Bio-efficacy of Hedyotis sithiravaraiensis callus extract: A stepwise evaluation through phytochemical profiling, antioxidant activity, molecular docking, and cytotoxicity

Abstract

This study investigates the bioactive potential of Hedyotis sithiravaraiensis S. Muruganandam, Devanath., S. Ravikumar & D. Naras. Sp. nov. callus extract through Phytochemical Profiling, Antioxidant activity, Molecular Docking, ADME analysis, and Cytotoxicity in A549 lung cancer cells. The highest levels of total phenolic content (45.96 ± 0.80 mg GAE/g) and total flavonoid content (45.28 ± 0.53 mg QE/g) were found in 75 % acidified methanol. The DPPH scavenging activity (IC₅₀ = 617.81 ± 0.54 μg/mL) and the ABTS⁺ radical cation scavenging activity (IC₅₀ = 554.41 ± 0.25 μg/mL) both demonstrated that this extract exhibits significant antioxidant activity. Seventeen bioactive components were found in the H. sithiravaraiensis callus extract by GC-MS analysis. Absorption, distribution, metabolism, and excretion studies of 17 compounds also demonstrated favorable drug-like properties and pharmacokinetic characteristics. These bioactive compounds demonstrated significant binding affinities to the KRAS G12C and Bcl-2 proteins, as shown in molecular docking experiments. These affinities were comparable to those of common standard drugs, such as Adagrasib and Obatoclax. The callus extract showed 31.45 ± 0.04 % cytotoxicity in A549 cells at the highest dose, indicating selective cytotoxicity toward lung cancer cells. These findings highlight H. sithiravaraiensis callus extract as a promising anticancer source targeting the MAPK pathway and demonstrate the value of integrated phytochemical and biological analyses in drug discovery and conservation of medicinal plants.