From Molecular Design to In Vivo Validation: Integrated Efficacy and Safety Profiling of Orotic Acid–1,2,4-Triazole Hybrids with Wound-Healing Activity

Chronic and difficult-to-heal wounds require topical agents that combine reparative activity with favorable local disposition and an acceptable preliminary safety profile. This study evaluated three orotic acid–1,2,4-triazole hybrids (1–3) using an integrated computational and experimental workflow. Accelerated weight histogram simulations were performed in an atomistic human stratum corneum lipid model; acute toxicity was assessed by in silico rat LD50 prediction, 96 h Danio rerio LC50 testing, and OECD 425 oral testing in rats. Compounds selected on the basis of toxicity were incorporated into electrospun polyvinylpyrrolidone K-30 dressings and evaluated in a rat excisional wound model. Compound 1 showed pronounced lipid partitioning and potential intralamellar retention, whereas compounds 2 and 3 encountered broad membrane-interior free-energy barriers. Compound 2 had the least favorable acute-toxicity profile and was excluded. By day 8, wound area decreased by 89.0% with compound 1 and 78.6% with compound 3, versus 70.5% with the drug-free vehicle dressing (p < 0.001 for both comparisons). Neither treatment caused clinically evident deterioration or a consistent biochemical pattern of marked hepatic or renal dysfunction, although hematological and compound-specific biochemical changes were observed. The compound 1-loaded dressing therefore warrants further mechanistic, histological, and longer-term safety evaluation as the lead formulation.