Systemic Chemotherapy of Melanoma: NMR Studies of Lonidamine & N-Mustard Activity

黑色素瘤的全身化疗：洛尼达明的 NMR 研究

• 批准号: 8304740
• 负责人: JERRY D GLICKSON
• 金额: $ 47.27万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-21 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8304740
• 关键词: Acids; Affect; Aftercare; Alkylating Agents; Bioenergetics; Blood; Body Weight decreased; Bone Marrow; Brain; Cancer Patient; Cardiac; Cell Respiration; Cell membrane; Chlorambucil; Choline; Clinic; Clinical; Clinical Trials; Complete Blood Count; Cyclophosphamide; DNA Repair; Detection; Diffusion; Disease; Dose; Drug Kinetics; Early Diagnosis; Electrocardiogram; Electrolytes; Exhibits; Funding; Future; Glucose; Glutathione S-Transferase; Glycolysis; Goals; Growth; Heart; High Pressure Liquid Chromatography; Histopathology; Homeostasis; Human; Hyperglycemia; Imidazole; Individual; Infusion procedures; Injection of therapeutic agent; Ions; Lactic acid; Life; Liver; Lonidamine; Magnetic Resonance Imaging; Malignant Neoplasms; Maximum Tolerated Dose; Measures; Melphalan; Metastatic Melanoma; Methods; Mitochondria; Modeling; Monitor; Monocarboxylic Acid Transporters; Mus; Mustard; Mustard Agent; Nitrogen; Normal Cell; Normal tissue morphology; Nude Mice; Organ; Outcome; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacotherapy; Physiologic pulse; Preparation; Procedures; Propionic Acids; Protocols documentation; Pyruvate; Recovery; Regimen; Relative (related person); Renal function; Resolution; Respiration; Schedule; Serum; Skeletal Muscle; Skin Cancer; Staging; Stimulus; Systemic Therapy; Therapeutic; Therapeutic Index; Time; Toxic effect; Transferase; Tumor Volume; Ursidae Family; Virulent; Xenograft Model; Xenograft procedure; aminopropylphosphonate; base; cancer cell; cell killing; chemotherapy; extracellular; inhibitor/antagonist; liver function; meetings; melanoma; mortality; neoplastic cell; non-invasive monitor; pH gradient; pre-clinical; programs; research clinical testing; response; response marker; temozolomide; transport inhibitor; tumor

DESCRIPTION (provided by applicant): Melanoma remains one of the deadliest of human cancers with no effective method for treating the disseminated disease. We propose to develop an effective method for systemic chemotherapy of melanoma by [tumor] selective [intracellular] acidification to sensitize melanomas to N-mustard alkylating agents. Our strategy is to coadminister glucose with the monocarboxylic acid transport (MCT) inhibitor lonidamine LND in order to trap lactic acid produced by glycolysis in the tumor and to inhibit oxidative metabolism by impeding pyruvate trans- port into mitochondria. This approach depends on the MCT being the key pathway for tumor intracellular pH (pHi) homeostasis, which has been confirmed in our melanoma models. Previous treatment with LND under hyperglycemic conditions led to some mortality. We were funded on this 2yr R01 to develop a safe reproducible method for acidification of human DB1 melanoma xenografts with LND. By administering LND 100 mg/kg i.p. without glucose, we have achieved acidification of the tumor to a pHi of 6.4¿0.1 for at least hr during which time the NTP/Pi ratio of the tumor decreased by 50%; there was no mortality, no significant loss of weight and minimal toxicity associated with this treatment. [LND induced minimal changes in extracellular pH (pHe) of normal tissue, but modified the pH gradient across the plasma membrane of the tumor from pretreatment values of pHi 6.9, pHe 7.0 or !pH=~0 to a post-treatment values of pHi=6.4, pHe=6.85 or !pH=-0.45. Steady state lactate in the tumor increased 3-fold relative to pre-LND levels.] Respiration, EKG, electrolyte levels and blood oxygenation did not change, while blood analysis is still in progress. The pHi and ATP/Pi of skeletal muscle did not change, and there was only a small transient decrease in pHi and ATP/Pi of the liver. Melphalan (LPAM 7.5 mg/kg i.v.)+ LND exhibited a growth delay of 10.5¿0.5 d vs. LND alone, 0.6¿0.7 d. and LPAM alone 1.4¿0.1 d. The [enhanced] response of LPAM to acidification is attributed to an increase in the concentration of the active intermediate (aziridinium ion), a decrease in GSH due to decreased activity of glutathione-S-transferase and a decrease in DNA repair resulting from acid inhibition of O6-alkyl-transferase. Tumor volume decreased by 52.5 ¿12.5%, consistent with the estimated log10 cell kill=0.3012 = log10(2.0). These preliminary studies set the stage for implementation of multidose systemic treatment of xenograft models of human melanoma in preparation for eventual clinical trials. The Aims of this renewal proposal are: 1) To maximize tumor acidification while still avoiding life threatening toxicity by coadministration of low levels of glucose with LND, and to also extend these studies to the more glycolytic DB8 melanoma human xenograft model. 2) Since other N-mustard agents might exhibit similar enhancements of activity at acidic pHi values but perhaps with lower toxicity, LPAM will be compared with cyclophosphamide, chlorambucil and bendamustine under single and multidose administration. 3) To evaluate potential 1H NMR markers for noninvasive early detection of response--lactate and total choline by MRS and apparent diffusion constant (ADC) and T2 measured by MRI. PUBLIC HEALTH RELEVANCE: Melanoma, the most virulent form of skin cancer, is not responsive to chemotherapy. This project proposes to develop a method to sensitize melanomas to chemotherapy by [selective intracellular acidification of] the cancer cells while minimally affecting normal cells. As a basis for future clinic trials, we will identify the optimal drug and optimal conditions for its administration in multiple doses; we will also develop noninvasive NMR methods for early detection of therapeutic response, which will effectively tailor-fit the therapy to the needs of the individual patient.

