Synthesis and Characterization of an Aspirin-fumarate Prodrug that Inhibits NFκB Activity and Breast Cancer Stem Cells

Abstract

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 anticancer 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 CD44 + CD24-immunophenotype. These results establish a viable strategy to develop improved anti-inflammatory drugs for chemoprevention and cancer therapy.

Ameliorative role of aspirin in paraquat‐induced lung toxicity via mitochondrial mechanisms

“…ASA inhibits cyclooxygenase, which can remove hydroxy radical's production due to antioxidant and anti‐inflammatory properties of ASA . It also reduces the oxidative stress and inhibits NF‐κB by inhibiting TNF‐α . Furthermore, studies have confirmed the inhibition of lipid peroxidation and inhibit inflammatory mediators of IL‐4 and activator protein‐1 after aspirin administration .…”

Aspirin Actions in Treatment of NSAID-Exacerbated Respiratory Disease

“…Esophageal, prostate, breast, gastric, and gastro-intestinal cancers (153,(169)(170)(171)(172)(173) Downregulates atherothrombosis, inactivates platelet aggregation and cancer metastasis in esophageal and gastro-intestinal cancers. Inhibits angiogenesis in gastric cancer.…”

“…After its own activation, NF-kB can activate the transcription of various proinflammatory cytokines, chemokines and adhesion molecules. In cancer, a deregulated NF-kB pathway promotes tumor cell survival, proliferation, migration, invasion, angiogenesis, and resistance to therapy (169). Increased expression of NF-kB is associated with nasal polyposis (207).…”

<p>Self-Renewal Signalling Pathway Inhibitors: Perspectives on Therapeutic Approaches for Cancer Stem Cells</p>

“…These findings established a viable strategy for development of improved anti-inflammatory agents for chemoprevention and cancer treatment. 113 To improve drug efficacy and minimize toxicity, one study found that the fumarate-based ASA prodrug GTCpFE represents a prototype for development of novel anti-inflammatory and anti-CSC drugs for treatment of potentially aggressive breast cancers. 114…”