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: 10208481
• 负责人: ERIC V SHUSTA
• 金额: $ 38.18万
• 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/10208481
• 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; 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; 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; vector

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.

抽象的 全世界有数百万人患有神经系统疾病，例如阿尔茨海默氏病，中风和 脑癌。蛋白质/基因分析技术和高通量药物筛查技术的进步 已经产生了许多新毒品。但是，血脑屏障（BBB）阻碍了 通过限制大脑的吸收，这种新一代神经疗法的开发和临床实现 在大多数小分子疗法中，并且禁止脑摄取蛋白质和基因的药物。一个 有希望的无创脑输送策略利用内源性BBB传输机制作为一种 意味着从血液到大脑的毒品货物。这种受体介导的传输系统可以是 使用抗体的独家特异性针对的，而抗体又与药物有效载荷相关联 小分子，蛋白质或DNA疗法。与血侧的受体结合后，抗体 - 药物共轭物充当转运蛋白的人造底物，并从血液中转移，跨过 BBB，进入大脑。但是，目前的方法产生了有限的大脑吸收 转运蛋白无处不在，靶向试剂的抗体渗透性较低。 因此，该提案的重点是识别和验证新的抗体和 可以介导BBB治疗交叉的同源转运蛋白。通过开采确定抗体 大型噬菌体显示针对包括人类诱导多能干细胞的BBB模型的抗体文库 衍生的脑微血管内皮样细胞（BMEC）。这些BMEC的障碍特征是良好的 适用于能够在体外越过BBB的抗体筛选大型抗体库。而且，在 转录组水平，这些细胞中的转运蛋白谱与新鲜分离的人非常相关 Bmecs。最后，人类采购确保确定的抗体识别人类BBB转运蛋白。 使用此创新平台，我们已经确定了一组高价值抗体，这些抗体表明 能够结合人类和鼠BBB并提供与药物相关的数量 吸毒货物到鼠大脑。我们建议通过识别这些大脑靶向抗体进一步验证 他们的同源转运蛋白。接下来，我们将确定它们的药代动力学特性，生物分布和大脑 重新特定。最后，将测试抗体的介导大脑吸收共轭药物的能力 货物到正常的大脑，并以鼠卒中模型的形式出现疾病。那些展示的抗体 重要和选择性的大脑吸收将代表具有潜力的新的无创药物载体 在许多神经系统疾病环境中应用。