title: Bioactivity guided fractionation and hypolipidemic property of a novel HMG-CoA reductase inhibitor from Ficus virens Ait.
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.
authors.affiliation: Department of Biosciences, Clinical Biochemistry & Natural Product Research Lab, Integral University, Lucknow, 226026, India. danishik786@gmail.com.
authors.forename: Danish
authors.initials: D
authors.lastname: Iqbal
authors.affiliation: Department of Biosciences, Clinical Biochemistry & Natural Product Research Lab, Integral University, Lucknow, 226026, India. contactskhan@gmail.com.
authors.forename: M Salman
authors.initials: MS
authors.lastname: Khan
authors.affiliation: Department of Biosciences, Clinical Biochemistry & Natural Product Research Lab, Integral University, Lucknow, 226026, India. sajid_987@rediffmail.com.
authors.forename: Mohd Sajid
authors.initials: MS
authors.lastname: Khan
authors.affiliation: Department of Biosciences, Clinical Biochemistry & Natural Product Research Lab, Integral University, Lucknow, 226026, India. ahmadsaheem@gmail.com.
authors.forename: Saheem
authors.initials: S
authors.lastname: Ahmad
authors.affiliation: Department of Pharmacy, Integral University, Lucknow, 226026, India. sarfarajpharma@gmail.com.
authors.forename: Md Sarfaraj
authors.initials: MS
authors.lastname: Hussain
authors.affiliation: Department of Pharmacognosy & Phytochemistry, Faculty of Pharmacy, Jamia Hamdard, New Delhi, 110062, India. maliphyto@gmail.com.
authors.forename: Mohd
authors.initials: M
authors.lastname: Ali
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000302:isolation & purification / Q000494:pharmacology; D005591:Chemical Fractionation; D008076:Cholesterol, HDL / Q000097:blood; D008078:Cholesterol, LDL / Q000097:blood; D004187:Disaccharides / Q000302:isolation & purification / Q000494:pharmacology*; D030681:Ficus / Q000737:chemistry*; D006017:Glycolipids / Q000302:isolation & purification / Q000494:pharmacology*; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000302:isolation & purification / Q000494:pharmacology*; D006949:Hyperlipidemias / Q000188:drug therapy; D000960:Hypolipidemic Agents / Q000302:isolation & purification / Q000494:pharmacology*; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D008297:Male; D024301:Plant Bark / Q000737:chemistry; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014280:Triglycerides / Q000097:blood
chemical_list: D000975:Antioxidants; D008076:Cholesterol, HDL; D008078:Cholesterol, LDL; D004187:Disaccharides; D006017:Glycolipids; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D000960:Hypolipidemic Agents; D010936:Plant Extracts; D014280:Triglycerides; C000604280:n-octadecanyl-O-glucopyranosyl-(6'-1')-O-glucopyranoside; D006903:Hydroxymethylglutaryl CoA Reductases