Fragment-Based Methodology for the Identification of Phosphatase Inhibitors

描述（由申请人提供）：本提案旨在开发基于片段的方法来鉴定磷酸酶抑制剂。该方法包括三个步骤：(1)筛选具有不同低分子量o -芳基基团的o -芳基磷酸酯库，使用简单的分光光度法鉴定磷酸酶底物；(2)通过快速模拟合成和评价对鉴定出的o -芳基磷酸酯底物进行优化；(3)通过用已知的磷酸同分酯直接取代磷酸盐，将优化后的底物转化为抑制剂。为了开发该方法，将针对蛋白酪氨酸1B (PTP1B)，一种与糖尿病有关的磷酸酶。磷酸酶已被证明可以改善几种疾病的疾病状态，包括骨质疏松症、免疫疾病、癌症、阿尔茨海默病、糖尿病和肥胖症。特别是，PTP1B是治疗2型糖尿病的有效靶点。抑制PTP1B已被证明有助于治疗小鼠糖尿病而无副作用。虽然PTP1B是作为初始靶点进行研究，但该方法将适用于与各种疾病有关的大量磷酸酶。使用常用的组合方法寻找PTP1B抑制剂并非没有困难。特别麻烦的是在PTP1B筛查中获得的大量假阳性。提出的方法将直接解决与其他方法对磷酸酶抑制剂设计发现的陷阱。所提出的方法将使对磷酸酶家族抑制剂的产生成为可能。磷酸酶与许多疾病有关，包括骨质疏松症、免疫疾病、癌症、阿尔茨海默病、糖尿病和肥胖。最初的努力将集中于开发一种与2型糖尿病有关的磷酸酶抑制剂。从这种方法中产生的磷酸酶抑制剂可以为针对许多严重疾病的药物开发提供起点。

DESCRIPTION (provided by applicant): This proposal aims to develop fragment-based methodology toward the identification of phosphatase inhibitors. The method to be developed consists of three steps: (1) a library of O-aryl phosphates with diverse, low molecular weight O-aryl groups is screened to identify phosphatase substrates using a simple spectrophotometric-based assay, (2) the identified O-aryl phosphate substrates are optimized by rapid analogue synthesis and evaluation, and (3) the optimized substrates are converted to inhibitors by direct replacement of the phosphate with known phosphate isosteres. To develop the methodology, protein tyrosine 1B (PTP1B), a phosphatase implicated in diabetes, will be targeted. Phosphatases have been shown to enhance disease states in several maladies including osteoporosis, immune diseases, cancer, Alzheimer's disease, diabetes, and obesity. In particular, PTP1B is a well- validated target in the treatment of type 2 diabetes. Inhibition of PTP1B has been shown to aid in the treatment of diabetes in mice without side effects. While PTP1B is being pursued as an initial target, the methodology will be applicable to a large number of phosphatases involved in a variety of diseases. Finding inhibitors of PTP1B using commonly adopted combinatorial methods has not been without difficulty. Particularly troublesome is the high number of false positives obtained in screens against PTP1B. The methodology proposed will directly address the pitfalls found with alternative methodologies toward phosphatase inhibitor design. The proposed methodology will enable the generation of inhibitors toward families of phosphatase enzymes. Phosphatases have been implicated in a number of diseases including osteoporosis, immune diseases, cancer, Alzheimer's disease, diabetes, and obesity. Initial efforts will be focused on development of inhibitors toward a phosphatase involved in type 2 diabetes. Phosphatase inhibitors generated from this methodology could provide starting points for the development of Pharmaceuticals against a number of serious maladies.