Acetylcholine esterase inhibitor and applications thereof
An acetylcholinesterase and inhibitor technology, applied in the field of acetylcholinesterase inhibitors, can solve problems such as poor therapeutic effect of AChE inhibitors, and achieve the effect of obvious neuroprotective effect
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Embodiment 1
[0022] Embodiment 1 The influence of compound on AChE enzymatic activity
[0023] Acetylcholinesterase (AChE), thioacetylcholine iodide (ATC) as a substrate, and 5,5-dithiobis(2-nitrobenzoic acid) (DTNB) as a chromogenic reagent were purchased from Sigma. The determination of AChE inhibitory activity was carried out with reference to the method reported by Ellan et al. (Ellman, G. L. et al. Biochem. Pharmacol. 1961, 7, 88.). Add 40 μL of phosphate buffer (pH = 8.0) to each well of a 96-well plate, and then add 0.39, 0.78, 1.56, 3.125, 6.25, 12.5, 25, 50 and 100 μM of 10 μL of the test compound solution or blank control into Then add 10 μL of AChE to the corresponding air and incubate at 37°C for 5 min on a shaker. Add 20 μL of DTNB solution and incubate on a shaker at 37°C for 5 min, then add 10 μL of substrate ATC and incubate on a shaker at 37°C for 3 min, measure the absorbance at 412 nm with a microplate reader, and calculate Inhibition rate of test compound on AChE. ...
Embodiment 2
[0026] Example 2 Compounds on Aβ 1-42 Induced protective effect of SH-SY5Y nerve cell injury
[0027] SH-SY5Y was purchased from the National Cell Bank of the United States, and was routinely cultured in DMEM medium containing 10% fetal bovine serum in an incubator at 37°C and 5% CO2 saturated humidity. Take 2-2.5×10 5 Cells / mL were seeded in 96-well plates, and experiments were performed after 24 h of culture. Replace with fresh culture medium, and divide the cells into normal group, Aβ 1-42 Injury model group, test compound pretreatment group. Compounds of formulas I, II and III (10 μM) were added to the test compound treatment group, EGCG (tea polyphenols) was added to the EGCG positive control group, and no addition was made to the Aβ group and the normal group. After culturing for 2 h, Aβ at a final concentration of 10 μM was added to each group except the normal group 1-42 , continue to culture for 24h, add MTT (final concentration 0.5 mg / ml), after culture for 3 ho...
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