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

基本信息

批准号：
10414970
负责人：
Rolf A Brekken
金额：
$ 46.63万
依托单位：
BAYLOR UNIVERSITY
依托单位国家：
美国
项目类别：
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/10414970
关键词：
4T1 Acute Antibodies Antibody-drug conjugates Antimitotic Agents Antineoplastic Agents Binding 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 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

项目摘要

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。项目摘要/摘要对于高度选择性和有效治疗转移性和三重阴性乳腺癌的治疗剂。该提议推进了创新且有前途的策略重点是首次综合和评估Betabody-grug共轭（BDC），其具有靶向磷脂酰丝氨酸（PS）的工程蛋白（称为betabodies）。 PS 选择性地暴露在癌细胞和内皮细胞的质膜外叶上肿瘤微环境，但仅限于正常细胞质膜的内部小叶。这为选择性靶向有效的小分子抗癌有效载荷提供了一个精致的机会（kgp18 和kgp156）到肿瘤微环境。 β型较小，可以更好地渗透肿瘤与先前评估的PS选择性抗体相比，该抗体表现出较差的分布脉管系统。 BDC旨在选择性地释放KGP18和KGP156酶细胞外酶（组织蛋白酶B，尿激酶型纤溶酶原激活剂和纤溶酶）上调和/或证明乳腺癌细胞和相关肿瘤脉管系统的活性增强。 KGP18和KGP156有效微管蛋白聚合的抑制剂，并表现出作用双重机理，其功能非常活跃抗增生剂（低NM至PM范围内的细胞毒性），并且具有深刻有效的血管破坏药物，对与肿瘤相关的脉管系统造成不可逆转的损害最终导致肿瘤坏死。这些基于合成苯贝苯的有效载荷（KGP18和KGP156）将在比较研究中评估到单甲基雌激素E（MMAE），这是许多临床相关抗体 - 药物的有效载荷共轭。应当指出的是，VDA在机械上与研究良好的血管生成不同抑制剂。将研究两种个人交付策略。 KGP18和KGP156将是合成的用独特的蛋白酶选择性肽的链接器功能化，从而呈现药物 - 链链前药构建体在生物学上惰性，直到被肿瘤微环境中高水平的特定蛋白酶裂解，因此有选择地释放有效的有效载荷（KGP18或KGP156）。在第二个策略中，将准备BDC 从最好的药物保护选择性接头结构中。假设是适当设计的药物链接器结构及其相应的BDC将这些有效的有效载荷束缚（KGP18或KGP156）将表现出对肿瘤脉管系统和肿瘤细胞的高选择性。药代动力学研究将评估从其相应的构建体和偶联物中释放效应药物。的功效和选择性这些药物链构建体和BDC将使用基于细胞的研究评估，并确定了鼠原位乳腺癌模型（MDA-MB-231人异种移植物和同烯型4T1肿瘤）。总而言之预计每种策略将导致肿瘤特异性药物输送和有效的抗肿瘤活性。