Identification of Cyclic Peptide Antagonists of an Anti-Opioid G Protein-Coupled Receptor

抗阿片G蛋白偶联受体环肽拮抗剂的鉴定

基本信息

批准号：
10256110
负责人：
Sid Ahmed Labed
金额：
$ 25.51万
依托单位：
EVODENOVO, INC.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-15 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10256110
关键词：
Absence of pain sensation Address Adverse effects Agonist American Analgesics Animal Model Bacteria Behavior Behavioral Biological Assay Biological Models Brain region Bypass Caenorhabditis elegans Cell Membrane Permeability Chronic Clinical Trials Complex Coupled Cyclic Peptides Cyclization Dependence Detection Development Divorce Drug Prescriptions Eligibility Determination Engineering Escherichia coli Evaluation Event FDA approved Family Florida Foundations G-Protein-Coupled Receptors Goals Government Habenula Health Human Hydrolysis Institutes Investigation Knock-out Knockout Mice Lead Libraries Locomotion Mammalian Cell Mammals Medial Modeling Morphine Movement Mus Nematoda Neurons Opioid Opioid Analgesics Opioid Receptor Orphan Outcome Pain Pain management Peptides Persons Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Phase Physiological Positioning Attribute Predisposition Property Receptor Signaling Recombinants Research Institute Rewards Safety Signal Transduction Small Business Innovation Research Grant System Technology Testing Therapeutic Time Validation Withdrawal abuse liability antagonist base clinically relevant clinically significant cost design drug discovery experimental study feeding follow-up high throughput screening in vivo industry partner inhibitor innovation locus ceruleus structure member mu opioid receptors neural circuit neurotoxicity novel opioid overdose opioid use disorder peptide drug receptor recombinant peptide screening side effect small molecule therapeutic candidate therapeutically effective trafficking tv watching

项目摘要

Identification of Cyclic Peptide Antagonists of an Anti-Opioid G Protein-Coupled Receptor RFA-DA-19-019 R43 Phase I SBIR PI: Sid A. labed Project Summary The highly addictive properties of opioid drugs, coupled with routine over-prescription, have resulted in a national crisis. Accidental overdose by opioids has increased over 400% in the last 15 years, from ~8,000 opioid- related overdoses to more than 30,000 in 2015. The staggering cost of the opioid health crisis to the American public exceeds $50 billion annually, highlighting the need for safer therapeutics for pain management. Drs. Kirill Martemyanov and Brock Grill at the Scripps Institute, Florida, have engineered a model system expressing a functional mammalian opioid receptor (MOR) in the model nematode Caenorhabditis elegans for discovery of opioid modulators. Using this system, they identified an orphan G Protein Coupled Receptor, GPR139, as a negative regulator of MOR signaling. In mammals, GPR139 is co-expressed with MOR in opioid-sensitive brain regions and influences MOR trafficking and signaling. Deletion of GPR139 in mice enhanced opioid-induced inhibition of neuronal firing, increased the analgesic and rewarding effects of morphine, and reduced withdrawal. Previous efforts to target GPR139, which have largely focused on identification of small molecule agonists, has failed to produce a therapeutic candidate antagonist with favorable pharmacological properties. To address this unmet need, EvoDenovo will screen an innovative library of cyclic peptides (CPs) for new GPR139 antagonists by combining two state-of-the-art C. elegans technologies exclusive to EvoDenovo: 1) InVivo Display: a high-throughput screening technology invented by EvoDenovo that can be used for peptide-based drug discovery that bypasses the necessity of peptide purification by directly feeding live recombinant E. coli clones, each expressing a different cyclic peptide, directly to nematodes expressing mammalian MOR and human GP139. This drastically reduces the cost and time for screening. 2) The anti-opioid behavior platform developed at Scripps by Drs. Martemyanov and Gril that assesses the behavioral effects of opioids and identifies pharmacological outcomes of different drugs. The combination of both platforms, assisted by an automated C. elegans movement tracking system, will yield a powerful high throughput screening engine capable of interrogating thousands of recombinant peptides within a few days, all in a unique in vivo setup. GPR139 antagonists identified using this assay would likely be missed by conventional screening protocols. EvoDenovo uses CPs instead of linear peptides. The cyclization of peptides increases gut stability by eliminating vulnerable N- and C-termini, reduces susceptibility to proteolytic hydrolysis, and enhances membrane permeability. There are 2 specific aims: Aim 1: Perform a behavioral screen for cyclic peptide antagonists of GPR139 in C. elegans. Aim 2: Validate hits generated from the C. elegans platform in mammalian cell-based assays. This Phase I proposal will provide the foundation for a Phase II SBIR which will include a larger-scale screen and follow-up on prioritizing CP hits, mammalian testing, and IND enabling studies. EvoDenovo RFA-DA-19-019/ PI: Sid A. Labed Project Summary - Page 1 of 1

