Small molecule targeting an epigenetic regulator for the treatment of osteoarthritis

靶向表观遗传调节剂的小分子治疗骨关节炎

• 批准号: 10508839
• 负责人: Maddalena Adorno
• 金额: $ 123.99万
• 依托单位: DORIAN THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10508839
• 关键词: Adult; Affect; Aging; Analgesics; Apoptosis; BMI1 gene; Bacterial Artificial Chromosomes; Biochemical; Biological Assay; Bone Marrow; CDKN2A gene; Cartilage; Catabolism; Cell Aging; Cells; Chondrocytes; Chromatin; Chromatin Remodeling Factor; Chromosome 21; Chronic; Clinical Trials; Collagen; Congenital Disorders; Data; Degenerative polyarthritis; Deposition; Development; Disease; Dose; Down Syndrome; Down-Regulation; Drug Kinetics; Early Onset Alzheimer Disease; Enzymes; Epigenetic Process; Formulation; Gene Expression; Genes; Genetic; Goals; Health; Histology; Histone H2A; Histones; Human; Immune System Diseases; In Vitro; Injections; Intra-Articular Injections; Kinetics; Knee; Knee Osteoarthritis; Lead; Link; Luciferases; Lysine; Maintenance; Measures; Mediating; Mesenchymal Stem Cells; Microarray Analysis; Mitochondria; Modeling; Musculoskeletal Diseases; Natural regeneration; Obesity; Operative Surgical Procedures; Osteoporosis; Pathology; Patients; Peptide Hydrolases; Permeability; Persons; Pharmaceutical Preparations; Pharmacodynamics; Phase; Physiological; Plasma; Population; Prevalence; Proliferation Marker; Property; RNA marker; Rattus; Recombinants; Regulation; Role; Safety; Series; Site; Small Business Innovation Research Grant; Small Interfering RNA; Solubility; Specificity; Stains; Stratification; Synovial Fluid; Testing; Therapeutic Index; Time; Tissues; Toxic effect; Toxicity Tests; Toxicology; Treatment Efficacy; Ubiquitin; Variant; analog; antagonist; burden of illness; cell regeneration; cytotoxicity; differential expression; drug candidate; drug development; efficacy testing; genetic variant; genomic locus; human tissue; improved functioning; in vivo; innovation; joint destruction; knee replacement arthroplasty; knock-down; lead candidate; lead optimization; lead series; mouse model; novel; osteoarthritis pain; patient biomarkers; patient population; patient stratification; pharmacokinetics and pharmacodynamics; preclinical efficacy; prevent; promoter; response; scaffold; senescence; side effect; small molecule; small molecule inhibitor; stem cell function; stem cell self renewal; stem cells; systemic toxicity; treatment response; vector

