Characterization of A Novel Proteasome Inhibitor

新型蛋白酶体抑制剂的表征

• 批准号: 10597711
• 负责人: Jun O. Liu
• 金额: $ 52.16万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10597711
• 关键词: Adverse effects; Animal Model; Antineoplastic Agents; Apoptotic; Archaea; Bacteria; Binding; Binding Sites; Biological Availability; Bortezomib; CRISPR screen; Cell Line; Cell physiology; Cells; Chemicals; Clinic; Complex; Cryoelectron Microscopy; Development; Disease; Drug Kinetics; Drug Targeting; Drug resistance; Eukaryota; Exhibits; Garbage; Genetic Transcription; Human; Hydrolysis; Immunoglobulins; Immunosuppressive Agents; Induction of Apoptosis; Lead; Libraries; Lymphoma; Malignant Neoplasms; Mediating; Mediator; Multiple Myeloma; Names; Normal Cell; Patients; Penetration; Peptide Hydrolases; Peptide Library; Periodicity; Peripheral Nervous System Diseases; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Photoaffinity Labels; Plasma Cell Neoplasm; Play; Prodrugs; Property; Proteasome Inhibition; Proteasome Inhibitor; Proteins; Regulation; Resistance; Resolution; Role; Route; Safety; Signal Transduction; Sirolimus; Site; Structure; Structure-Activity Relationship; Tacrolimus Binding Proteins; Therapeutic; Toxic effect; Work; analog; anti-cancer; cancer cell; cancer type; combinatorial; design; genome-wide; improved; in vivo; inhibitor; insight; multicatalytic endopeptidase complex; novel; novel anticancer drug; potential biomarker; predictive marker; prolyl oligopeptidase; proteostasis; refractory cancer; success; synergism; tool; tumor

The human proteasome plays an essential role in both protein homeostasis and in the regulation of multiple cellular processes from signal transduction to transcription. It has also become a proven target for developing drugs for treating multiple diseases including multiple myeloma and lymphoma. Several inhibitors of the proteasome, including bortezomib, carfilzomib and ixazomib, have been used in the clinic, responsible for prolonging lives of multiple myeloma patients. However, the existing proteasome-targeted drugs suffer from severe toxicity and rapid emergence of drug resistance, which also limits their potential in treating other types of diseases. We have developed a rapamycin-inspired macrocycle library known as rapafucins by fusing the FKBP-binding domain (FKBD) of rapamycin with a combinatorial peptide library. A screen of the rapafucin library in multiple myeloma cell line NCI-H929 led to the identification of a potent inhibitor, Rapaprotin that induces apoptosis in NCI-H929 cells. Rapaprotin exhibited selective toxicity to cancer cells over normal cells. It also has a unique mechanism of action, requiring activation by an intracellular protease through cleavage of the macrocycle into a linear form, Rapaprotin-L, which inhibits all three types of protease activities of the proteasome. Moreover, Rapaprotin is synergistic with bortezomib and is capable of resensitizing bortezomib-resistant cancer cells to the drug. In this application, we will investigate the mechanism of activation of Rapaprotin by the cellular protease, validating Rapaprotin-L as the active species for its cellular activity. We will obtain a high-resolution cryo-EM structure of the complex between proteasome and Rapaprotin-L. We will optimize the potency and pharmacokinetic property of Rapaprotin through design and synthesis of new analogs. The optimized analogs of Rapaprotin will be assessed for their efficacy in animal models of multiple myeloma and other diseases. The newly developed Rapaprotin analogs will serve as useful chemical probes to facilitate the study of the function and pharmacology of the proteasome and promising leads for developing a new class of anticancer and immunosuppressive drugs with lower adverse effects.

人类蛋白酶体在蛋白质稳态和蛋白质代谢中发挥着重要作用。 从信号转导到细胞凋亡的多个细胞过程的调节 转录。它也已成为开发治疗药物的一个被证明的目标 包括多发性骨髓瘤和淋巴瘤在内的多种疾病。的多种抑制剂 已使用蛋白酶体，包括硼替佐米、卡非佐米和伊沙佐米 在临床上，负责延长多发性骨髓瘤患者的生命。 然而，现有的蛋白酶体靶向药物具有严重的毒性， 耐药性的迅速出现，这也限制了它们在治疗 其他类型的疾病。我们开发了一种雷帕霉素大环化合物 通过融合FKBP-结合结构域（FKBD）， 雷帕霉素与组合肽库的比较。Rapafucin文库的屏幕， 多发性骨髓瘤细胞系NCI-H929导致了有效抑制剂的鉴定， Rapaprotin诱导NCI-H929细胞凋亡。Rapaprotin显示 对癌细胞的选择性毒性超过正常细胞。它还有一个独特的机制 的作用，需要通过细胞内蛋白酶的裂解激活， 大环化合物转化为线性形式，Rapapartin-L，其抑制所有三种类型的 蛋白酶体的蛋白酶活性。此外，Rapaprotin与 并且能够使硼替佐米抗性癌细胞对硼替佐米再敏感。 药在本申请中，我们将研究激活的机制， 通过细胞蛋白酶测定Rapaprotin，验证Rapaprotin-L为活性物质 因为它的细胞活性。我们将获得一个高分辨率的冷冻电镜结构， 蛋白酶体与Rapaprotin-L之间的复合物。我们将优化效力， 通过设计和合成新Rapaprotin的药代动力学性质 类似物将评估Rapaprotin的优化类似物在以下方面的功效： 多发性骨髓瘤和其他疾病的动物模型。新开发的 Rapaprotin类似物将作为有用的化学探针，以促进对细胞内蛋白质的研究。 蛋白酶体的功能和药理学， 开发一类新的抗癌和免疫抑制药物， 不良影响