Small Molecule KLF15 Agonists for Kidney Disease

治疗肾脏疾病的小分子 KLF15 激动剂

• 批准号: 10553107
• 负责人: Sandeep K Mallipattu
• 金额: --
• 依托单位: NORTHPORT VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10553107
• 关键词: Actins; Activities of Daily Living; Adult; Agonist; Albuminuria; American; Attenuated; Biological Assay; Cardiovascular Diseases; Cells; Cellular Stress; Centers for Disease Control and Prevention (U.S.); Characteristics; Chemicals; Chronic Kidney Failure; Clinical Research; Cytoskeleton; Data; Diabetic Nephropathy; Disease; Dose; Drug Kinetics; Epithelial Cells; Excretory function; Filtration; Focal and Segmental Glomerulosclerosis; Functional disorder; Gene Expression; General Population; Glucocorticoids; Goals; Heart Hypertrophy; Human; In Vitro; Injury; Injury to Kidney; Kidney; Kidney Diseases; Laboratories; Lead; Luciferases; Maintenance; Mediating; Metabolism; Modeling; Morphology; Mus; Neurodegenerative Disorders; Organoids; Pathway interactions; Patients; Pre-Clinical Model; Predisposition; Prevalence; Property; Regulator Genes; Renal glomerular disease; Reporting; Research; Role; Signal Transduction; Specificity; Structure; Structure-Activity Relationship; Testing; Therapeutic; Therapeutic Effect; Toxic effect; Treatment Efficacy; United States; United States Department of Veterans Affairs; Veterans; absorption; analog; cardiovascular risk factor; clinically relevant; common treatment; design; ginsenoside M1; glomerular filtration; high throughput screening; improved; in vivo; interest; knock-down; lead optimization; lipid biosynthesis; loss of function; military veteran; mouse model; novel; novel lead compound; novel therapeutics; pharmacokinetics and pharmacodynamics; podocyte; pre-clinical; preclinical study; prevent; promoter; repository; scaffold; small molecule; targeted treatment; therapeutic evaluation; transcription factor; transcriptome sequencing

The Centers for Disease Control and Prevention estimates more than 15% of adults in the United States, over 30 million Americans have chronic kidney disease (CKD). Podocytes are epithelial cells in the glomerulus whose major function is the maintenance of the kidney filtration barrier to prevent CKD. Furthermore, the prevalence of CKD in the Veteran population is a third higher than in the general population. Podocyte injury is implicated in diseases such as Focal Segmental Glomerular Sclerosis (FSGS). In these diseases, the podocyte loses characteristic morphologic features and the functional capacity to maintain the glomerular filtration barrier. In several recent studies, we reported the essential role of Krüppel-Like Factor 15 (KLF15), a kidney-enriched transcription factor, in maintaining podocyte actin cytoskeleton under cell stress. For instance, loss of function studies in preclinical proteinuric murine models demonstrated that KLF15 is required to prevent podocyte injury and the salutary benefits of glucocorticoids (GCs), the most common treatment for primary glomerulopathies, are mediated by KLF15. As well, the responsiveness to GCs in human primary glomerulopathies is associated with podocyte-specific expression of KLF15. Furthermore, induction of human KLF15 in podocytes attenuated kidney injury in proteinuric murine models, without the adverse sequelae of GCs. Collectively, these preclinical and clinical studies on the renoprotective effects of KLF15 induction motivated us to identify novel small molecule KLF15 agonists for kidney disease. We initially generated and conducted a cell-based high-throughput screening (HTS) to screen small molecules that induce KLF15 activity. Subsequent dose-escalating studies identified novel lead compounds with a half maximal effective concentration (EC50), in the optimal therapeutic window, required to induce KLF15 activity. Based on EC50 and druggability, we conducted Structure-Activity Relationship (SAR) on the lead compound K-7 and generated 16 lead analogues, of which BT501, BT502, BT503, BT514, and BT412 induced KLF15 promoter activity with or without cell stress. We also performed intial pharmacokinetic studies for K-7 in mice and also showed that human podocytes treated with K-7 and lead analogues attenuated podocyte injury in the setting of cell stress. Furthemore, RNA-seq of K-7 treated human podocytes shows inhibition of pathway IL-17RA-mediated actin cytoskeleton destabilization, thereby providing the rationale to utilize a mechanistic approach to optimize selectivity of KLF15 agonists. Finally, we observed that K-7 attenuated albuminuria and restored podocyte markers in a preclinical proteinuric murine model. Based on these compelling preliminary data and strong scientific rigor of prior research, we hypothesize that optimization of lead KLF15 agonists in preclinical studies will serve as a key therapeutic in proteinuric kidney diseases. We propose to test our hypothesis by (1) improving the pharmacodynamic and pharmacokinetic properties of lead KLF15 agonists, (2) utilizing a mechanistic approach to optimize selectivity of KLF15 agonists, and (3) testing the therapeutic role of lead KLF15 agonists in mitigating and/or reversing kidney injury in preclinical proteinuric models. This proposal will address a current gap in the field by developing an integrated framework to optimize lead novel KLF15 agonists and test their therapeutic role in preclinical proteinuric models. The long-term goal of our project is to identify the optimal KLF15 agonist that can be advanced for IND studies for the treatment of primary glomerulopathies. Identification of novel targets for the treatment of proteinuric diseases is of major interest to the VA, given the high burden of CKD among U.S. Veterans. Furthermore, therapeutic strategies that mitigate the long-term use of GCs will have a tremendous impact on the complications associated with GCs in U.S. Veterans. Finally, the therapeutic role of KLF15 induction might extend beyond kidney disease, as other laboratories have demonstrated the beneficial effects of KLF15 in cardiac hypertrophy, neurodegenerative disease, and adipogenesis.

