Engineering a long-acting relaxin agonist to treat liver fibrosis

设计长效松弛素激动剂来治疗肝纤维化

• 批准号: 10383001
• 负责人: IRINA AGOULNIK
• 金额: $ 30.87万
• 依托单位: ZEBRA BIOLOGICS, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-17 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10383001
• 关键词: Acute; Address; Agonist; Amino Acids; Animal Model; Animals; Anti-Inflammatory Agents; Antibodies; Applications Grants; BZLF1 gene; Biological; Birth; Blood; Carbon Tetrachloride; Cardiovascular Diseases; Cardiovascular system; Cellular Assay; Centrilobular hepatic necrosis; Choline; Chronic; Cirrhosis; Clinical; Clinical Research; Clinical Trials; Collagen; Complementarity Determining Regions; Congestive Heart Failure; Continuous Intravenous Infusion; Diet; Disease; Dose; Drug Kinetics; Engineering; Exhibits; Extracellular Matrix; Fatty acid glycerol esters; Fibrosis; Frequencies; G-Protein-Coupled Receptors; Goals; Growth Factor; Half-Life; Heart; Heart failure; Hepatic; High Fat Diet; Hormones; Human; Immunoglobulin G; Inbred BALB C Mice; Injections; Intoxication; Intravenous infusion procedures; Joints; Kidney; Liver; Liver Fibrosis; Liver diseases; Lung; Measurable; Medical; Modeling; Molecular; Mus; Organ; Outcome; Pathologic; Pathology; Peptides; Pharmacodynamics; Pharmacologic Substance; Pharmacology; Phase; Plasma; Pregnancy; Production; Property; Protein Engineering; Proteins; Reagent; Recombinants; Relaxin; Research; Research Proposals; Rodent Model; Safety; Serum; Skin; Small Business Technology Transfer Research; Subcutaneous Injections; Technology; Testing; Therapeutic Index; Tissues; Toxic effect; Validation; Vertebral column; Work; Zebra; antibody engineering; antifibrotic treatment; base; bone; clinical investigation; drug candidate; efficacy evaluation; efficacy study; efficacy testing; experience; hemodynamics; immunogenicity; in vivo; mouse model; nonalcoholic steatohepatitis; peptide hormone; pharmacodynamic biomarker; pharmacokinetics and pharmacodynamics; receptor; relaxin receptor; reproductive; reproductive organ; research clinical testing; response; safety assessment; skin disorder; subcutaneous; success

Project Summary/Abstract Relaxin is a heterodimeric 53 amino acid peptide hormone that induces cardiovascular compliance and reproductive tissue remodeling during pregnancy and parturition. In addition to reproductive organs, the relaxin receptor, RXFP1, is also expressed in the liver, heart, lung, kidney, bone and skin. This broad tissue localization has led to the recognition that relaxin is a pleiotropic hormone with vasodilatory, antifibrotic, tissue remodeling, antiapoptotic, and anti-inflammatory properties in animal models. The efficacy of relaxin has been tested in human clinical trials in diseases ranging from acute and chronic heart failure, to fibrotic diseases of skin, lung, and liver; however, relaxin's short half-life and need for continual intravenous infusion have limited its clinical utility. While the rationale to treat fibrosis and cardiovascular diseases with relaxin remains high, better RXFP1 agonists must be developed that display longer serum half-lives and measurable pharmacodynamic readouts, while maintaining a safety profile commensurate with chronic RXFP1 agonist treatment. We have used Zebra Biologics' proprietary Protein-in-Protein (PiP) antibody technology to insert a single-chain relaxin construct into the complementarity-determining region (CDR) of an immunoglobulin G backbone. This technology has been used previously to engineer proteins and peptides with half-lives of days vs. minutes for the native molecules. A relaxin-PiP, H2-PiP, has now been engineered with an intrinsic potency comparable to recombinant relaxin in cellular assays. The objective of this proposal is to elucidate pharmacokinetic and pharmacodynamic properties of H2-PiP. We hypothesize that this long-acting relaxin-PiP agonist molecule will be a superior antifibrosis drug candidate compared to relaxin, allowing proof-of-concept efficacy and safety assessment with less frequent subcutaneous injections, thus obviating the need for continuous intravenous infusion. We submit a Phase 1 STTR grant application addressing the following Specific Aims (SA): SA1: Pharmacokinetic-pharmacodynamic (PK-PD) analysis of H2-PiP. The goal of SA1 is to determine the half-life and maximal active serum concentration of H2-PiP, and to correlate blood exposure with known pharmacodynamic markers of RXFP1 activation in the context of acute CCl4 toxicity. This correlation will allow us to establish estimates of dose and dose frequency for determining efficacy in models of liver fibrosis. SA2: Efficacy of H2-PiP in mouse models of hepatic fibrosis. The goal of SA2 is to determine the efficacy of H2-PiP in two mouse models of chronic liver fibrosis that display key molecular and histopathological features of human liver fibrosis: 1) CCl4 exposure and 2) high fat diet (HFD) treatment. Successful completion of these studies will set the stage for eventual clinical studies in a range of diseases where tissue fibrosis and hemodynamic pathologies are manifest.

