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Application of lambda interferon to preparation of anti-Hantaan virus drugs

A technology of Hantaan virus and interferon, which is applied in the direction of antiviral agents, drug combinations, blood diseases, etc., can solve the problems of lowering the fatality rate, not reducing complications, and long antipyretic time, achieving obvious therapeutic effect and reducing nucleic acid Level of expression, fast-acting effects

Active Publication Date: 2015-07-22
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are also reports comparing the efficacy of interferon combined with ribavirin and ribavirin alone. It is believed that interferon in the combined treatment group can shorten the symptoms of congestion and oozing and percussion pain in the kidney area in patients with early HFRS, but the rate of expiration is high. , but it cannot reduce complications and mortality. On the contrary, due to the toxic side effects of interferon, the average time for fever reduction in the combined treatment group is longer, headache and kidney pain symptoms disappear slowly, and the platelet count returns to normal slowly. Therefore, interferon is used in combination. The advantages and disadvantages of ribavirin and ribavirin in the treatment of HFRS still need further experimental research
Recently, a new member has been added to the interferon family: type III interferon, or IFN-λ, has been found to have the same antiviral effect as other types of interferon, such as: hepatitis B virus (HBV), hepatitis C virus (HCV) , human cytomegalovirus (HCMV), human immunodeficiency virus (HIV), etc., and IFN-λ anti-HCV research has entered the phase II clinical stage, but the anti-HTNV effect of IFN-λ has not been reported by scientific and technological research

Method used

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  • Application of lambda interferon to preparation of anti-Hantaan virus drugs
  • Application of lambda interferon to preparation of anti-Hantaan virus drugs
  • Application of lambda interferon to preparation of anti-Hantaan virus drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1MTT method detects the cytotoxic effect of IFN-λ1 / λ2

[0033] A549 cells cultured in DMEM (containing 10% fetal bovine serum, V / V) were digested with trypsin and diluted to 1~3×10 5 cells / mL, 100 μL per well was inoculated in a 96-well plate. After the cells grew into a single layer, different concentrations (1ng / mL, 10ng / mL, 100ng / mL, 1000ng / mL) of IFN-λ1, IFN- λ2, cultured for 72 hours, and the cytotoxicity of IFN-λ1 and IFN-λ2 was detected by MTT method. The results showed that the cytotoxicity of IFN-λ1 and IFN-λ2 from 1ng / mL to 1000ng / mL was less than 10% on A549 cells, and statistics There was no significant difference in the analysis, so it can be considered that IFN-λ1 and IFN-λ2 have no toxicity to A549 cells ( figure 1 ).

Embodiment 2I

[0034] Example 2 IFN-λ anti-HTNV infection in vitro has time effect and dose effect

[0035] (1) IFN-λ has a time effect on HTNV infection in vitro

[0036] A549 cells cultured in DMEM (containing 10% fetal bovine serum, V / V) were digested with trypsin and diluted to 1~3×10 5 cells / mL, 400 μL per well was inoculated in a 24-well plate. After the cells grow into a single layer, add IFN-λ1 and IFN-λ2 (100ng / mL) diluted with DMEM (containing 2% fetal bovine serum, V / V) to pretreat A549 cells for 24h, and HTNV 76-118 strain infection for 2h ( MOI=5), and washed three times with PBS, then added DMEM (containing 2% fetal bovine serum, V / V) medium, 37 ° C 5% CO 2 (V / V) culture; cells were collected on the 1st day, 4th day, 7th day and 10th day of infection, and real-time quantitative RT-PCR was used to detect the expression of HTNV S gene, GAPDH was used as an internal reference, and IFN-λ was not added treated as the control group. Primers are as follows:

[0037] HTNV S gene u...

Embodiment 3I

[0044] Example 3 Antiviral Factors Induced by IFN-λ

[0045] A549 cells cultured in DMEM (containing 10% fetal bovine serum, V / V) were digested with trypsin and diluted to 1~3×10 5 cells / mL, 400 μL per well was inoculated in a 24-well plate. After the cells grow into a single layer, add IFN-λ1 and IFN-λ2 (100ng / mL) diluted with DMEM (containing 2% fetal bovine serum, V / V) to pretreat A549 cells, and then treat them 3h, 6h, and 12h after treatment, respectively. and 24h, the cells were collected, and the expression of antiviral factors OAS-1, ISG-56, MxA and MxB was detected by real-time quantitative RT-PCR. GAPDH was used as an internal reference, and those without interferon treatment were used as a control group. Primers are as follows:

[0046] OAS-1 upstream primer: 5'-AGAAGGCAGCTCACGAAACC-3',

[0047] OAS-1 downstream primer: 5'-CCACCACCCAAGTTTCCTGTA-3';

[0048] ISG-56 upstream primer: 5'-GCTGAAGTGTGGAGGAAAGA-3',

[0049] ISG-56 downstream primer: 5'-AGCAAAGAAAATGGC...

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Abstract

The invention discloses an application of lambda interferon to the preparation of anti-Hantaan virus drugs and belongs to the field of anti-virus drugs. The invention discovers that lambda interferon can induce generation of various anti-virus factors and inhibit Hantaan virus infection in vitro and in vivo, does not have toxic or side effects, can protect cells from being infected with Hantaan viruses in vitro and increase the survival rate and mean survival days of animals infected with the Hantaan viruses in vivo, relieves pathological lesion of tissues, and reduces viral nucleic acid level. The discovery shows that lambda interferon can be used for preparing the anti-Hantaan virus drugs and also can be used for preparing drugs for treating and preventing hemorrhagic fever with renal syndrome caused by the Hantaan viruses.

Description

technical field [0001] The invention belongs to the field of antiviral drugs, in particular to the application of novel interferon-λ interferon (interferon lambda, IFN-λ) in the preparation of anti-Hantaan virus drugs. Background technique [0002] The main clinical features of hemorrhagic fever with renal syndromes (HFRS) are fever, hemorrhage, and kidney damage. my country is the country most affected by HFRS. HFRS patients account for more than 90% of the world, and its pathogens are mainly Han Hantaan virus (HTNV), people can be infected with the virus through aerosol inhalation, wounds, digestive tract and other ways. For a long time, people have been working on the research of antibodies and cytotoxic T cells, hoping to develop a vaccine to curb HTNV infection, but no significant breakthrough has been seen. At present, the clinical treatment of HFRS is still mainly symptomatic treatment, combined with antiviral treatment. The antiviral drug is mainly ribavirin, and th...

Claims

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Application Information

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IPC IPC(8): A61K38/21A61P31/14A61P13/12A61P7/04
Inventor 侯炜李宁陈清宙罗凡
Owner WUHAN UNIV
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