Development of Modified Caveolin-1 Scaffolding Domain Peptides with Improved Pharmacological Properties as Therapeutic Agents for Scleroderma Skin Disease

开发具有改善药理特性的修饰的 Caveolin-1 支架结构域肽作为硬皮病皮肤病的治疗剂

• 批准号: 10544238
• 负责人: STANLEY R HOFFMAN
• 金额: $ 25.96万
• 依托单位: FIBROTHERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10544238
• 关键词: Active Sites; Adipocytes; Adipose tissue; Affect; Amino Acids; Animals; Automobile Driving; Biological; Biological Models; Bleomycin; Cancer Center; Cells; Center for Translational Science Activities; Collagen; Dependence; Deposition; Dermal; Dermis; Detection; Development; Disease; Dose; Enzymes; Evaluation; Excretory function; FDA approved; Fatty acid glycerol esters; Fibrosis; Future; Goals; High Pressure Liquid Chromatography; Implant; In Vitro; Inflammation; Inflammatory; Injections; Laboratories; Lead; Lipoatrophy; Literature; Lung; Maximum Tolerated Dose; Messenger RNA; Metabolism; Methods; Modeling; Modification; Mus; Myofibroblast; Names; Occupations; Oral Administration; Patients; Peptides; Pharmaceutical Preparations; Pharmacology; Phase; Phosphotransferases; Pirfenidone; Property; Proteins; Proteolysis; Protocols documentation; Pulmonary Fibrosis; Regulation; Safety; Scleroderma; Side; Skin; Specificity; Subcutaneous Injections; Systemic Scleroderma; Tertiary Protein Structure; Testing; Therapeutic; Therapeutic Agents; Therapeutic Index; Thick; Toxic effect; Toxicology; Transforming Growth Factor beta; Water; absorption; base; caveolin 1; clinical development; design; digital; drug development; effective therapy; experimental study; improved; in vivo; lead candidate; mouse model; nintedanib; novel; novel lead compound; osmotic minipump; phase 2 study; scaffold; side effect; skin disorder; skin fibrosis; subcutaneous; success; therapeutic target; uptake

Abstract Our long-term objective is to develop an effective treatment for scleroderma (systemic sclerosis, SSc) skin fibrosis. Caveolin-1 is a promising therapeutic target in fibrotic diseases. The profibrotic effects of caveolin-1 deficiency in cells and in mouse models is suppressed by a peptide equivalent to its active site (caveolin-1 scaffolding domain, CSD). However, CSD lacks suitable pharmacologic properties for drug development. To overcome this problem, we developed novel, modified versions of CSD. We first divided CSD into three subregions and found that all three suppressed bleomycin-induced skin fibrosis. To improve, the pharmacological properties, we then modified CSD and each subregion to be water soluble and protected from proteolysis. This modification greatly enhanced the uptake by cells of all four modified peptides and also greatly increased their ability to inhibit several purified kinases in vitro. We have so far tested only one of the four modified peptides in vivo and it was outstandingly active in inhibiting bleomycin-induced dermal fibrosis as well as the associated loss of the intradermal adipose layer (lipoatrophy). These initial studies justify and outstandingly support our proposal to identify a Lead Compound from among the four candidates, then evaluate its Therapeutic Index (ratio between toxic and beneficial doses). Our studies and the literature also suggest that our peptides will be more effective and have fewer side effects than the blockbuster FDA- approved drug pirfenidone (brand name Esbriet). In summary, to proceed with drug development we must identify a Lead Compound. Due to their distinct pharmacological and functional differences, we must do a side- by-side comparison of our four modified peptides. 1) Select a Lead Compound using two model systems: Systemic Bleomycin Treatment and Subcutaneous TGFβ Injection. We will identify a Lead Compound, then demonstrate its specificity and activity by comparing it to a control peptide (scrambled Lead) and to nintedanib. Peptides will be delivered s.c. in a Therapeutic Protocol, beginning one week after fibrosis is induced. Primary Readouts will be dermal fibrosis and lipoatrophy. Secondary Readouts will be the levels of markers for myofibroblasts, adipocytes, and inflammatory cells. Success will be defined as >50% reversal of the deleterious effects of bleomycin and TGFβ on the Primary and Secondary Readouts. 2) Determine the Therapeutic Index of the Lead Compound. The dose-dependence of the beneficial effects of the Lead Compound will be determined using doses above and below our current standard dose. The toxicity of the Lead Compound will be evaluated in a Single-Treatment Maximum Tolerated Dose (MTD) Experiment using 1X, 5X, 25X, and 125X our current standard dose. We will consider these studies to be a success if the Therapeutic Index is >50. In summary, these studies will provide a novel Lead Compound that meets our Criteria for Success, both in terms of suppression of skin disease and of safety.

摘要我们的长期目标是为硬皮病（全身性硬化症， SSC）皮肤纤维化。 Caveolin-1是纤维化疾病中有前途的治疗靶标。纤维化效应 小鼠1细胞和小鼠模型中的缺陷被等效于其活性部位的肽抑制 （Caveolin-1脚手架域，CSD）。但是，CSD缺乏药物的合适的药学特性 发展。为了克服这个问题，我们开发了CSD的新颖，修改的版本。我们首先将CSD分开 分为三个区域，发现这三个抑制了博来霉素诱导的皮肤纤维化。为了改善 药理学特性，然后我们修改了CSD，每个子区域都固体并保护 蛋白水解。这种修饰大大增强了所有四个改性胡椒的细胞的吸收 大大提高了它们在体外抑制几种纯化激酶的能力。到目前为止，我们仅测试了一个 体内四个改性肽，它在抑制博来霉素诱导的真皮纤维化方面非常活跃为 以及相关的皮内脂肪层（脂质）的损失。这些最初的研究证明了合理性和 在四个候选人中识别我们的提议，以支持我们的建议，然后 评估其治疗指数（有毒和有益剂量之间的比率）。我们的研究和文学 表明我们的宠物比大片FDA-更有效，副作用更少 已批准的药物Pirfenidone（品牌Esbriet）。总而言之，为了进行药物开发，我们必须 识别铅化合物。由于它们明显的药理和功能差异，我们必须做一个侧面 我们四个改良肽的副比较。 1）使用两个模型系统选择铅化合物： 全身性博来霉素治疗和皮下TGFβ注射。我们将确定铅化合物，然后 通过将其与对照肽（炒铅）和nyntedanib进行比较来证明其特异性和活性。 肽将交付S.C.在治疗方案中，诱导纤维化后一周开始。基本的 读数将是真皮纤维化和脂肪植物。次要读数将是标记的级别 肌纤维细胞，脂肪细胞和炎症细胞。成功将定义为> 50％的逆转 博来霉素和TGFβ对初级和次要读数的有害影响。 2）确定 铅化合物的治疗指数。铅的有益作用的剂量依赖性 化合物将使用高于我们当前标准剂量的剂量确定。毒性的毒性 铅化合物将在单一处理的最大耐受剂量（MTD）实验中进行评估 1倍，5x，25倍和125倍我们当前的标准剂量。如果这些研究是成功的 治疗指数> 50。总而言之，这些研究将提供一种新颖的铅化合物，使我们 在抑制皮肤病和安全方面，成功的标准。