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: 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
• 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

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.

项目摘要： 现在，市场上有8个FDA批准的抗体 - 毒剂偶联物（ADC）。 ADC技术广泛 追求肿瘤学，自身免疫性，眼科和传染病应用。尽管有巨大的 然而，成功，ADC开发过程充满了与ADC聚合，接头 - 稳定性问题，意外毒性和药代动力学不良。虽然不可能为此蒙上盲人 绝大多数ADC共享的一个现象的脚是一个共同的因素 疏水性胡椒接头（即瓣膜PABC）在血浆中稳定，但被溶酶体组织蛋白酶迅速裂解。 最近的各种研究表明，该连接器在循环中比以前认为的更不稳定，是 易受血浆酯酶（CES1C）和淋巴细胞相关蛋白酶（中性脑酯酶）的裂解。 现在人们普遍认为，中性粒细胞弹性蛋白酶的裂解是剂量限制性中性粒细胞和 对于许多ADC而言，常见的血小板减少症。迫切需要新一代的可切合接头 克服这些责任。在此，我们提出了对“重新设计” ADC链接器的优化和评估 含有天冬酰胺（ASN）基序，这些基序被天冬酰胺基内蛋白酶（也称为豆类）迅速切割。我们的 团队已经确定了几个含有ASN的序列，这些序列在保留的同时被溶酶体豆科素迅速切割 小鼠和人血浆中的稳定性。我们的初步数据表明，包含这些接头的ADC展示了 与经典瓣膜-PABC接头可比的体外细胞毒性可比 - 但不容易被CES1C或 中性粒细胞弹性蛋白酶。我们设计了三个特定的目的，以优化该链接器并证明模型ADCS 与传统的Valcit-PABC接头相比，使用这些接头具有提高的治疗潜力。 AIM＃1是 专注于肽接头本身的优化。我们将准备一个基于FRET的ASN的库 肽是为了识别可纳入各种癌症ADC中的稳健接头。 AIM＃2是 专注于开发适合豆药裂解胡椒粉的合适的自使性垫片。垫片会 为了促进溶酶体中的快速切割和血浆中的高稳定性。 AIM＃3将建立 在胰腺癌模型中，豆类可转化的ADC的治疗效用。具体来说，我们将合并 我们进入抗高潮ADC的新型接头，这些接头对治疗胰腺癌感兴趣。 ADC将是 将正面对比与传统ADC（TAK-264）进行了比较，该ADC因剂量限制而失败了2期临床试验 中性粒细胞减少。我们预计我们的新颖的连接器将在维护该ADC的同时提高对此ADC的可耐受性 效率。这些目标的实现将首先确定豆科治疗的适用性 应用 - 并将为临床应用的技术增压。开发新颖的胰腺癌靶向 ADC将在巨大未满足医疗需求的领域铺平临床评估的道路。