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Bioactivity guided fractionation and hypolipidemic property of a novel HMG-CoA reductase inhibitor from Ficus virens Ait.

Iqbal D, Khan MS, Khan MS, Ahmad S, Hussain MS, Ali M - Lipids Health Dis (2015)

Bottom Line: The current perspective for the search of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitor has been shifted towards a natural agent also having antioxidant property.This inhibitor exhibited not only higher free radical scavenging activity but also marked HMG-CoA reductase inhibitory activity with an IC50 value of 84±2.8 ng/ml.The in vivo study revealed that administration of FVBM extract (at higher dose, 100 mg/rat) and the inhibitor (1 mg/rat) to Triton WR-1339-induced hyperlipidemic rats significantly ameliorated the altered levels of plasma lipids and lipoproteins including hepatic HMG-CoA reductase activity; this effect was comparable to the effect of standard drug atorvastatin.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosciences, Clinical Biochemistry & Natural Product Research Lab, Integral University, Lucknow, 226026, India. danishik786@gmail.com.

ABSTRACT

Background: The current perspective for the search of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitor has been shifted towards a natural agent also having antioxidant property. Thus, this study was intended to isolate and identify the bioactive compounds from methanolic extract of Ficus virens bark (FVBM) and to evaluate their antioxidant, HMG-CoA reductase inhibitory and hypolipidemic activity.

Methods: Bioactivity guided fractionation and isolation of bioactive compound from FVBM extract has been done to isolate and characterize the potent HMG-CoA reductase (HMGR) inhibitor with antioxidant activity by using repeated extensive column chromatography followed by spectroscopic methods, including Infrared (IR), 1H & 13C nuclear magnetic resonance (NMR) and Mass spectrum analysis. The in vitro HMGR inhibition and enzyme kinetic assay was determined using HMG-CoA as substrate. In addition, antioxidant activity of the new isolated compound, was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and FRAP value. In-silico molecular informatics of HMGR enzyme type inhibition and pharmacokinetics data of the new compound was further evaluated through molecular docking and ADME-T studies. Further, in-vivo hypolipidemic property of FVBM extract and newly isolated compound was also analyzed in triton-WR 1339 induced rats.

Results: Thereby, we report the discovery of n-Octadecanyl-O-α-D-glucopyranosyl(6'→1″)-O-α-D-glucopyranoside (F18) as a novel HMG-CoA reductase inhibitor with strong antioxidant property. This inhibitor exhibited not only higher free radical scavenging activity but also marked HMG-CoA reductase inhibitory activity with an IC50 value of 84±2.8 ng/ml. This inhibitory activity concurred with kinetic study that showed inhibition constant (K i) of 84 ng/ml via an uncompetitive mode of inhibition. The inhibition was also corroborated by molecular docking analysis and in silico pharmacokinetics data. The in vivo study revealed that administration of FVBM extract (at higher dose, 100 mg/rat) and the inhibitor (1 mg/rat) to Triton WR-1339-induced hyperlipidemic rats significantly ameliorated the altered levels of plasma lipids and lipoproteins including hepatic HMG-CoA reductase activity; this effect was comparable to the effect of standard drug atorvastatin.

Conclusions: The in vitro, in silico and in vivo results clearly demonstrated the antioxidant potential and therapeutic efficacy of the inhibitor as an alternate drug against hyperlipidemia.

No MeSH data available.


Related in: MedlinePlus

Binding pattern studies of standard (a) and n-Octadecanyl-O-α-D-glucopyranosyl (6′ → 1″)-O-α-D-glucopyranoside (b) with HMG-CoA reductase.
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Fig7: Binding pattern studies of standard (a) and n-Octadecanyl-O-α-D-glucopyranosyl (6′ → 1″)-O-α-D-glucopyranoside (b) with HMG-CoA reductase.

Mentions: To further corroborate our findings, the molecular interaction studies were carried out to find out the plausible ligand-receptor interactions between the inhibitor and HMG-CoA reductase. The docked structures of pravastatin-HMG-CoA reductase were found to be surrounded by Ser684, Asp690, Arg590, Lys735 and Glu559 amino acid residues, which play important role in stabilizing the complex, whereas n-Octadecanyl-O-α-D-glucopyranosyl(6′ → 1″)-O-α-D-glucopyranoside-HMG-CoA reductase complex involved contributions from residues Asn755, Asp765, Lys691, Glu559, Gly560, Ala525, Arg590, which are not from the catalytic domain of HMG-CoA reductase. This observation indicated an uncompetitive mode of inhibition. The non-catalytic site accommodated the compound (Table 3) with a dominant nonelectrostatic energy contribution. The inhibitor showed binding energy of–5.58 Kcal/mol indicating high affinity for the binding site, though pravastatin was a more stronger ligand (ΔG:–6.25 kcal/mol) (Figure 7 and Table 3).Table 3


Bioactivity guided fractionation and hypolipidemic property of a novel HMG-CoA reductase inhibitor from Ficus virens Ait.

