Anti-Cancer Activities of PTPase Inhibitors

PTPase 抑制剂的抗癌活​​性

• 批准号: 6745187
• 负责人: Taolin Yi
• 金额: $ 33.95万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-06 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6745187
• 关键词: 3T3 cells; SDS polyacrylamide gel electrophoresis; active sites; antimony; antineoplastics; antiprotozoal agents; athymic mouse; biological products; biological signal transduction; biotherapeutic agent; enzyme activity; gluconate; interferon alpha; mass spectrometry; pharmacokinetics; phosphatase inhibitor; polymerase chain reaction; protein tyrosine phosphatase; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): Protein tyrosine phosphatases (PTPases) are key signaling molecules and potential targets for developing inhibitors as novel therapeutics. Our preliminary studies demonstrate for the first time that sodium stibogluconate (SS) and glucantime (GT), antimony (Sb) based anti-leishmania drugs of unknown mechanism, are inhibitors of selective PTPases with activity against cancer cells in vitro and in mouse models at nontoxic doses and in synergy with IFNs. We propose the following specific aims: Aim 1. To test the hypothesis that SS anti-cancer activity functions via inactivating PRLs and SHPs, we will: a), assess the role of PRLs in SS activity against cancer cell lines using SS-insensitive PRL mutants; b), assess the role of SHPs in SS synergy with IFNa in cell growth inhibition using SHP-deficient cell lines and SHP transfectants; c), define the pharmacodynamics of SS in inhibiting PRLs/SHPs in mouse models; d), determine the effects of SS on PRLs-regulated signaling molecules in cancer cells; e), determine the effects of SS on IFNa signaling molecules in WM9 melanoma cells against which SS and IFNa showed a synergistic effect. Aim 2. To test the hypothesis that SS inactivates target PTPases via covalent modification of a catalytic cysteine residue in the enzymes through the Sb in the drugs while the gluconic acid conjugated to Sb facilitates Sb/cysteine interactions and defines the drugs' PTPase specificity, we will: a), define the nature and significance of SS-induced mass increase in PRL-2 by mass spectrometry and mutational analysis; b), assess the ability of SS to modify MKP1 and PRL-1R86 mutant insensitive to SS inhibition; c), determine the effects of Sb and gluconic acid on PTPases sensitive or insensitive to SS inhibition; d), determine the PTPase inhibitory activity and anti-cancer activity of different molecular weight fractions of SS, which is mixture of polymers of Sb/gluconic acid complexes of 100-4000 Da. Aim 3. To test the hypothesis that GT has anti-cancer activity via targeting PTPases, we will: a), determine its inhibitory activity against a panel of PTPases; b), define its inhibitory mechanism by mass spectrometry and mutational analysis; c), assess its anti-cancer activity against human cancer cell lines in vitro and in mouse models; d), assess the role of its target PTPases in mediating its anti-cancer activity.

描述（由申请人提供）：蛋白质酪氨酸磷酸酶（PTPases）是关键信号分子，也是开发抑制剂作为新疗法的潜在靶标。我们的初步研究首次证明了stibogluconate钠（SS）和葡萄糖酸钠（GT），基于二氧化抗合性（SB）的抗leishmania药物，具有未知机制的抗 - 利什曼原虫药物，是对与iFN和IFNS中氧毒性剂量的小鼠模型中对癌细胞活性的选择性PTPase的抑制剂。我们提出以下具体目的：目标1。测试以下假设：SS抗癌活性通过失活的PRL和SHP函数，我们将：a），a），评估PRL使用SS不敏感的PRL突变体针对癌细胞系的SS活性的作用； b），使用SHP缺陷型细胞系和SHP转染剂评估SHP在IFNA与IFNA在细胞生长抑制中的作用； c），在抑制小鼠模型中抑制PRLS/SHP时定义SS的药效学； d），确定SS对癌细胞中PRLS调节的信号分子的影响； e），确定SS对WM9黑色素瘤细胞中IFNA信号分子的影响，SS和IFNA表现出协同作用。 Aim 2. To test the hypothesis that SS inactivates target PTPases via covalent modification of a catalytic cysteine residue in the enzymes through the Sb in the drugs while the gluconic acid conjugated to Sb facilitates Sb/cysteine interactions and defines the drugs' PTPase specificity, we will: a), define the nature and significance of SS-induced mass increase in PRL-2通过质谱和突变分析； b），评估SS修改MKP1和PRL-1R86突变体对SS抑制不敏感的能力； c），确定SB和葡萄糖酸对PTPases对SS抑制敏感或不敏感的影响； d），确定SS不同分子量分数的PTPase抑制活性和抗癌活性，该分子量是100-4000 DA的SB/葡萄糖酸配合物的聚合物的混合物。目的3。为了测试GT通过靶向PTPases具有抗癌活性的假设，我们将：a），确定其对PTPase小组的抑制活性； b），通过质谱和突变分析来定义其抑制机制； c），在体外和小鼠模型中评估其针对人类癌细胞系的抗癌活性； d），评估其靶PTPases在介导其抗癌活性中的作用。