Novel Protein Engineered Drug Conjugates Targeting Phosphatidylserine for the Treatment of Breast Cancer

靶向磷脂酰丝氨酸的新型蛋白质工程药物缀合物用于治疗乳腺癌

• 批准号: 10183201
• 负责人: Rolf A Brekken
• 金额: $ 47.56万
• 依托单位: BAYLOR UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-05 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10183201
• 关键词: 4T1; Acute; Antibodies; Antibody-drug conjugates; Antimitotic Agents; Antineoplastic Agents; Binding; Biological; Biological Assay; Biology; Blood Vessels; Breast Cancer Cell; Breast Cancer Model; Breast Cancer Treatment; Breathing; Cathepsin L; Cathepsins B; Cell Death; Cell division; Cell membrane; Cells; Characteristics; Cleaved cell; Comparative Study; Cytotoxic agent; Diffuse; Drug Delivery Systems; Drug Design; Drug Kinetics; Endothelial Cells; Enzymes; Evaluation; Exposure to; Future; Growth; Half-Life; Home; Human; Immune; Inbred BALB C Mice; Individual; Libraries; MDA MB 231; Malignant Neoplasms; Mammary Neoplasms; Mediating; Modality; Monoclonal Antibodies; Mouse Mammary Tumor Virus; Mus; Myeloid-derived suppressor cells; Necrosis; Normal Cell; Patients; Penetration; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Phosphatidylserines; Plasma; Plasmin; Prodrugs; Protein Engineering; Research; Rest; SCID Mice; Safety; Signal Transduction; Therapeutic Agents; Time; Toxic effect; Transglutaminases; Treatment Efficacy; Tumor Immunity; Tumor Markers; Tumor-Associated Vasculature; Tumor-associated macrophages; Urokinase; Validation; Vascular Endothelial Cell; Xenograft procedure; analog; angiogenesis; anti-cancer; anti-cancer therapeutic; anti-tumor immune response; antiproliferative agents; base; beta 2-glycoprotein I; bioluminescence imaging; cancer cell; catalyst; chemical stability; chemical synthesis; clinically relevant; cytokine; cytotoxicity; design; drug development; drug discovery; extracellular; improved; in vitro Assay; in vivo; innovation; mouse model; nanomolar; neoplastic cell; next generation; novel; orthotopic breast cancer; side effect; small molecule; targeted treatment; therapeutically effective; triple-negative invasive breast carcinoma; tubulin polymerization inhibitor; tumor; tumor growth; tumor microenvironment; tumor specificity; tumor-immune system interactions

7. Project Summary/Abstract There remains an urgent, and largely unmet need for the advancement of highly selective and effective therapeutic agents for the treatment of metastatic and triple negative breast cancer. This proposal advances an innovative and promising strategy focused on the first time synthesis and evaluation of betabody-drug conjugates (BDCs), which feature engineered proteins (referred to as betabodies) that target phosphatidylserine (PS). PS is selectively exposed on the outer leaflet of the plasma membrane of cancer cells and endothelial cells in the tumor microenvironment, but is confined to the inner leaflet of the plasma membrane of normal cells. This provides an exquisite opportunity for selective targeting of potent small-molecule anticancer payloads (KGP18 and KGP156) to the tumor microenvironment. Betabodies are smaller allowing for better tumor penetration compared with previously evaluated PS-selective antibodies, which demonstrated poor distribution beyond the vasculature. BDCs are designed to selectively release KGP18 and KGP156 extracellularly by enzymes (cathepsin B, urokinase-type plasminogen activator, and plasmin) that are upregulated and/or demonstrate enhanced activity by breast cancer cells and associated tumor vasculature. KGP18 and KGP156 are potent inhibitors of tubulin polymerization and demonstrate dual mechanism of action functioning as remarkably active antiproliferative agents (cytotoxicity in low nM to pM range) and as profoundly effective vascular disrupting agents, which impart irreversible damage to tumor-associated vasculature ultimately leading to tumor necrosis. These synthetic benzosuberene-based payloads (KGP18 and KGP156) will be evaluated in comparative studies to monomethyl-auristatin E (MMAE), which is a payload of choice in many clinically relevant antibody-drug conjugates. It should be noted that VDAs are mechanistically distinct from the well-studied angiogenesis inhibiting agents. Two individual delivery strategies will be investigated. KGP18 and KGP156 will be synthetically functionalized with distinct, protease-selective peptide-based linkers rendering the drug-linker prodrug constructs biologically inert until cleaved by specific proteases that are present at high levels in the tumor microenvironment, thus selectively releasing the potent payload (KGP18 or KGP156). In a second strategy, BDCs will be prepared from the best drug-protease selective linker constructs. The hypothesis is that appropriately designed drug-linker constructs and their corresponding BDCs tethered to these highly potent payloads (KGP18 or KGP156) will demonstrate high selectivity for tumor vasculature and tumor cells. Pharmacokinetic studies will evaluate the release of the effector drugs from their corresponding constructs and conjugates. The efficacy and selectivity of these drug-linker constructs and BDCs will be evaluated using cell-based studies and established murine orthotopic breast cancer models (MDA-MB-231 human xenografts and syngeneic 4T1 tumors). In summary, it is expected that each strategy will result in tumor-specific drug delivery and potent anti-tumor activity.

