Application of interferon lambda in treatment of novel coronavirus (2019-nCoV) infection

A technology for coronavirus and virus infection, applied in antiviral agents, medical preparations containing active ingredients, peptide/protein components, etc., can solve problems such as hematology and nervous system suppression side effects

Pending Publication Date: 2021-09-24
HANGZHOU SCIWIND BIOSCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no receptor distribution in bone marrow-derived hematopoietic stem cells (such as NK cells), so there is no hematological and nervous system inhibition and other side effects caused by interferon α treatment.

Method used

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  • Application of interferon lambda in treatment of novel coronavirus (2019-nCoV) infection
  • Application of interferon lambda in treatment of novel coronavirus (2019-nCoV) infection
  • Application of interferon lambda in treatment of novel coronavirus (2019-nCoV) infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Implementation example 1: Construction of interferon lambda expression engineering bacteria

[0048] Human interferon λ1 / λ2 / λ3 / λ4 gene fragments (NM_172140.2 / NM_172138.2 / NM_172139.2) were obtained by chemical synthesis, and inserted into the prokaryotic expression plasmid pET- 30a and verified by sequencing. Resulting expression plasmids for transformation assays.

[0049] The plasmid containing the target gene obtained above was transformed into BL21(DE3) competent cells (Invitrogen), 50 μl of BL21 competent cells were placed on an ice bath to melt, the target DNA was added, shaken gently, and placed in an ice bath for 30 minute. Then heat shock in a 42°C water bath for 30 seconds, and then quickly transfer the centrifuge tube to an ice bath for 2 minutes without shaking the centrifuge tube. Add 500 μl of sterile LB medium (without antibiotics) to the centrifuge tube, mix well, place at 37° C., and incubate at 180 rpm for 1 hour to recover the bacteria. Pipette 200...

Embodiment 2

[0050] Implementation example 2: expression and purification of interferon lambda

[0051] Add 50 μl of bacterial solution (bacterial solution expressing interferon lambda) to 50 ml of LB medium, and add 50 μl of kanamycin at the same time, mix well, place in a constant temperature shaker at 30°C, and inoculate overnight. Take 10ml of the overnight inoculated bacterial solution and add it to 1000ml LB medium, and add 1000μl kanamycin at the same time. Shake well and place in a shaker at 37°C at 200rpm, cultivate until the OD600 of the bacterial solution is 0.4-0.6, add 0.5mM IPTG for induction, continue to cultivate for 4 hours, and collect the bacterial cells. The expressed IL29 mutant accounts for about 30-50% of the total bacterial protein, and mainly exists in the form of inclusion bodies.

[0052] The fermented cells were washed with TE (10mmol / L Tris-HCl, 1mmol / L EDTA, pH 6.5) solution (m:V=1:10) for 3 times, then 60Mpa high-pressure homogenate was crushed, after homoge...

Embodiment 3

[0056] Example 3: In vitro drug efficacy of novel coronavirus

[0057] Cells: VeroE6 cells, preserved by the Pathogen Center, Institute of Medical Experimental Animals, Chinese Academy of Medical Sciences.

[0058] Virus: 2019-nCoV, titer 10 5 TCID 50 / ml, stored at -80°C by the Pathogen Center, Institute of Medical Experimental Animals, Chinese Academy of Medical Sciences. Use a viral titer of 100TCID 50 .

[0059] In a sterile 96-well culture plate, add 200 μl to each well with a concentration of 5×10 4 cell / ml Vero E6 cells, 37°C 5% CO 2 Cultivate for 24 hours;

[0060](2) The test drug was diluted to 2 concentrations, each concentration was replicated into 5 wells, and 100 μl was added to each well for 24 hours;

[0061] (3) Add 100 μl of 100 TCID to each well 50 Virus;

[0062] (4) Set up cell control, blank control (solvent control) and virus control (negative control) at the same time;

[0063] (5) Cells at 37°C, 5% CO 2 Incubation in the incubator for 4-5 da...

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Abstract

The invention relates to application of IFN-lambda in preparation of a drug for treating a patient infected with a novel coronavirus. The level of the novel coronavirus in the infected patient can be effectively reduced, and the IFN-lambda shows more excellent performance in the aspects of controlling cytokine storm caused in the treatment process and relieving respiratory distress syndrome caused by the cytokine storm.

Description

Technical field: [0001] The present invention relates to the application of interferon lambda in the treatment of novel coronavirus (2019-nCoV) infection. Background technique: [0002] Novel coronavirus pneumonia, as an acute respiratory infectious disease, has been included in the Class B infectious disease stipulated in the "Law of the People's Republic of China on the Prevention and Control of Infectious Diseases", and it is managed as a Class A infectious disease. [0003] WHO officially named the novel coronavirus pneumonia as COVID-19 (corona virus disease 2019). The International Committee on Taxonomy of Viruses named the virus as severe acute respiratory syndrome coronavirus 2 (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2). [0004] Coronaviruses are a class of enveloped, single-stranded positive-sense RNA viruses that can infect many mammals, including humans. Clinically, it was found that the virus first caused lung lesions, and then caused multip...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/21A61P31/14
CPCA61K38/21A61P31/14
Inventor 杨柳李岩冯静郭万军潘海
Owner HANGZHOU SCIWIND BIOSCI CO LTD
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