项目概要/摘要 松弛素是一种异二聚体 53 个氨基酸肽激素，可诱导心血管顺应性和 怀孕和分娩期间生殖组织的重塑。除了生殖器官外，松弛素 受体 RXFP1 也在肝脏、心脏、肺、肾脏、骨骼和皮肤中表达。这种广泛的组织 定位已导致人们认识到松弛素是一种多效性激素，具有血管舒张、抗纤维化、组织 动物模型中的重塑、抗凋亡和抗炎特性。松弛素的功效 在人体临床试验中对从急性和慢性心力衰竭到纤维化疾病等疾病进行了测试 皮肤、肺和肝脏；然而，松弛素的半衰期短以及需要持续静脉输注限制了其作用 其临床效用。虽然用松弛素治疗纤维化和心血管疾病的理由仍然很充分， 必须开发更好的 RXFP1 激动剂，其血清半衰期更长且可测量 药效读数，同时保持与慢性 RXFP1 激动剂相称的安全性 治疗。我们使用 Zebra Biologics 专有的蛋白中蛋白 (PiP) 抗体技术来插入 将单链松弛素构建体插入免疫球蛋白 G 的互补决定区 (CDR) 骨干。该技术之前已被用于设计半衰期为数天的蛋白质和肽 与天然分子的分钟相比。松弛素-PiP，H2-PiP，现已被设计为具有内在效力 在细胞测定中与重组松弛素相当。该提案的目的是阐明 H2-PiP 的药代动力学和药效学特性。我们假设这种长效松弛素-PiP 与松弛素相比，激动剂分子将成为更好的抗纤维化候选药物，从而可以进行概念验证 通过较少的皮下注射频率进行疗效和安全性评估，从而消除了需要 持续静脉输注。 我们提交第一阶段 STTR 拨款申请，解决以下具体目标 (SA)： SA1：H2-PiP 的药代动力学-药效 (PK-PD) 分析。 SA1 的目标是确定 H2-PiP 的半衰期和最大活性血清浓度，并将血液暴露与已知的 急性 CCl4 毒性背景下 RXFP1 激活的药效学标志物。这种相关性将允许 我们建立剂量和剂量频率的估计，以确定肝纤维化模型的疗效。 SA2：H2-PiP 在小鼠肝纤维化模型中的功效。 SA2 的目标是确定功效 H2-PiP 在两种慢性肝纤维化小鼠模型中的表达，显示出关键的分子和组织病理学特征 人类肝纤维化的特征：1）CCl4暴露和2）高脂肪饮食（HFD）治疗。 这些研究的成功完成将为一系列疾病的最终临床研究奠定基础 组织纤维化和血流动力学病理明显。