Method for minim hepatic tissue in vitro incubation and detecting CYP450 enzymatic activity
A technology of liver tissue and enzyme activity, which is applied in the field of detection of CYP450 enzyme activity by in vitro incubation of trace liver tissue, which can solve the problems of expensive equipment, large amount of liver tissue, and long centrifugation time, and achieve the effect of saving cost and time
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0032] 1. Solution preparation
[0033] 1.1 Homogenate: add KCl 6g Tris 3.0g to 500ml distilled water;
[0034] 1.2 Incubation solution: add K to 500ml distilled water 2 HPO 4 11g, then adjust the pH to 7.4 with HCl;
[0035] 1.3 Precipitating agent: TCA was added with distilled water to make a 10% (W / V) solution for later use.
[0036] 1.4 Stock solutions of probe substrates and metabolites: Weigh an appropriate amount of PN and ACE respectively, dissolve them in methanol and dilute to a methanol solution with a required concentration of 1-10 mg / ml.
[0037] 2. Sample
[0038] 2.1 20 healthy Kunming rats, male, were randomly divided into four groups, 5 rats in each group were weighed, fed to the stomach after fasting for 12 hours on the first day, the control group was given normal saline, and the other three groups were given drug C (such as berberine derivatives ) High, medium and low doses of drugs, low dose group: 0.3mg / ml, medium dose group: 0.6mg / ml, high dose gro...
Embodiment 2
[0074] Confirmation experiment of the change of CYP1A2 enzyme activity after gavage of high, medium and low doses of drug C to mice:
[0075] 1. Thirty-two healthy Kunming rats, male, were randomly divided into four groups, with 8 rats in each group, and weighed.
[0076] 2. Blank control group (physiological saline), 0.64 mg / ml low dose group, 1.6 mg / ml medium dose group, and 4 mg / ml high dose group.
[0077] 3. Gastrointestinal administration after fasting for 12 hours on the first day, the control group was given normal saline, and the other groups were given high, medium and low doses of drugs.
[0078] 4. Gastrointestinal administration every day and fasting at night for 8 days. On the 8th day, the mice were sacrificed and the liver was taken out.
[0079] 5. The liver was incubated with phenacetin probe drug in vitro, and the effect of drug C on liver CYP1A2 was evaluated by LC-MS / MS detection.
[0080] Solution preparation and incubation methods, and LC-MS / MS detectio...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com