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Design of a bioactive mimetic of soluble klotho for the treatment of chronic kidney disease

用于治疗慢性肾病的可溶性 klotho 生物活性模拟物的设计

基本信息

批准号：
10483849
负责人：
Christopher Yanucil
金额：
$ 25.61万
依托单位：
ALPHA YOUNG LLC
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：
2022-07-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10483849
关键词：
Affinity Animal Model Animals Binding Biological Biological Assay Biological Markers Blood Blood Circulation Cardiac Cardiac Myocytes Cardiovascular Diseases Cardiovascular system Caring Cause of Death Cell Culture Techniques Cells Cessation of life Chronic Kidney Failure Clinical Trials Dialysis procedure Drug Targeting Excretory function FGFR4 gene Fibroblast Growth Factor Fibroblast Growth Factor Receptor 1 Fibroblast Growth Factor Receptors Fibroblasts Fibrosis Functional disorder Future Generations Genetic Goals Half-Life Heart Heart Hypertrophy Heparin Heparin Binding Hormones Hypertrophy Individual Inflammation Inhibition of Apoptosis Injections Integral Membrane Protein Kidney Kidney Transplantation Label Laboratories Length Liver Longevity Lysine Mass Spectrum Analysis Measures Mediating Metabolism Methods Minerals Modeling Mus Mutate Mutation Organ Oxidative Stress Pathologic Pathology Patients Phage Display Phase Physiological Point Mutation Procedures Production Property Proteins Rattus Recombinants Serum Severity of illness Signal Transduction Site Source Technology Testing Therapeutic Time Tissues Tubular formation Variant Vascular calcification Virus base bone cardiovascular injury cardiovascular risk factor cell type cost design drug candidate drug development fibroblast growth factor 23 glycosylation heart damage high risk improved in vivo evaluation inorganic phosphate klotho protein mimetics mortality mouse model mutant novel novel therapeutic intervention novel therapeutics overexpression paracrine preclinical study preservation prevent protective effect receptor receptor binding renal damage senescence therapeutic evaluation tissue injury tool transplantation therapy uptake

项目摘要

PROJECT SUMMARY Over 37 million individuals in the U.S. have chronic kidney disease (CKD) and are at high risk to die from cardiovascular complications. While great strides have been made to improve CKD care and dialysis access, minimal advances have been made in drug development to stall or reverse kidney damage and associated pathologies. Currently, therapeutic options to prevent cardiovascular damage in CKD do not exist, and the only cure for CKD is kidney transplantation. Elevations in serum levels of phosphate and fibroblast growth factor (FGF) 23 are a hallmark of CKD and associated with an increased risk of cardiovascular death. Expression levels of klotho, a regulator of phosphate metabolism in the kidney, are reduced in CKD. Klotho can be released from the kidney as soluble klotho (sKL) that circulates in the blood and acts as a binding partner for FGF receptors (FGFR) on various tissues. Reductions in serum sKL levels have been shown to contribute to CKD-associated pathologies. sKL seems to protect tissues by substituting for renal klotho thereby promoting FGF23/FGFR1- induced renal phosphate excretion and lowering systemic phosphate levels, as well blocking the direct pathologic actions of FGF23 and of paracrine FGFs. While elevating klotho expression has shown therapeutic potential in animal models of CKD, further advances have been stymied by sKL’s short half-life and technical difficulties to produce the recombinant sKL protein in sufficient amounts, along with a lack of tools to measure sKL activity. Alpha Young LLC has developed a novel method to produce the recombinant sKL protein as well as a novel assays to determine the bioactivity of sKL based on its ability to bind FGF23 and FGFR1. We have generated an early-stage mimetic protein, and here we will introduce additional point mutations to increase sKL’s stability and bioactivity. In Phase 1, we will modify sKL’s glycosylation sites and heparin binding domain, and we will screen for mutant variants with increased binding affinities for FGF23 and FGFR1 to improve bioactivity, and decreased heparin binding affinity to increase half-life. In Phase 2, we will optimize our identified sKL variants by utilizing a phage display-based approach to introduce mutations into sKL’s FGFR binding domain with the goal to increase FGFR1 binding affinity. Candidates with the desired changes in binding properties will be tested for their biological activity using cell culture models that can determine the effect of sKL on FGF23-regulated signaling, renal phosphate uptake, cardiac hypertrophy and on fibroblast activation induced by paracrine FGFs. The half-life of the most promising candidates will be tested by injection studies in in rats. Finally, the most active and stable sKL variant will be injected into mouse models of CKD, followed by the analysis of renal phosphate excretion and cardiovascular damage. We propose that the administration of our sKL mimetic can serve as a novel therapeutic approach in CKD to lower serum phosphate levels and to protect from the damaging actions of FGFs. A successful completion of our project would provide us with a potent drug candidate and the opportunity to pursue early-stage partners for advancing and validating its potential for future clinical trials.

项目总结