Small Molecule KLF15 Agonists for Kidney Disease

治疗肾脏疾病的小分子 KLF15 激动剂

• 批准号: 10359057
• 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/10359057
• 关键词: Actins; Activities of Daily Living; Address; 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 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; base; 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.

疾病控制和预防中心估计了联合成年人的15％以上 州，超过3000万美国人患有慢性肾脏疾病（CKD）。足细胞是上皮细胞 肾小球的主要功能是维持肾脏过滤屏障以防止CKD。 此外，退伍军人人口中CKD的患病率比普通人群高三分之一。 足细胞损伤隐含在诸如局灶性节段性肾小球硬化症（FSGS）之类的疾病中。在这些 疾病，足细胞失去了特征的形态特征和维持功能的能力 肾小球滤过屏障。在最近的几项研究中，我们报道了类似Krüppel样因子15的基本作用 （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，临床前蛋白尿鼠模型中功能研究的丧失表明KLF15是 预防足细胞损伤和糖皮质激素（GCS）的有益益处（最常见）所需 原发性肾小球病的治疗是由KLF15介导的。同样，人类对GC的响应能力 原发性肾小球病与KLF15的足细胞特异性表达有关。此外，诱导 人类KLF15在蛋白尿模型中减弱肾脏损伤中的人KLF15，没有逆境 GC的后遗症。总的来说，这些关于KLF15的伦名保护作用的临床前和临床研究 诱导促使我们确定新型的小分子KLF15激动剂用于肾脏疾病。 我们最初生成并进行了基于细胞的高通量筛选（HTS）以筛选小 影响KLF15活性的分子。随后的剂量提升研究确定了新型铅化合物 最大有效浓度（EC50）在最佳治疗窗口中诱导KLF15所需 活动。根据EC50和可药用，我们在铅上进行了结构活性关系（SAR） 化合物K-7并产生了16个铅类似物，其中BT501，BT502，BT503，BT514和BT412诱导 KLF15启动子活性，有或没有细胞应激。我们还为K-7进行了Intial药代动力学研究 小鼠，还表明用K-7处理的人足细胞和类似物治疗了足细胞损伤 在细胞应力的环境中。此外，K-7处理的人足细胞的RNA-Seq显示了途径的抑制 IL-17RA介导的肌动蛋白细胞骨架不稳定，从而提供了利用机械的理由 优化KLF15激动剂的选择性的方法。最后，我们观察到K-7减弱了蛋白尿和 在临床前蛋白尿模型中恢复的足细胞标记。基于这些引人入胜的初步 数据和强烈的先前研究的科学严谨性，我们假设优化铅klf15激动剂在 临床前研究将作为蛋白尿肾脏疾病的关键治疗方法。我们建议测试我们的 通过（1）改善铅KLF15激动剂的药效和药代动力学特性的假设，（2） 使用机械方法来优化KLF15激动剂的选择性，（3）测试治疗作用 临床前蛋白尿模型中铅KLF15激动剂的缓解和/或逆转肾损伤的激动剂。 该建议将通过开发一个集成框架以优化现场的当前差距 铅新型KLF15激动剂，并在临床前蛋白尿模型中测试其治疗作用。长期目标 我们的项目的是确定最佳的KLF15激动剂，可以进行IND研究以治疗 原发性肾小球病。确定治疗蛋白尿疾病的新目标是主要的 鉴于美国退伍军人的CKD负担很高，VA的兴趣。此外，治疗策略 减轻GC的长期使用将对与之相关的并发症产生巨大影响 美国退伍军人的GC。最后，KLF15诱导的治疗作用可能超出肾脏疾病，如 其他实验室已经证明了KLF15在心脏肥大中的有益作用， 神经退行性疾病和脂肪形成。