Efficient identification of Acetylcholinesterase and Hyaluronidase inhibitors from Paeonia parnassica extracts through a HeteroCovariance Approach.
Λέξεις-κλειδιά
Αφηρημένη
BACKGROUND
On the basis of the relevant reference in the poem Theriaca of the ancient Greek physician Nicander and its traditional use, Paeonia parnassica was selected for the evaluation of two extracts obtained from the roots and aerial parts to inhibit hydrolytic enzymes involved in snake envenomation. The secondary metabolites which contribute to these activities were detected through a novel HeteroCovariance NMR based approach. Afterwards these ingredients were isolated, identified and evaluated for their inhibitory potency.
OBJECTIVE
The identification of acetylcholinesterase and hyaluronidase inhibitors from Paeonia parnassica extracts was used as a case study for the introduction of a recently developed methodology to evaluate ethnopharmacological data and exploit them for the discovery of bioactive natural compounds. This process is based on the fractionation of the selected extracts and the simultaneous phytochemical analysis and biological assessment of the resulting fractions, which permits the rapid detection of the specified secondary metabolites prior to any laborious and time-consuming purification.
METHODS
The roots and aerial parts of P. parnassica were extracted using methanol:water 50:50 and the two resulted extracts were fractionated by Centrifugal Partition Chromatography. The obtained fractions were evaluated in-vitro for their ability to inhibit acetylcholinesterase and hyaluronidase enzymes and their 1H-NMR spectra were recorded. The biological activity was statistically correlated with the spectral data through the HeteroCovariance Approach (HetCA). Finally the purification, identification and biological evaluation of targeted secondary metabolites were carried out.
RESULTS
The general chemical structures and some explicit secondary metabolites which contribute (e.g. gallotannins, gallic acid derivatives) or not (characteristic "cage-like" monoterpenes of the genus, glycosylated flavonoids) to the anti-acetylcholinesterase and anti-hyaluronidase activities were detected through HetCA. The consequent isolation and biological evaluation of targeted compounds were performed in order to validate the effectiveness and precision of the methodology. This procedure revealed the most active ingredients of both extracts obtained from roots and aerial parts in the above mentioned biological targets, as well as other compounds possessing moderate activity.
CONCLUSIONS
The results of this study contributed to the verification of the ancient text Theriaca regarding the use of Paeonia parnassica to treat the snake bite symptoms. Furthermore, the ingredients of the Paeonia parnassica extracts, which were responsible for their anticholinesterase and anti-hyaluronidase activities, were determined applying a HetCA methodology before their isolation. Therefore, the current work provides clear evidence that HetCA could consist an efficient tool for the exploitation of traditional medicine information in order to discover bioactive natural compounds and develop new pharmacotherapies which serve the needs of contemporary medicine.
