Novel method for researching phase II metabolism of flavone compound by using model organism zebra fish

A compound and zebrafish technology, applied in the field of compound metabolism research, can solve problems such as blanks, achieve high accuracy, guarantee accuracy, and strong operability

Inactive Publication Date: 2012-10-03
JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recent studies have shown that zebrafish still have phase II metabolic enzymes [Y.Morcillo, G.Janer, S.C.M.O'Hara, D.R.Livingstone, C.Porte.Environmental Toxicology and Chemistry, 23(4), 990-996(2004); D.V.Almeida, B.F.Nornberg, L.A.Geracitano, D.M.Barros, J.M.Monserrat, L.F.Marins.Fish Physiol Biochem.36(3):347-53(2010)], which provides a basis for the phase II metabolism of drugs in zebrafish, Domestic research in this area is still blank

Method used

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  • Novel method for researching phase II metabolism of flavone compound by using model organism zebra fish
  • Novel method for researching phase II metabolism of flavone compound by using model organism zebra fish
  • Novel method for researching phase II metabolism of flavone compound by using model organism zebra fish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Studying the Metabolism of 5-Hydroxyflavones with the Model Organism Zebrafish

[0038] 1. Materials and Instruments

[0039] Test drug: 5-hydroxyflavone

[0040] Preparation method: weigh 1-2 mg equivalent to 5-hydroxyflavone, dissolve in 1 ml dimethyl sulfoxide (DMSO), add 250 ml Robust purified water, and shake well.

[0041] Animal: Zebrafish, provided by the Institute of Model Biology, Nanjing University.

[0042] Reagents: acetonitrile and formic acid are chromatographically pure (Germany, Merck Company), dimethyl sulfoxide (DMSO, Sinopharm Chemical Reagent Co., Ltd.), ultrapure water Robust purified water.

[0043] Instruments: Agilent 1100 series high performance liquid chromatography, including G1311A quaternary gradient pump, G1313A autosampler, G1316A column thermostat, G1315B DAD detector; Chemstation 6.01 chromatographic workstation (Agilent, USA). Waters Alliance 2695-ZQ 2000 liquid chromatography-mass spectrometry instrument, including dual h...

Embodiment 2

[0058] Example 2 Studying the Metabolism of 7-Hydroxyflavones with the Model Organism Zebrafish

[0059] 1. Materials and Instruments

[0060] Drug tested: 7-hydroxyflavonoids

[0061] Preparation method: weigh 1-2 mg equivalent to 7-hydroxyflavone, dissolve in 1 ml dimethyl sulfoxide (DMSO), add 250 ml Robust purified water, and shake well.

[0062] Animals, reagents, instruments: the same as in Example 1.

[0063] 2. Experimental method

[0064] 2.1 Administration and sampling methods:

[0065] Zebrafish were taken, placed in brown bottles containing 30mL solution, and divided into 2 groups on average, with 5 fish in each group. One group was 0.4% DMSO purified water (blank fish group), and the other group was 0.4% DMSO pure aqueous solution of 7-hydroxyflavone (drug fish group). At the same time, a blank solvent group (0.4% DMSO purified water) and a blank drug group (0.4% DMSO 7-hydroxyflavone pure aqueous solution) were set up. In the drug fish group, 0h, 2h, 4h, 6h, ...

Embodiment 3

[0074] Example 3 Studying the metabolism of chrysin with the model organism zebrafish

[0075] 1. Materials and Instruments

[0076] Drug tested: Chrysin

[0077] Preparation method: Weigh 1-2mg equivalent to chrysin, dissolve in 1ml dimethyl sulfoxide (DMSO), add 250ml Robust purified water, and shake well.

[0078] Animals, reagents, instruments: the same as in Example 1.

[0079] 2. Experimental method

[0080] 2.1 Administration and sampling methods:

[0081] Zebrafish were taken, placed in brown bottles containing 30mL solution, and divided into 2 groups on average, with 5 fish in each group. One group was 0.4% DMSO purified water (blank fish group), and the other group was 0.4% DMSO pure aqueous solution of chrysin (medicated fish group). At the same time, a blank solvent group (0.4% DMSO purified water) and a blank drug group (0.4% DMSO pure aqueous solution of chrysin) were set up. In the drug fish group, 0h, 2h, 4h, 6h, 8h, 12h, 18h, and 24h after the zebrafish ...

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Abstract

The invention discloses a novel method for researching the phase II metabolism of a flavone compound by using a model organism zebra fish. In the novel method, the zebra fish is selected as an object of drug metabolism research, experiment grouping design is carried out through an optimized experiment grouping method, drug administration is carried out by adopting an optimized drug administration mode, a metabolite of the flavone compound metabolized by the zebra fish is extracted by adopting an optimized method, the metabolite is analyzed by especially adopting an optimized analysis method, thus, the integral experimental method is high in operability and high in accuracy degree of an experimental result, the real conditions of the phase II metabolism of drugs in vivio and especially the real conditions of the phase II metabolism of the flavone compound in vivio can be objectively reflected, and the defect that a general in-vitro metabolism experiment is difficult to reflect a comprehensive result of in-vivo metabolism and the defects of large used drug quantity and high labor intensity of a general in-vivo metabolism experiment can be overcome. The experimental method has the advantages of high repeatability, small quantity of needed experimented drugs, low cost, low labor intensity, wide application range and high work efficiency by carrying out experiment research in a batched way.

Description

technical field [0001] The invention relates to a method for studying the metabolism of a class of compounds, in particular to a new method for studying phase II metabolism of flavone compounds by using the model organism zebrafish. Background technique [0002] Flavonoids are compounds with a 2-phenylchromone (flavone) structure. Flavonoids are usually combined with sugars in plants to form glycosides or free states (aglycones). Most plants contain flavonoids, which have important biological activities, such as protecting the cardiovascular system, antibacterial and anti-virus, anti-tumor activity, anti-inflammatory, anti-oxidation and liver protection, etc., but the bioavailability of flavonoids Low, it is difficult to reach the effective concentration in the body. The phase II metabolism of flavonoids combined with glucuronidation and sulfation reactions in vivo is an important reason for the low bioavailability of flavonoids. [0003] Drug metabolism is usually divide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 韦英杰李萍贾晓斌齐炼文王长梅孙娥陈斌
Owner JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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