Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2010 Apr;77(4):559-66.
doi: 10.1124/mol.109.062141. Epub 2010 Jan 11.

Computer-aided discovery, validation, and mechanistic characterization of novel neolignan activators of peroxisome proliferator-activated receptor gamma

Nanang Fakhrudin  1 , Angela Ladurner, Atanas G Atanasov, Elke H Heiss, Lisa Baumgartner, Patrick Markt, Daniela Schuster, Ernst P Ellmerer, Gerhard Wolber, Judith M Rollinger, Hermann Stuppner, Verena M Dirsch
Affiliations

Computer-aided discovery, validation, and mechanistic characterization of novel neolignan activators of peroxisome proliferator-activated receptor gamma

Nanang Fakhrudin et al. Mol Pharmacol. 2010 Apr.

Abstract

Peroxisome proliferator-activated receptor gamma (PPAR gamma) agonists are used for the treatment of type 2 diabetes and metabolic syndrome. However, the currently used PPAR gamma agonists display serious side effects, which has led to a great interest in the discovery of novel ligands with favorable properties. The aim of our study was to identify new PPARgamma agonists by a PPAR gamma pharmacophore-based virtual screening of 3D natural product libraries. This in silico approach led to the identification of several neolignans predicted to bind the receptor ligand binding domain (LBD). To confirm this prediction, the neolignans dieugenol, tetrahydrodieugenol, and magnolol were isolated from the respective natural source or synthesized and subsequently tested for PPAR gamma receptor binding. The neolignans bound to the PPAR gamma LBD with EC(50) values in the nanomolar range, exhibiting a binding pattern highly similar to the clinically used agonist pioglitazone. In intact cells, dieugenol and tetrahydrodieugenol selectively activated human PPAR gamma-mediated, but not human PPAR alpha- or -beta/delta-mediated luciferase reporter expression, with a pattern suggesting partial PPAR gamma agonism. The coactivator recruitment study also demonstrated partial agonism of the tested neolignans. Dieugenol, tetrahydrodieugenol, and magnolol but not the structurally related eugenol induced 3T3-L1 preadipocyte differentiation, confirming effectiveness in a cell model with endogenous PPAR gamma expression. In conclusion, we identified neolignans as novel ligands for PPAR gamma, which exhibited interesting activation profiles, recommending them as potential pharmaceutical leads or dietary supplements.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Chemical structures of the compounds selected for pharmacological investigation.
Fig. 2
Fig. 2
PPARγ ligand binding potential of neolignans. Serial dilutions of the tested compounds were prepared in DMSO and then mixed with a buffer solution containing the hPPARγ LBD tagged with GST, terbiumlabeled anti-GST antibody, and fluorescently labeled PPARγ agonist. After 1 h of incubation, the ability of the test compounds to bind to the PPARγ LBD and thus displace the fluorescently labeled ligand was estimated from the decrease of the emission ratio 520 nm/495 nm upon excitation at 340 nm. Each data point represents the mean ± S.D. from three independent experiments performed in duplicate.
Fig. 3
Fig. 3
Influence of the neolignans on the hPPARγ-mediated reporter gene transactivation. HEK-293 cells, transiently cotransfected with a plasmid encoding full-length hPPARγ, a reporter plasmid containing PPRE coupled to a luciferase reporter, and EGFP as internal control, were stimulated with the indicated concentrations of the respective compounds for 18 h. Luciferase activity was normalized by the EGFP-derived fluorescence, and the result was expressed as fold induction compared with the negative control (DMSO vehicle treatment). The data shown are means ± S.D. of three independent experiments each performed in quadruplet.
Fig. 4
Fig. 4
Influence of neolignans on PPARγ coactivator recruitment. The ability of the hPPARγ-ligand complex formed with the test compounds to recruit the TRAP220/DRIP-2 coactivator peptide was measured as described in detail under Materials and Methods. Serial dilutions of the tested compounds were prepared in DMSO and then mixed with a buffer solution containing the hPPARγ LBD tagged with GST, terbium-labeled anti-GST antibody, and fluorescein-labeled TRAP220/DRIP-2 coactivator peptide. After incubation for 1 h, the emission at 520 and 495 nm after excitation at 340 nm was measured, and the 520 nm/495 nm ratio was used as a measure for the TRAP220/DRIP-2 coactivator recruitment potential of the respective compounds. Each data point represents the mean ± S.D. from three independent experiments performed in duplicate.
Fig. 5
Fig. 5
Putative interactions between the hPPARγ binding pocket and the neolignans 1 (A), 2 (B), and 3 (C). The docking results were visualized using the LigandScout software with the following color code: hydrogen bond acceptor (red arrow), hydrogen bond donor (green arrow), hydrophobic interaction (yellow sphere) and aromatic interaction (blue rings). The ligand binding pocket was depicted as surface colored based on the hydrophilicity/lipophilicity.
Fig. 6
Fig. 6
Adipogenic activity of compounds 1 to 4. A, 3T3-L1 preadipocytes were differentiated to adipocytes as described in the Materials and Methods section. After 7 to 8 days of differentiation with the indicated test compounds (1 μM rosiglitazone, 50 μM BADGE, and 10 μM neolignans, respectively), Oil Red O staining was performed to clearly visualize the accumulated lipids. Representative photos of one experiment of three with consistent results are depicted. B, to get a quantitative measure, the dye accumulated in the cells (treated as described under A) was solubilized by 100% isopropanol and photometrically quantified at 550 nm. The data shown are means ± S.D. from three independent experiments. *, p < 0.05; ***, p < 0.001, as estimated by two-tailed paired t test.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Anghel SI, Wahli W. Fat poetry: a kingdom for PPAR gamma. Cell Res. 2007;17:486–511. - PubMed
    1. Baños G, Pérez-Torres I, El Hafidi M. Medicinal agents in the metabolic syndrome. Cardiovasc Hematol Agents Med Chem. 2008;6:237–252. - PubMed
    1. Bardot O, Aldridge TC, Latruffe N, Green S. PPAR-RXR heterodimer activates a peroxisome proliferator response element upstream of the bifunctional enzyme gene. Biochem Biophys Res Commun. 1993;192:37–45. - PubMed
    1. Bedu E, Wahli W, Desvergne B. Peroxisome proliferator-activated receptor beta/delta as a therapeutic target for metabolic diseases. Expert Opin Ther Targets. 2005;9:861–873. - PubMed
    1. Bensky D, Clavey S, Stöger E. Chinese Herbal Medicine–Materia Medica. Eastland Press, Inc.; Seattle, WA: 2004.

Publication types

LinkOut - more resources