Development of cell-permeable peptides and proteins

项目摘要 目前FDA批准的药物通常是小分子(MW&lt；500)或大蛋白质(MW&lt；500) &gt；5000)。小分子通常仅限于靶向含有以下物质的蛋白质(和其他生物分子) 深层结合口袋(例如，酶和GPCR)，占所有与疾病相关的人类的10% 蛋白质。另一方面，生物制品(例如，单抗)仅限于细胞外靶点， 占所有药物靶点的另外10%。剩下的~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)的蛋白质目前都不能被任何一种药物所抑制 接近。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……里面 此外，大约有7000种人类遗传病影响着大约10%的美国人口；只有非常 其中一小部分人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~80%的不可药物蛋白质并治疗人类遗传 疾病。实现这一目标需要将大的生物分子有效地输送到哺乳动物体内 手机。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 (CPPS)，它在体外将所有主要药物形式有效地输送到哺乳动物细胞的胞浆中，并 并阐明了它们的胞内摄取和胞体逃逸的机制。在接下来的五年里 几年后，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是如何 细菌毒素和一些人类蛋白如何通过质膜直接转运 内体进入胞浆，以及CPPs和一些蛋白质是如何离开哺乳动物细胞的 定义了“非常规蛋白质分泌”机制。第二，我们将使用机械知识 获得以开发性能改善的CPPs，例如，具有肿瘤组织特异性的CPPs，以及 设计一种哺乳动物的膜转位结构域(MTD)，用于细胞内蛋白质的输送。 最后，我们将利用环状cpp和mtds来开发细胞渗透肽和蛋白质，如 针对几个关键的“无法下药”的目标的化学探测器和潜在的治疗方法。

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.