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，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素瘤全身化疗方法，通过[肿瘤]选择性[细胞内]酸化使黑素瘤对n -芥菜烷化剂敏感。我们的策略是将葡萄糖与单羧酸转运（MCT）抑制剂lonidamine LND共同施用，以捕获肿瘤中糖酵解产生的乳酸，并通过阻碍丙酮酸转运到线粒体来抑制氧化代谢。这种方法依赖于MCT是肿瘤细胞内pH （pHi）稳态的关键途径，这已经在我们的黑色素瘤模型中得到证实。以前在高血糖条件下用LND治疗导致一些死亡。我们在这个为期2年的R01项目中获得资助，开发一种安全可重复的方法来酸化带有LND的人类DB1黑色素瘤异种移植物。通过给予LND 100 mg/kg，不含葡萄糖，我们已经实现了肿瘤的酸化至6.4¿0.1的pHi至少一小时，在此期间，肿瘤的NTP/Pi比率下降了50%；没有死亡，没有显著的体重减轻和最小的毒性与这种治疗相关。[LND诱导正常组织的细胞外pH （pHe）变化很小，但改变了肿瘤质膜上的pH梯度，从预处理值pHi为6.9，pHe为7.0或！pH=~0至处理后值pHi=6.4, pHe=6.85或pH=-0.45。相对于lnd前，稳态乳酸水平在肿瘤中增加了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只有短暂的小幅度下降。与单独使用LND相比，Melphalan (LPAM 7.5 mg/kg i.v)+ LND表现出10.5 ~ 0.5 d的生长延迟，0.6 ~ 0.7 d和LPAM单独使用1.4 ~ 0.1 d。LPAM对酸化的[增强]反应归因于活性中间体（aziridinium离子）浓度的增加，谷胱甘肽- s转移酶活性的降低导致GSH的降低，以及o6 -烷基转移酶的酸抑制导致DNA修复的减少。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。这些初步研究为实施人类黑色素瘤异种移植模型的多剂量全身治疗奠定了基础，为最终的临床试验做准备。该更新提案的目的是：1)通过与LND共同施用低水平葡萄糖，最大限度地提高肿瘤酸化程度，同时仍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 -芥菜剂可能在酸性pHi值下表现出类似的活性增强，但可能毒性较低，因此将LPAM与环磷酰胺、氯霉素和苯达莫司汀在单次和多次给药下进行比较。3)评价可能用于无创早期反应检测的1H NMR标记物——MRS测量乳酸和总胆碱，MRI测量表观扩散常数（ADC）和T2。