Chloroform Extraction, Phytochemical Screening, GC-HRMS Analysis and Computational Investigation of Ehretia Laevis Roxb. as Potential MELK Inhibitor for the Treatment of Cancer

Maternal embryonic leucine zipper kinase (MELK), a serine/threonine kinase, plays a vital role in cancer progression by regulating cell proliferation, apoptosis, and stem cell maintenance. It is often overexpressed in various cancers, including breast cancer and glioblastoma, making it an attractive target for cancer therapies. Ehretia laevis Roxb (Boraginaceae), commonly known as Ajan Vruksha, is a renowned medicinal plant in Ayurveda that is traditionally used for treating reproductive and digestive disorders. In this study, we aimed to explore the anticancer potential of Ehretia laevis Roxb against MELK. The chloroform extract was obtained by fractional extraction, followed by fractionation using column chromatography and thin-layer chromatography (TLC). Qualitative chemical analysis was conducted, and the extract was subjected to GC-HRMS analysis to identify potential phytoconstituents. A total of 34 compounds were identified and evaluated through computational analysis of the MELK protein (PDB ID: 5IH9). Several compounds exhibited excellent binding affinities, suggesting their potential as MELK inhibitors. Further in-depth ADMET analysis revealed five promising compounds as potent MELK inhibitors. Among these, Compound 12 (2-Methyl-2-phenyl-5-(1,4-dihydropyridin-4-ylidene)-1,3-dioxan-4,6-dione) demonstrated the highest binding affinity (-8.7 kcal/mol. It forms a stable complex with the target protein, establishing one carbon-hydrogen bond with Glu87 and multiple hydrophobic interactions with Leu27, Ile149, Tyr88, Cys89, Val25, Lys40, Ile17, Ala38, and Pro90. This study highlights the potential of compound 12 as a novel MELK inhibitor, paving the way for further investigation in various in vitro and in vivo models to assess its clinical applicability.