Drug Discovery and Development

核心B：项目摘要：这个多学科、多站点的PPG应用程序的总体目标是构建 细胞衰老发现科学的坚实基础，导致一系列治疗策略的流水线 延缓或预防与年龄相关的疾病。四个PPG项目中的每个项目都侧重于老年人的作用 特定年龄相关健康后果中的细胞：项目1，代谢功能障碍(Kirkland/Tchkonia)；项目2， 骨骼脆性(Khosla/Farr)；项目3，血管功能障碍(Miller)；项目4，骨骼肌丧失 和功能障碍(勒布拉瑟)。为了确定因果关系，每个项目都将使用遗传和药理学 去除衰老细胞的方法。重要的是，我们已经确定了感觉剂，药物能够 特异性诱导衰老细胞的凋亡，具有细胞类型特异性，如在脂肪细胞和 不存在于内皮细胞，反之亦然。药物发现和开发核心(DDDC)将与 其他三个核心，以协助所有四个项目，以确定最有效的治疗其 感兴趣的单元类型。DDDC已经开发了基于原代小鼠和人类细胞的多种检测方法 筛选和优化具有感官疗效的化合物和药物组合。感光剂和感光剂的例子 DDDC鉴定的生理活性物质(抑制衰老相关分泌组的药物)包括 BCL-2家族成员的抑制剂，达沙替尼与槲皮素的组合，HSP90抑制剂， 类黄酮非瑟素、雷帕霉素、IKK/NF-κB抑制剂、JAK抑制剂、多聚体白藜芦醇和线粒体- 以自由基清除者为目标。没有一种感官治疗化合物被证明能有效地清除或 调节所有类型的衰老细胞。此外，鉴于到目前为止鉴定的许多抗衰老药物都是FDA 批准的抗癌药物，它们的感光活性以及它们的细胞类型特异性可能是 通过药物化学改进的。因此，DDDC的基本作用是开发和实施分析 用于根据需要测量小鼠、大鼠和/或人类细胞类型的感官治疗活性 四个项目。小鼠、大鼠和/或人类细胞将通过不同的方式诱导衰老 机制，包括氧化和遗传毒性应激、复制衰老和衰老治疗 被认为最适合感兴趣的细胞类型及其正常生理的细胞条件培养液 活着。2D和3D衰老细胞培养分析将被开发并用于原代和 次级屏幕。这些检测方法将用于比较现有的老年治疗药物，筛选新的 化合物，并测试已知的老年疗法的类似物以提高活性。此外，DDDC将 为确定药物的稳定性和在年轻人体内的分布提供DMPK分析 和老年小鼠，帮助每个项目为他们提议的实验确定正确的剂量方案。 最后，Core将验证已识别的化合物在体内减轻衰老细胞负担的能力 采用小鼠加速衰老模型，携带p16INKA和p21Cip1报告细胞衰老。

Core B: Project Summary: The overall goal of this multi-disciplinary, multi-site PPG application is to build a firm foundation of discovery science in cellular senescence that leads to a pipeline of therapeutic strategies that slow or prevent age-associated diseases. Each of the four PPG Projects focus on the role of senescent cells in specific age-related health outcomes: Project 1, Metabolic Dysfunction (Kirkland/Tchkonia); Project 2, Skeletal Fragility (Khosla/Farr); Project 3, Vascular Dysfunction (Miller); and Project 4, Skeletal Muscle Loss and Dysfunction (LeBrasseur). To establish cause and effect, each project will use genetic and pharmacologic approaches to ablate senescent cells. Importantly, we have already established that senolytics, drugs able to induce apoptosis specifically of senescent cells, have cell-type specificity, such as working in adipocytes and not in endothelial cell, or vice versa. The Drug Discovery and Development Core (DDDC) will work with the other three cores to assist all four projects with the identification of the most effective senotherapeutics for their cell types of interest. The DDDC has developed multiple assays based on primary murine and human cells for screening and optimizing senotherapeutic compounds and drug combinations. Examples of senolytics and senomorphics (drugs that inhibit the senescence-associated secretome) identified by the DDDC include inhibitors of BCL-2 family members, the combination of dasatinib plus quercetin, HSP90 inhibitors, the flavonoid fisetin, rapamycin, inhibitors of IKK/NF-κB, JAK inhibitors, multimeric resveratrol and mitochondrial- targeted free radical scavengers. No one senotherapeutic compound has proven effective in clearing or modulating all senescent cell types. In addition, given that many of the senolytics identified to date are FDA approved anti-cancer drugs, it is likely that their senolytic activity as well as their cell-type specificity can be improved by medicinal chemistry. Thus, the essential role of the DDDC is to develop and implement assays for measuring senotherapeutic activity in the cells types of murine, rat and/or human cells as needed for the four projects. Mouse, rat and/or human cells will be induced to undergo senescence through different mechanisms, including oxidative and genotoxic stress, replicative senescence and treatment with senescent cell conditioned media, as deemed most appropriate to the cell type of interest and its normal physiology in vivo. 2D and 3D senescent cell culture assays will be developed and utilized for use in both primary and secondary screens. These assays will be used for comparing existing senotherapeutics, screening novel compounds, and testing analogs of known senotherapeutics for improved activity. In addition, the DDDC will provide DMPK analysis of senotherapeutic compounds for determining drug stability and distribution in young and old mice to assist each project in determining the right dosing regimen for their proposed experiments. Finally, the Core will validate the ability of the identified compounds to reduce the senescent cell burden in vivo using mouse models of accelerated aging carrying p16INKA and p21Cip1 reporters of cellular senescence.