Tumor-specific drug activation by pericellular proteases

细胞周蛋白酶激活肿瘤特异性药物

• 批准号: 10554408
• 负责人: Anthony John O'Donoghue
• 金额: $ 17.76万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10554408
• 关键词: Antibodies; Antibody-drug conjugates; Antineoplastic Agents; Binding; Biological; Biological Assay; Blood; Cardiotoxicity; Cells; Circulation; Complex; Coupling; Data; Data Set; Disabled Persons; Disease; Dose Limiting; ERBB2 gene; Endosomes; Engineering; Enzyme Stability; Enzymes; Epithelial Cells; Epitopes; Grant; Growth; Human; Kinetics; Knowledge; Liver; Lysosomes; MCF7 cell; Malignant - descriptor; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Organelles; Organoids; Patients; Peptide Hydrolases; Peptides; Peripheral Nervous System Diseases; Pharmaceutical Preparations; Plasma; Proteolysis; Resistance; SKBR3; Sampling; Serum; Site; Specificity; Substrate Specificity; Testing; Therapeutic; Therapeutic Effect; Toxic effect; Tumor-Derived; anti-cancer; cancer cell; comparative efficacy; design; drug development; extracellular; improved; malignant breast neoplasm; neoplastic cell; premature; protein aminoacid sequence; rational design; restoration; screening; tumor; tumor microenvironment

ABSTRACT Dysregulated pericellular proteolysis is a major driver of the malignant transformation of normal epithelial cells to cancer cells. The increased proteolytic activity in the tumor microenvironment has been exploited for drug development purposes. For example, antibody-drug conjugates (ADCs) consist of a highly potent anti-cancer payload conjugated to a tumor-targeting antibody. This coupling renders the cargo inactive until it is released by proteolytic cleavage in the pericellular space or upon degradation of the antibody within the target cell. ADCs are still handicapped by dose-limited toxicities, most likely due to off-target cell binding or premature drug release by proteases in circulation. The off-target binding can be overcome by engineering protease-activated pro- antibodies (pADCs) that can bind only after being activated at the site of disease by a protease while the premature drug release can be improved by utilizing peptide linker sequences that are stable in the presence of blood proteases but are efficiently cleaved by proteases at the site of disease. Our hypothesis is that pADCs that require two proteolytic activation steps, may improve our ability to selectively target anti-cancer drugs to the tumor microenvironment. The pADC will be converted into an ADC by a pericellular protease and subsequently bind to the tumor cell. The payload will be released by either an extracellular protease or an intracellular (endosomal or lysosomal) protease. The objectives of this proposal are to design ADCs and pADCs that have superior plasma stability and improved release kinetics by cancer-associated proteases. Our long-term objectives are to develop pADCs with enhanced anti-cancer efficacy compared to current therapeutics. In Aim 1, we will generate a substrate specificity profile for proteases secreted by patient-derived organoids of breast cancer. In addition, we will isolate lysosomes and endosomes and characterize the proteolytic activity in these organelles. Finally, we will evaluate protease activity in serum and plasma to understand the circulating proteolytic activity. In Aim 2, we will develop ADCs and pADCs that are selectively activated by cancer proteases and stable in plasma and serum.

抽象的 细胞周围蛋白水解失调是正常上皮细胞恶性转化的主要驱动力 到癌细胞。肿瘤微环境中蛋白水解活性的增加已被利用用于药物 开发目的。例如，抗体 - 药物结合物（ADC）由高度有效的抗癌体组成 有效载荷结合到靶向肿瘤的抗体中。这种耦合使货物不活跃，直到被释放 靶细胞内的细胞周围空间或抗体降解后的蛋白水解裂解。 ADC 仍然受到剂量限制毒性的残障 通过循环中的蛋白酶。可以通过工程蛋白酶激活的促进靶向来克服脱靶结合 抗体（PADC）仅在蛋白酶在疾病部位激活后才可以结合，而 可以通过利用在存在下稳定的肽接头序列来改善过早药物释放 血液蛋白酶，但在疾病部位有效地被蛋白酶裂解。我们的假设是PADC 需要两个蛋白水解激活步骤，可以提高我们有选择性靶向抗癌药物的能力 肿瘤微环境。 PADC将通过周围蛋白酶转换为ADC，随后将其转换为 与肿瘤细胞结合。有效载荷将由细胞外蛋白酶或细胞内释放 （内体或溶酶体）蛋白酶。该提案的目标是设计具有的ADC和PADC 优质的血浆稳定性和通过癌症相关蛋白酶改善释放动力学。我们的长期 与当前的治疗剂相比，目标是开发具有增强抗癌功​​效的PADC。 在AIM 1中，我们将生成由患者衍生的类器官分泌的蛋白酶的底物特异性曲线 乳腺癌。此外，我们将分离溶酶体和内体，并表征 这些细胞器。最后，我们将评估血清和血浆中的蛋白酶活性以了解循环 蛋白水解活性。在AIM 2中，我们将开发由癌症蛋白酶选择性激活的ADC和PADC 并在血浆和血清中稳定。