Application of carbonic anhydrase inhibitors to increase the penetration of doxorubicin and its liposomal formulation into 2D and 3D triple negative breast cancer cell cultures

The aim of our study was to assess the influence of two carbonic anhydrase (CA) inhibitors (methazolamide (MTZ)) and U-104 on weakly basic anticancer drug doxorubicin (DOX) and pegylated liposomal doxorubicin (PLD) delivery into monolayer-cultured 4T1 murine breast cancer cells (2D cultures) and tumor spheroids (3D cultures) at pH 6.0 and 7.4. The effect of compounds on cell viability was evaluated by MTT assay. Spheroids were formed using 3D Bioprinting method. The penetration of DOX and PLD into cells and spheroids was evaluated using fluorescence microscopy. Both MTZ and U-104 increased the DOX (5 µM) and PLD (concentration corresponding to 5 µM DOX) penetration into monolayer-cultured cells at acidic conditions but did not enhance drug delivery at physiological pH. Pretreatment with U-104 inhibitors also increased DOX and PLD delivery into tumor spheroids. Thus, U-104 may be worthy of further studies as possible transport modulator of weakly basic drugs.