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A pharmaceutical composition for preventing and treating chronic hepatitis B and its application

A technology for chronic hepatitis B and composition, applied in the directions of drug combination, drug delivery, active ingredients of heterocyclic compounds, etc., can solve problems such as large toxic and side effects, and achieve the effects of low toxic and side effects, avoiding degradation and high stability

Active Publication Date: 2017-09-15
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to overcome the shortcomings of existing hepatitis B treatment drugs with large toxic and side effects, the present invention provides a novel pharmaceutical composition for preventing and treating chronic hepatitis B

Method used

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  • A pharmaceutical composition for preventing and treating chronic hepatitis B and its application
  • A pharmaceutical composition for preventing and treating chronic hepatitis B and its application
  • A pharmaceutical composition for preventing and treating chronic hepatitis B and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: Culture of human liver cancer cell line HepG2.2.15 transfected with HBV

[0014] After thawing the HepG 2.2.15 cells transfected with HBV, inoculate into 25cm 2 Culture flask, the concentration is 1×10 5 cells / mL, after the cells are confluent, add 0.25% (w / v) trypsin solution to digest at 37°C for 3-5min. Add culture medium by pipetting, subculture at 1:3, collect supernatant every day for 2 consecutive weeks, and refrigerate for inspection. The cell culture medium is MEN medium, every 100mL contains 10mL of fetal bovine serum, 1mL of 3% (w / v) glutamine solution, G418 380μg / mL, gentamicin 50U / mL; cell digestion solution is 0.25% ( w / v) trypsin solution, prepared with Hanks solution.

Embodiment 2

[0015] Embodiment 2: Toxicity test of theanine bromide to HepG2.2.15 cells

[0016] Get the logarithmic growth phase HepG2.2.15 cells cultured in Example 1, adjust the cell concentration with cell culture medium to be 1 × 10 5 mL -1 , seed the cells in a 96-well plate, 200 μL per well, 37 °C 5% CO 2 Cultivate in an incubator, and discard the original culture medium after it adheres to the wall. Add 200 μL of culture solution with theanine bromide concentration of 16, 64, 125, 250, 500 μmol / L to each well of the theanine bromide group respectively, and set the cell culture wells without drug addition as the blank control group , each group repeated 6 wells. Place each group of cells in CO 2 Incubate in an incubator for 72 hours, add 10 μL of 5 μg / mL MTT to each well of cells, keep 100 μL of supernatant in each well, and store at 37°C in 5% CO 2 After culturing in the incubator for 4 hours, measure the optical density (OD) value of each well with a microplate reader at a re...

Embodiment 3

[0017] Embodiment 3: The inhibitory test of theanine bromide aromatic amide to HepG2.2.15 cell secretion HBsAg and HBeAg

[0018] Get the logarithmic growth phase HepG2.2.15 cells cultured in Example 1, adjust the cell concentration with cell culture medium to be 1 × 10 5 mL -1 , seed the cells in a 96-well plate, 200 μL per well, 37 °C 5% CO 2 Cultivate in an incubator, and discard the original culture medium after it adheres to the wall. In each well of the theanine bromide group with different doses, 200 μL of culture solution with theanine bromide concentration of 8, 16, 32, and 64 μmol / L was added respectively. Groups were replicated for 6 wells. Place each group of cells in CO 2 After culturing in the incubator for 72 hours, the supernatant of cell culture in each group was collected for enzyme-linked immunosorbent assay. The determination method was carried out according to the operating instructions of HBeAg kit and HBsAg kit (Huamei Bioengineering Co., Ltd.). Th...

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Abstract

The invention discloses a pharmaceutical composition for preventing and treating hepatitis B and an application thereof. The pharmaceutical composition is prepared from active ingredients and pharmaceutically acceptable auxiliary materials, wherein the active ingredients include theanine bromine amide. The pharmaceutical composition has high biological activity on resisting hepatitis B virus, and is small in side effects; and the injection provided is high in stability and expected to be developed into an anti-HBV first-line drug.

Description

technical field [0001] The invention belongs to the technical field of natural medicines, in particular to a pharmaceutical composition for preventing and treating chronic hepatitis B and its application. Background technique [0002] Hepatitis B is a worldwide infectious disease caused by the hepatotropic hepatitis B virus (HBV). About 350 million people in the world are infected with HBV, and my country accounts for about half. Hepatitis B has caused great harm to human health, and it is urgent to seek high-efficiency and low-toxicity anti-HBV natural medicines (Liu Jian. New progress in combined antiviral therapy for chronic hepatitis B[J]. Journal of Clinical Rational Drug Use, 2013, 6 (26): 174-176.). [0003] At present, the methods for treating chronic hepatitis B on the market mainly include interferon, adefovir dipivoxil, entecavir and other drugs and injection of hepatitis B vaccine. However, the above-mentioned drugs or vaccines have relatively large shortcoming...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/40A61K31/37A61K9/08A61K9/19A61P31/20A61P1/16
Inventor 律海峡朱斌姚冰洋卢慧娟翟德胜
Owner XINXIANG MEDICAL UNIV
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