The main purpose of this work is to investigate the phytochemical composition of Marrubium alysson L. non-polar fraction. GC/MS analysis was used to evaluate the plant extract’s saponifiable and unsaponifiable matter. Although M. alysson L. lipoidal matter saponification produced 30.3% of fatty acid methyl esters and 69.7% of unsaponifiable matter. Phytol was the most dominant substance in the unsaponifiable materials. Notably, marrubiin which is one of the most prominent metabolites of Marrubium alysson L. was not detected through our adopted GC/MS technique. Thus, further characterization was proceeded through simple and rapid HPTLC analysis which successfully managed to identify marrubiin. Based on the regression equation, the concentration of marrubiin in M. alysson L. extract was 14.09 mg/g of dry extract. Concerning acetylcholinesterase inhibitory activity, both the crude M. alysson L. total methanolic extract and the non-polar fraction displayed reasonable inhibitory activity against acetylcholinesterase (AChE), whereas the pure compound marrubiin was considered to be the most effective and potent AChE inhibitor, with an IC₅₀ value of 52.66 (µM). According to the molecular docking studies, potential sites of interaction between the pure chemical marrubiin and AChE were examined. The results show that Tyr124 on AChE residue was critical to the activity of the aforementioned drug. Based on the depicted marrubin AChE inhibition activity and reported safety profile, this chemical metabolite is considered as a promising lead compound for further pre-clinical investigation as well as drug development and optimization.

Herbal medicine is considered a natural source of a variety of beneficial active constituents. Since ancient times, herbal medicines have been used to cure different types of diseases. The World Health Organization (WHO) reports that the usage of herbal medicines has surpassed that of conventional pharmaceuticals by a factor of two to three worldwide due to it showed a remarkable safety profile when compared to conventional medicine. The use of herbal plants for primary healthcare is expanding tremendously. This isn't just because they are less expensive, but also because they are more widely accepted culturally, work better with the human body, and have less adverse effects. The process of extracting medicinal plants involves separating active plant extracts or secondary metabolites using the proper solvent and various extraction techniques. The Labiatae (Lamiaceae) family is one of the largest and most distinctive families of flowering plants, with about 220 genera and nearly 4000 species worldwide. Genius Marrubium consists of 97 flowering plants belonging to the Labiatae family that are native to the temperate regions of Europe, North Africa and Asia. Some other species grow in North and South America. This genus also contains a wide range of compounds including terpenes, oils, iridoids, sterols, glycosides, saponins, phenolic compounds and flavonoids. The species of this genus are characterized through having potential therapeutic activities as antispasmodic, hypolipidemic, hypotensive, hypoglycemic, anti-inflammatory and analgesic properties. This chemical review shows genus Marrubium plants extract contains different classes of bioactive chemical classes as terpenes, flavonoids, essential oil and sterols.