Targeting vulnerabilities in copper metabolism in the development of cancer therapies

项目总结 尽管在癌症的检测和治疗方面已经有了重要的进展，但仍然有 迫切需要开发新的治疗策略。铜(铜)是人体必需的微量营养素。 与正常组织相比，癌细胞的数量更多。几种在癌症中起关键作用的酶需要 铜的活性，包括赖氨酸氧化酶和几种致癌的激酶(例如，MEK1；ULK1)。在临床前阶段 在肿瘤模型中，许多研究表明铜络合剂抑制肿瘤的生长和转移 添加到饮食中。在临床试验中，通过口服铜络合剂来消耗铜被发现可以显著延缓疾病。 间皮瘤患者的进展，并显著延长乳腺癌患者的生存期。当它 很明显，耗竭铜是一种很有前途的抗癌策略，因此有必要开发出 特异性靶向铜致癌酶的转运途径。在目前的提案中，我们追求高度的 针对铜转运蛋白ATP7A的创新方法。 我们广泛的初步研究证实ATP7A是一种治疗靶点，包括：1)肠道特异性 小鼠ATP7A基因的缺失降低了全身铜状态，达到了癌症患者的治疗水平； 2)ATP7A是向赖氨酰氧化酶家族输送铜所必需的，赖氨酸氧化酶家族在 乳腺癌和肺癌细胞系中ATP7A的靶向缺失可减少原发肿瘤的生长 4)ATP7A的高表达与肿瘤的低生存期显著相关 病人。基于这些发现，我们假设ATP7A的一种小分子抑制剂将是 治疗癌症。利用计算机辅助药物设计，我们已经识别出一种名为MKV3的命中分子 以纳摩尔亲和力与ATP7A结合，抑制癌细胞中ATP7A的活性。用MKV3治疗的小鼠 显示血清铜生物标志物铜蓝蛋白活性降低，肿瘤生长减少。在这 建议，我们将进行结构指导优化，以确定具有更高效力的MKV3类似物 类药物特性(目标1)；进行药代动力学研究，以确定适合于 药效学研究(目标2)；评估最有利的MKV3类似物在小鼠乳腺模型中的应用 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).