Drug Discovery Pipeline Targeting Pathologically Leaky Calcium Release Channels in Age-Related Indications

针对年龄相关适应症中病理性渗漏钙释放通道的药物研发管线

• 批准号: 10157143
• 负责人: RAZVAN LIVIU CORNEA
• 金额: $ 49.94万
• 依托单位: PHOTONIC PHARMA, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-15 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10157143
• 关键词: Aging; Alzheimer' s Disease; Alzheimer' s disease model; Animals; Binding; Biological Assay; Brain; Calcium; Calcium Signaling; Calmodulin; Cell physiology; Cells; Chemicals; Chicago; Clinical; Collaborations; Complement; Development; Dose; Endoplasmic Reticulum; Ensure; FKBP1B gene; Family suidae; Fluorescence; Fluorescence Resonance Energy Transfer; Functional disorder; Funding; Future; Grant; Heart; Human; In Situ; Individual; Interruption; Label; Lead; Legal patent; Letters; Libraries; Licensing; Membrane; Monitor; Muscle; Myocardium; Neurons; Organelles; Pathogenesis; Pathologic; Pathology; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Protein Isoforms; Reader; Research; Rest; RyR1; RyR2; RyR3; Ryanodine Receptor Calcium Release Channel; Sarcoplasmic Reticulum; Skeletal Muscle; Small Business Innovation Research Grant; Source; Specificity; Structure; Structure-Activity Relationship; System; Systems Analysis; Tacrolimus Binding Proteins; Technology; Therapeutic; Translating; Universities; Validation; Vesicle; Work; age related; age related neurodegeneration; analog; base; commercialization; design; drug discovery; experience; follow-up; high throughput screening; inhibitor/antagonist; interest; novel therapeutic intervention; photonics; prevent; receptor function; response; screening; small molecule; targeted agent; therapeutic target

Project Summary Photonic Pharma LLC (PP) proposes a Phase I SBIR project to launch high-throughput screening (HTS) for discovery of small-molecule allosteric inhibitors of pathologically leaky ryanodine receptor (RyR) calcium (Ca) release channels, to arrest or reverse age-related neurodegeneration, focused on Alzheimer’s disease (AD). In AD, dysregulated intracellular Ca fuels a pathophysiological vicious cycle. RyRs are responsible for triggered Ca release from intracellular storage organelles (sarcoplasmic reticulum – SR; endoplasmic reticulum – ER). The resulting Ca signaling is essential for many cellular processes in muscle and non-muscle cells. Dysregulated RyR function, producing a Ca-leaky state of the resting ER/SR, has been identified as a key contributor to age-related pathologies, particularly AD. In this proposal, we target RyR2, the predominant brain isoform, which has been clearly identified as a target for AD pathogenesis and therapy. We have translated our academic research at UMN toward fluorescence-based HTS structural assays targeting RyR2 (US patent 10,281,476). This FRET assay uses fluorescently labeled RyR2 modulators, calmodulin (CaM) and FKBP12.6 (FKBP), to monitor RyR2-CaM binding. Using small pilot screens, we have demonstrated an inverse correlation between compound effects on our FRET readout and resting RyR2 activity (Ca leak). This established an HTS platform to identify allosteric regulators with therapeutic potential to alleviate RyR2 Ca leak. In our academic work at UMN, we have a funded R01 grant to apply this technology to studying RyR2 function and therapy in the heart; PP has obtained an exclusive commercial license for this UMN patent. In this project, we will apply this technology for therapeutic discovery to correct intracellular Ca signaling in Alzheimer’s disease. In Phase I, we will establish a screening funnel for (a) a large-scale HTS campaign and (b) subsequent lead development via structure-activity relationship (SAR) studies. In Phase II, we will expand SAR, and begin cell-based and animal studies using models of AD. To achieve the objectives in Phase I, PP (1) has demonstrated experience applying this FRET system for RyR2-targeted HTS, (2) will use its proprietary high-precision, high-throughput FLT plate reader (FLT-PR), (3) has functionally validated Hits yielded by this HTS platform, and (4) has identified leak inhibitors of RyR2 in heart. The Phase I Specific Aims: Aim 1 – Primary HTS of a 50,000-compound CNS-selected library, to identify compounds that correct the leaky RyR state. Aim 2 – Determine functional effects of Hits from Aim 1. We have assembled an outstanding interdisciplinary team, including academic experts from several universities (medicinal chemist Aldrich at UMN; RyR2 AD expert Stutzmann at Rosalind Franklin University, human RyR2 expert Zima at Loyola University Chicago, complementing the Photonic team’s expertise on AD-targeted HTS. These collaborations, and our track record of successful collaboration with a major drug company, combine to ensure the successful future commercialization of this project.

项目摘要 Photonic Pharma LLC（PP）提案I期SBIR项目，启动高通量筛查（HTS） 发现小分子变构抑制剂的病理渗漏ryanodine受体（RYR）钙（CA） 释放通道，以阻止或逆转与年龄相关的神经变性，重点是阿尔茨海默氏病 （广告）。在AD中，细胞内CA的失调为病理生理的恶性循环提供。 Ryrs负责 从细胞内存储细胞器（肌质网 - SR;内质网）中触发了Ca释放 - ER）。所得的CA信号对于肌肉和非肌肉细胞中的许多细胞过程至关重要。 RYR功能失调，产生静息ER/SR的Ca-Leaky状态，已被确定为钥匙 与年龄有关的病理，特别是AD的贡献者。在此提案中，我们靶向主要的大脑RYR2 同工型，已被清楚地识别为AD发病机理和治疗的靶标。我们有 将我们在UMN的学术研究转换为基于荧光的HTS HTS结构性测定RYR2 （美国专利10,281,476）。该FRET分析使用荧光标记的RYR2调节剂，钙调蛋白（CAM）和 FKBP12.6（FKBP），以监视RYR2-CAM结合。使用小型飞行员屏幕，我们证明了一个 化合物对我们的FRET读数和静息RYR2活性（CA泄漏）之间的逆相关性。这 建立了一个HTS平台，以确定具有治疗潜力的变构调节器，以减轻RYR2 CA 泄露。在UMN的学术工作中，我们有一项资助的R01赠款，将这项技术应用于研究RYR2 心脏的功能和治疗； PP已获得该UMN专利的独家商业许可证。在这个 项目，我们将将这项技术应用于治疗发现以纠正细胞内CA信号传导 阿尔茨海默氏病。在第一阶段，我们将为（a）大规模HTS广告系列建立筛选漏斗 （b）随后通过结构活性关系（SAR）研究进行铅发展。在第二阶段，我们将 扩展SAR，并使用AD模型开始基于细胞和动物研究。在阶段实现对象 I，pp（1）已经证明了将此fret系统应用于RYR2靶向HTS的经验，（2）将使用其 专有高精度，高通量FLT读取器（FLT-PR），（3）已在功能上验证了命中 由该HTS平台产生的，（4）已确定了RYR2的泄漏抑制剂。第一阶段的特定目的： AIM 1 - 50,000个混合CNS选择库的主要HTS，以识别纠正该化合物的化合物 泄漏的里尔州。 AIM 2 - 确定AIM 1的命中功能效应。我们已经组装了一个 出色的跨学科团队，包括来自几所大学的学术专家 阿尔德里奇（Aldrich）在Umn； Ryr2 AD专家Stutzmann，Rosalind Franklin University，Human Ryr2专家Zima 芝加哥洛约拉大学（Loyola University），完成了光子团队在针对广告的HTS方面的专业知识。这些 合作，以及我们与一家大型制药公司成功合作的往绩记录， 确保该项目的未来成功商业化。