Novel application of aurantiamide alcohol ester as MMP-9 inhibitor
A golden amido alcohol ester, MMP-9 technology, applied in medical preparations containing active ingredients, organic active ingredients, bone diseases, etc., can solve problems such as unclear new targeting mechanism, and achieve the effect of great clinical application value
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Embodiment 1
[0033] Example 1 Molecular docking binding experiment of golden amido alcohol ester and MMP-9 protein
[0034] Such as figure 1 As shown, a computer-assisted molecular docking experiment between the drug ligand and the protein receptor was carried out on the golden amido alcohol ester and the MMP-9 protein to further analyze the ligand-receptor interaction mechanism; the binding energy and interaction energy were calculated, and Binding pattern analysis was performed.
[0035] After molecular docking and binding energy calculation, compared with the original ligand inhibitor, the key site of docking is consistent, and the binding energy is -72.0206kcal / mol. The results show that golden amido alcohol ester and MMP-9 protein have better drug-protein Molecular docking binding capacity.
Embodiment 2
[0036] The action activity of embodiment 2 in vitro golden amido alcohol ester
[0037] 1. In vitro osteoclastogenesis assay
[0038] RAW 264.7 cells use DMEM medium, containing 10% fetal bovine serum, 5% penicillin-streptomycin, temperature 37°C, CO 2 The culture was carried out under the condition of 5%. To induce osteoclasts, RAW264.7 cells were cultured on 96-well plates in the presence of RANKL concentration of 50 ng / mL and 5, 10, 25 μM gold amidoalcohol ester. On the 7th day, the cells were fixed, and then TRAP activity was stained according to the protocol of the kit. TRAP-positive multinucleated cells with more than 3 nuclei were considered osteoclasts using a light microscope (IX71; Olympus).
[0039] 2. Western blot analysis
[0040] Western blot was used to determine the level of protein expression. RAW 264.7 cells were seeded into 60mm dishes, 5×10 5 Cells / dish were incubated overnight and then treated with RANKL (50 ng / mL) for 48 hours in the absence or pres...
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