Targeting TGF-βR1 Signaling Pathway in Pancreatic Cancer: A Potential Approach with Synthetic Flavanols

Introduction: Pancreatic adenocarcinoma is a highly aggressive cancer with a poor prognosis and a five-year survival rate of just 13%. Its asymptomatic onset, rapid progression, and resistance to therapy make it challenging to treat. Transforming Growth Factor-β (TGF-β) signaling, particularly through TGF-β Receptor 1 (TGF-βR1/ALK-5), plays a major role in tumor progression by inducing Epithelial-Mesenchymal Transition (EMT), immune evasion, and apoptosis resistance. Targeting ALK-5 is a promising strategy for therapeutic intervention. Methods: Twenty-nine synthetic flavonols were designed to inhibit ALK-5 and docked using Schrodinger’s Glide XP. The compounds were synthesized via a green, one-pot method and characterized using 1H-NMR, 13CNMR, Mass Spectrometry, CHN analysis, and IR spectroscopy. The anti-cancer activity was evaluated against MiAPaCa-2 pancreatic cancer cells by measuring GI50, TGI, and LC50. ALK-5 inhibition was quantified using the ADP-Glo® Kinase Assay, assessing ATP transfer. Results: RFL-1 showed the strongest binding affinity (–9.38 kcal/mol) at ALK-5’s active site and the highest kinase inhibition (ATP transfer: 3.67%), outperforming quercetin (9.22%). It also demonstrated an IC50 of 14.92 ± 3.54 μM. Ten flavonols exhibited strong cytotoxicity (GI50 < 10 μM), while four others showed moderate activity (GI50 = 23-26 μM). Discussion: RFL-1 and related flavonols (RFL-12, RFL-20, RFL-25, RFL-28) effectively inhibited ALK-5 and suppressed the growth of pancreatic cancer cells. Their dual activity supports further development as targeted anti-cancer agents. Conclusion: Synthetic flavonols, particularly RFL-1, show promise as ALK-5 inhibitors and potential therapies for pancreatic adenocarcinoma, warranting further in vivo validation. conclusion: A) Flavonols were designed, synthesized and ADP-Glo™ Kinase Inhibition Assay was performed on Transforming Growth factor-β Receptor 1 (TGF- βR1)/ ALK -5, which was found to be up-regulated in Pancreatic cancer. B) The Synthesized novel flavonols were explored for their anti-pancreatic potential on MiaPaCa 2 cell lines, resulting in 10 flavonols having GI50 10 C) Flavonol RFL-1 exhibited stronger inhibitory effects on ALK-5, lower IC50 value and excellent anti-pancreatic cancer potential, compared to the standard Adriamycin and Quercetin.