The goal of the project is to develop a disease-modifying treatment for Osteoarthritis (OA) by targeting USP16, a chromatin modifier involved in regulation of senescence and stem cell self-renewal. The treatment will consist of an intra-articular injection of a small molecule inhibiting USP16 in patients with moderate OA. USP16 is a deubiquitinase (DUB) enzyme that removes ubiquitin from histone H2A on lysine 119, a critical mark for the maintenance of multiple somatic tissues. Interestingly, triplication of USP16 is associated with Down Syndrome (DS), a congenital disorder characterized by triplication of chromosome 21 (HSA21). Patients with DS show signs of accelerated aging, including early-onset Alzheimer’s, immune dysfunction and osteoporosis. We have previously shown that reducing the levels of USP16 in DS mouse models or human tissues results in improved function of somatic stem cells and reduction in senescence, therefore alleviating the conditions associated with DS. This evidence makes USP16 an attractive target to ameliorate some of the aging-related pathologies. Notably, USP16 expression is highly upregulated in OA chondrocytes and synovial tissues. Moreover, the genetic locus of USP16 contains a SNP strongly associated with familial OA. We found that genetic downregulation of USP16 in patient-derived articular chondrocytes promotes cellular and mitochondrial health and reduces senescence associated markers, like SA-ꞵ-gal and p16Ink4a. Moreover, microarray analyses of OA chondrocytes treated with siRNA targeting USP16 showed an increase in markers of proliferation and collagen deposition, and a reduction of apoptosis and catabolism markers. Furthermore, USP16 knockdown in human bone marrow-derived mesenchymal stem cells promoted differentiation into mature chondrocytes and increased matrix deposition. Using a biochemical assay testing the enzymatic activity of recombinant human USP16, we identified small molecule inhibitors. We validated 19 hits with low IC50 and chose small molecule scaffolds with IC50 between 0.06 μM and 9.4 μM. During Phase I of the project we will optimize them to increase potency, specificity and solubility. During Phase II we will move the two best compound series into Lead Optimization, followed by preclinical efficacy in OA models and early toxicity testing of the two best lead compounds. We will assess their safety, pharmacokinetics (PK), ability to engage target and modulate chromatin as a pharmacodynamic (PD) measure, and efficacy in preventing or reversing loss of cartilage in a surgical rat OA model, with the goal of establishing a relationship between time on target, dose and efficacy. We will also study the role of the rs6516886 SNP and understand if specific variants of this genomic locus are linked to differential expression of USP16 and different responses to treatment, potentially leading to stratification of the target patient population. The proposed studies will inform subsequent GLP studies to support an IND and clinical trial.

该项目的目标是通过针对 USP16 开发一种缓解骨关节炎 (OA) 疾病的疗法， 参与衰老和干细胞自我更新调节的染色质修饰剂。治疗包括 中度 OA 患者关节内注射抑制 USP16 的小分子。 USP16 是 去泛素酶 (DUB) 酶，可去除组蛋白 H2A 赖氨酸 119 上的泛素，赖氨酸是 维持多个体细胞组织。有趣的是，USP16的三倍体与唐氏综合症有关 (DS)，一种以 21 号染色体 (HSA21) 三倍体为特征的先天性疾病。 DS患者表现 加速衰老的迹象，包括早发性阿尔茨海默病、免疫功能障碍和骨质疏松症。我们有 先前表明，降低 DS 小鼠模型或人体组织中 USP16 的水平可改善 体干细胞的功能和减少衰老，从而减轻与 DS。这一证据使 USP16 成为改善某些衰老相关病症的有吸引力的靶点。 值得注意的是，USP16 表达在 OA 软骨细胞和滑膜组织中高度上调。此外， USP16 的遗传位点包含与家族性 OA 密切相关的 SNP。我们发现遗传 患者来源的关节软骨细胞中 USP16 的下调可促进细胞和线粒体健康 并减少衰老相关标记，如 SA-ꞵ-gal 和 p16Ink4a。此外，OA 的微阵列分析 用靶向 USP16 的 siRNA 处理的软骨细胞显示增殖和胶原标记物增加 沉积，以及细胞凋亡和分解代谢标记物的减少。此外，USP16 在人类体内的敲除 骨髓间充质干细胞促进分化为成熟软骨细胞并增加 基质沉积。使用生化测定法测试重组人 USP16 的酶活性，我们 鉴定出小分子抑制剂。我们验证了 19 个具有低 IC50 的命中，并选择了具有 IC50 介于 0.06 μM 和 9.4 μM 之间。在项目的第一阶段，我们将优化它们以提高效力， 特异性和溶解度。在第二阶段，我们将把两个最好的化合物系列转移到先导化合物优化中， 其次是两种最佳先导化合物在 OA 模型中的临床前疗效和早期毒性测试。我们将 评估其安全性、药代动力学 (PK)、参与靶标和调节染色质的能力 药效学 (PD) 测量以及预防或逆转手术大鼠 OA 软骨损失的功效 模型，目的是建立目标时间、剂量和功效之间的关系。我们也会学习 rs6516886 SNP 的作用并了解该基因组位点的特定变异是否与差异相关 USP16 的表达和对治疗的不同反应，可能导致目标患者的分层 人口。拟议的研究将为后续 GLP 研究提供信息，以支持 IND 和临床试验。