Application of phillyrin in preparation of digestants
A technology for forsythin and digestion aid, which is applied in the application field of forsythin in the preparation of digestive aid drugs, and can solve the problems of affecting curative effect, high price of enzyme preparations, and decreased activity
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Embodiment 1
[0015] Take 100 μL of PL solution (0.5 mg / mL) and place it in a 1.5 mL centrifuge tube, add 20 μL of 5 mg / mL forsythin solution, dilute to 900 μL with pH=7.5 Tris-HCl buffer, mix well, and incubate at 37°C for 15 min. Then add 100 μL of pNPB solution (10 mmol / L), shake well, and quickly transfer to a 96-well microplate. Measure the absorbance value at 400nm with a microplate reader, read the number every 3 minutes, and record the data within 15 minutes. Take time as the abscissa and the absorbance value as the ordinate, and calculate the change rate K of the absorbance value with time. The inhibition rate was calculated according to the following formula:
[0016] PL inhibition rate (%) = (K 正常组 -K 实验组 ) / K 正常组 ×100%
[0017] In the reaction system, no enzyme was used as the blank control, no forsythin was used as the normal group, and orlistat was used as the positive control, and each experiment was repeated 3 times.
[0018] Experimental results: when 20 μL of 5 mg / mL ...
Embodiment 2
[0020] Adding 20 μL of 20 mg / mL forsythin solution, that is, when the final concentration is 400 μg / mL, the activity of pancreatic lipase can be increased to 134.11%. Other conditions are with embodiment 1.
Embodiment 3
[0022] Adding 20 μL of 40 mg / mL forsythin solution, that is, when the final concentration is 800 μg / mL, the activity of pancreatic lipase can be increased to 146.43%. Other conditions are with embodiment 1.
[0023] The summary results of each embodiment are as figure 1 shown.
[0024] 2. Mechanism experiment of forsythin to promote pancreatic lipase activity:
[0025] Forsythin solutions with a concentration approximately equal to the IC50 value were prepared respectively. With no forsythin as the control group and forsythin as the experimental group, the enzyme reaction rate was measured when the pNP concentration was 0.25, 0.50, 0.75, 1.00, 1.25 mmol / L respectively. According to the Lineweaver-Burk double reciprocal plotting method. The type of inhibition was judged according to the position of the intersection of two straight lines in the control group and the experimental group.
[0026] Experimental results such as figure 2 As shown, it is proved that forsythin ha...
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