Identification of hydroxyquinazoline alkaloids from Justicia adhatoda L. leaves, a traditional natural remedy with NF- κ B and AP-1-mediated anti-inflammatory properties and antioxidant activity

Ethnopharmacological relevance: Justicia adhatoda L. is used as traditional medicine in Nepal to treat cough, asthma, and inflammatory disorders, and is indicated as "Asuro ". Leaves are used worldwide as herbal medicine due to cardiotonic, expectorant, anti-asthmatic, and bronchodilatory properties. The aim of this work was to study the phytochemical composition of leaves of Nepalese J. adhatoda and assess their anti-inflammatory and antioxidant properties in vitro . Materials and methods: Secondary metabolites were extracted from dried leaves using methanol (JAME: J. adhatoda methanol extract). They were analysed by means of liquid chromatography coupled with multiplestage mass spectrometry (LC-MS n ). Anti-inflammatory potential was determined by the NF- kappa B and AP -1 inhibition assay, and DPPH, ABTS, and beta-carotene bleaching assays were performed to assess its antioxidant properties. Results: JAME is a rich source of secondary metabolites, especially quinazoline alkaloids such as vasicine, vasicinone, vasicoline, and adhatodine. 7-Hydroxy derivatives of peganidine, vasicolinone, and adhatodine were also identified by means of MS n data and are here reported in J. adhatoda for the first time. JAME inhibited NF- kappa B and AP -1 expression in THP-1 cells to a greater extent than the positive control prednisolone. A moderate radicalquenching property was observed in DPPH and ABTS assays, but the anti-carotene bleaching activity was significantly higher than the reference BHT. Conclusions: To the best of our knowledge, this is the first insight into the phytochemical composition of Asuro leaves from Nepal and their bioactivity. Our results will contribute to the valorisation of this medicinal species still widely used in the traditional and complementary medicine.