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| | Inhibition of CB1, CB2 and A-Glucosidase by Kuwanon G and Albanin G Purified from Morus alba |
| Description: | The CB1 binding assay, described in Example 2, was used to test the Kuwanon G and Albanin G compounds isolated and identified in Example 5. The Kuwanon G Albanin G compounds were tested at concentrations ranging between 0.04 μg/mL and 20 μg/mL, to obtain a dose-response curve for each compound. The sample concentration was plotted against the percent inhibition and the IC50 (defined as the concentration at which 50% inhibition of binding activity is achieved in relation to the control) was determined. CB1 assay data for each compound are shown in Table 5.Inhibition of CB2 receptor-ligand binding activity of the purified Kuwanon G and Albanin G compounds was also examined using methods similar to those described in Example 2 for the CB1 receptor, with some modifications. Briefly, human cannabinoid CB2 receptor protein expressed in CHO-K1 cells were used in modified HEPES buffer (pH 7.0). A 30 μg aliquot of CB2-membrane was mixed with tritium labeled nonspecific CB1 agonist [3H] WIN-55,212-2 (2.4 nM) and test samples of Kuwanon G and Albanin G compounds, or just the nonspecific ligand R (+)-WIN-55,212-2 (10 μM) (positive control) were incubated in incubation buffer (20 mM HEPES (pH 7.0), 0.5 mg/ml BSA) for 90 minutes at 37° C. After incubation, the membranes were filtered and washed; the filters were then counted to determine the amount of radiolabeled [3H] WIN-55,212-2 that was specifically bound to the CB2-membrane. |
| Affinity data for this assay | |
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