term depression (LTD) that is expressed by CP-AMPAR internalization and, through this mGlu1- LTD, reduce cocaine craving. In 2020, the PI obtained a NIDA U18 grant that has taken our project to the Assay Development stage. In this Phase I STTR application, we propose two Aims that will result in the identification of novel chemotypes and position us to apply for Phase II STTR funding. Classic in vitro screens for mGlu1 activation rely on detecting Ca2+ mobilization in cell lines. We have developed a novel assay to selectively detect activation of the signaling pathway downstream of mGlu1 that mediates craving reduction, namely the rapid translation of the LTD protein oligophrenin-1 (OPHN1). Briefly, HiBiT-tagged OPHN1 (stably transfected into an mGlu1 over-expressing cell line) generates a luminescent signal upon mGlu1 stimulation. Aim 1 will optimize this assay so it can be used in high throughput screening in Aim 2. The assay will be optimized by reduction of baseline signal to achieve an optimal signal window and Z’ ≥ 0.6 as a benchmark for success. Reproducibility at scale will be evaluated during a pilot screen of 1430 unique compounds tested in triplicate at a single dose. In Aim 2, the optimized assay will be used for a quantitative high throughput primary screen of 4382 bioactive compounds at Oregon State University’s High-Throughput Screening Services Laboratory. Hits from this screen and additional compounds chosen for similarity to our mGlu1 PAM tool compound will be sourced fresh for dose curves and cytotoxicity assessment, and medicinal chemistry prioritization conducted. If we are successful in achieving our Phase I goals (optimize and execute the primary screen, identify and validate hit scaffolds), we will submit a Phase II application to assess prioritized compounds in secondary assays, optimize their activity and PK properties during iterative SAR studies, and conduct in vivo testing (‘incubation of craving’ model) to identify lead compounds. To achieve these goals, the PI has assembled a strong team with expertise in drug screening, medicinal chemistry, business development, and the needs of substance use disorder patients.

项目摘要 治疗可卡因使用障碍的一个主要挑战是易受线索诱导的渴望和复发的影响 即使是在长期禁欲之后。目前还没有FDA批准的药物来减轻对可卡因的渴望。 Eleutheria Pharmaceuticals LLC的目标是开发代谢型谷氨酸受体1（mGlu 1）阳性 作为治疗剂的别构调节剂（PAM），以减少食欲并由此促进戒断， 患有可卡因使用障碍的人。靶标mGlu 1已由PI实验室进行了全面验证， 公司创始人，并通过其他工作。我们的研究使用了“药物渴望的孵化”模型，其中 在大鼠中，线索诱导的渴望在戒断期间逐渐增强（“潜伏”），然后在戒断期间保持较高水平。 个月人类也会产生渴望。我们发现孵化依赖于 谷氨酸突触在一个关键的大脑区域的药物渴望（核）通过纳入非典型高 电导AMPA型谷氨酸受体（CP-AMPAR）。mGlu 1 PAMs通过引发一种 一种长期抑郁症（LTD），由CP-AMPAR内化表达，通过这种mGlu 1- LTD，减少对可卡因的渴望。2020年，PI获得了NIDA U18资助，使我们的项目进入了检测阶段。 发展阶段在第一阶段的STTR应用中，我们提出了两个目标，这将导致识别 新的化学型和立场，我们申请第二阶段STTR资金。mGlu 1的经典体外筛选 激活依赖于检测细胞系中的Ca 2+动员。我们开发了一种新的检测方法， 激活mGlu 1下游的信号通路，介导渴求减少，即快速 LTD蛋白寡聚蛋白-1（OPHN 1）的翻译。简言之，将HiBiT标记的OPHN 1（稳定转染到转染细胞中）转染到细胞中。 mGlu 1过表达细胞系）在mGlu 1刺激时产生发光信号。目标1将优化 该测定因此可用于Aim 2中的高通量筛选。将通过减少 基线信号以实现最佳信号窗口，并且Z' ≥ 0.6作为成功的基准。时的重现性 将在以单剂量一式三份测试的1430种独特化合物的中试筛选期间评估规模。在 目的2、优化后的方法可用于4382生物活性物质的高通量筛选 化合物在俄勒冈州州立大学的高通量筛选服务实验室。来自此屏幕的点击数 选择与我们的mGlu 1 PAM工具化合物相似的其他化合物将新鲜来源于剂量 曲线和细胞毒性评估，并进行药物化学优先级排序。如果我们成功地 为了实现我们的第一阶段目标（优化和执行初级筛选，识别和验证命中支架），我们 我将提交第二阶段申请，以评估二级试验中的优先化合物，优化其活性， 和PK特性，并进行体内试验（“渴望培养”模型）， 识别先导化合物。为了实现这些目标，PI组建了一支强大的药物专业团队， 筛查、药物化学、业务发展和物质使用障碍患者的需求。