Inhibition of Angiogenesis by TNP 470 and Ovalicin

TNP 470 和 Ovalicin 抑制血管生成

• 批准号: 7231691
• 负责人: Jun O. Liu
• 金额: $ 36.43万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-05 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7231691
• 关键词: Affect; Angiogenesis Inhibition; Angiogenesis Inhibitors; Antineoplastic Agents; Binding; Biochemical; Biological Factors; Biological Models; Biological Process; Biotin; Budgets; Calcium-Binding Proteins; Catalytic Domain; Cell Cycle; Cell Cycle Arrest; Cell Cycle Progression; Cell Cycle Regulation; Cell Proliferation; Collection; DNA Microarray Chip; DNA Microarray format; Development; Dissection; Endothelial Cells; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Eukaryota; Eukaryotic Cell; Excision; Family; G1 Phase; Genes; Genome; Human; In Vitro; Investigation; Knock-out; Knowledge; Lead; Link; Mammalian Cell; Mediating; Mediator of activation protein; Methionine; Mitosis; Molecular Probes; Molecular Profiling; N-terminal; Phase II Clinical Trials; Physiological; Play; Process; Prokaryotic Cells; Protein Biosynthesis; Protein Isoforms; Proteins; Role; Specificity; TNP470; TP53 gene; Testing; Tumor Suppressor Genes; Work; Yeasts; analog; angiogenesis; cancer cell; fumagillin; high throughput screening; inhibitor/antagonist; methionine aminopeptidase 2; methionyl aminopeptidase; mutant; neoplastic cell; novel; ovalicin; small molecule; small molecule libraries

DESCRIPTION (provided by applicant): The post-translational processing of initiator methionine during protein synthesis is a universal biological process that is conserved from prokaryotes to eukaryotes. Methionine aminopeptidases are the enzymes catalyzing the removal of initiator methionine. One gene is known in prokaryotes while two genes are known in eukaryotes encoding methionine aminopeptidases. The importance of initiator methionine processing is underscored by the fact that deletion of the methionine aminopeptidase genes in either prokaryotes or eukaryotes is lethal. Of the two methionine aminopeptidase genes in eukaryotes, the type 2 enzyme (MetAP2) has been shown to be the direct target for the fumagillin family of angiogenesis inhibitors, including its analog TNP-470. Work in the past several years has provided strong evidence that MetAP2 is a physiologically relevant target for TNP-470. It has also been found that inhibition of endothelial cell proliferation by TNP-470 is mediated by the tumor suppressor gene p53. Thus, TNP-470 is capable of activating p53, which induces the expression of p21that is responsible for the cell cycle blockade of endothelial cells. These studies reveal a unique role of MetAP2 in the progression of the endothelial cell cycle. Recently, a novel anticancer drug entering Phase II clinical trial known as bengamide was found to inhibit both MetAP2 and MetAP1 in vitro and to block cell cycle in both G1 and G2/M phase. It is hypothesized that MetAP1 may play a role in the cell cycle at the G2/M phase. The major objective of the current proposal is to further delineate the physiological functions of MetAP1 and MetAP2 employing yeast as a model system and to identify isoform-specific inhibitors for MetAP1 and MetAP2 by high throughput screens. The functions of the different domains in MetAP1 and MetAP2 will be investigated by creating various yeast mutants expressing different domains of these enzymes and determining the phenotypic changes using DNA microarray. The different known activators of p53 will be systematically examined to identify potential mediators of p53 activation by TNP-470. Molecular probes of bengamide will be prepared to confirm its interaction with MetAP1 and MetAP2. High throughput screens will be conducted to identify specific inhibitors for MetAP1 and MetAP2. The newly identified inhibitors for MetAP1 will be employed to assess the physiological role of MetAP1 in the cell cycle progression in the G2/M phase.

描述（由申请人提供）：蛋白质合成过程中引发剂甲硫氨酸的翻译后加工是从原核生物到真核生物都保守的通用生物过程。甲硫氨酸氨基肽酶是催化除去引发剂甲硫氨酸的酶。原核生物中已知一种基因，而真核生物中已知两种编码甲硫氨酸氨肽酶的基因。原核生物或真核生物中甲硫氨酸氨基肽酶基因的缺失都是致命的，这一事实强调了起始物甲硫氨酸加工的重要性。在真核生物的两种蛋氨酸氨肽酶基因中，2 型酶 (MetAP2) 已被证明是血管生成抑制剂夫马洁林家族（包括其类似物 TNP-470）的直接靶标。过去几年的工作提供了强有力的证据，证明 MetAP2 是 TNP-470 的生理相关靶标。还发现TNP-470对内皮细胞增殖的抑制是由抑癌基因p53介导的。因此，TNP-470 能够激活 p53，从而诱导 p21 的表达，p21 负责内皮细胞的细胞周期阻断。这些研究揭示了 MetAP2 在内皮细胞周期进展中的独特作用。最近，进入II期临床试验的新型抗癌药物bengamide在体外可同时抑制MetAP2和MetAP1，并阻断细胞周期在G1和G2/M期。据推测，MetAP1 可能在细胞周期的 G2/M 期发挥作用。当前提案的主要目标是利用酵母作为模型系统进一步描述 MetAP1 和 MetAP2 的生理功能，并通过高通量筛选鉴定 MetAP1 和 MetAP2 的异构体特异性抑制剂。将通过创建表达这些酶的不同结构域的各种酵母突变体并使用 DNA 微阵列确定表型变化来研究 MetAP1 和 MetAP2 中不同结构域的功能。将系统地检查不同的已知 p53 激活剂，以确定 TNP-470 激活 p53 的潜在介质。将制备苯甲酰胺的分子探针以确认其与MetAP1和MetAP2的相互作用。将进行高通量筛选以确定 MetAP1 和 MetAP2 的特定抑制剂。新鉴定的 MetAP1 抑制剂将用于评估 MetAP1 在 G2/M 期细胞周期进程中的生理作用。