Effects of treatments with CPBMF-223 (50 mg/kg/day, 5 days/week, 3 weeks) and 5-fluorouracil (5-FU; 30 mg/kg/day, every other day), by intraperitoneal (i.p.) route, on tumor relative volume (RTV) and body weight changes (%) in nude mice injected with human colorectal carcinoma (HCT-116). RTV curve during treatment for female (A) and male (B) mice. Body weight changes curve for female (C) and male (D) animals. Representative pictures to show the sizes of the tumors excised on day 21 (E). Chemical structure of 6-(((1,3-dihydroxypropan-2-yl)amino)methyl)-5-iodopyrimidine-2,4(1H,3H)-dione (CPBMF-223) (F). 5-FU treatment was used as a positive control drug for colorectal cancer. Data were expressed with the mean ± standard error of the mean. Statistical analysis was performed by two-way analysis of variance, followed by Bonferroni post-test. Data significant in relation to vehicle (negative control group) at *p < 0.05, **p < 0.01, and ***p < 0.001. q.o.d.: every other day.

Migration assay in HCT-116 cell over 48 h of exposure to CPBMF-223 and TPI (competitive inhibitor of hTP). Measurement of HCT-116 cell migration (%) over time (0, 4, 8, 12, 24 and 48 h) (A) and representative images of migration after 48 h of treatment (B). DMEM with 10% FBS was used as positive control of migration. Five independent measurements per picture were performed in order to calculate the percentage of migration, considering time 0 of each treatment with 0%. Data were expressed with the mean ± standard error of the mean. Statistical analysis was performed by two-way analysis of variance, followed by Bonferroni post-test. Data significant in relation to positive control (DMEM 10% FBS) at *p < 0.05 and ***p < 0.001.

Effects of treatment with CPBMF-223 (50 mg/kg/day, 5 days/week, 3 weeks) and 5-fluorouracil (5-FU; 30 mg/kg, every other day) by intraperitoneal (i.p.) route. Immunolabeling for vascular endothelial growth factor (VEGF; angiogenesis marker) (A and D) and Ki-67 (cell proliferation marker) (B and E); and hematoxylin-eosin (HE) for number of blood vessels (C and F) (angiogenesis marker) present in the tumor tissue for female and male nude mice. Representative images of HE in different treatment groups and sexes (G). 5-FU treatment was used as a positive control drug for colorectal cancer. Results are expressed as optical density or number of vessels and represent the mean ± standard error of the mean for five images captured per tumor (x200 for HE and VEGF; x400 for Ki-67). Statistical analysis was determined by one-way analysis of variance, followed by Dunnet’s post-test. *p < 0.05, **p < 0.01, and ***p < 0.001 significantly from vehicle group (negative control). n = 5–7 animals per group.

Effect of CPBMF-223 (50–500 µM) acute treatment in early stages of heartbeat, exploratory behavior, and neurotoxicity in zebrafish larvae. The heartbeat was evaluated in 2 and 5 days postfertilization (dpf) (A and B), while distance (C), velocity (D), angular velocity (E) and frequency outside area (F) were evaluated in 5 dpf. Data are expressed mean ± standard error of the mean from 18 (Heartbeat) or 30 animals tested individually for each group and were analyzed by one-way analysis of variance, followed by Tukey post-test. *p < 0.05, **p < 0.01, and ***p < 0.001 significantly from vehicle group (negative control).