Identifying Novel Senolytic Agents and Molecular Targets

识别新型抗衰老药物和分子靶点

• 批准号: 10229301
• 负责人: Guangrong Zheng
• 金额: $ 37.81万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10229301
• 关键词: Adverse effects; Aging; Animal Model; Apoptosis; Apoptotic; Capsicum; Cell Aging; Cells; Cellular biology; Chronic; Clinical; Deterioration; Development; Diet; Disease; Dose; Elderly; Evaluation; Future; Genetic; Human; Inflammation; Lead; Longevity; Methods; Molecular; Molecular Target; Mus; Natural Products; Oral; Organ; Oxidative Stress; Pathway interactions; Pharmaceutical Preparations; Phenotype; Piper; Process; Property; Proteins; Proteomics; RNA Interference; Reporting; Research; Risk Factors; Safety; Structure-Activity Relationship; Technology; Therapeutic; Therapeutic Agents; Tissues; Toxic effect; Transgenic Mice; age related; aged; analog; anti aging; antioxidant enzyme; base; chemoradiation; design; drug development; healthspan; improved; in vitro activity; in vivo; in vivo evaluation; insight; irradiation; lead candidate; novel; oxidation; premature; prevent; scale up; screening; senescence; small molecule; success; tissue stem cells

PROJECT SUMMARY/ABSTRACT Cellular senescence is an important fundamental mechanism of aging and age-related diseases. It has been demonstrated that genetic elimination of senescent cells (SCs) can dramatically alleviate aging-related phenotypes and extend both healthspan and lifespan in mice. Senolytics, referring to small-molecules that can selectively induce apoptosis in SCs, thus have the potential to be developed as “anti-aging” drugs that extend human healthspan by preventing or treating age-related diseases. We and others have recently discovered a number of senolytics that target various SC anti-apoptotic pathways (SCAPs). Subsequent studies on these senolytics in animal models have shown that clearance of SCs can treat several age-related diseases and chemo-/radio-therapy caused adverse effects in mice, thus demonstrating the therapeutic potential of senolytics. However, most of the reported senolytics have potential on-target and off-target toxicities. In addition, SCs are not all the same; different SCs may use different SCAPs to resist apoptosis. Thus, it is important to continue the search of new SCAPs/protein targets and new senolytics. Piperlongumine (PL), a dietary natural product, was identified as a novel senolytic agent through our targeted screening campaign. PL appears to be safe, without observable toxicity in vivo and having a high maximum tolerated oral dose in mice. However, its mechanism of action (MOA) against SC is undefined, raising the potential of discovering novel molecular targets for senolytic drug development. Therefore, we hypothesize that PL can be used to identify new molecular targets in SCs, which can then be exploited for the development of novel and specific senolytics. Our preliminary structure–activity relationship (SAR) studies have identified PL analogues (piperlogues) with senolytic profile comparable to that of ABT263, one of the most potent and selective senolytics identified to date, and also designed and synthesized PL-based probe molecules for the pulldown of target proteins in SCs. With proteomics analyses and RNAi technology, we identified, validated, and characterized oxidation protein 1 (OXR1) as one of the protein targets of PL in SCs. The objectives of this application are to continue the discovery and evaluation of piperlogues as potent and selective senolytics, identification of their molecular targets, and elucidation of the MOA of these novel agents. Specifically, we will pursue the following specific aims: 1) design, synthesize, and evaluate novel piperlogues with improved senolytic activity and druglike properties; 2) identify and validate the protein targets of PL and piperlogues in SCs, and elucidate their MOA; and 3) evaluate the in vivo efficacy and safety of lead piperlogues. Success in these proposed aims will not only produce anti-aging drug development candidates but also identify and validate additional novel molecular target(s) to enable future design and target-based discovery of novel senolytics and to gain more insights into the biology of cellular senescence and the MOA of PL and piperlogues.

项目概要/摘要 细胞衰老是衰老和年龄相关疾病的重要基本机制。它一直 证明通过基因消除衰老细胞（SC）可以显着缓解与衰老相关的问题 表型并延长小鼠的健康寿命和寿命。 Senolytics，指的是能够 选择性诱导干细胞凋亡，因此有潜力开发为延长寿命的“抗衰老”药物 通过预防或治疗与年龄相关的疾病来延长人类的健康寿命。我们和其他人最近发现了一个 针对各种 SC 抗凋亡途径 (SCAP) 的 senolytics 数量。后续对这些的研究 动物模型中的 senolytics 表明清除 SC 可以治疗多种与年龄相关的疾病 化疗/放疗对小鼠造成了不良影响，从而证明了其治疗潜力 senolytics。然而，大多数报道的 senolytics 都具有潜在的靶向和脱靶毒性。在 此外，SC 并不完全相同；不同的SC可能使用不同的SCAP来抵抗细胞凋亡。因此，它是 继续寻找新的 SCAP/蛋白质靶点和新的 senolytics 非常重要。胡椒长明 (PL) 膳食天然产品，通过我们的针对性筛选活动被确定为一种新型的衰老剂。 PL 似乎是安全的，体内没有可观察到的毒性，并且在小鼠中具有较高的最大耐受口服剂量。 然而，其针对 SC 的作用机制 (MOA) 尚未确定，这提高了发现新颖的潜力 senolytic 药物开发的分子靶点。因此，我们假设PL可以用来识别 SC 中的新分子靶标，然后可用于开发新型和特定的 senolytics。 我们的初步构效关系 (SAR) 研究已确定 PL 类似物 (piperlogues) 与 ABT263 相当的 senolytic 特性，ABT263 是公认的最有效和选择性的 senolytics 之一 日期，并还设计和合成了基于 PL 的探针分子，用于在 SC 中下拉目标蛋白。 通过蛋白质组学分析和 RNAi 技术，我们鉴定、验证和表征了氧化蛋白 1 (OXR1) 作为 SC 中 PL 的蛋白质靶标之一。该应用程序的目标是继续 作为有效和选择性的衰老剂的胡椒系物的发现和评估，其分子的鉴定 目标，并阐明这些新型药物的作用机制。具体来说，我们将采取以下具体措施 目标：1) 设计、合成和评估具有改善的 senolytic 活性和药物样作用的新型哌啶衍生物 特性; 2) 鉴定并验证SCs中PL和piperlogues的蛋白靶标，并阐明它们的MOA； 3) 评估先导哌啶衍生物的体内功效和安全性。这些拟议目标的成功不会 不仅生产抗衰老药物开发候选药物，还识别和验证其他新分子 目标，以实现新型 senolytics 的未来设计和基于目标的发现，并获得更多见解 细胞衰老的生物学以及 PL 和 Piperogues 的 MOA。