Application of a kind of plant lectin pha-1 in the preparation of anti-coronavirus medicine
A technology of phytohemagglutinin and coronavirus, applied in the field of preparation of anti-coronavirus drugs, phytohemagglutinin PHA-L, which can solve problems such as adverse reactions and urgent research and development of anti-highly pathogenic coronavirus drugs
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Examples
Embodiment 1
[0067] Example 1 Phytohemagglutinin detection of SARS-CoV-2 S protein binding activity and antiviral activity
[0068] 1. Experimental materials
[0069] Lentil lectin, wheat germ agglutinin (WGA), sessile elderberry agglutinin (SSL), peanut lectin (Peanut lectin), Huaihuai agglutinin (MAL) were purchased from Wako (Japan) Datura agglutinin (DSL), succinyl-concanavalin A (succ-Con A), galactus agglutinin (GNL), phytohemagglutinin (PHA-E), leukolectin (PHA-L ), phytohemagglutinin-M (PHA-M) and phytohemagglutinin-P (PHA-P) from red kidney bean were purchased from Sigma Aldrich; SARS-COV-2 S trimer expressed in HEK293 cells was purchased from at ACRO Biosystems (China, Beijing); 293T cells were provided by the American Type Culture Collection (ATCC); Huh7 cells were provided by the Japanese Research Biological Resource Collection (JCRB); the SARS-CoV-2spike (Wuhan-Hu- 1, MN908947; hCoV-19 / South Africa / KRISP-K007869 / 2020, B.1.351, EPI_ISL_860630; hCoV-19 / England / QEUH-F56F0F / 2021...
Embodiment 2
[0082] Example 2 Detection of the Inhibitory Effect of Plant Hemagglutinin on the Infection of SARS-CoV-2 S Protein Mutants and SARS-CoV-2 Epidemic Mutants
[0083] In this example, the inhibitory effects of various plant lectins on the infection of SARS-CoV-2 S protein mutant strains and SARS-CoV-2 epidemic mutant strains, including the British epidemic mutant strain B.1.1.7, were detected. , South African popular mutant strain B.1.351, Brazilian popular mutant strain P.1, combined mutant strain D614G+I472V, combined mutant strain D614G+A435S, A222V, K417N, S477N, E484K, N501Y, combined mutant strain A222V+D614G, combined mutant strain Mutant strains such as strain K417N+D614G, combined mutant strain S477N+D614G, combined mutant strain E484K+D614G, combined mutant strain N501Y+D614G;
[0084] Lectins are mainly aimed at the glycosyl part of glycoproteins. In this example, the inventors studied whether the change of the glycation site of SARS-CoV-2 S protein would affect its n...
Embodiment 3
[0094] Example 3 Detection of Hemagglutination Activity and Cytotoxicity of Plant Hemagglutinin
[0095] Since many natural lectins can cause hemagglutination, in this example, the inventors studied the hemagglutination activity of different lectins, and studied the cytotoxicity of Huh7 and 293T cells.
[0096] 1. Detection of hemagglutination activity of phytohemagglutinin
[0097] Chicken blood samples were used to detect the hemagglutination activity of phytohemagglutinin, the rooster red blood cells were washed with PBS, and resuspended at a final concentration of 1% (v / v), and 50 μL of 2-fold serially diluted phytohemagglutinin solution was mixed with an equal volume of red blood cells The solution was mixed in a 96-well round-bottom plate, and a 2-fold serial dilution of influenza antigen (B / Maryland / 15 / 2016, NIBSC-UK-EN63QG, NIBSC code: 18 / 104, HA: 69 μg / mL) was performed and served as a positive control , PBS solution was used as a negative control, the plate was incu...
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Description
Claims
Application Information
- IPC
- A61K38/16; A61P31/14
- CPC
- A61K38/168; A61P31/14
- Inventors
- 王佑春; 王兰