Development of cell-permeable peptides and proteins

细胞渗透性肽和蛋白质的开发

• 批准号: 10609048
• 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/10609048
• 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; Engineering; Enzymes; FDA approved; G-Protein-Coupled Receptors; Genetic Diseases; Goals; Human; Human Genetics; In Vitro; Investigation; Knowledge; Mammalian Cell; Membrane; Modality; Monoclonal Antibodies; Penetration; Peptides; Periodicity; Permeability; Pharmaceutical Preparations; Pharmacological Treatment; Population; Property; Protein Secretion; Proteins; Research; Therapeutic; Tumor Tissue; cellular targeting; 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.

项目摘要 目前FDA批准的药物通常是小分子（MW <500）或大蛋白（MW >5000）。小分子通常限于靶向蛋白质（和其他生物分子）， 深装订袋（例如，酶和GPCR），占所有人类疾病相关疾病的约10% proteins.另一方面，生物制剂（例如，单克隆抗体）限于细胞外靶， 占所有药物靶点的约10%。其余约80%的药物靶点，主要是 参与细胞内蛋白质-蛋白质相互作用（PPI）的蛋白质，目前通过以下任一种药物都不可用： approach.同样的限制也适用于使用小分子和蛋白质作为研究工具。在 此外，约有7000种人类遗传疾病影响约10%的美国人口;只有一个非常 其中一小部分目前正在接受药物治疗。我研究的总体目标是 开发一种通用的方法来靶向约80%的不可药物化的蛋白质，并治疗人类遗传性疾病。 疾病实现这一目标需要将大生物分子有效地递送到哺乳动物体内。 cell.在过去的十年里，我的团队发现了一类新的环状细胞穿透肽 （CPP），其在体外有效地将所有主要药物形式递送到哺乳动物细胞的胞质溶胶中， 并阐明了它们的内吞摄取和内体逃逸机制。未来五 今年，我们将继续进行三个方面的调查。首先，我们将研究线性和循环CPP 直接穿过质膜，细菌毒素和一些人类蛋白质如何逃脱 内体进入胞质溶胶，以及CPPs和一些蛋白质如何通过一个很差的 定义了“非常规蛋白质分泌”机制。其次，我们将利用机械性知识 获得开发具有改进性能的CPP，例如，对肿瘤组织具有特异性的CPP，以及 工程化哺乳动物膜易位结构域（MTD）用于蛋白质的细胞内递送。 最后，我们将利用环状CPP和MTD开发细胞渗透性肽和蛋白质， 化学探针和潜在的治疗剂针对几个关键的“不可药”的目标。