Targeted protein stabilization using Protein Rescue Targeting Chimeras (PRESTACs)

使用蛋白质救援靶向嵌合体 (PRESTAC) 实现靶向蛋白质稳定

基本信息

批准号：
10081989
负责人：
Kumar Suresh
金额：
$ 29.74万
依托单位：
PROGENRA, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-05 至 2022-08-04
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10081989
关键词：
Affect Agonist Animal Model Attenuated Binding Bioavailable Biochemical Biological Assay Biophysics Cell physiology Cells Chemicals Chimera organism Clinical Trials Complex Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Deubiquitinating Enzyme Development Disease Enzymes Future Gene Expression Genes Genetic Transcription Goals In Vitro Insulin Resistance Ligand Binding Ligands Ligase Link Malignant Neoplasms Mediating NIH 3T3 Cells Nerve Degeneration Non-Insulin-Dependent Diabetes Mellitus Nuclear Receptors Oral PPAR gamma Pathologic Pathology Pathway interactions Peptide Hydrolases Peroxisome Proliferator-Activated Receptors Pharmaceutical Preparations Phase Physiological Play Process Protac Protease Inhibitor Proteasome Inhibitor Protein Family Proteins Proteolysis Reporter Role Safety System TP53 gene Therapeutic Therapeutic Effect Thiazolidinediones Transcriptional Regulation Tumor Suppressor Proteins Ubiquitin Ubiquitination adiponectin base design innovation interest multicatalytic endopeptidase complex novel preservation prevent protein degradation protein expression protein function recruit reduce symptoms rosiglitazone small molecule success therapeutic target tool ubiquitin ligase ubiquitin-protein ligase

项目摘要

Targeting the ubiquitin pathway is a potential option for treating a variety of devastating diseases, including cancers and neurodegeneration. Recent focus on heterobifunctional proteolysis targeting chimeras, (PROTACs), which recruit E3 ligases to targets of interest for ubiquitination and degradation of the target, is a promising therapeutic avenue. PROTACs have been developed to degrade a wide range of protein families, and new targets are continually being investigated. In cells, ubiquitin conjugation by ligases is reversed by de- conjugating proteases (DUBs), and the dynamics of these two enzyme classes regulates protein turnover, compartmentation, and other processes. Some cell pathologies are driven by excessive degradation of a key protein, for example, a tumor suppressor, and the pathological state can be corrected by attenuating degradation, thereby sparing the critical protein. Formally, DUBs could be recruited by a heterobifunctional molecule to deubiquitinate target proteins and prevent their degradation, and it is therefore hypothesized that heterobifunctional molecules called Protein Rescue Targeting Chimeras (PRESTACs) can be generated by chemically linking a DUB ligand to a target ligand and evaluated for their physiological function. Progenra has identified and characterized novel small molecule USP7 ligands that bind USP7 and activate its DUB function. These molecules, along with ligands for targets of interest, will be used to design and synthesize PRESTACs, which will represent the first tethering of DUB activity to disease-relevant targets and provide a novel tool for expansion of UPS-based therapies. As proof of concept, bifunctional molecules will be synthesized to recruit the DUB USP7 to stabilize the nuclear receptor PPAR, for which several agonists are known. Thiazolidinediones (TZDs) targeting PPAR treat type 2 diabetes by inducing genes that decrease insulin resistance. Ligand binding of TZDs to PPAR not only regulates transcription, but also leads to PPAR ubiquitination and proteasomal degradation. Thus, stabilizing PPAR will prolong its activation and augment the effects of these drugs. PRESTACs will be designed using Progenra’s USP7 activators and rosiglitazone, a TZD whose interaction with and effects on PPAR are well-characterized. PRESTAC candidates will be evaluated biochemically, biophysically, and in NIH 3T3 cells for their ability to form ternary complexes (USP7-PRESTAC-PPAR), to stabilize PPAR, and to affect downstream targets of PPAR activation (for example, PPAR reporter assays, levels of adiponectin).

针对泛素通路是治疗多种破坏性疾病的潜在选择，包括癌症和神经退行性疾病最近关注的是针对嵌合体的异双功能蛋白水解，（PROTAC）将 E3 连接酶招募到感兴趣的靶标上，以实现靶标的泛素化和降解，是一种 PROTAC 已被开发用于降解多种蛋白质家族，并且新的靶点正在不断被研究，在细胞中，连接酶的泛素结合被去逆转。缀合蛋白酶（DUB），这两种酶的动态调节蛋白质周转，一些细胞病理是由关键的过度降解引起的。蛋白质，例如肿瘤抑制因子，可以通过减弱降解来纠正病理状态，形式上，DUB 可以被异源分子招募，从而形成双功能分子。去泛素化目标蛋白并防止其降解，因此发现称为蛋白质救援靶向嵌合体 (PRESTAC) 的异双功能分子可以通过以下方式生成 Progenra 将 DUB 配体与目标配体进行化学连接并评估其生理功能。鉴定并表征了结合 USP7 并激活其 DUB 功能的新型小分子 USP7 配体。这些分子以及感兴趣靶标的配体将用于设计和合成 PRESTAC，这将代表 DUB 活性与疾病相关靶标的首次结合，并为作为概念证明，将合成双功能分子以招募基于 UPS 的疗法。 DUB USP7 可稳定核受体 PPAR，已知有几种噻唑烷二酮类激动剂。 (TZD) 靶向 PPAR 通过诱导降低胰岛素抵抗的基因来治疗 2 型糖尿病。 TZD 与 PPAR 的结合不仅调节转录，还导致 PPAR 泛素化和蛋白酶体化因此，稳定 PPAR 将延长其激活时间并增强这些药物的作用。 PRESTAC 将使用 Progenra 的 USP7 激活剂和罗格列酮（一种 TZD，其与对 PPAR 的影响已得到充分表征，PRESTAC 候选物将进行生化评估。在生物物理上，以及在 NIH 3T3 细胞中，它们能够形成三元复合物 (USP7-PRESTAC-PPAR)， PPAR，并影响 PPAR 激活的下游靶点（例如，PPAR 报告基因检测、脂联素水平）。