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Application of 23-hydroxybetulinic acid in preparation of medicines for preventing and treating influenza

A technology of betulinic acid and hydroxyl group, applied in the field of preparation of 23-hydroxy betulinic acid in the preparation of influenza prevention and treatment drugs, can solve the problems of no patent disclosure, no report on the anti-influenza virus application of 23-hydroxy betulinic acid, etc., and the preparation process is mature. Effect

Inactive Publication Date: 2018-07-24
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] There is no report about the anti-influenza virus effect of 23-hydroxybetulinic acid and its application in the preparation of anti-influenza drugs, and there is no related patent publication

Method used

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  • Application of 23-hydroxybetulinic acid in preparation of medicines for preventing and treating influenza
  • Application of 23-hydroxybetulinic acid in preparation of medicines for preventing and treating influenza
  • Application of 23-hydroxybetulinic acid in preparation of medicines for preventing and treating influenza

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Inhibitory Effect of 23-Hydroxybetulinic Acid on Proliferation of H1N1 Subtype Influenza Virus in A549 Cells

[0025] After the A549 cells grow to a single layer, add the H1N1 strain virus liquid (containing 100 TCID50) diluted with the basal medium, 100 μL / well, incubate at 37°C for 2 hours, discard the virus supernatant, and phosphate buffered saline (PBS solution) ) was washed once, and 3.75, 7.5 and 15 μg / ml of 23-hydroxybetulinic acid were added respectively, 100 μL / well. In the experiment, a normal control group (without adding test drug, without adding influenza virus liquid) and an influenza virus control group (without adding test drug) was set up at the same time, and each test concentration was set in three parallels. The cells continued to be cultured, and the culture was terminated at 8h, 24h, 48h, and 72h respectively. After observing the pathological changes, the cell plate was repeatedly frozen and thawed three times at -80°C and 4°C, so that ...

Embodiment 2

[0027] Embodiment 2: The test of 23-hydroxy betulinic acid to A549 cytotoxicity

[0028] After the A549 cells grow to a single layer in the 96-well cell culture plate, the medium is discarded, washed twice with PBS (phosphate buffered saline), and 100 μL of 23-hydroxybetulinic acid diluted with different concentrations in DMEM cell maintenance solution is added , six replicate wells were made for each concentration. Place at 37°C, 5% CO 2 Incubate for 48 hours in an ambient incubator protected from light. Discard 20 μl of supernatant, add 20 μl of MTT solution to each blank, and continue to culture in the cell culture incubator for 4 hours. Terminate the culture, carefully absorb the culture medium in the well, and discard it. 150 μl of dimethyl sulfoxide (DMSO) was injected into each well, and the shaker was shaken at a low speed for 10 min to fully dissolve the formazan crystals. Absorbance was measured at 490 nm using an enzyme-linked immunosorbent assay. from figure...

Embodiment 3

[0029] Example 3: Inhibitory Effect of 23-Hydroxybetulinic Acid on Influenza Virus RNA Proliferation in A549 Cells

[0030] After the A549 cells grow to a monolayer in the 6-well cell culture plate, discard the medium, wash with PBS (phosphate buffer saline) twice, and add 100TCID diluted with DMEM cell maintenance solution. 50H5N1 virus, 1ml / well, incubated at 37°C for 2h, discarded the virus supernatant, washed 2 times with PBS, and added 15, 7.5, and 3.75 μg / mL of 23-hydroxybetulinic acid, 2mL / well, respectively. At the same time, a normal control group (no test drug, no H5N1) and H5N1 control group (no test drug) were set up in the experiment, and three parallels were set for each test concentration. The cells were cultured at 37°C until 48 h after infection. After observing the lesions, freeze and thaw the cell plate at -80°C and 4°C for three times to fully lyse the cells, so that all the viruses in the cells are released into the cell supernatant, and then the supernat...

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Abstract

The invention discloses application of a traditional Chinese medicine anemone root extract 23-hydroxybetulinic acid in preparation of medicines for preventing and treating influenza. Pharmacological experiments prove that the 23-hydroxybetulinic acid is capable of effectively inhibiting reproduction of influenza virus in cells, has an obvious effect of protecting mice infected with the influenza virus, and can serve as a medicinal composition to be matched with an auxiliary agent so as to prepare injection or an oral preparation to prevent and treat the influenza.

Description

technical field [0001] The invention relates to the field of medicines, in particular to the use of 23-hydroxy betulinic acid in preparing medicines for preventing and treating influenza. Background technique [0002] Influenza is a respiratory infectious disease caused by influenza virus that seriously threatens human beings and causes huge losses to human society. Influenza virus has a wide range of hosts, is easy to mutate, and can spread across species, and its prevention and control has become a difficult problem in the medical field. Influenza outbreaks have occurred frequently in recent years. In 2009, the swine-derived new type A H1N1 influenza caused tens of thousands of deaths worldwide; in 1997, highly pathogenic influenza A virus H5N1 appeared in Hong Kong, which can be directly transmitted from birds to humans, and has a high risk of infection. Fatality rate; the H7N9 highly pathogenic avian influenza virus distributed in East my country in 2013 has also caused ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/585A61P31/16
CPCA61K31/585
Inventor 陈建新杨霞廖明亓文宝焦培荣汤有志靳珍吴倩倩
Owner SOUTH CHINA AGRI UNIV
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