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

• 批准号: 10561636
• 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
• 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/10561636
• 关键词: Address; Alcohols; Amines; Antibody-drug conjugates; Area; Asparagine; Autoimmunity; Behavioral; Biological Assay; Biological Markers; Cancer cell line; Carbamates; Catabolism; Cathepsins; Circulation; 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; Marketing; Measures; Medical; Modeling; Mus; Neutropenia; Oncology; Ophthalmology; Peptide Hydrolases; Peptides; Pharmacologic Substance; Phase II Clinical Trials; Plasma; Predisposition; Process; Rapid screening; Rattus; Resistance; Safety; Site; System; Technology; Testing; Therapeutic; Thrombocytopenia; Toxic effect; Work; Xenograft Model; asparaginylendopeptidase; 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

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： 目前市场上有8种FDA批准的抗体药物缀合物（ADC）。ADC技术正被广泛应用于 用于肿瘤学、自身免疫、眼科学和传染病应用。尽管巨大的 然而，成功的ADC开发过程充满了与ADC聚集，连接体- 稳定性问题、意外毒性和不良药代动力学。虽然不可能把责任归咎于 虽然这是一种单一现象，但绝大多数ADC的一个共同点是依赖于 疏水肽接头（即ValCit-PABC），其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，这种接头在循环中比以前认为的更不稳定， 易受血浆酯酶（Ces 1C）和淋巴细胞相关蛋白酶（中性粒细胞弹性蛋白酶）的切割。 现在人们普遍认为，中性粒细胞弹性蛋白酶的切割是导致剂量限制性中性粒细胞减少症的原因， 血小板减少症是许多ADC常见的。迫切需要新一代的可裂解连接体， 克服这些债务。在此，我们提出了“重新设计的”ADC接头的优化和评估， 含有天冬酰胺（Asn）基序，其被天冬酰胺酰内切蛋白酶（也称为豆荚蛋白）快速切割。我们 研究小组已经确定了几个含有Asn的序列，这些序列被溶酶体豆荚蛋白快速切割， 在小鼠和人血浆中的稳定性。我们的初步数据显示，结合这些连接子的ADC表现出 与经典ValCit-PABC接头相当的体外细胞毒性-但不易被Ces 1C或 中性粒细胞弹性蛋白酶我们设计了三个具体目标来优化该连接体，并证明模型ADC 与传统的ValCit-PABC接头相比，使用这些接头的药物具有改善的治疗潜力。目标#1 重点在于肽接头本身的优化。我们将准备一个基于FRET的含Asn的文库， 本发明的目的是提供一种新的肽，以鉴定可以掺入各种癌症靶向ADC中的稳健接头。目标#2 集中于开发用于豆荚蛋白可切割肽的合适的自分解间隔区。间隔件将 为了促进在溶酶体中的快速裂解和在血浆中的高稳定性，目标#3将建立 豆类蛋白可裂解ADC在胰腺癌模型中的治疗效用。具体来说，我们将把 将我们的新型接头转化为抗GCC ADC，其对于治疗胰腺癌是有意义的。ADC将 与由于剂量限制而导致II期临床试验失败的传统ADC（TAK-264）进行头对头比较 中性粒细胞减少症。我们预期，我们的新型接头将赋予这种ADC改善的耐受性，同时维持ADC的生物相容性。 功效这些目标的实现将坚定地建立豆类切割用于治疗的适用性。 应用-并将平衡临床应用的技术。新型胰腺癌靶向治疗的研究进展 ADC将为在一个巨大的未满足的医疗需求领域进行临床评估铺平道路。