High-throughput screen to identify modulators of CendR-mediated cellular uptake

高通量筛选以鉴定 CendR 介导的细胞摄取调节剂

• 批准号: 8262597
• 负责人: ERKKI RUOSLAHTI
• 金额: $ 4.88万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8262597
• 关键词: Affect; Agonist; Amino Acids; Antineoplastic Agents; Arginine; Binding; Binding Sites; Biochemical; Biological; Biological Assay; C-terminal; Cell Line; Cell Surface Receptors; Cell surface; Cells; Chemicals; Chemotherapy-Oncologic Procedure; Collaborations; Coupled; Development; Dose; Elements; Fluorescence Polarization; Genomics; Homing; In Vitro; Infectious Agent; Inflammatory; Lead; Libraries; Lysine; Mediating; Molecular; Molecular Weight; Morphologic artifacts; Names; Neuropilin-1; Neuropilins; Pathway interactions; Penetration; Peptides; Positioning Attribute; Proteins; Proteolytic Processing; System; Tissue Sample; Tissues; Toxin; Tumor Tissue; Validation; Venoms; Work; base; carboxyl group; high throughput screening; innovation; nanoparticle; novel; novel diagnostics; novel therapeutics; small molecule; small molecule libraries; tool; tumor; uptake

DESCRIPTION (provided by applicant): This application proposes high-throughput screening for small molecular weight compounds that mimic the internalizing and tissue-penetrating peptide motif R/KXXR/K (R=arginine; K=lysine; X=any amino acid). Peptides and proteins containing this motif activate a pinocytotic transport pathway into cells and through tissues by binding to a cell surface receptor, neuropilin-1. The R/KXXR/K motif has to be in a C-terminal position to bind to neuropilin; even blocking the C-terminal carboxyl group of the arginine residue eliminates the activity. Because of this feature, the motif has been designated the C-end Rule or CendR motif. Tumor- specific homing peptides that are internalized into cells and spread within tumor tissue contain both a specific tumor-homing sequence and a cryptic CendR motif, which is activated in tumors by proteolytic processing. A payload can either be covalently coupled to a CendR peptide or simply co-administered with it to achieve tissue penetration. Thus, the CendR pathway provides a way of overcoming one of the main limitations of cancer chemotherapy, poor penetration of anticancer drugs into tumor tissue. This pathway can also transport nanoparticles, enabling deep tissue delivery of nanoparticle payloads. Many inflammatory proteins, infectious agents and venoms possess proteins with CendR elements that may mediate entry into cells and spreading through tissues. Novel high-throughput assays that are based on the binding of a CendR peptide to a fragment of neuropilin-1 have been developed in collaboration with the Sanford-Burnham Center for Chemical Genomics, and a pilot screen using the assay has been successfully carried out. Larger screens are now proposed to assemble a panel of "hits". These molecules will be validated as specific modulators of the CendR pathway and their biological activity as agonists or antagonists of the CendR pathway will be determined assessed. The compounds resulting from this work that resemble the CendR peptides in their activity will be useful tools for introducing materials into cells and tissues, as well as for exploring the molecular basis of the CendR tissue penetration phenomenon. They may also serve as lead compounds in the development of novel diagnostic and therapeutic delivery systems. The screens may also yield antagonists of the CendR system, which could be useful in inhibiting the transport of toxins and infectious agents through the CendR transport system.

描述（由申请人提供）：本申请提出高通量筛选小分子量化合物，模拟内化和组织穿透肽基序R/KXXR/K （R=精氨酸；K=赖氨酸；X=任何氨基酸）。含有该基序的肽和蛋白质通过与细胞表面受体neuropilin-1结合，激活进入细胞并通过组织的胞质转运途径。R/KXXR/K基序必须位于c端才能与neuropilin结合；即使阻断精氨酸残基的c端羧基也能消除活性。由于这一特点，该基序被称为c端规则或CendR基序。肿瘤特异性归巢肽被内化到细胞中并在肿瘤组织中传播，它包含一个特定的肿瘤归巢序列和一个隐式的CendR基序，它在肿瘤中通过蛋白水解过程被激活。有效载荷既可以与CendR肽共价偶联，也可以简单地与它共同施用以实现组织渗透。因此，CendR途径提供了一种克服癌症化疗的主要限制之一的方法，即抗癌药物难以渗透到肿瘤组织中。这种途径也可以运输纳米颗粒，使纳米颗粒有效载荷能够在组织深处传递。许多炎症蛋白、感染因子和毒液都含有含有CendR元素的蛋白，这些蛋白可能介导进入细胞并在组织中扩散。与斯坦福-伯纳姆化学基因组学中心合作开发了基于CendR肽与neuropilin-1片段结合的新型高通量检测方法，并成功地进行了使用该检测方法的试点筛选。现在，更大的屏幕被提议组装一个“热门”面板。这些分子将被验证为CendR通路的特异性调节剂，它们作为CendR通路的激动剂或拮抗剂的生物活性将被确定评估。从这项工作中得到的与CendR肽活性相似的化合物将成为将材料引入细胞和组织以及探索CendR组织渗透现象的分子基础的有用工具。它们也可以作为先导化合物用于开发新的诊断和治疗递送系统。筛选也可能产生CendR系统的拮抗剂，这可能有助于抑制毒素和感染因子通过CendR运输系统的运输。