Development of caveolae-targeted antibody-drug conjugates

开发小凹靶向抗体药物偶联物

基本信息

批准号：
9974488
负责人：
Bogdan Olenyuk
金额：
$ 44.12万
依托单位：
PROTEOGENOMICS RESEARCH INSTIT/SYS/ MED
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-08 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9974488
关键词：
Address Affinity Annexin A1 Antibodies Antibody-drug conjugates Antineoplastic Agents Attenuated BT 474 Binding Biodistribution Biological Biometry Blood Blood Vessels Breast Cancer Cell Breast Cancer Treatment Buffers Cancer Model Carboplatin Caveolae Cell Wall Cell surface Chemistry Cisplatin Cleaved cell Clinic Complex Data Data Analyses Development Dextrans Disseminated Malignant Neoplasm Dose Drug Delivery Systems Drug Kinetics Drug resistance Endothelial Cells Endothelium Goals Half-Life Histology Human Image Immunoconjugates In Vitro Injections Intravenous Kinetics MDA MB 231 Mammary Neoplasms Maximum Tolerated Dose Measures Metastatic Neoplasm to the Lung Metastatic breast cancer Modeling Modernization Monitor Multi-Drug Resistance Multimodal Imaging Mus Neoplasm Metastasis Normal tissue morphology Pathway interactions Patients Penetration Pharmaceutical Preparations Phase Plasma Platinum Pre-Clinical Model Property Proteins Publishing Pump Quality Control Rattus Risk Rodent Safety Services Side Solid Solid Neoplasm Solubility Specificity Speed System Testing Therapeutic Toxic effect Translating Translations Treatment Efficacy Vascular Endothelial Cell Vascular Endothelium X-Ray Computed Tomography Xenograft procedure animal imaging base cGMP production cancer therapy carboxylate carcinogenicity chemotherapy clinical candidate clinical efficacy clinical translation design dosage drug efficacy humanized antibody improved in vivo innovation intravital microscopy lung metastatic malignant breast neoplasm neoplastic cell novel novel therapeutics oxaliplatin pre-clinical replication factor C research clinical testing response side effect single photon emission computed tomography systemic toxicity targeted delivery targeted treatment therapeutic effectiveness therapy outcome treatment response tumor uptake vascular endothelium permeability

项目摘要

PROJECT 2 SUMMARY The overall objective of the Project 2 is to utilize a newly discovered, active transendothelial transport pathway, the caveolae pumping system, in order to provide an effective solution to the delivery and toxicity problem of chemotherapeutics in metastatic breast cancer treatment. In order to address these problems and significantly improve therapeutic outcome we propose to develop novel antibody-drug conjugates (ADCs) that exploit our newly discovered endothelial cell (EC) caveolae targeting system in order to sidestep passive delivery problem of modern chemotherapeutics. We have established that EC caveolae in preclinical models can rapidly and specifically pump anti-Annexin A1 antibodies and attached cargo across the vascular endothelial barrier directly into solid tumors. Based on this discovery, we propose to design novel EC caveolae-targeted ADCs for greatly enhanced efficacy in tumor destruction in breast cancer. Our main hypothesis is that immunoconjugates that fully utilize the advantages of the EC caveolae-pumping system will dramatically increase delivery of chemotherapeutic drugs into tumors, thereby requiring much lower dosages and dramatically enhancing efficacy of treatment. This hypothesis will be tested by the following specific aims: In Aim 1, we will design and synthesize Pt(II)-carboxymethyl dextran (CMD) ADCs and optimize carboxylate derivatization of CMD, conjugation chemistry, and Pt(II) loading in order to maximize binding affinity toward human Annexin A1 protein. We will test and optimize physicochemical properties, stability on storage, in human plasma, and Pt(II) release kinetics in the tumor interstitium. In Aim 2 we will assess therapeutic efficacy of the ADCs in metastatic cancer models. The efficacy of our targeted delivery system will be examined in MDA-231-LM2-4, and rat tumor models with 13762 breast cancer cells metastasized to lung using whole-body animal imaging with X-ray/CT. In Aim 3 we will examine the therapeutic efficacy of the ADCs in tumors with human blood vessels using intravital microscopy (IVM). We will monitor tumor size in response to therapy in human-on-human IVM model using BT-474 tumor spheroids, and in patient-derived xenografts (PDX) IVM model. In this Aim we will also translate our select preclinical candidate for clinical testing, where successful targeted therapeutic will be selected as clinical candidates for cGMP production (Core B) for subsequent testing in a Phase 1 safety trial in Project 3. Antibodies for conjugation to form ADCs will be provided by Core B. Core B will also provide quality control analysis of ADCs and confirmation of antibody binding affinity following conjugation. Tumor targeting, delivery and accumulation of these novel therapies will be assessed in rodents using multi-modality imaging services provided by Core C, which includes X-ray/CT, SPECT-CT, IVM, and histology. Core E will handle the biostatistical analysis of data generated during the course of the project. The long-term goal is to translate our key basic discoveries into unique, innovative delivery platform in order to improve therapeutic efficacy of human cancer therapy.

