PHOTOCHEMICAL AND ACTIVITY GUIDED ISOLATION STUDIES OF NEPETA BAYTOPII HEDGE ET LAMOND PLANT

Nepeta species are the most important members of the Lamiaceae family, rich iniridoids and secondary metabolites and, have antioxidant, antimicrobial, diuretic, antiasthmatic, antiseptic, analgesic, anticancer, anti-inflammatory, insecticidal properties. Molecules were purified showing antioxidant, enzyme inhibitor, antibacterial, and DNA protection activities by performing activity-directed isolation studies of the aerial part of the endemic Nepeta baytopii Hedge & Lamond collected in Bingöl. The crude extract of the N. baytopii (NB) plant was obtained with a methanol:chloroform (MK) solvent mixture. The this extract applying silica gel column chromatography and first fractions were collected using n-hexane (H), chloroform (K), ethyl acetate (E), and methanol (M) solvents, respectively. Phytochemical analyzes and bioactivities of crude extract and first fractions were determined using spectroscopic techniques. Second fractions were obtained applying column chromatography the first fractions and were applied activity tests. Active fractions were determined by bioactivity guided fractions. Then, the compounds were purified using column chromatography or preparative-HPLC to bioactive fractions, and their molecular structures were elucidated by different NMR techniques. The activity results of the extract, fractions, and pure molecules were evaluated by principal component and SPSS analyses. The crude extract's total phenol and flavonoid amounts were determined as 20.24±0.15 mg GAE/g extract and 12.86±0.05 mg CE/g extract, respectively. It was noted that the main components of the extract were rosmarinic acid, cis-11- eicosanoate, and palmitate. The main component of the NBH fraction is 9,12,15- octadecatrienoic acid, its methyl ester and octadecanoic acid, methyl ester, while the NBK, NBE, and NBM is quercetin-3-glycoside. NBH fractions; showed high reducing power, H2O2 and superoxide anion scavenging, BChE, α-amylase, and tyrosinase inhibition. NBHFr4, NBHFr5, and NBHFr9 are active fractions. NBK fractions; showed effective antioxidant, BChE, and lipase inhibition, and NBKFr5 is the active fraction. NBE fractions; showed active in H2O2 scavenging and metal chelating activities, and NBEFr10 is the active fraction. NBM fractions; showed active reducing power and H2O2 scavenging activities and were recorded as active fractions of NBMFr3, NBMFr4, and NBMFr7. From the active fractions, apigenin, ixoroside, daucosterol, ursolic acid, β-sitosterol, and Molecule L were purified. The molecules' bioactivities, enzyme kinetics, and molecular docking structures were determined. As a result, it was determined that the molecules isolated by bioactivity-directed isolation studies were antioxidants against oxidative damages, regulating the activities of metabolic enzymes and protective molecules against DNA damages.