Targeting vulnerabilities in copper metabolism in the development of cancer therapies

项目摘要 尽管在癌症的检测和治疗方面已经有了重要的进展，但仍然存在一个问题。 迫切需要开发新的治疗策略。铜（Cu）是一种必需的微量营养素， 相对于正常组织，癌细胞的含量更高。几种在癌症中起关键作用的酶需要 Cu的活性，包括赖氨酰氧化酶和几种致癌激酶（例如，MEK1; ULK1）。在临床前 癌症模型，许多研究表明，肿瘤生长和转移抑制铜螯合剂 添加到饮食中。在临床试验中，通过口服铜螯合剂消耗铜被发现可以显着减缓疾病 在间皮瘤患者中的进展，并显著延长乳腺癌患者的生存期。虽然 显然，铜消耗是一种有前途的抗癌策略，需要开发 特异性地靶向Cu递送至致癌酶的途径。在目前的提案中，我们追求一个高度 通过靶向铜转运蛋白ATP 7A的创新方法。 我们广泛的初步研究验证了ATP 7A作为治疗靶点，包括：1）肠特异性 鼠ATP 7A的缺失将全身铜状态降低至显示在癌症患者中具有治疗性的水平; 2)ATP 7A是将铜传递给赖氨酰氧化酶家族所必需的，赖氨酰氧化酶家族在以下方面具有充分记载的作用： 3）乳腺癌和肺癌细胞系中ATP 7A的靶向缺失减少原发性肿瘤生长 4）ATP 7A表达升高与癌症中较低的存活率显著相关 患者基于这些发现，我们假设ATP 7A的小分子抑制剂将是一种有效的抑制剂。 治疗癌症。使用计算机辅助药物设计，我们已经确定了一种名为MKV 3的命中分子， 以纳摩尔亲和力结合ATP 7A并抑制癌细胞系中的ATP 7A活性。用MKV 3处理的小鼠 显示血清Cu生物标志物铜蓝蛋白的活性降低和肿瘤生长减少。在这 我们将进行结构指导优化，以鉴定具有改善效力的MKV 3类似物， 药物样性质（目的1）;进行药代动力学研究，以确定适用于 药效学研究（目的2）;并评估最有利的MKV 3类似物在小鼠乳腺癌模型中的作用 癌症（目标3）。

PROJECT SUMMARY Although there have been important advances in the detection and treatment of cancer, there remains an urgent need to develop new therapeutic strategies. Copper (Cu) is an essential micronutrient that is required in higher amounts by cancer cells relative to normal tissues. Several enzymes with key roles in cancer require Cu for their activity, including lysyl oxidases and several oncogenic kinases (e.g., MEK1; ULK1). In pre-clinical models of cancer, many studies have shown that tumor growth and metastasis is suppressed by Cu chelators added to the diet. In clinical trials, Cu depletion via an oral Cu chelator was found to significantly slow disease progression in patients with mesothelioma, and significantly extend survival in breast cancer patients. While it is clear that Cu depletion is a promising anticancer strategy, there is a need to develop therapies that specifically target pathways of Cu delivery to oncogenic enzymes. In the current proposal, we pursue a highly innovative approach by targeting the Cu transporter, ATP7A. Our extensive preliminary studies validate ATP7A as a therapeutic target including: 1) Intestine-specific deletion of murine ATP7A lowers systemic copper status to levels shown to be therapeutic in cancer patients; 2) ATP7A is required to deliver copper to the family of lysyl oxidases, which have well-documented roles in metastasis; 3) Targeted deletion of ATP7A in breast and lung cancer cell lines reduces primary tumor growth and metastasis in mice; 4) Elevated ATP7A expression is significantly correlated with lower survival in cancer patients. Based on these findings, we hypothesize that a small molecule inhibitor of ATP7A will be therapeutic in cancer. Using computer-aided drug design, we have identified a hit molecule called MKV3 that binds to ATP7A with nanomolar affinity and inhibits ATP7A activity in cancer cell lines. Mice treated with MKV3 showed reduced activity of the serum Cu biomarker, ceruloplasmin, and reduced tumor growth. In this proposal, we will conduct structure guided optimization to identify MKV3 analogs with improved potency and drug like properties (Aim 1); conduct pharmacokinetic studies to identify MKV3 analogs that are suitable for pharmacodynamic studies (Aim 2); and evaluate the most favorable MKV3 analog in mouse models of breast cancer (Aim 3).