New Human Antibodies for CNS Drug Delivery

用于中枢神经系统药物输送的新型人类抗体

• 批准号: 10376351
• 负责人: ERIC V SHUSTA
• 金额: $ 38.21万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10376351
• 关键词: Acquired Immunodeficiency Syndrome; Alzheimer' s Disease; American; Antibodies; Antibody Specificity; Antibody-drug conjugates; Binding; Biodistribution; Biological Products; Blood; Blood - brain barrier anatomy; Blood Circulation; Brain; Brain Diseases; Brain-Derived Neurotrophic Factor; Cells; Cerebrum; Characteristics; Clinical; Coupling; Data; Development; Diffuse; Disease; Disease model; Dose; Drug Delivery Systems; Drug Efflux; Drug Kinetics; Drug Transport; Endothelium; Ensure; Exhibits; Fc Receptor; Gene Proteins; Generations; Genes; Hematological Disease; Human; Immunoprecipitation; In Vitro; Insulin; Insulin Receptor; Libraries; Link; Liposomes; Low Density Lipoprotein Receptor; Malignant neoplasm of brain; Mediating; Medicine; Mining; Modeling; Monitor; Mus; Neuropeptides; Neurotensin; Parkinson Disease; Patients; Persons; Phage Display; Pharmaceutical Preparations; Pharmacodynamics; Pharmacologic Substance; Pharmacology; Pharmacotherapy; Property; Proteins; Reagent; Research; Route; Side; Source; Specificity; Stroke; System; TFRC gene; Techniques; Technology; Testing; Therapeutic; Transferrin; Translating; Validation; antibody libraries; base; blood-brain barrier crossing; blood-brain barrier permeabilization; clinical translation; dalton; delivery vehicle; drug candidate; drug development; efflux pump; high-throughput drug screening; human tissue; in vivo; induced pluripotent stem cell; innovation; intravenous administration; intravenous injection; lipophilicity; nanoparticle; natural hypothermia; nervous system disorder; novel; novel therapeutics; pre-clinical; programs; receptor; response; screening; small molecule; small molecule therapeutics; stroke model; tandem mass spectrometry; targeted delivery; targeted treatment; therapeutic DNA; transcriptome; transcytosis; uptake

ABSTRACT Millions of people worldwide suffer from neurological diseases such as Alzheimer’s disease, stroke, and brain cancer. Advances in protein/gene profiling techniques and high throughput drug screening technologies have spawned many new drug candidates. However, the blood-brain barrier (BBB) has impeded the development and clinical realization of this new generation of neurotherapeutics by restricting the brain uptake of most small molecule therapeutics, and prohibiting brain uptake of protein- and gene-based medicines. A promising noninvasive brain delivery strategy takes advantage of endogenous BBB transport mechanisms as a means to shuttle drug cargo from the blood to the brain. Such receptor-mediated transport systems can be targeted using the exquisite specificity of antibodies that are in turn linked to a drug payload that can include small molecules, proteins, or DNA therapeutics. After binding to the receptor on the blood side, the antibody- drug conjugate acts as an artificial substrate for the transporter and is transcytosed from the blood, across the BBB, and into the brain. However, current approaches have yielded limited brain uptake because the targeted transporters are ubiquitously expressed and the antibody targeting reagents have low BBB permeability. Therefore, this proposal is focused on the identification and validation of a new panel of antibodies and cognate transporters that can mediate BBB crossing of therapeutics. The antibodies were identified by mining a large phage display antibody library against a BBB model comprising human induced pluripotent stem cell- derived brain microvascular endothelial-like cells (BMECs). The barrier characteristics of these BMECs are well- suited to screen large antibody libraries for those antibodies capable of crossing the BBB in vitro. Moreover, at the transcriptome level, the transporter profiles in these cells correlate quite well to freshly isolated human BMECs. Finally, the human sourcing ensures that identified antibodies recognize human BBB transporters. Using this innovative platform, we have identified a panel of high value antibodies that preliminary data indicate are capable of binding both the human and murine BBB and delivering pharmacologically relevant amounts of drug cargo to the murine brain. We propose to further validate these brain targeting antibodies by identifying their cognate transporters. Next, we will determine their pharmacokinetic properties, biodistribution and brain regiospecificity. Finally, the antibodies will be tested for their ability to mediate brain uptake of conjugated drug cargo to normal brain and to diseased brain in the form of a murine stroke model. Those antibodies that exhibit significant and selective brain uptake would represent new, noninvasive drug delivery vectors with potential application in many neurological disease settings.

摘要 全球数百万人患有神经系统疾病，如阿尔茨海默病，中风， 脑癌蛋白质/基因谱分析技术与高通量药物筛选技术研究进展 催生了许多新的候选药物然而，血脑屏障（BBB）阻碍了 通过限制大脑摄取， 大多数小分子治疗药物，并禁止大脑摄取蛋白质和基因为基础的药物。一 有前途的非侵入性脑递送策略利用内源性血脑屏障转运机制作为一种有效的治疗方法。 将毒品从血液运送到大脑这种受体介导的转运系统可以是 使用抗体的精确特异性进行靶向，这些抗体又与药物有效载荷连接， 小分子、蛋白质或DNA治疗剂。在与血液侧的受体结合后，抗体- 药物偶联物作为转运蛋白的人工底物，从血液中跨膜转运， BBB，进入大脑。然而，目前的方法产生了有限的脑摄取，因为靶向的 转运蛋白普遍表达，并且抗体靶向试剂具有低的BBB渗透性。 因此，该提案集中于一组新抗体的鉴定和验证， 可以介导治疗药物穿过BBB的相关转运蛋白。抗体是通过挖掘 针对包含人诱导多能干细胞的BBB模型的大型噬菌体展示抗体库- 衍生的脑微血管内皮样细胞（BMEC）。这些BMEC的屏障特性良好- 适用于筛选大的抗体库中能够在体外穿过血脑屏障的抗体。而且在 这些细胞中的转录组水平、转运蛋白谱与新鲜分离的人类 BMEC。最后，人源确保鉴定的抗体识别人BBB转运蛋白。 使用这个创新平台，我们已经确定了一组高价值的抗体，初步数据表明， 能够结合人和鼠BBB，并递送与人和鼠BBB相关量的 药物运输到小鼠大脑。我们建议进一步验证这些脑靶向抗体， 它们的同源转运蛋白接下来，我们将确定其药代动力学特性，生物分布和脑 区域特异性最后，检测抗体介导结合药物脑摄取的能力 以鼠中风模型的形式将货物运输至正常脑和患病脑。这些抗体表现出 显著的和选择性的脑摄取将代表新的、非侵入性的药物递送载体， 在许多神经系统疾病中的应用。