INHIBITION OF THE ANTICANCER TARGET GLYOXALASE I

描述：在此更新应用程序中，我们建议继续开发 通过可逆或可逆作用发挥作用的新型抗肿瘤药物 不可逆地抑制乙二醛酶途径的酶。在之前的 资助期间，我们已经证明烯二醇类似物抑制剂 途径中第一个酶的（GSC(O)N(OH)R，其中 GS = 谷胱甘肽）， 乙二醛酶 I，在体外和体内抑制小鼠和人类肿瘤的生长 体内。生长抑制似乎是由于细胞毒性物质的积累 甲基乙二醛，乙二醛酶途径的底物。我们还有 发现了一种有效的“亚砜前药”，可快速生成烯二醇 肿瘤细胞内的类似物。此外，我们在以下方面也取得了重大进展： 了解结构生物化学和作用机制 乙二醛酶 I 和途径中的第二种酶，乙二醛酶 II。我们现在将 利用这些信息来开发更有效的抗肿瘤药物。具体目标 1.) 合成亚砜前药 CH3(CH)nS(O)C(O)N(OH)C6H4Cl (n = 1,5,8,10)，旨在快速扩散到肿瘤细胞中并产生 烯二醇类似物通过与细胞内的酰基交换反应 谷胱甘肽。 2.) 测试结构之间的相关性 亚砜及其将烯二醇类似物递送至 L1210 白血病的能力 B16 黑色素瘤细胞，并在培养物中抑制这些肿瘤的生长。 3.) 评估最有前途的药物的药代动力学和抗肿瘤特性 亚砜前药治疗患有 B16 黑色素瘤的小鼠。 4.) 确定 GlxI 活性位点定向不可逆失活的分子基础 S-(4-溴-2,3-二氧丁基)谷胱甘肽。 5.) 合成亲和标签 人类 GlxI，其目标是活性位点中的 Glu172 或 Cys60。一些 亲和标记被设计为可被 GlxII 水解，GlxII 是一种酶活性， 正常细胞中较高，但肿瘤细胞中较低。

DESCRIPTION: In this renewal application we propose to continue the development of novel new classes of antitumor agents that function by either reversibly or irreversibly inhibiting the enzymes of the glyoxalase pathway. In the previous funding period, we have demonstrated that enediol analog inhibitors (GSC(O)N(OH)R, where GS = glutathionyl) of the first enzyme in the pathway, glyoxalase I, inhibit the growth of murine and human tumors in vitro and in vivo. Growth inhibition appears to be due to the buildup of cytotoxic methylglyoxal, the substrate for the glyoxalase pathway. We have also discovered an efficient 'sulfoxide prodrug' for rapidly generating the enediol analog inside tumor cells. In addition, we have made major advances in our understanding of the structural biochemistry and mechanisms-of-action of glyoxalase I and the second enzyme in the pathway, glyoxalase II. We will now use this information to develop more potent antitumor agents. The specific aims are: 1.) To synthesize the sulfoxide prodrugs CH3(CH)nS(O)C(O)N(OH)C6H4Cl (n = 1,5,8,10), designed to rapidly diffuse into tumor cells and give rise to the enediol analogs via an acyl-interchange reaction with intracellular glutathione. 2.) To test for correlations between the structures of the sulfoxides and their ability to deliver enediol analog into L1210 leukemia and B16 melanoma cells, and to inhibit the growth of these tumors, in culture. 3.) To evaluate the pharmacokinetics and antitumor properties of the most promising sulfoxide prodrugs in mice bearing B16 melanotic melanoma. 4.) To determine the molecular basis of active-site directed irreversible inactivation of GlxI by S-(4-bromo-2,3-dioxobutyl)glutathione. 5.) To synthesize affinity labels of human GlxI, which target either Glu172 or Cys60 in the active site. Some of the affinity labels are designed to be hydrolyzed by GlxII, an enzyme activity that is high in normal cells but low in tumor cells.