Competitive protein tyrosine phosphatase 1B (PTP1B) inhibitors, prenylated caged xanthones from Garcinia hanburyi and their inhibitory mechanism

Protein tyrosine phosphatase 1B (PTP1B) plays important role in diabetes, obesity and cancer. The methanol extract of the gum resin of Garcinia hanburyi (G. hanburyi) showed potent PTP1B inhibition at 10µg/ml. The active compounds were identified as prenylated caged xanthones (1-9) which inhibited PTP1B in dose-dependent manner. Carboxybutenyl group within caged motif (A ring) was found to play a critical role in enzyme inhibition such as 1-6 (IC₅₀s=0.47-4.69µM), whereas compounds having hydroxymethylbutenyl 7 (IC₅₀=70.25µM) and methylbutenyl 8 (IC₅₀>200µM) showed less activity. The most potent inhibitor, gambogic acid 1 (IC₅₀=0.47µM) showed 30-fold more potency than ursolic acid (IC₅₀=15.5µM), a positive control. In kinetic study, all isolated xanthones behaved as competitive inhibitors which were fully demonstrated with K_m, V_max and K_ik/K_iv ratio. It was also proved that inhibitor 1 operated under the enzyme isomerization model having k₅=0.0751µM^-1S^-1, k₆=0.0249µM^-1S^-1 and K_i^app=0.499µM. To develop a pharmacophore model, we explored the binding sites of compound 1 and 7 in PTP1B. These modeling results were in agreement with our findings, which revealed that the inhibitory activities are tightly related to caged motif and prenyl group in A ring.