Cabazitaxel

R1-AD1, R1-D567 and R1-I567 [16 (link)], CWR22Rv-1 and PC3 cells were cultured in RPMI 1640 medium with L-glutamine (Corning #10-040-CV) supplemented with 10% fetal bovine serum (Thermo Fisher Scientific #10437-036) and penicillin/streptomycin (Corning #30-002-Cl) in a humidified incubator at 37°C with 5% CO2. JMJD3/UTX inhibitor GSK-J4 (Sigma #SML0701) and Cabazitaxel (LC Laboratories #C-2581) were dissolved in DMSO (Sigma #D2650) to prepare 20mM and 20μM stock correspondingly.

Soluble recombinant human TRAIL (rhsTRAIL) was obtained from PeproTech Inc. (Rocky Hill, NJ, USA). Semisynthetic paclitaxel and docetaxel were purchased from Sigma Aldrich Chemical Company (St. Louis, MO, USA). The purity of paclitaxel and docetaxel was ≥97% (HPLC). Cabazitaxel was purchased from LC Laboratories (Woburn, MA, USA). The purity of Cabazitaxel was ≥99% (HPLC). Tested compounds were dissolved in DMSO (dimethyl sulfoxide) to obtain the working concentrations.

Synthesized polymers were dissolved in CH₂Cl₂ (Avantor Performance Materials Poland S.A., Gliwice, Poland) and then mixed with docetaxel or cabazitaxel (LC Laboratories, Woburn, MA, USA). The concentration of each solution prepared for electrospinning was optimized in order to obtain a structure with well-developed fibres. Different nonwovens were obtained with the various amount of the drug: 3–5% (w/w) (Table 1) using a TL-Pro-BM electrospinning unit (Tong Li Tech, Shenzen, China). The device was equipped with two high voltage power supplies. One of them, that generates positive electrical potential, was applied to the spinneret, while the second one, that generates negative potential, was applied to the fibers collector, in form of a steel mandrel of 27 mm diameter (rotating collector). Polymer solutions were dosed to the spinning nozzle through a PTFE capillary, by using a PHD Ultra 4400 syringe pump (Harvard Apparatus, Holliston, MA, USA). Nonwoven mats were produced according to process parameters, presented in Table 1.