Mechanism of Activity of Lonidamine

氯尼达明的活性机制

• 批准号: 8584841
• 负责人: JERRY D GLICKSON
• 金额: $ 56.74万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8584841
• 关键词: Address; Alkylating Agents; Anthracyclines; Antineoplastic Agents; Biochemical Pathway; Biochemistry; Bioenergetics; Bioreactors; Cardiac; Cell membrane; Cells; Characteristics; Clinical; Collaborations; Data; Detection; Development; Disease; Doxorubicin; Electron Transport; Electrons; Energy Metabolism; Funding; Glycolysis; Glycolysis Inhibition; Goals; Grant; Heart; Hexokinase 2; Human; Isolated limb perfusion; Isotopes; Kinetics; Label; Lactic acid; Liver; Liver Mitochondria; Lonidamine; Magnetic Resonance Spectroscopy; Malignant Neoplasms; Measures; Mechlorethamine; Mediating; Melanoma Cell; Melphalan; Metabolic; Metabolism; Methods; Mitochondria; Monitor; Monocarboxylic Acid Transporters; Mustard; Nude Mice; Oxidation-Reduction; Oxidative Phosphorylation; Pharmaceutical Preparations; Pharmacologic Substance; Phospholipid Metabolism; Production; Protein Isoforms; Pyruvate; Radiation Tolerance; Rana; Respiration; Skin Cancer; Staging; Systemic Therapy; Techniques; Testing; Therapeutic Agents; Time; Tissues; Translations; Tumor Oxygenation; Universities; Virulent; Warburg Effect; Xenograft procedure; Xenopus laevis; base; cancer cell; cancer therapy; chemotherapeutic agent; chemotherapy; clinical practice; design; egg; glucose metabolism; improved; in vivo; inhibitor/antagonist; liquid chromatography mass spectrometry; melanoma; metabolomics; monolayer; neoplastic cell; network models; novel; public health relevance; pyruvate carrier; research clinical testing; response; tumor; tumor metabolism; tumor microenvironment; tumor xenograft

DESCRIPTION (provided by applicant): Melanoma remains one of the deadliest of human cancers with no effective method for treating the disseminated disease. While targeted therapies have shown some efficacy, they have thus far proven to be noncurative, which points to the need for other forms of systemic therapy. This proposal is part of a long-term effort to develop methods for systemic therapy of melanoma. We have recently shown that administration of lonidamine (LND, 100 mg/kg) substantially enhances the activity of melphalan and have now extended this finding to doxorubicin treatment. As a key step towards eventual clinical translation, we will now examine the detailed mechanism of LND activity. We and others have shown that LND produces tumor specific intracellular acidification and a substantial decrease in tumor energy status (NTP/Pi). A number of mechanisms have been proposed for LND: 1) inhibition of hexokinase II, 2) interference with mitochondrial electron transport, 3) inhibition of cellular lactate export through the monocarboxylate transporters (MCT). We are proposing a fourth mechanism to explain the bioenergetic decline of the tumor following treatment with LND: 4) inhibition of the putative mitochondrial pyruvate carrier (MPC), which would deplete the tumor cells of a key substrate for oxidative phosphorylation and induce an increase in glycolytic metabolism to compensate for decreased oxidative ATP production. We have recently validated a novel extension of isotopomer analysis called cumomer analysis and applied it to perfused DB1 melanoma cells. We believe that this is the first validated metabolic network model of tumor energy metabolism. We propose to test the hypotheses that 1) selective tumor acidification results from inhibition of MCT1, and 2) tumor de-energization is caused by inhibition of the MPC. In Aim 1 of this proposal, we propose to use 13C magnetic resonance spectroscopy (MRS) and liquid chromatography-mass spectrometry (LC-MS) in conjunction with bonded cumomer analysis to test these hypotheses in perfused DB1 and DB8 melanoma cells and in in vivo xenografts of these tumor lines. In Aim 2 we will directly measure the inhibitory effect of LND on MCT1 and MCT4 expressed in Xenopus laevis and will also evaluate the effect of LND on isolated liver and cardiac mitochondria and on permeabilized DB1 and DB8 cells. All of the criticisms of the previous review have been addressed including the critical issue of clinical translation for which we assembled a team of leading experts on melanoma who recommended: 1) define the mechanism of LND (the aim of this proposal), 2) first incorporate LND into hyperthermic isolated limb perfusion of melanoma with melphalan (a method already in clinical practice), and 3) then proceed to systemic therapy of cancers that are already treated with N-mustards or doxorubicin. This project will have a major impact on elucidating the mechanism of activity of a new class of drugs that like LND inhibit MCTs and other key transporters in tumor cells and thereby modify the tumor microenvironment to augment the activity of conventional anticancer agents. It will also develop novel metabolomic methods for the study of tumor metabolism and mechanisms of cancer drug activity.

描述（由申请人提供）：黑色素瘤仍然是最致命的人类癌症之一，并且没有有效的方法来治疗这种播散性疾病。虽然靶向治疗已显示出一定的疗效，但迄今为止已被证明是非治愈性的，这表明需要其他形式的全身治疗。该提案是开发黑色素瘤全身治疗方法的长期努力的一部分。我们最近表明，给予洛尼达明（LND，100 mg/kg）可显着增强美法仑的活性，并且现已将这一发现扩展到阿霉素治疗。作为最终临床转化的关键一步，我们现在将研究 LND 活性的详细机制。我们和其他人已经证明，LND 会产生肿瘤特异性细胞内酸化，并显着降低肿瘤能量状态 (NTP/Pi)。 LND 的多种机制已被提出：1) 抑制己糖激酶 II，2) 干扰线粒体电子传递，3) 抑制通过单羧酸转运蛋白 (MCT) 的细胞乳酸输出。我们提出第四种机制来解释 LND 治疗后肿瘤生物能下降：4）抑制假定的线粒体丙酮酸载体（MPC），这会耗尽肿瘤细胞氧化磷酸化的关键底物并诱导糖酵解代谢来补偿氧化 ATP 产生的减少。我们最近验证了同位素异构体分析的一种新扩展，称为累积异构体分析，并将其应用于灌注的 DB1 黑色素瘤细胞。我们相信这是第一个经过验证的肿瘤能量代谢代谢网络模型。我们建议测试以下假设：1）抑制 MCT1 导致选择性肿瘤酸化，2）抑制 MPC 导致肿瘤失能。在本提案的目标 1 中，我们建议使用 13C 磁共振波谱 (MRS) 和液相色谱-质谱 (LC-MS) 结合键合累积体分析，在灌注的 DB1 和 DB8 黑色素瘤细胞和体内测试这些假设这些肿瘤系的异种移植物。在目标 2 中，我们将直接测量 LND 对非洲爪蟾中表达的 MCT1 和 MCT4 的抑制作用，还将评估 LND 对离体肝脏和心脏线粒体以及透化 DB1 和 DB8 细胞的影响。之前审查的所有批评都已得到解决，包括临床转化的关键问题，为此我们组建了一个由黑色素瘤领先专家组成的团队，他们建议：1）定义 LND 的机制（本提案的目的），2）首先将 LND 纳入使用美法仑对黑色素瘤进行离体高温灌注（一种已在临床实践中的方法），3) 然后对已经用 N-芥末或多柔比星治疗的癌症进行全身治疗。该项目将对阐明 LND 等新型药物的活性机制产生重大影响，这些药物可抑制 MCT 和肿瘤细胞中的其他关键转运蛋白，从而改变肿瘤微环境以增强传统抗癌药物的活性。它还将开发新的代谢组学方法来研究肿瘤代谢和癌症药物活性机制。