美国疾病控制与预防中心估计，美国超过 15% 的成年人 美国有超过 3000 万美国人患有慢性肾病 (CKD)，足细胞是足细胞中的上皮细胞。 肾小球的主要功能是维持肾脏滤过屏障以预防 CKD。 此外，退伍军人人群中 CKD 的患病率比普通人群高三分之一。 足细胞损伤与局灶节段性肾小球硬化症 (FSGS) 等疾病有关。 疾病、豆荚细胞特征形态特征和维持功能的能力 在最近的几项研究中，我们报道了 Krüppel 样因子 15 的重要作用。 (KLF15)，一种富含肾脏的转录因子，在细胞应激下维持足细胞肌动蛋白细胞骨架。 例如，临床前蛋白尿小鼠模型的功能丧失研究表明，KLF15 预防足细胞损伤和糖皮质激素 (GC) 的有益益处所必需的，糖皮质激素是最常见的 原发性肾小球病的治疗也是由 KLF15 介导的。 原发性肾小球病与 KLF15 的足细胞特异性表达有关。 足细胞中的人 KLF15 可减轻蛋白尿小鼠模型中的肾损伤，且不会产生不良反应 总的来说，这些关于 KLF15 肾脏保护作用的临床前和临床研究。 诱导促使我们鉴定出治疗肾脏疾病的新型小分子 KLF15 激动剂。 我们最初生成并进行了基于细胞的高通量筛选（HTS）来筛选小分子 随后的剂量递增研究发现了诱导 KLF15 活性的新型先导化合物。 在最佳治疗窗内，诱导 KLF15 所需的最大有效浓度 (EC50) 为一半 根据 EC50 和成药性，我们对先导化合物进行了结构活性关系 (SAR)。 化合物 K-7 并生成 16 个先导类似物，其中 BT501、BT502、BT503、BT514 和 BT412 诱导 在有或没有细胞应激的情况下，我们还对 K-7 进行了初步药代动力学研究。 小鼠，还表明用 K-7 和铅类似物处理的人足细胞可减轻足细胞损伤 此外，K-7 处理的人足细胞的 RNA-seq 显示了通路的抑制。 IL-17RA 介导的肌动蛋白细胞骨架不稳定，从而提供了利用机制的基本原理 优化 KLF15 激动剂选择性的方法最后，我们观察到 K-7 可以减轻白蛋白尿和 基于这些必要的初步研究，在临床前蛋白尿小鼠模型中恢复了足细胞标记。 数据和强大的科学研究，我们追求先导 KLF15 激动剂的优化 我们建议测试我们的临床前研究将作为蛋白尿肾病的关键治疗方法。 通过 (1) 改善先导 KLF15 激动剂的药效学和药代动力学特性，(2) 利用机械方法优化 KLF15 激动剂的选择性，以及 (3) 测试治疗作用 主要 KLF15 激动剂在临床前蛋白尿模型中减轻和/或逆转肾损伤的作用。 该提案将通过开发一个综合框架来优化该领域当前的差距 开发新型 KLF15 激动剂并在临床前蛋白尿模型中测试其治疗作用。 我们项目的目的是确定最佳的 KLF15 激动剂，可用于 IND 研究，用于治疗 确定治疗蛋白尿疾病的新靶点至关重要。 鉴于美国退伍军人 CKD 负担沉重，退伍军人管理局对此很感兴趣。 减轻GC的长期使用将对与相关并发症产生巨大影响 最后，KLF15 诱导的治疗作用可能不仅仅限于肾脏疾病。 其他实验室已经证明了 KLF15 对心脏肥大的有益作用， 神经退行性疾病和脂肪生成。