抗阿片G蛋白偶联受体环肽拮抗剂的鉴定 RFA-DA-19-019 R43 I 期 SBIR PI：Sid A.Labed 项目概要阿片类药物的高度成瘾性，加上常规的过度处方，导致了国家危机。过去 15 年中，阿片类药物意外服用过量增加了 400% 以上，从约 8,000 件阿片类药物 2015 年，相关过量用药人数超过 30,000 人。阿片类药物健康危机给美国人造成了惊人的损失公众每年花费超过 500 亿美元，凸显了对更安全的疼痛管理疗法的需求。博士。基里尔佛罗里达州斯克里普斯研究所的 Martemyanov 和 Brock Grill 设计了一个模型系统，表达了线虫模型中的功能性哺乳动物阿片受体（MOR）发现阿片类调节剂。使用该系统，他们鉴定了一个孤儿 G 蛋白偶联受体 GPR139，作为 MOR 信号的负调节因子。在哺乳动物中，GPR139 在阿片类药物敏感的大脑中与 MOR 共表达区域并影响 MOR 贩运和信号传导。小鼠中 GPR139 的缺失增强了阿片类药物诱导的抑制神经元放电，增加吗啡的镇痛和奖赏作用，并减少戒断。之前针对 GPR139 的努力主要集中在小分子激动剂的鉴定上，未能生产出具有良好药理学特性的治疗候选拮抗剂。致地址为了满足这一未满足的需求，EvoDenovo 将筛选创新的环肽 (CP) 库以寻找新的 GPR139 通过结合 EvoDenovo 独有的两种最先进的线虫技术来对抗拮抗剂：1) InVivo Display：EvoDenovo发明的一种高通量筛选技术，可用于基于肽的筛选通过直接饲喂活重组大肠杆菌来绕过肽纯化必要性的药物发现每个表达不同环肽的克隆直接作用于表达哺乳动物 MOR 的线虫和人类 GP139。这大大减少了筛选的成本和时间。 2）抗阿片类药物行为平台由 Drs 在斯克里普斯开发。 Martemyanov 和 Gril 评估阿片类药物的行为影响并确定不同药物的药理结果。两个平台的结合，在自动化 C 的协助下。 elegans 运动跟踪系统将产生强大的高通量筛选引擎，能够在几天内询问数千种重组肽，所有这些都在独特的体内设置中进行。探地雷达139 传统的筛选方案可能会漏掉使用该测定法鉴定的拮抗剂。进化进化使用 CP 代替线性肽。肽的环化通过消除脆弱的肽来增加肠道稳定性 N 端和 C 端，降低对蛋白水解的敏感性，并增强膜通透性。有 2 个具体目标：目标 1：对 GPR139 的环肽拮抗剂进行行为筛选 C.线虫。目标 2：在基于哺乳动物细胞的检测中验证线虫平台生成的命中结果。这第一阶段提案将为第二阶段 SBIR 提供基础，其中将包括更大规模的屏幕和后续优先考虑 CP 命中、哺乳动物测试和 IND 启用研究。 EvoDenovo RFA-DA-19-019/ PI：Sid A. Labed 项目摘要 - 第 1 页，共 1 页