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A method for in vitro evaluation of pulmonary metabolic characteristics of drugs

A lung and drug technology, applied in the field of biomedicine, can solve the problems that the personalized research plan is not universal, the drug metabolism cannot be correctly evaluated, and the probability of false positive and false negative is high, so as to facilitate promotion and save manpower Material resources, the effect of reducing the probability of false positive or false negative

Active Publication Date: 2021-09-21
瑞德肝脏疾病研究(上海)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, inhalation preparations are administered through the lungs, avoiding the first-pass effect of the liver, and acting directly on the lung tissue. Therefore, it is not entirely suitable to use liver cells and their subcellular tissues for the study of drug metabolism characteristics. The study of its metabolic characteristics and drug interaction is limited to the study of the molecular mechanism of the metabolic pathway of specific compounds. The personalized research plan is not universal, and the stability of the method is poor, and the probability of false positive and false negative is high. There is still a lack of perfect technical solutions for in vitro studies of characteristics
[0004] (Aune T, et al.Deacetylation to 2-aminofluorene as a major initial reaction in the microsomal metabolism of 2-acetylaminofluorene to mutagenic products in preparations from rabbit lung and liver[J].Cancer Research,1985,45(11Pt2):5859.) It is disclosed that the metabolic activity of rabbit lung microsomes for 2-acetylaminofluorene and 2-aminofluorene is different, and the metabolic activity of the lung is higher than that of the liver, which indicates that some drugs may be more vigorously metabolized in the lung, and for such The hepatic metabolism evaluation criteria for drugs are often inaccurate, and the metabolism of drugs in the body cannot be correctly evaluated

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  • A method for in vitro evaluation of pulmonary metabolic characteristics of drugs
  • A method for in vitro evaluation of pulmonary metabolic characteristics of drugs

Examples

Experimental program
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Embodiment 1

[0065] Example 1 Preparation of Animal Lung Subcellular Tissue

[0066] Animals were fasted overnight, anesthetized with isoflurane or 3% pentobarbital sodium aqueous solution 30mg / kg intraperitoneal injection, surgical scissors were used to open the abdominal cavity and thorax, the hemostatic forceps clamped the abdominal aorta, cut open the left ventricle of the heart, and inserted with normal saline Or a 50mL syringe of 0.15M potassium chloride solution (the solution is pre-cooled), quickly inject the solution to rinse until the lungs are milky white, and take the lung tissue in cold 0.15M potassium chloride (or 0.9% sodium chloride) solution In the beaker, soak and wash for 2-3 times according to the ratio of solid-liquid volume ratio of 2:3, and then wash twice with buffer A, then take the cleaned lung tissue and buffer A and transfer to centrifuge at the ratio of solid-liquid volume ratio of 2:3. tube, shredded, homogenized with a high-speed homogenizer, and collected th...

Embodiment 2

[0067] Example 2 Testosterone in vitro evaluation method of lung metabolic characteristics

[0068] (1) After thawing Beagle lung S9 in a water bath at 37°C, dilute lung S9 to 3 mg / mL with Tris buffer;

[0069](2) Prepare positive quality control system, negative quality control system, control group reaction system and test group reaction system respectively, wherein the positive quality control system includes mixing 2-aminofluorene and lung S9 and pre-incubating for 5 minutes, then adding NADPH coenzyme, According to the volumes of 2-aminofluorene, lung S9 and NADPH coenzyme being 25 μL, 25 μL and 50 μL, respectively, mix evenly, the final concentration of 2-aminofluorene is 0.05 μM, the final concentration of NADPH coenzyme is 1 mM, and the protein content of lung S9 is 3mg / mL; Negative quality control system includes mixing 2-aminofluorene and lung S9 and pre-incubating for 5min, then adding 0.1mM Tris buffer, according to 2-aminofluorene, lung S9 and Tris buffer respecti...

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Abstract

The present invention provides a method for in vitro evaluation of drug lung metabolism characteristics, said method comprising the following steps: (1) preparing lung subcellular tissue; (2) using the lung subcellular tissue obtained in step (1) to prepare positive quality control system, negative quality control system, control group reaction system and test group reaction system, incubated, centrifuged to get the supernatant; The parent content of the positive substrate, and the parent content of the analyte in the control group reaction system and the test group reaction system supernatant; The positive substrate of the positive quality control system includes 2-aminofluorene, 4-methoxy Any one or a combination of at least two of 1,8-naphthalimide, phenacetin or 4-sweetyramediol. The evaluation method of the invention has a stable effect, is a perfect evaluation method for pulmonary administration, and provides methodological support for new drug development and drug interaction.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to an evaluation method for drug safety, in particular to an in vitro evaluation method for the pulmonary metabolic characteristics of drugs, in particular to an in vitro evaluation method for the pulmonary metabolic characteristics of inhaled drugs. Background technique [0002] According to the "Guiding Principles for Drug Interactions" (2012) of the State Food and Drug Administration of the People's Republic of China (CFDA), the metabolism of new drugs should be determined during the drug development process, and the interaction between the drug and other drugs should be used as The study was conducted as part of the safety and efficacy evaluation. Since the elimination of oral and intravenous drugs or their metabolites is usually metabolized in the liver, most of the tissues used for in vitro research on drug metabolism are the liver and its subcellular tissues. [0003] With the conti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 胡卓汉孙易李国栋郝祝兵荣义
Owner 瑞德肝脏疾病研究(上海)有限公司