Ispitivanje sekundarnih metabolita i farmakološke aktivnosti odabranih vrsta roda Hieracium L. (Asteraceae) sa Balkanskog poluostrva

Abstract

hibridogenih). Semipreparativnim LC-MS metodama, iz MeOH ekstrakta cvasti H. calophyllum izolovana su četiri seskviterpenska laktona (krepizid E i tri nova jedinjenja: 8-epi-ikserisamin A, kalofilamin A i kalofilamin B), a iz MeOH ekstrakta podzemnih organa H. scheppigianum pet fenolkarboksilnih (mono- ili di-O-kafeoilhina) kiselina (identifikacija na osnovu NMR i MS spektara). Ukupno četiri seskviterpenska laktona, sedam mono- ili di-O-kafeoilhina kiselina i 19 flavonoida (luteolin, apigenin, diosmetin, 15 njihovih heterozida i heterozid kvercetina) kvantifikovani su u suvim MeOH ekstraktima ispitivanih podzemnih organa dve vrste i herbi 28 vrsta LC-MS metodama, i pet triterpena (derivata ursana, oleana i lupana) u suvim CH2Cl2 ekstraktima herbi 28 vrsta GC-FID-MS metodom (identifikacija pomoću standarda, NMR, MS i/ili UV spektara). Hemosistematski značaj jedinjenja identifikovanih u herbama 28 vrsta utvrđen je multivarijantnim statističkim metodama (PCA, nMDS i UPGMA). Suvi MeOH ekstrakti herbi su in vitro pokazali značajan ukupni antioksidantni potencijal i sposobnost neutralizacije slobodnih DPPH•, ABTS•+ i OH• radikala (28 vrsta) i inhibicije acetilholinesteraze (27 vrsta) i butirilholinesteraze (7 vrsta). Među testiranih 12 jedinjenja, najbolju antiholinesteraznu aktivnost ispoljila su tri flavonoidna aglikona i 8-epi-ikserisamin A. Odabrani suvi MeOH i CH2Cl2 ekstrakti herbi šest vrsta pokazali su antimikrobnu aktivnost prema 18, a 8- epi-ikserisamin A i krepizid E prema 4 mikroorganizma. Isti odabrani izolati ispoljili su citotoksični efekat prema tri, odnosno jednoj tumorskoj ćelijskoj liniji, uz zanemarljivu inhibiciju rasta zdravih ćelija. Tri odabrana MeOH i CH2Cl2 ekstrakta su u modelu inflamacije šape pacova izazvane karageninom pokazala antihiperalgezijsku, ali ne i antiedematoznu aktivnost; bezbednost primene dva ekstrakta koji su ispoljili značajnu antihiperalgezijsku aktivnost pokazana je in vivo rotarod i testom akutne toksičnosti. Ključne reči:

nonhybridogenous and 16 hybridogenous). Using semipreparative LC-MS methods, four sesquiterpene lactones (crepiside E and three new compounds: 8-epiixerisamine A, calophyllamine A and calophyllamine B) were isolated from the MeOH extract of H. calophyllum flowering heads, and five phenolic (mono- or di-Ocaffeoylquinic) acids from the MeOH extract of H. scheppigianum underground parts (identification based on NMR and MS spectra). Total of four sesquiterpene lactones, seven mono- or di-O-caffeoylquinic acids and 19 flavonoids (luteolin, apigenin, diosmetin, their 15 glycosides and one quercetin glycoside) were quantified in the dried MeOH extracts of the investigated underground parts of two species and herbs of 28 species, using LC-MS methods, and five triterpenes (ursane, oleane and lupane derivatives) in the dried CH2Cl2 extracts of the herbs of 28 species, using GC-FID-MS method (identification using standards, NMR, MS and/or UV spectra). Chemosystematic significance of the compounds identified in the herbs of 28 species was determined using multivariate statistics (PCA, nMDS and UPGMA). Dried MeOH herb extracts in vitro exhibited significant total antioxidant activity and scavenging of DPPH•, ABTS•+ and OH• radicals (28 species), as well as the ability to inhibit acetylcholinesterase (27 species) and butyrylcholinesterase (7 species). Among 12 tested compounds, three flavonoid aglycones and 8-epiixerisamine A exhibited the highest anticholinesterase activity. Selected dried MeOH and CH2Cl2 herb extracts of six species showed antimicrobial activity against 18, whereas 8-epiixerisamine A and crepiside E against four microorganisms. The same selected isolates exhibited cytotoxic activity against three and one cancer cell lines, respectively, with negligible inhibition of the normal cells growth. In the carrageenan-induced localized inflammation model in rats, three selected MeOH and CH2Cl2 extracts exhibited antihyperalgesic, but not antiedematous activity; safety of the oral administration of two extracts that exhibited significant antihyperalgesic activity was demonstrated in vivo in rotarod test and acute oral toxicity study.