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: 9909608
• 负责人: W Vallen Graham
• 金额: $ 22.43万
• 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 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9909608
• 关键词: Actomyosin; Acute; Acute Disease; Adverse effects; Affect; Affinity; Anti-Tumor Necrosis Factor Therapy; Binding; Biology; Biopsy; Cardiac Muscle Contraction; Celiac Disease; Cell Adhesion Molecules; Cell Line; Cellular Assay; Characteristics; Chemicals; Chronic; Chronic Disease; Clinical Drug Development; Clinical Trials; Crystallization; Data; Development; Diarrhea; Disease; Disease model; Docking; Endocytosis; Endothelium; Enteral; Enterocolitis; Enzymes; Epithelial; Epithelial Cells; Epithelium; Event; Experimental Animal Model; Experimental Models; Food Hypersensitivity; Foundations; Functional disorder; Gastrointestinal Diseases; Health; Homeostasis; Human; Hydrophobicity; Hypotension; Immune; Immunoglobulins; Immunosuppressive Agents; In Vitro; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Interleukin-10; Intestinal Diseases; Intestines; Knockout Mice; Lead; Libraries; Light; Measures; Mediating; Medicine; Methods; Molecular; Mucous Membrane; Mus; Muscle; Muscle Contraction; Myosin Alkali Light Chains; Myosin Light Chain Kinase; Myosin Type II; Nature; Nutrient; Outcome; Permeability; Pharmaceutical Preparations; Phase; Phosphorylation; Phosphotransferases; Physics; Physiological; Positioning Attribute; Property; Publishing; RNA Splicing; Regulation; Reporting; Rho-associated kinase; Role; Signal Transduction; Site; Skeletal Muscle; Small Business Innovation Research Grant; Smooth Muscle; Specificity; Stimulus; Structure; Systemic disease; T-Lymphocyte; TNF gene; Tertiary Protein Structure; Therapeutic; Therapeutic Agents; Tight Junctions; Tissues; Toxic effect; Variant; Waste Products; Water; Work; Wound Healing; active method; base; clinical application; clinical candidate; cytokine; design; graft vs host disease; immune activation; immunomodulatory therapies; improved; in vivo; infection risk; insight; intestinal epithelium; lead candidate; lead optimization; non-muscle myosin; novel strategies; novel therapeutics; nutrient absorption; occludin; pathogen; phase 1 study; pre-clinical; preservation; prevent; recruit; repaired; restoration; screening; simulation; small molecule; standard of care; systemic toxicity; therapeutic target; tool

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. One 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. Although useful as a tool compound, Divertin lacks characteristics required for a lead compound. This proposal will overcome that obstacle to clinical application of Divertin by discovering lead compounds with similar activities. Cutting edge, physics-based rational design and molecular dynamic simulation methods has already been used to probe an expansive compound library and identify those with predicted high-affinity binding to the MLCK1 IgCAM3 crystal structure. Those molecules will be subjected to a 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. These lead compounds will be suitable for optimization and development to facilitate IND-enabling studies for a first-in-class barrier-restorative therapy to manage gastrointestinal and systemic diseases.

概括 肠道屏障功能在肠道和全身性疾病（包括传染性疾病）中受到损害 小肠结肠炎、食物过敏、乳糜泻、移植物抗宿主病 (GvHD) 和炎症 肠道疾病（IBD）。 Thelium Therapeutics 的联合创始人发现了 肌球蛋白轻链激酶 (MLCK) 在屏障调节中的作用，并证明靶向肠道 上皮 MLCK 抑制限制实验性 IBD 和 GvHD。不幸的是，严重的毒性 与上皮和其他组织（例如平滑肌）中屏障无关的 MLCK 功能相关 肌肉，排除 MLCK 酶活性的治疗靶向。我们最近报道了 (Graham et al., Nature Medicine, 2019) 特定的 MLCK 剪接变体 MLCK1 是 屏障调节并依赖于免疫球蛋白-细胞粘附介导的相互作用 分子结构域 3 (IgCAM3)。我们解析了 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 个药物样分子的库。一 化合物，Divertin，结合 IgCAM3，防止细胞因子诱导的 MLCK1 募集，肌球蛋白 II 调节性轻链磷酸化和屏障功能障碍。重要的是，Divertin 并没有抑制 MLCK 酶功能、上皮伤口愈合或平滑肌收缩以及体内 毒性研究未能确定不良反应。 Divertin 预防急性 TNF 诱导的屏障 体内（小鼠）和离体（人肠道活检）损失和恢复免疫介导 体内屏障丧失（IL-10 敲除小鼠）。最后，Divertin 延迟了发病并预防了 实验性免疫介导（T 细胞转移）IBD 的进展，如屏障所示 保存和恢复，减少粘膜免疫激活，并提高生存率。 虽然 Divertin 可用作工具化合物，但缺乏先导化合物所需的特性 化合物。该提案将克服 Divertin 临床应用的障碍 发现具有类似活性的先导化合物。尖端的、基于物理的理性设计 分子动力学模拟方法已被用于探索广泛的 化合物库并鉴定那些预测与 MLCK1 IgCAM3 具有高亲和力结合的化合物 晶体结构。这些分子将接受排序筛选漏斗来识别 具有合适的 MLCK1 结合亲和力的化合物，在保护和恢复上皮细胞方面的功效 屏障功能，并且缺乏酶抑制活性。这些先导化合物将 适合优化和开发，以促进一流的 IND 研究 用于治疗胃肠道和全身疾病的屏障恢复疗法。