In vitro anti-inflammatory and in silico anti-viral assessment of phytoconstituents in polyherbal Ayurvedic formulation ‘Arogyamrita Kwath’

Abstract

Background

Arogyamrita Kwath (AMK) is a polyherbal decoction comprising ten medicinal plants, viz., Albizia lebbeck, Andrographis paniculata, Tinospora cordifolia, Adhatoda vasica, Solanum xanthocarpum, Curcuma longa, Glycyrrhiza glabra, Terminalia bellirica, Withania somnifera and Trachyspermum ammi. The plants of the AMK formulation are traditionally used for the treatment of inflammation and respiratory ailments, but no scientific evidence has been reported so far for this formulation.

Objectives

To evaluate anti-inflammatory activity of AMK formulation in vitro and its fractions and to predict in silico anti-viral activity of identified potential phytoconstituents.

Materials and methods

The MTT cell cytotoxicity assay, nitric oxide (NO) inhibition assay and cytokines assay were carried out at concentrations 100 and 200 μg/mL. The phytoconstituents were identified by UPLC-PDA and UPLC-HRMS analyses. For pharmacoinformatics study molecular docking and molecular dynamics methods were used.

Results

The study revealed that AMK significantly inhibited NO in comparison to dexamethasone (100 μg/mL) and pro-inflammatory cytokines in RAW264.7 cells. The three fractions, n-hexane, EtOAc and n-BuOH prepared from the AMK formulation were non-cytotoxic against RAW264.7 murine macrophage cells during MTT cytotoxicity assay and showed satisfactory results during cytokines assay. Ethyl acetate fraction contains active phytoconstituents in appreciable quantities. 16 phytoconstituents have been identified by UPLC-HRMS analysis in the formulation and four phytocompounds were quantified by UPLC-PDA. Molecular dynamics study helped in identifying two macromolecular targets (viral replicase and the membrane protein), which are relatively more important.

Conclusion

In the present study, anti-inflammatory activity of AMK was evaluated and the claimed anti-viral property was re-confirmed by molecular modelling in this work. The results clearly established that AMK showed remarkable anti-inflammatory and anti-viral activities.