Abstract Background: This study aimed to predict the pharmacokinetic and toxicological properties of lead flavonoids from the roots of T. egregia [praecansone A (1) and pongachalcone (2)], and to assess the gastroprotective effects and possible underlying mechanisms of the root extract in mice.Methods: Quantitative and qualitative data for in silico absorption, distribution, metabolism, and excretion (ADME) analyses of the two flavonoids were acquired from the SwissADME database. Toxicity assessment was performed with the ProTox-II server. To evaluate the putative interactions of both flavonoids with opioid receptors and NO protein, we acquired structures of the targets (μ, κ, and δ-opioid receptors, and iNOS) in Homo sapiens from <ext-link ext-link-type="uri" xlink:href="https://www.rcsb.org/">https://www.rcsb.org/</ext-link>. For docking studies, AutoDock 4.2 was used for ligand and target arrangement, and AutoDock Vina was used for calculations. For in vivo assays, mice were pretreated (per os) with T. egregia (2, 20, or 200 mg/kg). After 60 min, 99.9% ethanol (0.2 mL) was injected (per os). At 30 min after ethanol injection, the mice were euthanized, and the gastric damage, gastric levels of hemoglobin, glutathione content, and activity of superoxide dismutase and catalase were evaluated. To elucidate T. egregia mechanisms, we used misoprostol, a prostaglandin analog; indomethacin, an inhibitor of prostaglandin synthesis; L-arginine, an NO precursor; L-NAME, an antagonist of NO synthase; naloxone, an opioid antagonist; and morphine, an opioid agonist.Results: in silico results showed that flavonoids (1) and (2) had favorable ADME properties and toxicity profiles, and exhibited satisfactory binding energies data (below −6.0 kcal/mol) when docked into their targets (μ, κ, and δ-opioid receptors, and iNOS). T. egregia decreased the ethanol-induced gastric damage and hemoglobin levels, and increased the glutathione content, and activity of superoxide dismutase and catalase. Naloxone and L-NAME, but not indomethacin, prevented T. egregia’s effects, thus suggesting that opioid receptors and NO are involved in T. egregia’s efficacy.Conclusions: Flavonoids (1) and (2) exhibited favorable pharmacokinetic properties, showing high lethal dose, 50% (LD₅₀; 3,800 and 2,500 mg/kg, respectively) values. Neither flavonoid was found to be hepatotoxic, carcinogenic, or cytotoxic to human cells. in vivo assays indicated that T. egregia ameliorated oxidative stress levels, and its mechanism is at least partially based on opioid receptors and NO. T. egregia may therefore be considered as a new gastroprotective strategy.