Fig. 6: Peptide 2012 induces robust anti-cancer effects in both in vitro and in vivo cancer models.
Dose-response curves reporting mean ± SD decrease in cell viability and corresponding average IC50 values of cancer cell lines SK-MEL-147, WM88, and ONDA7, as well as normal cell lines LX-2, Hep-G2, and NHEM, treated with serially diluted a 2012, b defactinib, and c 2020 (neg. control) for 96 h. Values reported for: SK-MEL-147 and LX-2 were derived from 4 biological replicates (n = 4) in (a–c); WM88 were derived from 4 biological replicates (n = 4) in (a) and 3 biological replicates in (b, c); ONDA7 were derived from 5 biological replicates (n = 5) in (a, c) and 3 biological replicates (n=3) in (b); Hep-G2 were derived from 3 biological replicates (n = 3) in (a–c); and NHEM were derived from 3 biological replicates (n = 3) in (a) and 4 biological replicates (n = 4) in (b, c). d Selectivity profile of 2012 and defactinib at 1 µM against 97 kinases measured in the KINOMEscan assay (Eurofins) where the size of red circles represents the percent of remaining kinase activity relative to DMSO control. e Annexin V Apoptosis EC50 curves reporting mean luminescence relative to DMSO (vehicle control) ± SD following treatment with 2012 (n = 7); 2020 (n = 5); or defactinib (n = 5) obtained after 96 h of drug treatment. All n’s represent independent biological replicates. f Invasion IC50 curves reporting mean ± SD reduction of invasion relative to DMSO (vehicle control) obtained from transwell invasion chamber assays after 48 h treatment of 2012 (n = 6), 2020 (neg. control) (n = 2), and defactinib (n = 4) in SK-MEL-147. All n’s represent independent biological replicates. g Pharmacokinetic evaluation via i.p. administration of 2012 in Balb/cJ mice revealing Cmax and plasma half-life. The concentration-time profile reports the remaining mean concentration of 2012 (µg/mL) ± SD at 0, 0.5,1, 2, 4, 8, and 24 h derived from 3 biological replicates (n = 3). h Effects on tumor growth in B16F10 syngeneic melanoma mouse model by 2012 treatment (20 mg/kg i.p. QD, n = 8 mice per group). Treatment started at day 12. Curves report mean tumor volume ± SD derived from 8 biological replicates per group (n = 8). Two-way ANOVA analysis and Dunnett’s multiple comparison test were performed, and statistically significant changes in tumor volume were denoted with corrected p values. Bolded p values indicate statistically significant values where p < 0.05. All source data are provided as a Source Data file.
