Identification of a Kavain Analog with Efficient Anti-inflammatory Effects

Olivier Huck,Hannah Mulhall,James Panek,Xiaxian Han,Bin Cai,Salomon Amar,Radha Iyer,Iryna Gumenchuk

doi:10.1038/s41598-019-49383-8

Abstract

Kavain, a compound derived from Piper methysticum, has demonstrated anti-inflammatory properties. To optimize its drug properties, identification and development of new kavain-derived compounds was undertaken. A focused library of analogs was synthesized and their effects on Porphyromonas gingivalis (P. gingivalis) elicited inflammation were evaluated in vitro and in vivo. The library contained cyclohexenones (5,5-dimethyl substituted cyclohexenones) substituted with a benzoate derivative at the 3-position of the cyclohexanone. The most promising analog identifed was a methylated derivative of kavain, Kava-205Me (5,5-dimethyl-3-oxocyclohex-1-en-1-yl 4-methylbenzoate.) In an in vitro assay of anti-inflammatory effects, murine macrophages (BMM) and THP-1 cells were infected with P. gingivalis (MOI = 20:1) and a panel of cytokines were measured. Both cell types treated with Kava-205Me (10 to 200 μg/ml) showed significantly and dose-dependently reduced TNF-α secretion induced by P. gingivalis. In BMM, Kava-205Me also reduced secretion of other cytokines involved in the early phase of inflammation, including IL-12, eotaxin, RANTES, IL-10 and interferon-γ (p < 0.05). In vivo, in an acute model of P. gingivalis-induced calvarial destruction, administration of Kava-205Me significantly improved the rate of healing associated with reduced soft tissue inflammation and osteoclast activation. In an infective arthritis murine model induced by injection of collagen-antibody (ArthriomAb) + P. gingivalis, administration of Kava-205Me was able to reduce efficiently paw swelling and joint destruction. These results highlight the strong anti-inflammatory properties of Kava-205Me and strengthen the interest of testing such compounds in the management of P. gingivalis elicited inflammation, especially in the management of periodontitis.

Highlights

Periodontitis is an inflammatory disease leading to the destruction of tooth-supporting tissues, that may be induced by dysbiosis[1]
In macrophages infected with P. gingivalis, the TNF-α decrease observed after treatment with this compound was associated with a reduced activation of Toll-like receptor (TLR)-related pathways and lipopolysaccharide-induced TNF factor (LITAF)[18]
To identify compounds with strong anti-inflammatory properties, kavain analogs were tested in a screening assay based on TNF-α inhibition in bone marrow macrophage (BMM) infected with P. gingivalis (Fig. 1B)

Summary

Introduction

Periodontitis is an inflammatory disease leading to the destruction of tooth-supporting tissues, that may be induced by dysbiosis[1]. A compound extracted from the Piper methysticum plant, has recently been described as a promising anti-inflammatory agent and has been evaluated in several in vitro and animal models[12,13,14,15]. Kava-241, is a synthesized kavain analog that has already demonstrated anti-TNF-α properties and the ability to reduce both P. gingivalis-induced periodontitis and arthritis[17,18]. In macrophages infected with P. gingivalis, the TNF-α decrease observed after treatment with this compound was associated with a reduced activation of Toll-like receptor (TLR)-related pathways and lipopolysaccharide-induced TNF factor (LITAF)[18]. The present study aimed to identify additional new kavain-derived analogs and to evaluate their anti-inflammatory properties in vitro and in vivo. The most promising analog identified was Kava-205Me, a methylated kavain analog

Methods

Results

Conclusion