Abstract
Natural product scaffolds can be combined with repurposed drugs, so called conjugate molecules may exhibit improved pharmacology. We designed a conjugate (XN-MT) of xanthohumol (a prenylated chalcone with anticancer properties) and metformin (an antidiabetic drug with reported anticancer effects) and evaluated its potential against breast cancer (BC) using network-based pharmacology, docking, molecular dynamics (MD) simulation and density functional theory (DFT) Studies. Network pharmacology revealed targets relevant to cell proliferation, apoptosis, CDK4/2/1 signaling, and metabolic regulation and identified pathways were crucial for breast tumorigenesis and chemoresistance. Molecular docking against high-priority targets, like CDK4, PTGS2, and CDK1, predicted favourable binding affinities and plausible interaction modes for the conjugate. MD simulations (100 ns) for the conjugate showed stable ligandprotein interactions and persistent hydrogen bond networks, supporting the conjugates predicted binding stability. ADME, toxicity and DFT analyses provided comprehensive insights into the pharmacokinetic profile and electronic behaviour of xanthohumol, metformin and their conjugate. Moreover, DFT calculations on conjugates showed a lower HOMO-LUMO energy gap, average local ionization, decreased electrostatic potential, lower electron affinity, and higher chemical potential than xanthohumol, metformin, and Palbociclib, indicating greater reactivity and stronger receptor interactions. This integrative in-silico pipeline supports the XN-MT conjugate as a potential candidate for further in-vitro studies and preclinical development in breast cancer therapy targeting CDK4.