Evaluation of USP30 small molecule inhibitors in models relevant to Cardiac Aging

USP30 小分子抑制剂在心脏衰老相关模型中的评价

基本信息

批准号：
10546047
负责人：
Bahareh Behrouz
金额：
$ 29.95万
依托单位：
VINCERE BIOSCIENCES, 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：
2022-08-15 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10546047
关键词：
Adult Aging Animal Model Autophagocytosis Back Binding Proteins Biological Assay Biological Sciences Biology Cardiac Cardiac Myocytes Cell Aging Cell model Cells Clinic Clinical Trials Data Deubiquitinating Enzyme Development Disease Evaluation Future Healthcare Systems Heart Heart Diseases Heart Injuries Heart failure Human In Vitro Inflammation Injections Lead Light Measures Methods Mitochondria Modeling Mus Muscle Cells Myocardial Myocardial Infarction Oligomycins PINK1 gene Parkin Permeability Persons Pharmacology Plant Roots Plasma Proteins Positioning Attribute Process Property Protein Kinase Quality Control Rattus Solubility Testing Therapeutic Tissues Ubiquitin age related antimycin cardiogenesis in vivo inflammatory marker inhibitor knock-down lead candidate mitochondrial dysfunction mortality multidisciplinary nanomolar neonatal mice novel therapeutics prevent senescence small molecule small molecule inhibitor ubiquitin ligase

项目摘要

ABSTRACT Vincere Biosciences has proprietary USP30 inhibitor small molecules being optimized in vivo towards development candidates for heart failure (HF) and cardiac aging. HF is an age-related disorder mechanistically rooted in the decline of mitochondrial quality control mechanisms in myocardial cells which leads to increased inflammation and senescence with decreased myocardial cell turnover. Despite strong rationale for the involvement of mitochondrial dysfunction and mitophagy, there have been no efforts in clinical trials to enhance mitophagy as a therapeutic approach for cardiac aging. Enhancement of mitophagy via knockdown of USP30 or increase in parkin reduces myocardial cell senescence in an aging model of heart disease both in primary cardiomyocytes and in adult rat and mouse myocardial cells in vivo. In light of this compelling evidence that increased mitophagy may provide benefit for cardiac aging, we are compelled to test our USP30 inhibitor compounds in relevant models. Our preliminary data demonstrate that we have developed proprietary compounds with low nanomolar in vitro potency for USP30 inhibition as measured using two orthogonal methods. The compounds are cell-penetrant and enhance mitophagy in the presence of antimycin/oligomycin (A/O) in human cells with endogenous expression of USP30, parkin, and substrates. Importantly, compounds do not damage or depolarize healthy mitochondria as measured by TMRE. Candidate lead-like compounds are highly selective when tested against a panel of over 40 deubiquitinating enzymes using two orthogonal assays. We have profiled compounds for various ADME properties and successfully optimized properties including solubility, permeability, microsomal stability, and plasma protein binding. Our lead compound has an excellent PK profile in rat and mouse and penetrates the heart at sustained levels well above 2x the compounds’ low nM IC50. The following Aims test the hypothesis that our selective USP30 inhibitors can reduce senescence in cell and animal models of cardiac aging. Aim 1: Determine whether USP30 inhibitors can reduce cell senescence in primary myocardial cells We will treat neonatal mouse myocardial cells with USP30 inhibitor/vehicle control in a D-Gal model to determine whether the small molecule inhibitors can reduce cellular senescence in culture. Aim 2: Assess the effects of USP30 inhibition on D-gal induced myocardial cell senescence in vivo . We will evaluate whether inhibition of USP30 can reduce the deficits induced by D-gal, including mitochondrial dysfunction, cellular senescence, and inflammatory markers. We will treat mice with daily D-gal injections, with or without co-administration of USP30 inhibitor for 8 weeks to induce senescence. We will evaluate the effect of compound on multiple markers of mitochondrial function, senescence, and inflammation.

抽象的 Vincere Biosciences 拥有专有的 USP30 抑制剂小分子，并在体内进行了优化心力衰竭（HF）和心脏衰老的发展候选者从机制上讲，心力衰竭是一种与年龄相关的疾病。根源在于心肌细胞线粒体质量控制机制的衰退，导致心肌细胞线粒体质量控制机制的下降。尽管有充分的理由，但炎症和衰老会导致心肌细胞更新减少。线粒体功能障碍和线粒体自噬的参与，临床试验中尚未做出努力来增强线粒体自噬作为心脏衰老的一种治疗方法，通过敲低 USP30 或增强线粒体自噬。 Parkin 的增加可减少原发性心脏病衰老模型中的心肌细胞衰老鉴于这一令人信服的证据表明，心肌细胞以及成年大鼠和小鼠体内的心肌细胞。增加线粒体自噬可能对心脏衰老有益，我们不得不测试我们的 USP30 抑制剂相关模型中的化合物。我们的初步数据表明，我们已经开发出体外低纳摩尔浓度的化合物使用两种正交方法测量的 USP30 抑制效力这些化合物具有细胞渗透性。并在抗霉素/寡霉素 (A/O) 存在的情况下增强具有内源性的人类细胞中的线粒体自噬 USP30、parkin 和底物的表达重要的是，化合物不会损害健康或使健康去极化。通过 TMRE 测量，候选先导化合物在测试时具有高度选择性。我们使用两种正交分析对超过 40 种去泛素化酶进行了分析。各种 ADME 特性并成功优化了特性，包括溶解度、渗透性、微粒体我们的先导化合物在大鼠和小鼠中具有出色的 PK 特性。以远高于化合物低 nM IC50 的 2 倍的持续水平渗透心脏。假设我们的选择性 USP30 抑制剂可以减少心脏病细胞和动物模型的衰老老化。目标 1：确定 USP30 抑制剂是否可以减少原代心肌细胞的细胞衰老我们将在 D-Gal 模型中用 USP30 抑制剂/载体对照处理新生小鼠心肌细胞，以确定小分子抑制剂是否可以减少培养中的细胞衰老。目标 2：评估 USP30 抑制对 D-gal 诱导的体内心肌细胞衰老的影响。我们将评估抑制 USP30 是否可以减少 D-gal 诱导的缺陷，包括线粒体我们将通过每天注射 D-gal 来治疗小鼠。或不联合使用USP30抑制剂8周来诱导衰老，我们将评估其效果。化合物对线粒体功能、衰老和炎症的多种标志物有影响。