A Biochemometric Approach for the Identification of In Vitro Anti-Inflammatory Constituents in Masterwort

Abstract

Peucedanum ostruthium (L.) Koch, commonly known as masterwort, has a longstanding history as herbal remedy in the Alpine region of Austria, where the roots and rhizomes are traditionally used to treat disorders of the gastrointestinal and respiratory tract. Based on a significant NF-κB inhibitory activity of a P. ostruthium extract (PO-E), this study aimed to decipher those constituents contributing to the observed activity using a recently developed biochemometric approach named ELINA (Eliciting Nature’s Activities). This -omics tool relies on a deconvolution of the multicomponent mixture, which was employed by generating microfractions with quantitative variances of constituents over several consecutive fractions. Using an optimized and single high-performance counter-current chromatographic (HPCCC) fractionation step 31 microfractions of PO-E were obtained. 1H NMR data and bioactivity data from three in vitro cell-based assays, i.e., an NF-ĸB reporter-gene assay and two NF-κB target-gene assays (addressing the endothelial adhesion molecules E-selectin and VCAM-1) were collected for all microfractions. Applying heterocovariance analyses (HetCA) and statistical total correlation spectroscopy (STOCSY), quantitative variances of 1H NMR signals of neighboring fractions and their bioactivities were correlated. This revealed distinct chemical features crucial for the observed activities. Complemented by LC-MS-CAD data this biochemometric approach differentiated between active and inactive constituents of the complex mixture, which was confirmed by NF-κB reporter-gene testing of the isolates. In this way, four furanocoumarins (imperatorin, ostruthol, saxalin, and 2’-O-acetyloxypeucedanin), one coumarin (ostruthin), and one chromone (peucenin) were identified as NF-κB inhibiting constituents of PO-E contributing to the observed NF-ĸB inhibitory activity. Additionally, this approach also enabled the disclose of synergistic effects of the PO-E metabolites imperatorin and peucenin. In sum, prior to any isolation an early identification of even minor active constituents, e.g. peucenin and saxalin, ELINA enables the targeted isolation of bioactive constituents and, thus, to effectively accelerate the NP-based drug discovery process.

A metabolomic approach to target antimalarial metabolites in the Artemisia annua fungal endophytes

“…Similar biochemometrics approach has been extensively applied in natural products research as a preliminary evaluation step in the process of drug discovery from natural sources 31 – 34 . Hence, further integration with in silico approaches like neural network-based virtual screening that was applied in this investigation, can increase the success rate of targeting the bioactive metabolites in complex crude extracts.…”

“…As a result of the previous extensive cheminformatic investigation, both physcion (1) and emodin (2) were validated this approach where they inhibited the in vitro plasmodial growth with IC 50 values of 0.9 and 1.9 µM, respectively. Similar biochemometrics approach has been extensively applied in natural products research as a preliminary evaluation step in the process of drug discovery from natural sources [31][32][33][34] . Hence, further integration with in silico approaches like neural network-based virtual screening that was applied in this investigation, can increase the success rate of targeting the bioactive metabolites in complex crude extracts.…”

Peucedanum ostruthium Inhibits E-Selectin and VCAM-1 Expression in Endothelial Cells through Interference with NF-κB Signaling

“…Only recently, we identified and isolated several bioactive compounds from PO by applying a novel biochemometric approach named Eliciting Nature’s Activities (ELINA; [ 18 , 130 ]) with an NF-ĸB reporter-gene assay plus two assays for E-selectin and VCAM-1 as biological read-out. However, the anti-inflammatory activity could only partially recovered (as compared to the total extract) by the combination of imperatorin and peucenin as a mixture, suggesting additional combinatorial effects [ 18 ].…”

Conservation Gaps in Traditional Vegetables Native to Europe and Fennoscandia

“…Some details on a few of the species: master-wort (belonging to the Apiaceae) has been cultivated in Fennoscandia and used to flavor beverages or for medicinal purposes [66]. Recent research has identified bioactive constituents as different furanocoumarins [67]. The plant is said to have been introduced to Fennoscandia in mediaeval time.…”