项目 2 总结项目 2 的总体目标是利用新发现的活跃跨内皮转运途径， Caveolae泵送系统，以提供有效的解决方案的输送和毒性问题转移性乳腺癌治疗中的化疗。为了解决这些问题并显着改善治疗结果，我们建议开发新型抗体药物偶联物（ADC），利用我们的新发现的内皮细胞（EC）小窝靶向系统以避免被动递送问题现代化疗的基础。我们已经确定临床前模型中的 EC 小窝可以快速且有效地特异性泵送抗膜联蛋白 A1 抗体和附着物穿过血管内皮屏障直接进入实体瘤。基于这一发现，我们建议设计新型 EC Caveolae 靶向 ADC 大大增强了乳腺癌肿瘤破坏的功效。我们的主要假设是充分利用 EC 细胞膜穴泵系统优势的免疫偶联物将显着增加化疗药物向肿瘤的输送，从而需要更低的剂量显着提高治疗效果。该假设将通过以下具体目标进行检验：目标 1，我们将设计和合成 Pt(II)-羧甲基葡聚糖 (CMD) ADC 并优化羧酸盐 CMD 衍生化、缀合化学和 Pt(II) 负载，以最大限度地提高与人膜联蛋白 A1 蛋白。我们将测试和优化理化性质、储存稳定性、人血浆和 Pt(II) 在肿瘤间质中的释放动力学。在目标 2 中，我们将评估治疗效果转移性癌症模型中的 ADC。我们的靶向递送系统的功效将在 MDA-231-LM2-4，以及全身13762个乳腺癌细胞转移至肺部的大鼠肿瘤模型使用 X 射线/CT 进行动物成像。在目标 3 中，我们将检查 ADC 对肿瘤的治疗效果使用活体显微镜（IVM）观察人体血管。我们将监测肿瘤大小对治疗的反应使用 BT-474 肿瘤球体的人对人 IVM 模型以及患者来源的异种移植物 (PDX) IVM 模型。在这个目标中，我们还将把我们选择的临床前候选药物转化为临床测试，如果成功的话靶向治疗将被选为 cGMP 生产（核心 B）的临床候选药物，用于后续在项目 3 的第一阶段安全试验中进行测试。用于缀合形成 ADC 的抗体将由 Core 提供 B. Core B还将提供ADC的质量控制分析和抗体结合亲和力的确认以下共轭。这些新疗法的肿瘤靶向、递送和积累将在使用 Core C 提供的多模态成像服务的啮齿动物，其中包括 X 射线/CT、SPECT-CT、IVM、和组织学。 Core E 将处理项目过程中生成的数据的生物统计分析。长期目标是将我们的关键基础发现转化为独特、创新的交付平台，以便提高人类癌症治疗的疗效。