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Liver microsome metabolism analysis of koumine and toxicokinetics of related animals

A technology of liver microsomes and kinetics, which is applied in the fields of biochemical analysis and biological metabolism, can solve the problems of providing references for clinical research and not being able to truly reflect drug metabolism, and achieve the effect of improving predictability

Inactive Publication Date: 2017-05-10
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, only conventional animals for drug metabolism research cannot reflect the most true drug metabolism, that is, it cannot provide a reliable reference for clinical research

Method used

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  • Liver microsome metabolism analysis of koumine and toxicokinetics of related animals
  • Liver microsome metabolism analysis of koumine and toxicokinetics of related animals
  • Liver microsome metabolism analysis of koumine and toxicokinetics of related animals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Liver Microsomal Metabolism Analysis of KM

[0044] Using ultra-high performance liquid chromatography (UPLC) to compare the distribution characteristics of metabolites of KM in liver microsomes of various genera and the kinetic characteristics of KM metabolism by liver microsomes of various genera, and to judge KM human-related animals.

[0045] 1 Experimental materials

[0046] 1.1 Liver microsomes

[0047] Liver microsomes of various species were purchased from Reid Liver Disease Research (Shanghai) Co., Ltd., protein content 20g L -1 , the batch numbers are: SUBK (human), RUIB (male minipig), BDVH (male SD rat), AWWJ (male beagle dog) and GPIU (female rhesus monkey), stored at -80°C.

[0048] 1.2 Reagents and Reagents

[0049] 1) Gelskinin hydrochloride, white powder, batch number 201310, purity 99%, provided by School of Pharmacy, Fujian Medical University.

[0050] 2) PBS (powder), 0.01M (2L / bag), pH 7.2-7.4, batch number 1225F022, Beijing Suo Laiba...

Embodiment 2

[0095] Example 2 KM rhesus monkey acute toxicity with toxicokinetics

[0096] 1 Experimental materials

[0097] 1.1 Animals

[0098] Ordinary-grade rhesus monkeys were provided by the Fujian Provincial Institute of Population and Family Planning Science and Technology, production license number: SCXK (Fujian) 2010-0002. 8 rhesus monkeys, male and female, aged 3-4 years, weighing 3.59-4.70kg.

[0099] 1.2 Reagents and Reagents

[0100] 1) Kiskinin hydrochloride (KM), white powder, batch number 201310, purity 99%, provided by the School of Pharmacy, Fujian Medical University, stored at 4°C.

[0101] 2) Internal standard substance (IS): kinesin A (GM), batch number MUST-16042112, purity 99.75%, Chengdu Master Biotechnology Co., Ltd., stored at 4°C.

[0102] 3) Methanol, lot number I512907948, Merck, Germany, chromatographically pure.

[0103] 4) Ethyl acetate, batch number I807568601, Merck, Germany, chromatographically pure.

[0104] 5) Ammonia water, batch number 20100406,...

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Abstract

The invention discloses liver microsome metabolism analysis of koumine (KM) and toxicokinetics of related animals. The method disclosed by the invention mainly comprises the following steps: 1, performing KM liver microsome metabolism analysis comprising the following sub-steps: (1) establishing a KM incubation system; (2) performing sample treatment; and (3) performing sample analysis, namely selecting KM human related animals by comparing the liver microsome metabolism product distribution characteristics of KM in various experimental animals and the kinetic characteristics of various kinds of KM on KM metabolism; 2, performing KM macaca rhesus acute toxicity associated toxicokinetics comprising the following sub-steps: (1) performing blood sample collection and plasma sample treatment on infected experimental animals; (2) performing quantitative determination; and (3) performing blood concentration data analysis. The method has the advantages that due to the in-vitro liver microsome metabolism analysis of KM in various experimental animnals, the human related animals are selected for performing associated toxicokinetics study, so that the predictability of toxicity assessment of the experimental animals on clinical study is improved.

Description

technical field [0001] The present invention relates to the fields of biochemical analysis and biological metabolism, in particular to KM liver microsomal metabolism analysis and related animal acute toxicity accompanied by toxicokinetics, providing scientific basis for later clinical research. Background technique [0002] Kumine (koumine, KM) is the main active alkaloid component of the Struggleaceae plant G. chinensis. Studies have shown that it has high efficiency and low toxicity. It has anti-tumor, analgesic and sedative, anxiolytic, therapeutic Rheumatoid arthritis and other pharmacological effects, and obtained the national patent of KM for the treatment of chronic pain and rheumatoid arthritis, so the research that has been carried out so far shows that KM has great pharmacodynamic application prospects. [0003] In the early stage, our research group invented the "Method for Separating and Preparing Gelkins Alkaloid Monomers from Gelkins by High-speed Countercurren...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/02
CPCG01N30/06G01N30/02
Inventor 林菁俞昌喜魏文增黄慧玲叶丽香
Owner FUJIAN MEDICAL UNIV
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