Evaluation of antiangiogenic role of EM011, a novel tubulin-binding agent

新型微管蛋白结合剂 EM011 的抗血管生成作用评估

• 批准号: 7531874
• 负责人: Ritu Aneja
• 金额: $ 13.11万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-12 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7531874
• 关键词: Affect; Angiogenesis Inhibitors; Antineoplastic Agents; Apoptosis; Binding; Biological Assay; Breast; Cell Cycle Progression; Cell Proliferation; Cytoskeleton; Data; Disease remission; Dose; Endothelial Cells; Evaluation; Goals; Human; Image; Immunologics; Invasive; Investigation; Link; Malignant neoplasm of prostate; Maximum Tolerated Dose; Measures; Membrane; Mentors; Microtubules; Morphogenesis; Outcome; Pathway interactions; Pharmaceutical Preparations; Phase; Preclinical Drug Evaluation; Prostate; Protein Dephosphorylation; Quality of life; Range; Role; Thalidomide; Therapeutic; Toxic effect; Transactivation; Transcriptional Activation; Tubular formation; Tubulin; Tubulin Binding Agent; Tyrosine Kinase Inhibitor; Vascular Endothelial Growth Factors; ZD-6474; angiogenesis; base; cancer cell; in vivo; inhibitor/antagonist; migration; neurotoxicity; novel; polymerization; prevent; time use; translational approach; tumor; tumor growth

DESCRIPTION (provided by applicant): Angiogenesis, necessary for tumor growth involves cell proliferation and directed migration. Thus, there is clearly a crucial role of cytoskeletal microtubule (MT) dynamics in angiogenesis; linking perturbations of MT dynamics to inhibition of tumor angiogeneisis. Furthermore, the PI3K/Akt survival pathway regulates HIF-la expression and activity. Our preliminary data strongly suggest that a semisynthetic tubulin-binding anticancer~ agent,(S)-3-((R)-9-bromo-4-methoxy-6-methyl-5,6,7,8-tetrahydro-[1,3]-dioxolo[4,5-g]-isoquinolin-5-yl)-6,7-dimethoxyisobenzofuran-1(3H)-one (EM011), has potent antiangiogenic activity (based upon NCI-DTP antiangiogenic drug screen). We also demonstrated that EM011 inhibits the P13K/Akt pathway through dephosphorylation of Akt. Our hypothesis is that EM011 will serve as an effective anticancer agent since it can target the MT cytoskeleton without causing any gross effects (over- or de- polymerization of MTs) with concomitant antiangiogenic effects. The specific aims for the mentored phase are: Aim 1. Evaluation of the antiangiogenic efficacy of EM011. This will be accomplished by a) measuring its ability to prevent invasion and migration of endothelial cells, b) determining its effect on protrusion and proliferation of microvessels by tubular morphogenesis assays, ex vivo explant cultures, and the chick chorloallantoic membrane (CAM) assay, c) evaluating EM011's ability to inhibit cellular proliferation, affect cell-cycle progression, and to induce apoptosis in endothelial cells. Aim 2. Determination of EM011's effect on the dynamic instability of MTs, HIF-1a expression and transactivation of downstream targets such as VEGF. We will also determine if EM011-medlated inhibition of Pl3KIAkt regulates HIF-1a expression and its transcriptional activation capability. Equally important, our preliminary data show that EM011 does not cause any hematologic, immunologic and neuronal toxicity. Based upon Its non-toxic attributes, we rationalize that combination of EM011 with other angiogenic inhibitors that function through independent mechanisms but show toxicity at their maximum tolerated doses (MTD5), presents a unique opportunity to reduce their doses to maximize therapeutic outcomes with decreased toxicity. The independent phase of the project will thus focus on the following two aims: Aim 3. Investigation of potential synergistic antiproliferative and antiangiogenic effects of combinations of EM011 and ZD6474 (a tyrosine kinase inhibitor), or thalidomide (endothelial cell proliferation inhibitor) in breast (MDA-MB-231) and prostate (PC-3) cancer cells. Aim 4. Determination of the in vivo efficacy of EM011 and its synergistic combinations with ZD6474 or thalidomide, as inhibitors of experimental primary and metastatic breast and prostate cancers in real-time using non-invasive bioluminescent imaging, As a practical and translational approach, the long-range goal of these studies would be to define and establish the usefulness of EM011 alone, and its synergistic combinations with other drugs, for remission of human breast and prostate cancers without compromising the quality of life.

描述（由申请人提供）：血管生成是肿瘤生长所必需的，涉及细胞增殖和定向迁移。因此，细胞骨架微管（MT）动力学在血管生成中显然起着至关重要的作用；将MT动力学的扰动与肿瘤血管生成的抑制联系起来。此外，PI3K/Akt存活通路调节HIF-la的表达和活性。我们的初步数据强烈表明，半合成的微管蛋白结合抗癌剂(S)-3-（(R)-9-溴-4-甲氧基-6-甲基-5,6,7,8-四氢-[1,3]-二氧唑[4,5-g]-异喹啉-5-基）-6,7-二甲氧基异苯并呋喃-1(3H)- 1（EM011）具有有效的抗血管生成活性（基于NCI-DTP抗血管生成药物筛选）。我们还证明了EM011通过Akt的去磷酸化抑制P13K/Akt通路。我们的假设是EM011将作为一种有效的抗癌剂，因为它可以靶向MT细胞骨架，而不会引起任何伴随抗血管生成作用的总体效应（MT的过度聚合或解聚）。指导阶段的具体目标是：目标1。EM011抗血管生成疗效评价。这将通过a)测量其阻止内皮细胞入侵和迁移的能力，b)通过管状形态发生实验、体外外植体培养和鸡肾尿囊膜（CAM）实验确定其对微血管突出和增殖的影响，c)评估EM011抑制细胞增殖、影响细胞周期进程和诱导内皮细胞凋亡的能力来实现。目标2。测定EM011对MTs动态不稳定性、HIF-1a表达及下游靶点如VEGF的转激活的影响。我们还将确定em011介导的Pl3KIAkt抑制是否调节HIF-1a表达及其转录激活能力。同样重要的是，我们的初步数据显示EM011不会引起任何血液学、免疫学和神经元毒性。基于EM011的无毒特性，我们认为EM011与其他血管生成抑制剂联合使用是合理的，这些抑制剂通过独立的机制起作用，但在其最大耐受剂量（MTD5）下显示毒性，提供了一个独特的机会来减少剂量，以最大限度地提高治疗效果，同时降低毒性。因此，项目的独立阶段将侧重于以下两个目标：EM011联合ZD6474（酪氨酸激酶抑制剂）或沙利度胺（内皮细胞增殖抑制剂）对乳腺（MDA-MB-231）和前列腺（PC-3）癌细胞的潜在协同抗增殖和抗血管生成作用的研究。目标4。利用非侵入性生物发光成像技术实时测定EM011及其与ZD6474或沙利度胺联合作为实验性原发性和转移性乳腺癌和前列腺癌抑制剂的体内疗效。作为一种实用和可转化的方法，这些研究的长期目标将是确定和建立EM011单独使用及其与其他药物协同使用的有效性。在不影响生活质量的情况下缓解人类乳腺癌和前列腺癌。