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

基本信息

批准号：
10300923
负责人：
Maddalena Adorno
金额：
$ 45.29万
依托单位：
DORIAN THERAPEUTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-30 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10300923
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 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 Pharmaceutical Preparations Pharmacodynamics Phase Physiological Plasma Population Presenile Alzheimer Dementia 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 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

项目摘要

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（一种参与衰老和干细胞自我更新调节的染色质修饰剂）开发一种针对骨关节炎 (OA) 的疾病缓解疗法。该疗法包括关节内注射少量药物。中度 OA 患者中抑制 USP16 的分子 USP16 是一种去泛素酶 (DUB)，可去除赖氨酸上组蛋白 H2A 上的泛素。 119 是维持多个体细胞组织的关键标志，提示 USP16 的三倍体与唐氏综合症 (DS) 相关，唐氏综合症是一种以 21 号染色体 (HSA21) 三倍体为特征的先天性疾病，患有 DS 的患者表现出加速衰老的迹象。我们之前已经证明，降低 DS 小鼠模型或人体组织中 USP16 的水平可改善成体干细胞的功能并减少。值得注意的是，USP16 的表达在 OA 软骨细胞和滑膜组织中高度上调。 SNP 与家族性 OA 密切相关，我们发现患者来源的关节软骨细胞中 USP16 的基因下调可促进细胞和线粒体健康。并减少衰老相关标记物，如 SA-ꞵ-gal 和 p16Ink4a。此外，针对 USP16 的 siRNA 对 OA 软骨细胞的微阵列分析显示，经过处理的增殖和胶原沉积标记物有所增加，并且细胞凋亡和分解代谢标记物有所减少。人骨髓来源的间充质干细胞的敲低促进分化为成熟软骨细胞并使用生化测定测试酶活性增加基质沉积。重组人 USP16 中，我们鉴定了 19 个具有低 IC50 的小分子抑制剂，并选择了 IC50 介于 0.06 μM 和 9.4 μM 之间的小分子支架。在该项目的第一阶段，我们将对其进行优化以提高效力、特异性和稳定性。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，然后对两个最好的先导化合物进行 OA 模型的临床前疗效和早期毒性测试。我们将评估它们的安全性、药代动力学 (PK)、作为药效学 (PD) 指标参与靶点和调节染色质的能力，以及在手术大鼠 OA 模型中预防或逆转软骨损失的功效，目的是建立一种我们还将研究 rs6516886 SNP 的作用，以及该基因组位点的特定变异是否与 USP16 的差异表达和对治疗的不同反应相关。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。