Evaluation of Stability and Anti-cancer activity of Copper(II) Tolfenamic Acid with an emphasis on Pancreatic

Purpose: Tolfenamic acid (TA) acts as an anti-cancer agent in several cancer models via down-regulating transcription factors Sp1 and Sp3, and an inhibitor of apoptotic protein, survivin. Copper (Cu) is an important element with multiple biological functions and has gained interest in medical applications. Recently, Cu-TA has been synthesized and tested for enhanced therapeutic activity. In this study, Cu-TA was investigated for its stability and anti-cancer activity using several cancer cell lines and mouse model for pancreatic cancer (PC). Method: Cu-TA was synthesized and characterized by UV visible spectroscopy and Fourier-transform infrared spectroscopy (FTIR). Anti-proliferative activity was evaluated against twelve cell lines representing six (breast, colon, glioblastoma, medulloblastoma, pancreatic and prostate) cancers using the CellTiter-Glo kit and compared with TA. Further studies were performed using PC cells. The expression of Sp1, Sp3 and survivin was determined by Western blot and qPCR. The stability of Cu-TA was determined using 8-12 month-old powder and six-month-old stock solution. Cardiomyocytes (H9C2) were used to test the cytotoxicity in non-malignant cells. Athymic mice were injected with PC cells and treated with vehicle (control) or 25 or 50 mg/kg of Cu-TA 3 times/week and the effect on tumor growth was monitored for 4 weeks Animals body weight changes were also observed to determine overt toxicity. Results: Cu-TA significantly more effective than TA against all tested cancer cells. The IC50 values of Cu-TA were 30 to 80% less when compared with TA. Comparison of the twelve-month-old powder and six-month-old stock solution using the Panc1 cells showed similar IC50values ( Conclusion: These in vitro and in vivo studies demonstrate that Cu-TA is more effective than TA and potentially useful as an effective anti-cancer agent.