The gram-negative, spiral-shaped bacterium Helicobacter pylori mostly inhabit the stomach of humans. It is linked to several digestive disorders, as peptic ulcers, chronic gastritis and has been identified as a major contributor to gastric cancer. Phytotherapy has been studied as potential alternative therapies for infections caused by H. pylori due to drug resistance and side effects. Laurus nobilis (bay leaf) has been traditionally utilized in culinary and medicinal applications due to its various health-promoting properties. This study aimed to explore the phytochemical profile of L. nobilis leaves using UPLC-ESI-MS/MS and assess its antioxidant, anti-Helicobacter pylori (H. pylori), and cytotoxic activities. The total phenolic and flavonoid contents of the methanol extract were measured, followed by antioxidant evaluation through DPPH and ABTS assays. The anti-H. pylori effect was tested via the micro-well dilution method, while the cytotoxicity was evaluated through the MTT assay on HepG2 and Caco2 cell lines. The methanol extract exhibited high phenolic content (91 ± 1.22 mg GAE/g), strong antioxidant activity (DPPH IC50 = 6.35 µg/ml), and moderate antibacterial activity against H. pylori (MIC = 7.81 µg/ml). The extract also showed significant cytotoxic activity against HepG2 and Caco2 cells, with IC50 values of 50.21 µg/ml and 61.17 µg/ml, respectively. These findings suggest that L. nobilis extract could act as a promising resource of natural anticancer and antioxidants compounds, as well as a complementary treatment for H. pylori infections.