Enhanced lung cancer cell inhibition using thymol nano-formulation: Design, physicochemical profiling, cytotoxic assessment, and computational interaction studies.

Lung cancer is one of the most prevalent and deadly malignancies globally, characterized by high mortality rates, frequent metastasis, and the development of resistance to conventional therapies. These challenges underscore the urgent need for novel and effective treatment strategies. Utilizing nano-formulation to enhance the delivery of hydrophobic, natural bioactive compounds is a promising approach to improve their aqueous solubility, bioavailability, and targeted therapeutic efficacy. This study developed a stable thymol nano-formulation (ThNF) using Tween-80, yielding monodisperse particles of 22.64 ± 1.47 nm with excellent long-term stability. The ThNF exhibited significantly enhanced in vitro anticancer activity against A549 lung carcinoma cells compared to free thymol, demonstrated by a lower IC50 value (38.87 μM vs. 53.59 μM), stronger time- and dose-dependent cytotoxicity, and a superior ability to inhibit metastatic behaviours like cell migration and clonogenic survival. Mechanistic investigation through computational studies revealed that thymol has a strong binding affinity for two established and effective targets in lung cancer pathogenesis: PKBα/Akt and VPS34/PI3K, with binding energies of -6.0 and -6.2 kcal/mol, respectively. Molecular dynamics simulations and MM-PBSA free energy calculations confirmed the stability of these complexes, which are primarily stabilized by van der Waals interactions. These findings collectively demonstrate that the nano-formulation strategy successfully elavates the anticancer activity of thymol, and its interaction with key oncogenic proteins PKBα and VPS34 provides preliminary evidence for its dual-target mechanism for lung cancer therapy.