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。我们建议开发一种有效的黑色素瘤全身化疗方法，通过[肿瘤]选择性[细胞内]酸化使黑色素瘤对N-芥子烷化剂敏感。我们的策略是将葡萄糖与单羧酸转运（MCT）抑制剂洛尼达明LND共同给药，以捕获肿瘤中糖酵解产生的乳酸，并通过阻止丙酮酸转运至线粒体来抑制氧化代谢。这种方法依赖于 MCT 作为肿瘤细胞内 pH (pHi) 稳态的关键途径，这一点已在我们的黑色素瘤模型中得到证实。先前在高血糖条件下使用 LND 治疗导致了一些死亡。我们在 2 年 R01 上获得资助，开发一种安全、可重复的方法，用于用 LND 酸化人类 DB1 黑色素瘤异种移植物。通过腹腔注射 LND 100 mg/kg在没有葡萄糖的情况下，我们已经将肿瘤酸化至 pHi 6.4±0.1 至少 hr，在此期间肿瘤的 NTP/Pi 比率下降了 50%；这种治疗没有导致死亡，体重没有明显减轻，而且毒性也很小。 [LND 引起正常组织细胞外 pH (pHe) 的微小变化，但改变了肿瘤质膜的 pH 梯度，从治疗前的 pHi 6.9、pHe 7.0 或 !pH=~0 到治疗后的 pHi=6.4、pHe=6.85 或 !pH=-0.45。肿瘤中的稳态乳酸相对于 LND 前的水平增加了 3 倍。] 呼吸、心电图、电解质水平和血氧没有变化，而血液分析仍在进行中。骨骼肌的pHi和ATP/Pi没有变化，肝脏的pHi和ATP/Pi仅有短暂的小幅下降。美法仑（LPAM 7.5 mg/kg i.v.）+ LND 的生长延迟为 10.5±0.5 天，而单独使用 LND 则为 0.6±0.7 天。单独使用 LPAM 1.4±0.1 d。 LPAM 对酸化的[增强]反应归因于活性中间体（氮丙啶鎓离子）浓度的增加、谷胱甘肽-S-转移酶活性降低导致的 GSH 减少以及 O6-烷基转移酶的酸抑制导致的 DNA 修复减少。肿瘤体积减少了 52.5 ¿12.5%，与估计的 log10 细胞杀伤=0.3012 = log10(2.0) 一致。这些初步研究为对人类黑色素瘤异种移植模型实施多剂量全身治疗奠定了基础，为最终的临床试验做准备。该更新提案的目标是：1) 通过低水平葡萄糖与 LND 联合给药，最大限度地酸化肿瘤，同时仍然避免危及生命的毒性，并将这些研究扩展到糖酵解性更强的 DB8 黑色素瘤人类异种移植模型。 2) 由于其他 N-芥末剂在酸性 pHi 值下可能表现出类似的活性增强作用，但毒性可能较低，因此将 LPAM 在单剂量和多剂量给药下与环磷酰胺、苯丁酸氮芥和苯达莫司汀进行比较。 3) 评估潜在的 1H NMR 标记物，用于无创早期检测反应——通过 MRS 检测乳酸和总胆碱，并通过 MRI 测量表观扩散常数 (ADC) 和 T2。 公众健康相关性：黑色素瘤是毒性最强的皮肤癌，对化疗没有反应。该项目提议开发一种方法，通过癌细胞[选择性细胞内酸化]使黑色素瘤对化疗敏感，同时对正常细胞的影响最小化。作为未来临床试验的基础，我们将确定最佳的 药物及其多剂量给药的最佳条件；我们还将开发用于早期检测治疗反应的无创核磁共振方法，这将有效地根据个体患者的需求定制治疗方案。