HPLC-LTQ orbitrap mass spectrometry-based molecular networking for identifying anthelmintic molecules in Morinda lucida Benth

Morinda lucida Benth (Rubiaceae) is a medicinal plant traditionally used to treat malaria and digestive parasitosis. In vitro pharmacological studies have shown that extracts of the plant were very active against nematodes. The aim of our work was to identify the compounds responsible for the anthelmintic activity of M. lucida. To achieve this, the leaf MeOH extract was fractionated and the anthelmintic activity of the fractions was evaluated on Haemonchus contortus and Caenorhabditis elegans and their cytotoxicity was evaluated on WI38 cells. The fractions were analyzed by HPLC-HRMS/MS and manual dereplication was coupled to molecular networking to identify the major compounds in the most active fractions. The results showed that three fractions, F8, F11 and F13 were the most active on both nematode models and less cytotoxic. In total, sixteen different compounds were putatively identified in these three fractions, 9 in F8, 12 in F11 and 12 in F13, as some are present in different fractions. Among these compounds, ten were identified for the first time in M. lucida. These are mainly iridoids including loganic acid (2) and isomer (4), methoxygaertneroside or isomer (14), dehydromethoxygaertneroside or isomer (18), borreriagenin (3) and isomer (5). We also identified flavonoids including rutin pentoside (10), kaempferol-3-O-rutinoside pentoside (12), and kaempferol-3-O-rutinoside (15). The Bioactivity-Based approach identified loganic acid or isomer I (2), asperulosidic acid or isomer (6), methoxygaertneroside or isomer (14), and kaempferol-3-O-rutinoside or isomer (15) as potential compounds responsible for the anthelmintic activity of the three fractions. However, it seems necessary to quantify them and evaluate the anthelmintic activity of each of these compounds in order to confirm the results of the Bioactivity-Based approach and to identify potential new anthelmintic molecules to prevent resistance development. Furthermore, possible synergy between them has also to be analyzed.& COPY; 2023 Published by Elsevier B.V. on behalf of SAAB.