5,7-dihydroxy-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy-2-(3,4,5-trihydroxyphenyl)chromen-4-one BDBM153266 Myricitrin (7)
3-[6-methyl-3,4,5-tris(oxidanyl)oxan-2-yl]oxy-5,7-bis(oxidanyl)-2-[3,4,5-tris(oxidanyl)phenyl]chromen-4-one BDBM115130 MYRICITRIN MLS000737865 SMR000528196 5,7-dihydroxy-3-[(3,4,5-trihydroxy-6-methyl-2-oxanyl)oxy]-2-(3,4,5-trihydroxyphenyl)-1-benzopyran-4-one cid_5352000 5,7-dihydroxy-3-(3,4,5-trihydroxy-6-methyl-tetrahydropyran-2-yl)oxy-2-(3,4,5-trihydroxyphenyl)chromone 5,7-dihydroxy-3-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxy-2-(3,4,5-trihydroxyphenyl)chromen-4-one
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- DPP III Enzyme Activity Assay The inhibitory activity of flavonoids toward human DPP III was assayed in a 50 mM Tris-HCl buffer, pH 7.4. In brief, recombinant human DPP III (0.29 nM) was preincubated with increasing concentrations of flavonoid first for 1 min at 25°C and then for 3 min at 37°C. The enzymatic reaction was started with Arg-Arg-NA (40 μM) as a substrate, and after the 15 min incubation at 37°C in a water bath, the reaction was stopped and the absorbance was measured using the spectrophotometric method described above[Abramić et al., Biol. Chem. Hoppe-Seyler., 369:29]. The stock solutions of flavonoids were prepared daily in dimethyl sulfoxide (DMSO, Sigma-Aldrich).
- Thioflavin-T (Th-T) Fluorescence Assay Fluorescence intensity was measured at 420 nm excitation and 485 nm emission using a microplate reader (MPR-A4 II; TOSOH, Tokyo, Japan, or Fluoroskan Ascent; Thermo Scientific, Rockford, IL). In brief, Aβ42 was dissolved in 0.1% NH4OH at 250 uM, and each flavonoid was dissolved in EtOH at 5 mM, followed by dilution with sodiumphosphate-buffered saline (PBS: 50 mM sodium phosphate and 100 mM NaCl, pH 7.4) at the desired concentration (Aβ42, 25 uM; flavonoids, 50 uM). NaIO4 or Tris(2-carboxyethyl)phosphine hydrochloride (TCEP-HCl) was initially dissolved in PBSat 100 mM, then diluted with PBS at 100 uM before use. Experiments under an anaerobic condition were performed in a desiccator evacuated by a diaphragm pump (about 8mmHg; KNF Lab LABOPORT vacuum pump, KNF Neuberger, NJ) at room temperature. Unless otherwise noted, the concentrations of Aβ42, flavonoids, and oxidant/reductant used in this study were 25, 50, and 100 uM, respectively.