Kastrati Irida, Delgado-Rivera Loruhama, Georgieva Gergana, Thatcher Gregory R J, Frasor Jonna
Physiology and Biophysics, College of Medicine, University of Illinois at Chicago;
Medicinal Chemistry and Pharmacognosy, College of Medicine, University of Illinois at Chicago.
J Vis Exp. 2017 Jan 18(119):54798. doi: 10.3791/54798.
Inflammation is a cancer hallmark that underlies cancer incidence and promotion, and eventually progression to metastasis. Therefore, adding an anti-inflammatory drug to standard cancer regiments may improve patient outcome. One such drug, aspirin (acetylsalicylic acid, ASA), has been explored for cancer chemoprevention and anti-tumor activity. Besides inhibiting the cyclooxygenase 2-prostaglandin axis, ASA's anti-cancer activities have also been attributed to nuclear factor ĸB (NFĸB) inhibition. Because prolonged ASA use may cause gastrointestinal toxicity, a prodrug strategy has been implemented successfully. In this prodrug design the carboxylic acid of ASA is masked and additional pharmacophores are incorporated. This protocol describes how we synthesized an aspirin-fumarate prodrug, GTCpFE, and characterized its inhibition of the NFĸB pathway in breast cancer cells and attenuation of the cancer stem-like properties, an important NFĸB-dependent phenotype. GTCpFE effectively inhibits the NFĸB pathway in breast cancer cell lines whereas ASA lacks any inhibitory activity, indicating that adding fumarate to ASA structure significantly contributes to its activity. In addition, GTCpFE shows significant anti-cancer stem cell activity by blocking mammosphere formation and attenuating the cancer stem cell associated CD44CD24 immunophenotype. These results establish a viable strategy to develop improved anti-inflammatory drugs for chemoprevention and cancer therapy.
炎症是一种癌症标志，是癌症发生、发展以及最终进展为转移的基础。因此，在标准癌症治疗方案中添加抗炎药物可能会改善患者的预后。其中一种药物阿司匹林（乙酰水杨酸，ASA）已被用于癌症化学预防和抗肿瘤活性的研究。除了抑制环氧合酶2 - 前列腺素轴外，ASA的抗癌活性还归因于对核因子κB（NFκB）的抑制。由于长期使用ASA可能会导致胃肠道毒性，一种前药策略已成功实施。在这种前药设计中，ASA的羧酸被掩盖，并引入了额外的药效基团。本方案描述了我们如何合成一种阿司匹林 - 富马酸酯前药GTCpFE，并表征其对乳腺癌细胞中NFκB途径的抑制作用以及对癌症干细胞样特性（一种重要的NFκB依赖性表型）的减弱作用。GTCpFE能有效抑制乳腺癌细胞系中的NFκB途径，而ASA则缺乏任何抑制活性，这表明在ASA结构中添加富马酸酯对其活性有显著贡献。此外，GTCpFE通过阻断乳腺球形成和减弱与癌症干细胞相关的CD44CD24免疫表型，显示出显著的抗癌干细胞活性。这些结果确立了一种可行的策略，用于开发改进的抗炎药物以进行化学预防和癌症治疗。