Iqbal D, Khan MS, Khan MS, Ahmad S, Hussain MS, Ali M - Lipids Health Dis (2015)

Binding pattern studies of standard (a) and n-Octadecanyl-O-α-D-glucopyranosyl (6′ → 1″)-O-α-D-glucopyranoside (b) with HMG-CoA reductase.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4352280&req=5

Fig7: Binding pattern studies of standard (a) and n-Octadecanyl-O-α-D-glucopyranosyl (6′ → 1″)-O-α-D-glucopyranoside (b) with HMG-CoA reductase.
Mentions: To further corroborate our findings, the molecular interaction studies were carried out to find out the plausible ligand-receptor interactions between the inhibitor and HMG-CoA reductase. The docked structures of pravastatin-HMG-CoA reductase were found to be surrounded by Ser684, Asp690, Arg590, Lys735 and Glu559 amino acid residues, which play important role in stabilizing the complex, whereas n-Octadecanyl-O-α-D-glucopyranosyl(6′ → 1″)-O-α-D-glucopyranoside-HMG-CoA reductase complex involved contributions from residues Asn755, Asp765, Lys691, Glu559, Gly560, Ala525, Arg590, which are not from the catalytic domain of HMG-CoA reductase. This observation indicated an uncompetitive mode of inhibition. The non-catalytic site accommodated the compound (Table 3) with a dominant nonelectrostatic energy contribution. The inhibitor showed binding energy of–5.58 Kcal/mol indicating high affinity for the binding site, though pravastatin was a more stronger ligand (ΔG:–6.25 kcal/mol) (Figure 7 and Table 3).Table 3

Bottom Line: The current perspective for the search of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitor has been shifted towards a natural agent also having antioxidant property.This inhibitor exhibited not only higher free radical scavenging activity but also marked HMG-CoA reductase inhibitory activity with an IC50 value of 84±2.8 ng/ml.The in vivo study revealed that administration of FVBM extract (at higher dose, 100 mg/rat) and the inhibitor (1 mg/rat) to Triton WR-1339-induced hyperlipidemic rats significantly ameliorated the altered levels of plasma lipids and lipoproteins including hepatic HMG-CoA reductase activity; this effect was comparable to the effect of standard drug atorvastatin.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosciences, Clinical Biochemistry & Natural Product Research Lab, Integral University, Lucknow, 226026, India. danishik786@gmail.com.

ABSTRACT

Background: The current perspective for the search of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase inhibitor has been shifted towards a natural agent also having antioxidant property. Thus, this study was intended to isolate and identify the bioactive compounds from methanolic extract of Ficus virens bark (FVBM) and to evaluate their antioxidant, HMG-CoA reductase inhibitory and hypolipidemic activity.

Methods: Bioactivity guided fractionation and isolation of bioactive compound from FVBM extract has been done to isolate and characterize the potent HMG-CoA reductase (HMGR) inhibitor with antioxidant activity by using repeated extensive column chromatography followed by spectroscopic methods, including Infrared (IR), 1H & 13C nuclear magnetic resonance (NMR) and Mass spectrum analysis. The in vitro HMGR inhibition and enzyme kinetic assay was determined using HMG-CoA as substrate. In addition, antioxidant activity of the new isolated compound, was measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and FRAP value. In-silico molecular informatics of HMGR enzyme type inhibition and pharmacokinetics data of the new compound was further evaluated through molecular docking and ADME-T studies. Further, in-vivo hypolipidemic property of FVBM extract and newly isolated compound was also analyzed in triton-WR 1339 induced rats.

Results: Thereby, we report the discovery of n-Octadecanyl-O-α-D-glucopyranosyl(6'→1″)-O-α-D-glucopyranoside (F18) as a novel HMG-CoA reductase inhibitor with strong antioxidant property. This inhibitor exhibited not only higher free radical scavenging activity but also marked HMG-CoA reductase inhibitory activity with an IC50 value of 84±2.8 ng/ml. This inhibitory activity concurred with kinetic study that showed inhibition constant (K i) of 84 ng/ml via an uncompetitive mode of inhibition. The inhibition was also corroborated by molecular docking analysis and in silico pharmacokinetics data. The in vivo study revealed that administration of FVBM extract (at higher dose, 100 mg/rat) and the inhibitor (1 mg/rat) to Triton WR-1339-induced hyperlipidemic rats significantly ameliorated the altered levels of plasma lipids and lipoproteins including hepatic HMG-CoA reductase activity; this effect was comparable to the effect of standard drug atorvastatin.

Conclusions: The in vitro, in silico and in vivo results clearly demonstrated the antioxidant potential and therapeutic efficacy of the inhibitor as an alternate drug against hyperlipidemia.

No MeSH data available.


Related in: MedlinePlus