Legumain to the rescue: A new ADC linker strategy to address the limitations of cathepsin cleavage

Legumain 来拯救：一种新的 ADC 连接策略，解决组织蛋白酶切割的局限性

基本信息

批准号：
10342525
负责人：
Lawrence Tumey
金额：
$ 30.31万
依托单位：
STATE UNIVERSITY OF NY,BINGHAMTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-10 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10342525
关键词：
Address Alcohols Amines Antibody-drug conjugates Area Asparagine Autoimmunity Behavioral Biological Assay Biological Markers Blood Circulation Cancer cell line Carbamates Catabolism Cathepsins Communicable Diseases Data Development Dose-Limiting Drug Kinetics Ensure Enzymes Evaluation Exatecan Exhibits FDA approved Fluorescence Resonance Energy Transfer Funding Generations Goals Head Hematology Human Hydrophobicity In Vitro Left Leukocyte Elastase Libraries Lymphocyte Lysosomes Malignant Neoplasms Malignant neoplasm of pancreas Manuscripts Measures Medical Modeling Mus Neutropenia Oncology Ophthalmology Peptide Hydrolases Peptides Pharmacologic Substance Phase II Clinical Trials Plasma Process Rapid screening Rattus Resistance Safety Site System Technology Testing Therapeutic Thrombocytopenia Toxic effect Work Xenograft Model asparaginylendopeptidase base clinical application cytotoxicity design efficacy evaluation enterotoxin receptor esterase foot head-to-head comparison improved interest member novel pancreatic cancer model research clinical testing screening side effect success unnatural amino acids

项目摘要

Project Summary: There are now 8 FDA-approved antibody-drug-conjugates (ADCs) on the market. ADC technology is being widely pursued for oncology, autoimmunity, ophthalmology, and infectious disease applications. In spite of tremendous success, however, the ADC development process is riddled with challenges associated with ADC aggregation, linker- stability concerns, unexpected toxicity, and poor pharmacokinetics. While it is impossible to lay the blame for this at the feet of a single phenomenon, one common factor shared by the vast majority of ADCs is the reliance on a hydrophobic peptide linker (i.e. ValCit-PABC) that is stable in plasma but rapidly cleaved by lysosomal cathepsins. Various studies have recently shown that this linker is more unstable in circulation was than previously believed, being susceptible to cleavage by both plasma esterases (Ces1C) and lymphocyte-associated proteases (neutrophil elastase). It is now widely believed that cleavage by neutrophil elastase is responsible for the dose-limiting neutropenia and thrombocytopenia so commonly seen for many ADCs. A new generation of cleavable linkers is urgently needed in order to overcome these liabilities. Herein, we propose the optimization and evaluation of “redesigned” ADC linkers that contain asparagine (Asn) motifs which are rapidly cleaved by asparaginyl endoprotease, also known as legumain. Our team has identified several Asn-containing sequences that are rapidly cleaved by lysosomal legumain while retaining stability in mouse and human plasma. Our preliminary data shows that ADCs that incorporate these linkers exhibit comparable in vitro cytotoxicity to classical ValCit-PABC linkers – but are not susceptible to cleavable by Ces1C or neutrophil elastase. We have designed three specific aims to optimize this linker and to demonstrate that model ADCs that use these linkers have improved therapeutic potential as compared to traditional ValCit-PABC linkers. Aim #1 is is focused on the optimization of the peptide linker itself. We will prepare a FRET-based library of Asn-containing peptides in order to identify robust linkers that can be incorporated into various cancer-targeting ADCs. Aim #2 is focused on the development of suitable self-immolative spacers for the legumain cleavable peptides. The spacer will be optimized in order to facilitate rapid cleavage in the lysosome and high stability in plasma. Aim #3 will establish the therapeutic utility of legumain-cleavable ADCs in a model of pancreatic cancer. Specifically, we will incorporate our novel linkers into anti-GCC ADCs that are of interest for the treatment of pancreatic cancer. The ADCs will be compared head-to-head with a traditional ADC (TAK-264) that failed phase 2 clinical trials due to dose limiting neutropenia. We anticipate that our novel linkers will impart an improved tolerability to this ADC while maintaining the efficacy. Accomplishment of these aims will firmly establish the suitability of legumain-cleavage for therapeutic applications – and will poise the technology for clinical application. Development of novel pancreatic-cancer targeting ADCs will pave the way towards clinical evaluation in an area of tremendous unmet medical need.

项目概要：目前市场上已有 8 种经 FDA 批准的抗体药物偶联物 (ADC) 技术得到广泛应用。尽管有巨大的应用，但仍被用于肿瘤学、自身免疫学、眼科和传染病。然而，ADC 开发过程却充满了与 ADC 聚合、链接器相关的挑战。稳定性问题、意外的毒性和不良的药代动力学，但不可能将其归咎于这一点。单一现象的脚，绝大多数 ADC 共有的一个共同因素是对疏水性肽接头（即 ValCit-PABC）在血浆中稳定，但被溶酶体组织蛋白酶快速裂解。最近的各种研究表明，这种连接体在循环中比以前认为的更不稳定，对血浆酯酶 (Ces1C) 和淋巴细胞相关蛋白酶（中性粒细胞弹性蛋白酶）裂解的敏感性。现在人们普遍认为中性粒细胞弹性蛋白酶的裂解是造成剂量限制性中性粒细胞减少症和血小板减少症在许多 ADC 中都很常见，因此迫切需要新一代可裂解接头。为了克服这些缺点，我们提出了“重新设计”的 ADC 连接器的优化和评估。含有天冬酰胺 (Asn) 基序，可被天冬酰胺酰内切蛋白酶（也称为“我们的 Legumain”）快速切割。研究小组已经鉴定了几个含有 Asn 的序列，这些序列可以被溶酶体 legumain 快速切割，同时保留我们的初步数据表明，包含这些连接子的 ADC 表现出稳定性。体外细胞毒性与经典 ValCit-PABC 连接体相当，但不易被 Ces1C 或我们设计了三个具体目标来优化该连接器并证明模型 ADC。与传统的 ValCit-PABC 连接体相比，使用这些连接体的治疗潜力有所提高。目标#1 是。我们的重点是肽接头本身的优化，我们将制备一个基于 FRET 的含 Asn 文库。目标#2 是为了识别可整合到各种癌症靶向 ADC 中的稳健连接体。专注于为legumain可裂解肽开发合适的自毁间隔物。进行优化以促进溶酶体中的快速裂解和血浆中的高稳定性。具体而言，我们将纳入legumain 可裂解 ADC 在胰腺癌模型中的治疗效用。我们的新型抗 GCC ADC 接头对治疗胰腺癌感兴趣。与因剂量限制而未能通过 2 期临床试验的传统 ADC (TAK-264) 进行了头对头比较我们预计我们的新型接头将提高该 ADC 的耐受性，同时保持这些目标的实现将坚定地确定legumain裂解对于治疗的适用性。应用——并将为临床应用开发新型胰腺癌靶向技术。 ADC 将为巨大的未满足医疗需求领域的临床评估铺平道路。