Identification of three coagulins as MMP inhibitors from Withania coagulans Dunal fruits

Abstract

Introduction and Aim: W. coagulans Dunal has been reported to contain an array of bioactive compounds. The present investigation was carried out to investigate MMP inhibitory molecules from W. coagulans Dunal fruit.   Materials and Methods: Isolation of active principle compounds and anti-ChC activity from methanol extract of W. coagulans Dunal fruit were carried out using chromatographic techniques and dot-blot assay on X-ray film. Active principle compounds were identified by ESI MS, 1H, and 13C NMR spectroscopy. The Bioefficacy of compounds was assessed by in vitro solution assay and gelatin zymography.   Results: Dot blot assay of methanol extract of W. coagulans Dunal fruit exhibited inhibitory activity against ChC (Clostridium histolyticum collagenase). Bioactivity assay guided chromatographic fractionation revealed the presence of five compounds out of which three were tentatively identified as Coagulin-H, Coagulin-L, and Coagulin-O by ESI MS, 1H, and 13C NMR spectroscopy. In vitro solution assay revealed Coagulin-H exhibits the highest MMP-2 and MMP-9  inhibitory activity. Coagulin-L and Coagulin-O exhibit 80.57 ± 2.1 % and 70.96 ± 2.8 % inhibitor activities against MMP-2 and 78.94 ± 3.6% and 63.15 ± 3.8% inhibitor activities against MMP-9 at 150 µg/ml respectively. In gel inhibition assay by gelatin zymography revealed that crude extracted residue of fruit exhibits dose-dependent inhibitory potential against MMPs of NIH3T3 fibroblast and HeLa cervical cells.   Conclusion: Our results advocate the anti-MMP potential of purified Coagulin-H, Coagulin-L, and Coagulin-O of W. coagulans Dunal fruits. These bioactive principles could be further investigated in detail for anti-cancer properties.