A Novel Approach to Restore Epithelial Barrier Homeostasis to Treat Inflammatory Bowel Disease

恢复上皮屏障稳态以治疗炎症性肠病的新方法

• 批准号: 10484275
• 负责人: W Vallen Graham
• 金额: $ 79.5万
• 依托单位: THELIUM THERAPEUTICS INC
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2022-11-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484275
• 关键词: Actomyosin; Acute; Acute Disease; Adverse effects; Affinity; Attenuated; Award; Binding; Biochemical; Biopsy; Celiac Disease; Cell Adhesion Molecules; Cell Line; Cellular Assay; Chronic; Chronic Disease; Clinical; Colitis; Crystallization; Data; Development; Disease; Dose; Drug Kinetics; Enteral; Enterocolitis; Epithelial; Epithelial Cells; Evaluation; Food Hypersensitivity; Formulation; Foundations; Functional disorder; Gastrointestinal Diseases; Health; Homeostasis; Human; Immune; Immunoglobulins; Immunomodulators; Immunosuppression; Immunosuppressive Agents; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Interleukin-10; Intestinal Diseases; Intestines; Knock-out; Knockout Mice; Lead; Libraries; Light; Mediating; Medicine; Modeling; Mucous Membrane; Mus; Muscle Contraction; Myosin Alkali Light Chains; Myosin Light Chain Kinase; Myosin Type II; Nature; Outcome; Pathologic; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacodynamics; Phase; Phosphorylation; Protein Isoforms; RNA Splicing; Regulation; Reporting; Risk; Role; Small Business Innovation Research Grant; Smooth Muscle; Stimulus; Structure; Structure-Activity Relationship; Systemic disease; T cell therapy; T-Lymphocyte; TNF gene; Therapeutic; Tight Junctions; Tissues; Toxic effect; Toxicity Tests; Variant; Waste Products; Water; Work; base; candidate selection; clinical application; cytokine; dosage; efficacy study; epithelial wound; experience; graft vs host disease; immune activation; improved; in vivo; intestinal barrier; intestinal epithelium; lead candidate; lead optimization; mouse model; novel; novel strategies; nutrient absorption; pathogen; phase 2 study; preservation; prevent; programs; recruit; repaired; restoration; scaffold; screening; small molecule; therapeutic target; tool; wound healing

SUMMARY Intestinal barrier function is compromised in enteric and systemic diseases, including infectious enterocolitis, food allergy, celiac disease, graft versus host disease (GvHD), and inflammatory bowel disease (IBD). The co-founders of Thelium Therapeutics discovered the central role of myosin light chain kinase (MLCK) in barrier regulation and demonstrated that targeted intestinal epithelial MLCK inhibition limits experimental IBD and GvHD. Unfortunately, severe toxicities associated with barrier-independent MLCK functions in epithelia and other tissues, e.g., smooth muscle, preclude therapeutic targeting of MLCK enzymatic activity. We recently reported (Graham et al., Nature Medicine, 2019) that a specific MLCK splice variant, MLCK1, is central to barrier regulation and depends on interactions mediated by immunoglobulin-cell adhesion molecule domain 3 (IgCAM3). We solved the IgCAM3 crystal structure, identified a drug binding pocket unique to IgCAM3, and screened a library of ~140,000 drug-like molecules. Our tool compound, Divertin, bound IgCAM3, prevented cytokine-induced MLCK1 recruitment, myosin II regulatory light chain phosphorylation, and barrier dysfunction. Critically, Divertin did not inhibit MLCK enzymatic function, epithelial wound healing, or smooth muscle contraction, and in vivo toxicity studies failed to identify adverse effects. Divertin prevented acute TNF-induced barrier loss in vivo (mice) and ex vivo (human intestinal biopsies), and restored immune-mediated barrier loss in vivo (IL-10 knockout mice). Finally, Divertin delayed onset and prevented progression of experimental immune-mediated (T cell transfer) IBD, as indicated by barrier preservation and restoration, reduced mucosal immune activation, and enhanced survival. In a Phase I SBIR, we advanced this program through the discovery of hit compounds with improved activities. Hit compounds were discovered through a rigorous rank order screening funnel to identify compounds with suitable MLCK1 binding affinities, efficacy in preserving and restoring epithelial barrier function, and absence of enzymatic inhibitory activity. In this Phase II proposal, these hit compounds will be optimized with iterative structure activity relationship studies and tested for toxicity and efficacy in mouse models of IBD. This work will facilitate IND-enabling studies for a first-in-class and best-in-class barrier-restorative therapy to manage gastrointestinal and systemic diseases.

概括 肠道屏障功能在肠和全身性疾病中受到损害，包括感染性 小肠结肠炎，食物过敏，腹腔疾病，移植物与宿主疾病（GVHD）和炎症性 肠病（IBD）。 Thelium Therapeutics的联合创始人发现了 肌球蛋白轻链激酶（MLCK）在屏障调节中，并证明了靶向肠 上皮MLCK抑制限制实验IBD和GVHD。不幸的是，严重的毒性 与无障碍的MLCK在上皮和其他组织中的功能相关，例如光滑 肌肉，排除MLCK酶活性的治疗靶向。我们最近报道 （Graham等人，自然医学，2019年），特定的MLCK剪接变体MLCK1对于 屏障调节，取决于免疫球蛋白细胞粘附介导的相互作用 分子结构域3（IgCAM3）。我们解决了IgCAM3晶体结构，鉴定了药物结合 IgCAM3独有的口袋，并筛选了约140,000个类似药物的分子的库。我们的工具 化合物，分离蛋白，结合的IgCAM3，防止了细胞因子诱导的MLCK1募集，肌球蛋白II 调节轻链磷酸化和屏障功能障碍。至关重要的是，分离蛋白没有抑制 MLCK酶功能，上皮伤口愈合或平滑肌收缩和体内 毒性研究未能鉴定出不良影响。分离蛋白阻止了急性TNF诱导的屏障 体内损失（小鼠）和外体（人肠活检），并恢复了免疫介导的 体内屏障损失（IL-10敲除小鼠）。最后，分离蛋白延迟发作并阻止 实验免疫介导的（T细胞转移）IBD的进展，如屏障所示 保存和恢复，粘膜免疫激活降低以及生存增强。在 第一阶段SBIR，我们通过发现改进的命中化合物提出了该程序 活动。通过严格的等级顺序筛选漏斗发现了命中化合物 识别具有合适MLCK1结合亲和力的化合物，保存和恢复的功效 上皮屏障功能，缺乏酶促抑制活性。在此第二阶段提案中， 这些命中化合物将通过迭代结构活动关系研究进行优化，并且 在IBD小鼠模型中测试了毒性和功效。这项工作将有助于索引 研究对管理的第一类和一流的障碍疗法 胃肠道和全身性疾病。