INHIBITION OF THE ANTICANCER TARGET GLYOXALASE I

描述：在此续订申请中，我们建议继续开发 新型抗肿瘤药物的可逆性或可逆性 不可逆地抑制乙醛酶途径的酶。在上一次 资助期内，我们已经证明了烯二醇类似物抑制剂 (GSC(O)N(OH)R，其中GS=谷胱甘肽)， 乙草酸酶I在体外和体内抑制小鼠和人肿瘤的生长 活着。生长抑制似乎是由于细胞毒素的积聚。 乙二醛，乙醛酶途径的底物。我们还有 发现了一种能快速生成二元醇的高效“亚砜前药” 肿瘤细胞内的类似物。此外，我们在我们的 对其结构生物化学和作用机制的认识 乙草酸酶I和途径中的第二种酶，乙草酸酶II。我们现在 利用这些信息来开发更有效的抗肿瘤药物。具体目标 有：1.)合成了亚砜前体药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并引起 通过与细胞内的酰基交换反应实现的苯二酚类似物 谷胱甘肽。2.)的结构之间的相关性 亚砜及其向L1210白血病和白血病细胞输送苯二酚类似物的能力 B16黑色素瘤细胞，并在培养中抑制这些肿瘤的生长。3.) 评价最有希望的药物的药代动力学和抗肿瘤性能 亚砜前体药物对荷B16黑色素瘤小鼠的治疗。4.)要确定 GLXi活性位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.)合成亲和标记物 人GLXi，靶向活性部位的Glu172或Cys60。其中一些 亲和标记的设计是为了被GlxII水解酶活性 在正常细胞中含量很高，但在肿瘤细胞中含量很低。

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.