Novel application of acetylcholine lipase activity inhibitor
An active inhibitor, the technology of acetylcholine, applied in the field of whitening agents, can solve the problems that the whitening effect cannot meet the needs of the public, and achieve excellent skin whitening effect, high safety, and the effect of inhibiting melanin production
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Embodiment 1
[0056] This example is an inhibitory test of acetylcholinesterase inhibitor on melanin production in melanocytes.
[0057] Treat melanocytes with acetylcholinesterase inhibitor (BW284c51) or acetylcholine, and then measure the content of melanin to detect whether its production is inhibited. The specific method is as follows:
[0058] Melan-A cells (from Ximbio, London) were cultured in DMEM medium supplemented with 10% FBS (fetal bovine serum), 1% penicillin / streptomycin, and placed in CO 2 Incubator (5% CO 2 , 37℃) for 24 hours, the Melan-A cells were made into a cell concentration of 3×10 5 A / mL suspension and seeded on a 6-well plate. Put in CO 2 Incubator (5% CO 2 , 37°C) for 24 hours. Divided into 3 groups, the ACh group was added with concentrations of 0mM, 0.1mM, 0.2mM, 1.0mM, 2.0mM, and the ACh+BW group was added with 10μMBW284c51, and the concentrations were 0mM, 0.1mM, 0.2mM, 1.0mM, 2.0mM ACh, DMSO solvent control was added to the blank group. After culturing for 24 h...
Embodiment 2
[0061] This example is an acetylcholinesterase inhibitor's melanin production inhibition test in the co-culture of keratinocytes and melanocytes.
[0062] Use acetylcholinesterase inhibitor (BW284c51) to co-culture keratinocytes and melanocytes, and then measure the content of melanin to detect whether its production is inhibited. The specific method is as follows:
[0063] The co-cultured Melan-A cells (from Ximbio, London, UK) and HaCaT (from AddexBio, San Diego, CA) cells were supplemented with 10% FBS (fetal bovine serum), 1% penicillin / streptomycin DMEM medium for culture, put CO 2 Incubator (5% CO 2 , 37℃) for 24 hours, the Melan-A cells were made into a cell concentration of 3×10 5 A / mL suspension and seeded on a 6-well plate. Put in CO 2 Incubator (5% CO 2 , 37°C) for 24 hours. Divided into 5 groups, among them, the control group-only add solvent DMSO, the light group-light for 10 minutes, the light + BW group-light for 10 minutes and add 10μM BW284c51, the light + ACh gr...
Embodiment 3
[0066] This example is an inhibitory test of acetylcholinesterase inhibitor (BW284c51) on melanin production in mouse skin.
[0067] Take a three-week-old C57BL / 6 mouse and put it in CO 2 Incubator (5% CO 2 , 37℃), and divided into 5 groups, of which, the control group-only add solvent DMSO, light group-light for 10 minutes, light + BW group-light for 10 minutes and add 10μM BW284c51, light + ACh Group-light for 10 minutes and add 0.1mM ACh, light + BW group-light for 10 minutes and add 10μM BW284c51, light + ACh+BW group-light for 10 minutes and add 10μM BW284c51 and 0.1mM Ach. After 24 hours of continuous cultivation under the same conditions, Fontana masson staining method was used to label melanin. Calculate the gray value of the black area of melanin and compare it with the control group.
[0068] Such as image 3 As shown, light significantly increased the production of melanin in the skin surface. The melanin production in the skin of mice treated with ACh or acetylcholin...
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