Smooth Muscle Myosin PROTACs as a Novel Treatment for Asthma

平滑肌肌球蛋白 PROTAC 作为哮喘的新型治疗方法

• 批准号: 10592797
• 负责人: Scott Alan Snyder
• 金额: $ 24.36万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-11-08 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10592797
• 关键词: Acetylcholine; Acute; American; Anesthesia procedures; Antineoplastic Agents; Asthma; Binding; Biological Assay; Bronchoconstriction; Bronchoconstrictor Agents; C57BL/6 Mouse; Cardiac; Cardiac Myocytes; Cells; Contracts; Development; Direct Costs; Disease; Dose; Economics; Epithelial Cells; Epithelium; Evaluation; Facilities and Administrative Costs; Goals; Half-Life; Human; Impairment; In Vitro; Incubated; Length; Lung; Measures; Mechanical ventilation; Morbidity - disease rate; Motor; Mus; Muscle; Muscle Cells; Muscle Contraction; Muscle Fibers; Myosin ATPase; Paralysed; Persons; Pharmaceutical Preparations; Play; Prevention; Process; Protac; Protein Isoforms; Proteins; Pulmonary Hypertension; Reporting; Residual state; Series; Side; Slice; Smooth Muscle; Smooth Muscle Myocytes; Smooth Muscle Myosins; Symptoms; Testing; Thalidomide; Trachea; Treatment Protocols; Tube; Ubiquitination; Western Blotting; airway muscle; analog; asthma exacerbation; bronchial epithelium; care costs; constriction; design; editorial; experimental study; in vivo; methacholine; multicatalytic endopeptidase complex; non-muscle myosin; novel; novel therapeutics; preclinical development; prevent; programs; prototype; recruit; respiratory smooth muscle; response; scaffold; skeletal; small molecule; ubiquitin-protein ligase

PROJECT SUMMARY/ABSTRACT In a 2007 editorial in NEJM, we proposed that depleting airway smooth muscle of myosin – the contractile mo- tor protein responsible for myocyte contraction – would be an effective strategy for preventing acute asthma attacks, but no drug strategy to accomplish this was available. However, a clever new class of compounds, Proteolysis Targeting Chimeras (PROTACs), has been developed that could achieve this goal. These small, bifunctional molecules catalyze proteasome degradation of a targeted protein by stimulating its ubiquitination. Each PROTAC contains one moiety that binds to the targeted protein, and is connected through a linker to a second “bait” moiety that binds an E3 ligase; when both sides are bound, the E3 ligase is brought close to the targeted protein, which it ubiquitinates and so targets for degradation. The process is catalytic because the PROTAC is released upon target degradation and is available for another cycle of target binding, E3 ligase re- cruitment, and target ubiquitination. Here, we propose to test the new idea that a PROTAC to degrade smooth muscle myosin within airway smooth muscle cells could be used as a new treatment for asthma. Smooth muscle myosin has a long half-life and slow turnover rate and so seems ideally suited for degradation by PROTACs. Depleting airway myocytes of myosin would drastically impair contraction, and PROTACs targeting smooth muscle myosin (SMM-PROTACs) should prevent acute airway constriction in eve- ryone with asthma. In this R21 proposal, we will pursue proof-of-principle experiments aimed at demonstrating SMM-PROTACs can inhibit bronchoconstriction, through the execution of four specific aims: 1) Synthesize and characterize a small series of prototype SMM-PROTACs 2) Demonstrate that SMM-PROTACs degrade smooth muscle myosin in human airway smooth muscle, but do not degrade cardiac, skeletal, or non-muscle myosins 3) Demonstrate that SMM-PROTACs inhibit constriction of airways in human lung slices 4) Demonstrate that SMM-PROTACs delivered into the airways blunt methacholine (MCh)-induced bron- choconstriction in mice Demonstration that SMM-PROTACs can inhibit bronchoconstriction by selectively stimulating degradation of smooth muscle myosin would justify their further preclinical development as a novel asthma treatment. This finding would also justify evaluation of whether SMM-PROTACs inhibit progression of other diseases in which smooth muscle contraction plays a key role.

项目概要/摘要 在 NEJM 2007 年的一篇社论中，我们提出，消耗气道平滑肌的肌球蛋白（收缩性分子） 负责心肌细胞收缩的tor蛋白——将是预防急性哮喘的有效策略 攻击，但没有可用的药物策略来实现这一目标。然而，一类巧妙的新型化合物， 蛋白水解靶向嵌合体 (PROTAC) 的开发可以实现这一目标。这些小， 双功能分子通过刺激目标蛋白的泛素化来催化目标蛋白的蛋白酶体降解。 每个 PROTAC 包含一个与目标蛋白结合的部分，并通过连接子连接到 结合 E3 连接酶的第二个“诱饵”部分；当两侧都结合时，E3 连接酶靠近 目标蛋白质，它泛素化并因此作为降解的目标。该过程具有催化作用，因为 PROTAC 在靶标降解后释放，可用于另一个靶标结合周期，E3 连接酶重新 crucrument 和目标泛素化。在这里，我们建议测试一下 PROTAC 降解的新想法 气道平滑肌细胞内的平滑肌肌球蛋白可作为一种新的治疗方法 哮喘。平滑肌肌球蛋白具有较长的半衰期和较慢的周转率，因此似乎非常适合 被 PROTAC 降解。消耗气道肌细胞的肌球蛋白会严重损害收缩，并且 靶向平滑肌肌球蛋白的 PROTAC（SMM-PROTAC）应能预防急性气道收缩。 瑞昂患有哮喘。在这个 R21 提案中，我们将进行原理验证实验，旨在证明 SMM-PROTAC 可以通过执行四个特定目标来抑制支气管收缩： 1) 合成并表征一小系列原型 SMM-PROTAC 2) 证明SMM-PROTACs降解人气道平滑肌中的平滑肌肌球蛋白， 但不会降解心脏、骨骼或非肌肉肌球蛋白 3) 证明SMM-PROTACs抑制人肺切片中的气道收缩 4) 证明 SMM-PROTAC 输送到气道中可以钝化乙酰甲胆碱 (MCh) 诱导的支气管 小鼠心肌收缩 证明 SMM-PROTAC 可以通过选择性刺激降解来抑制支气管收缩 平滑肌肌球蛋白将证明其作为一种新型哮喘治疗方法的进一步临床前开发是合理的。这 这一发现也证明了评估 SMM-PROTAC 是否能抑制其他疾病的进展是合理的。 平滑肌的收缩起着关键作用。