D2390

C57BL/6 mice (7 weeks old) were weighed and then immobilized for B16-F10 murine melanoma cells inoculation (2.5 × 10⁵ cells in 100 µL of phosphate buffered saline (PBS)) subcutaneously on the dorsal flank. After 5 days, all treatments were started via intraperitoneal injection for 10 consecutive days. The experimental groups used were PBS buffer (control group); Dacarbazine (DTIC) (40 or 80 mg/kg) (SIGMA D2390); MG-Pe (50 mg/kg); and the combination of DTIC+MG-Pe (80 mg/kg + 50 mg/kg). Animals were analyzed daily by measuring tumor diameters with a digital caliper (FORD). The volume was calculated using the formula: tumor volume (cm³) = (d). (D). (D). (0.52). Being: d = smaller diameter; D = larger diameter; 0.52 = ellipse volume calculation factor [76 (link)]. At the end of treatment, animals were anesthetized with ketamine (100 mg/kg) and xylazine (10 mg/kg) and euthanized.

Quininib (Q1), 1,4-dihydroxy quininib (Q7) (30 (link), 34 (link)), montelukast (Sigma #SML0101), HAMI 3379 (Cayman Chemical #10580), and dacarbazine (Sigma #D2390) were dissolved in 100% DMSO and stored as (10–50 mM) stock solutions. Working solutions (100 μM) were prepared fresh prior to each experiment in complete cell culture medium as described above. Drugs were made to final test concentrations by adding the required volume of the working solution to cells in complete media. 0.5% DMSO was used as vehicle control.