Chemical approaches for generating blood-brain barrier-permeable antibody conjugates

生成血脑屏障可渗透抗体缀合物的化学方法

• 批准号: 10028297
• 负责人: Kyoji Tsuchikama
• 金额: $ 39万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-05 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10028297
• 关键词: Antibodies; Antibody-drug conjugates; Area; Attenuated; Biodistribution; Blood - brain barrier anatomy; Blood Circulation; Brain; Brain Diseases; Brain Neoplasms; Cell Line; Cells; Central Nervous System Diseases; Chemicals; Clinical; Clinical Research; Disease; Drug Delivery Systems; Drug Kinetics; Drug usage; Extravasation; Foundations; Glioblastoma; In Vitro; Kinetics; Lead; Malignant neoplasm of brain; Measurable; Mediating; Modality; Molecular; Monoclonal Antibodies; Mus; Newly Diagnosed; Patients; Peptides; Pharmaceutical Preparations; Plasma; Property; Safety; Site; Specificity; Structure; Systemic Therapy; Technology; Therapeutic; Therapeutic Monoclonal Antibodies; Treatment Efficacy; Variant; Xenograft procedure; antibody conjugate; base; blood-brain barrier penetration; blood-brain barrier permeabilization; brain parenchyma; chemotherapeutic agent; design; drug development; effective therapy; epidermal growth factor receptor VIII; immunogenicity; improved; in vivo; intravital fluorescence microscopy; mouse model; novel; novel therapeutics; peptide structure; phase 3 study; receptor; stoichiometry; transcytosis; tumor

1. ABSTRACT The blood-brain barrier (BBB) restricts the influx of biomolecules from the vasculature to the brain parenchyma. This attenuates exposure levels of the brain to systemically administered drugs, especially large- size molecules such as antibodies. This issue also makes systemic treatment of glioblastoma (GBM), the most devastating brain cancer, ineffective in most cases. Recent clinical studies have demonstrated that a measurable number of GBM cells, in particular cells near the growing edge of the infiltrative tumor area, exist behind an intact BBB. Collectively, the features of the BBB create a special challenge for effective treatment of central nervous system (CNS) diseases, including brain cancer, using drugs that have proven efficacy in other diseases. Antibody-drug conjugates (ADCs) are an emerging drug class with prominent target specificity, durable therapeutic efficacy, and high translatability in drug development. While promising, clinical benefits of ADCs in the treatment of brain diseases, in particular GBM, remain unconfirmed. Unfortunately, recent interim analysis in a Phase 3 study using the anti-EGFRvIII ADC Depatux-M (formerly called ABT-414) revealed no survival benefit for patients with newly diagnosed GBM receiving this ADC. Thus, improvement in BBB penetrability for ADCs is critically needed to advance this promising molecular format toward truly effective and safe systemic therapy for CNS diseases. We have developed novel ADC linker technologies, including: 1) branched linkers for site-specific and simultaneous installation of two distinct molecules onto a single antibody and 2) enzymatically cleavable linkers with exceptional circulation stability. Using these technologies, we have successfully constructed homogeneous conjugates appended with peptides that facilitate traversing the BBB through receptor-mediated transcytosis. One of the homogeneous peptide conjugates, as compared to a conventional heterogeneous variant, showed greater accumulation into the brain parenchyma in healthy mice (2.7-fold) and orthotopic GBM tumors in a xenograft mouse model (3.6-fold). Based on these findings, we hypothesize that homogeneous conjugation of properly designed BBB-penetrating peptides with ADCs will be a promising approach for systemic drug delivery to the brain. In this project, we will prepare a variety of BBB-penetrating peptides and construct antibody conjugates with various conjugation modalities (linker attachment site, linker structure, and stoichiometry of the peptides and payloads). All conjugates will be evaluated in vitro and in vivo for plasma stability, receptor-mediated transcytosis efficiency, pharmacokinetics, biodistribution, tolerability, and immunogenicity profiles. We will then evaluate a panel of BBB-permeable ADCs for tumor targeting efficiency as well as therapeutic efficacy in cell line-based and patient-derived xenograft mouse models of orthotopic GBM. We will also perform intravital fluorescence microscopy to evaluate kinetics and dynamics of extravasation in both healthy and tumor-bearing mouse models. Successful completion of this project will clarify the effect of the peptide structure and conjugation modality on BBB penetrability of antibody conjugates as well as other drug properties. We also expect to identify rational molecular design to unleash the full therapeutic potential of monoclonal antibodies and ADCs for brain targeting, which may ultimately lead to novel drug development strategies toward a cure for difficult-to-treat CNS diseases, such as GBM.

1。摘要 血脑屏障（BBB）限制了从脉管系统到大脑的生物分子的涌入 实质。这减轻了大脑暴露于系统给药的药物，尤其是大型药物 大小分子，例如抗体。这个问题还使系统治疗胶质母细胞瘤（GBM），最多。 毁灭性的脑癌，在大多数情况下无效。最近的临床研究表明 可测量数量的GBM细胞，特别是浸润性肿瘤区域增长边缘附近的细胞，存在 完整的BBB后面。总体而言，BBB的特征对有效治疗的特殊挑战构成了特殊挑战 中枢神经系统（CNS）疾病，包括脑癌，使用在其他方面有效的药物 疾病。 抗体 - 药物缀合物（ADC）是具有突出目标特异性的新兴药物类别 治疗功效和药物开发中的高转化性。虽然有希望，但ADC在 脑部疾病的治疗，尤其是GBM，仍未得到证实。不幸的是，最近的临时分析 在使用抗EGFRVIII ADC DEPATUX-M（以前称为ABT-414）的3阶段研究中，没有生存 接受此ADC的新诊断为GBM的患者的好处。因此，提高了BBB的渗透性 迫切需要ADC以将这种有希望的分子格式推向真正有效且安全的系统性 中枢神经系统疾病的治疗。 我们已经开发了新颖的ADC连接器技术，包括：1）特定于网站的分支连接器 同时在单个抗体上同时安装两个不同的分子和2）酶上的连接器 具有出色的循环稳定性。使用这些技术，我们已经成功构建了 附加肽的均匀缀合物，可促进通过受体介导的BBB穿越BBB 转胞病。与传统的异质性相比 变体在健康小鼠（2.7倍）和原位GBM中显示出更大的积累到脑实质中 异种移植小鼠模型中的肿瘤（3.6倍）。基于这些发现，我们假设这种同质 合理设计的BBB渗透肽与ADC的结合将是一种有前途的方法 全身药物输送到大脑。在这个项目中，我们将准备各种BBB穿透肽和 构建与各种共轭方式（接头附件，接头结构和 肽和有效载荷的化学计量法）。所有结合物将在体外和体内评估血浆 稳定性，受体介导的转胞病效率，药代动力学，生物分布，耐受性和 免疫原性谱。然后，我们将评估一组BBB可渗透的ADC，以靶向肿瘤 以及基于细胞系和患者衍生的异种移植物模型的治疗功效 GBM。我们还将执行插入式荧光显微镜，以评估动力学和动力学 在健康和承担肿瘤的小鼠模型中渗出。 该项目的成功完成将阐明肽结构和结合方式的影响 关于抗体结合物以及其他药物特性的BBB渗透性。我们还希望确定理性 分子设计，以释放单克隆抗体和ADC的全部治疗潜力 靶向，这最终可能导致新型的药物开发策略，以治愈难以治疗 中枢神经系统疾病，例如GBM。