In Vitro and In Silico Assessment of Anticancer Activity of Venom-Derived and Biomimetic Neurotransmitter Inhibitor Pentapeptides.

Cancer, resulting from uncontrolled cell proliferation, is a leading cause of death worldwide and its incidence continues to rise. Challenges such as late diagnosis, aggressive tumor biology, and therapeutic resistance limit the effectiveness of current treatment strategies. Therefore, therapeutic peptides capable of penetrating cells and exhibiting high target specificity have emerged as an innovative approach in cancer therapy. Peptides derived from animal venoms have demonstrated antitumor activity and diagnostic potential in glioma, colon, ovarian, breast, and prostate cancers; for instance, chlorotoxin, crotamine, and eosinophil-derived neurotoxin are being investigated as biomarkers or target-specific anticancer agents. Critical signaling pathways in cancer cells, including PI3K/Akt/mTOR, EGFR, androgen receptor, PSMA, and CDK8, have been explored as targets for peptide-based therapeutics in both in silico and experimental studies. In this study, the anticancer potential of Vialox (Gly-Pro-Arg-Pro-Ala) derived from Tropidolaemus wagleri venom and Leuphasyl (Tyr-Ala-Gly-Phe-Leu) derived from botulinum neurotoxin as a biommimetic peptide were evaluated through molecular docking, MM-GBSA calculations, molecular dynamics simulations, ADMET analysis, and MTT cell assays. Docking studies revealed strong binding affinities of Vialox and Leuphasyl to key oncogenic targets, with docking scores ranging from -10.63 to -7.59 kcal/mol. The highest binding was observed for EGFR-Leuphasyl (-10.63 kcal/mol), followed by Vialox-EGFR (-9.24 kcal/mol). PI3K docking scores were also high, with -9.05 and -9.15 kcal/mol for Vialox and Leuphasyl, respectively. ADMET predictions indicated limited oral bioavailability and minimal BBB penetration. MM-GBSA calculations showed that Vialox and Leuphasyl interact differently with the EGFR binding pocket. Vialox binding is mainly driven by electrostatic interactions, with a binding energy of -65.29 kcal/mol. Leuphasyl relies on hydrophobic interactions but has a lower binding energy of -46.68 kcal/mol due to differences in charge distribution and polar interactions. In vitro MTT assays showed that both peptides exhibited the most pronounced cytotoxic effects in MDA-MB-231 and PC3 cells, supporting their potential selective anticancer activity. The findings suggest that both peptides may exhibit selective anticancer effects, and that combining in silico approaches with experimental validation represents an effective strategy for developing novel peptide-based cancer therapeutics.