7. 项目总结/摘要 对于推进高度选择性和有效的技术，仍然存在紧迫且基本上未得到满足的需求。 用于治疗转移性乳腺癌和三阴性乳腺癌的治疗剂。该提案提出了 创新且有前途的策略专注于β抗体-药物缀合物的首次合成和评估 （BDC），其特征是靶向磷脂酰丝氨酸（PS）的工程蛋白（称为β抗体）。聚苯乙烯 选择性地暴露在癌细胞和内皮细胞质膜的外层 肿瘤微环境，但仅限于正常细胞质膜的内层。这 为选择性靶向有效小分子抗癌有效负载（KGP18 和KGP156）对肿瘤微环境的影响。 Betabody 更小，可以更好地渗透肿瘤 与之前评估的 PS 选择性抗体相比，其分布较差 脉管系统。 BDC 旨在通过酶选择性地将 KGP18 和 KGP156 释放到细胞外 （组织蛋白酶 B、尿激酶型纤溶酶原激活剂和纤溶酶）上调和/或表现出 乳腺癌细胞和相关肿瘤血管系统的活性增强。 KGP18 和 KGP156 有效 微管蛋白聚合抑制剂，并表现出双重作用机制，具有非常活跃的作用 抗增殖剂（低 nM 至 pM 范围内的细胞毒性）和极其有效的血管破坏 药物，对肿瘤相关脉管系统造成不可逆的损害，最终导致肿瘤坏死。 这些基于苯并环庚烯的合成有效载荷（KGP18 和 KGP156）将在比较研究中进行评估 单甲基奥瑞他汀 E (MMAE)，它是许多临床相关抗体药物的有效负载选择 共轭物。应该指出的是，VDA 在机制上与经过充分研究的血管生成不同。 抑制剂。将研究两种单独的交付策略。 KGP18和KGP156将合成 用独特的、基于蛋白酶选择性肽的连接体进行功能化，从而形成药物-连接体前药结构 生物惰性，直到被肿瘤微环境中高水平存在的特定蛋白酶裂解为止， 从而选择性地释放有效负载（KGP18或KGP156）。在第二个战略中，BDC 将做好准备 来自最好的药物-蛋白酶选择性接头构建体。假设是适当设计的药物连接体 构建体及其相应的 BDC 与这些高效有效负载（KGP18 或 KGP156）相连 表现出对肿瘤血管和肿瘤细胞的高选择性。药代动力学研究将评估 效应药物从其相应的构建体和缀合物中释放。功效和选择性 这些药物接头结构和 BDC 将使用基于细胞的研究和已建立的小鼠模型进行评估 原位乳腺癌模型（MDA-MB-231 人类异种移植物和同基因 4T1 肿瘤）。概括起来就是 预计每种策略都会产生肿瘤特异性药物输送和有效的抗肿瘤活性。