Development of cell-permeable peptides and proteins

细胞渗透性肽和蛋白质的开发

• 批准号: 10405784
• 负责人: Dehua Pei
• 金额: $ 51.38万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10405784
• 关键词: Affect; Area; Bacterial Toxins; Binding; Biological Products; Cell membrane; Cells; Chemicals; Cytosol; DNA Sequence Alteration; Development; Disease; Drug Delivery Systems; Drug Targeting; Endosomes; Enzymes; FDA approved; G-Protein-Coupled Receptors; Genetic Diseases; Goals; Human; Human Genetics; In Vitro; Investigation; Knowledge; Mammalian Cell; Membrane; Modality; Monoclonal Antibodies; Peptides; Periodicity; Permeability; Pharmaceutical Preparations; Pharmacological Treatment; Population; Property; Protein Secretion; Proteins; Research; Therapeutic; Tissue Engineering; extracellular; human disease; improved; in vivo; novel; novel drug class; protein protein interaction; small molecule; tool; tumor specificity; uptake

Project Summary Current FDA-approved drugs are usually either small molecules (MW <500) or large proteins (MW >5000). Small molecules are generally limited to targeting proteins (and other biomolecules) that contain deep binding pockets (e.g., enzymes and GPCRs), which represent ~10% of all disease relevant human proteins. On the other hand, biologics (e.g., monoclonal antibodies) are restricted to extracellular targets, which represent another ~10% of all drug targets. The remaining ~80% drug targets, which are primarily proteins involved in intracellular protein-protein interactions (PPIs), are currently undruggable by either approach. The same limitations apply to the use of small molecules and proteins as research tools. In addition, there are ~7000 human genetic diseases affecting ~10% of the US population; only a very small fraction of them currently has pharmacologic treatment. The overall goal of my research is to develop a general approach to targeting the ~80% undruggable proteins and treating human genetic diseases. Accomplishing this goal requires effective delivery of large biomolecules into the mammalian cell. Over the past decade, my group has discovered a novel class of cyclic cell-penetrating peptides (CPPs), which efficiently deliver all major drug modalities into the cytosol of mammalian cells in vitro and in vivo, and elucidated their mechanism of endocytic uptake and endosomal escape. During the next five years, we will continue three areas of investigation. First, we will investigate how linear and cyclic CPPs directly translocate across the plasma membrane, how bacterial toxins and some human proteins escape the endosome into the cytosol, and how CPPs and some proteins exit the mammalian cell by a yet poorly defined “unconventional protein secretion” mechanism. Second, we will use the mechanistic knowledge gained to develop CPPs of improved properties, e.g., CPPs with specificity for tumor tissues, and engineer a mammalian membrane translocation domain (MTD) for intracellular delivery of proteins. Finally, we will leverage the cyclic CPPs and MTDs to develop cell-permeable peptides and proteins as chemical probes and potential therapeutics against several key “undruggable” targets.

项目概要 目前 FDA 批准的药物通常是小分子（MW <500）或大蛋白质（MW >5000）小分子通常仅限于含有的靶向蛋白质（和其他生物分子）。 深结合袋（例如酶和 GPCR），约占人类所有疾病相关的 10% 另一方面，生物制剂（例如单克隆抗体）仅限于细胞外靶标， 占所有药物靶点的另外约 10% 其余约 80% 的药物靶点主要是药物靶点。 参与细胞内蛋白质-蛋白质相互作用（PPI）的蛋白质目前无法通过以下任一方法进行成药： 同样的限制也适用于使用小分子和蛋白质作为研究工具。 此外，约有 7000 种人类遗传疾病影响着约 10% 的美国人口； 他们中的一小部分目前正在接受药物治疗。我研究的总体目标是 开发一种通用方法来靶向约 80% 不可成药的蛋白质并治疗人类遗传 实现这一目标需要将大生物分子有效地输送到哺乳动物体内。 在过去的十年中，我的团队发现了一类新型的环状细胞穿透肽。 （CPP），可在体外有效地将所有主要药物方式递送到哺乳动物细胞的胞质溶胶中， 在接下来的五年内，阐明了它们的内吞摄取和内体逃逸机制。 年内，我们将继续进行三个领域的研究，首先，我们将研究线性和循环 CPP 的性能。 直接穿过质膜，细菌毒素和一些人类蛋白质如何逃逸 内体进入细胞质，以及 CPP 和一些蛋白质如何以较弱的速度离开哺乳动物细胞 定义“非常规蛋白质分泌”机制 其次，我们将利用机械知识。 获得开发具有改进特性的CPP，例如对肿瘤组织具有特异性的CPP，以及 设计哺乳动物膜易位结构域（MTD）用于细胞内蛋白质递送。 最后，我们将利用环状 CPP 和 MTD 来开发细胞可渗透的肽和蛋白质，例如 针对几个关键“不可成药”目标的化学探针和潜在疗法。