Application of the small molecule Yadanziolide A in the preparation of drugs against the new coronavirus

Abstract

The present invention discloses the application of the drug small molecule Yadanziolide A in the preparation of anti-new coronavirus drugs, belonging to the field of medical technology. The small molecule drug Yadanziolide A (CAS No. 95258-14-3) screened by the present invention has the effect of significantly inhibiting the replication of the new coronavirus, which shows that the present invention further enriches the types of drugs against the new coronavirus.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to the application of the drug small molecule Yadanziolide A in the preparation of anti-new coronavirus drugs. Background Art

[0002] SARS-CoV-2 is a single-stranded positive-sense RNA virus with a genome length of approximately 30kb and is a member of the β-coronavirus genus.

[0003] The diameter of coronavirus particles is about 80-120nm, and its genome is the largest of all known RNA viruses. Like other coronaviruses, the SARS-CoV-2 viral capsid protein Spike glycoprotein is expressed on the viral capsid in the form of a trimer, and each monomer structure consists of an S1 subunit and an S2 subunit. When SARS-CoV-2 infects the human body, it binds to angiotensin converting enzyme 2 (ACE2) through the receptor binding domain (RBD) of the Spike protein S1 subunit on the viral capsid, inducing changes in the S2 subunit structure, thereby promoting the fusion of the virus and the host cell membrane, mediating the virus's invasion of host cells. Existing studies have shown that SARS-CoV-2 has a higher affinity for binding to the ACE2 receptor than severe acute respiratory syndrome coronavirus (SARS-CoV), which may be the reason why SARS-CoV-2 is more susceptible and spreads faster. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides the use of the drug small molecule Yadanziolide A in the preparation of anti-new coronavirus drugs.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] Application of the small molecule Yadanziolide A in the preparation of drugs against the new coronavirus.

[0007] Furthermore, the CAS number of the small molecule Yadanziolide A is 95258-14-3, and the structural formula is as follows:

[0008]

[0009] Furthermore, the small molecule Yadanziolide A is the only component in the drug.

[0010] Furthermore, the dosage form of the drug is any one of powder, tablet, granule, capsule, solution, emulsion and suspension.

[0011] Furthermore, the drug also contains a pharmaceutically acceptable carrier.

[0012] Compared with the prior art, the present invention has the following beneficial effects:

[0013] The small molecule drug Yadanziolide A screened by the present invention has the effect of significantly inhibiting the replication of the new coronavirus, providing a new idea for the prevention and treatment of the new coronavirus. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 This is a graph showing the replication activity results of the new coronavirus in Example 1.

[0015] Figure 2 This is a curve diagram of the inhibition of viral replication by the Yadanziolide A small molecule in Example 1. DETAILED DESCRIPTION

[0016] Example 1

[0017] 1. Referring to the biosafety level 2 SARS-CoV-2 trans-complementation cell culture system of Professor Ding Qiang and Professor Shi Peiyong, we constructed a SARS-CoV-2 trans-complementation cell culture system containing the Firefly gene. The specific method is as follows:

[0018] 1) Synthesize all the genes of the new coronavirus and the Firefly (Fluc) gene through a biological company;

[0019] 2) Using stable transfection technology, stably express the novel coronavirus orf3a and E genes in Caco2 cells (i.e., Caco2-3a-E cell line);

[0020] 3) Using molecular biology techniques to assemble in vitro the novel coronavirus genome SARS-CoV-2-△ORF3a-E-Firefly (Firefly gene inserted at the orf7a gene position) lacking orf7a, orf3a, and E genes;

[0021] 4) Using SARS-CoV-2-△ORF3a-E-Firefly as a template, transcribe the novel coronavirus genomic RNA using in vitro transcription technology, and electrophoretically transfer the RNA into the Caco2-3a-E cell line to produce the SARS-CoV-2-△ORF3a-E Fluc virus;

[0022] 5) SARS-CoV-2-△ORF3a-E Fluc virus can only replicate in Caco2-3a-E cell line, and the virus replication can be detected by detecting the expression of Firefly gene (such as Figure 1 shown).

[0023] 2. The Yadanziolide A small molecule drug was diluted with cell culture medium to concentrations of 80 μM, 40 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, 0.5 μM, 0.1 μM, 0.02 μM, 0.004 μM, 0.002 μM, and 0 μM (as a control).

[0024] 3. Inoculate Caco2-3a-E cells that have not been infected with the virus in a 96-well cell plate (4×10 4 After the cells grew to 90%, the culture medium in the culture plate was discarded, and the SARS-CoV-2-△ORF3a-E Fluc virus was infected at MOI = 0.01 (i.e., 50 μl of diluted virus was added to each well). After 1 hour of infection, the Yadanziolide A small molecule drug diluted in step 2 was added (i.e., 50 μl of diluted drug was added to each well). The cells were cultured in a cell culture incubator for 24 hours.

[0025] 4. Take out the 96-well cell culture plate and use the kit to detect the Luciferase value.

[0026] 5. Calculate the Luciferase value and draw a curve chart of Yadanziolide A inhibiting virus replication (such as Figure 2 ) and calculate the EC50.

[0027] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. The application of the small molecule drug Yadanziolide A in the preparation of anti-new coronavirus drugs, characterized in that: The CAS number of the small molecule Yadanziolide A is 95258-14-3.

2. The use according to claim 1, characterized in that: The small molecule Yadanziolide A is the only active ingredient in the drug.

3. The use according to any one of claims 1-2, characterized in that: The dosage form of the drug is any one of powder, tablet, granule, capsule, solution, emulsion and suspension.

4. The use according to any one of claims 1 to 2, characterized in that: The drug also contains a pharmaceutically acceptable carrier.

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