Application of Compound 3k in preparation of medicine for inhibiting porcine sapelovirus replication

Abstract

The invention belongs to the field of biology, and particularly relates to application of Compound 3k in preparation of a medicine for inhibiting porcine sapelovirus (PSV) from replication, in particular to application of Compound 3k in preparation of a medicine for inhibiting porcine sapelovirus (PSV) from replication. It is found that Compound 3k can efficiently inhibit the replication of PSV in cells, and the cytotoxicity is low. Therefore, the Compound 3k can be used for preparing antiviral drugs, can be applied to drugs for preventing or treating PSV infection, and has a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of biology, specifically relating to the application of Compound 3k in the preparation of drugs that inhibit the replication of porcine sapellovirus. Background Technology

[0002] Porcine sapelovirus (PSV) is a member of the genus PSV in the family Picornaviridae. It is a non-enveloped, single-stranded, positive-sense RNA virus. PSV was first discovered in the UK in 1960. Recent epidemiological surveys have shown that PSV has been detected in East my country, South China, Central China, and Western China, proving that the virus is widely distributed and continues to spread in my country.

[0003] PSV is considered one of the important pathogens causing diarrhea. Further research has found that, in addition to diarrhea, PSV can also cause multi-system diseases such as poliomyelitis, pneumonia, reproductive disorders, and myocarditis; mixed infections of PSV with other common viruses can severely aggravate the condition, leading to death and causing certain economic losses to the pig farming industry.

[0004] Currently, there are no antiviral drugs or commercially available PSV vaccines. Therefore, identifying specific drugs that can antagonize PSV replication is of great significance for the prevention and control of this disease.

[0005] Compound 3k is a pyruvate kinase M2 (PKM2) inhibitor. Its applications are primarily focused on the following areas: (1) PKM2-IN-1 can inhibit cancer cell proliferation: In cancer cell lines that highly express PKM2, such as HCT116, Hela, and H1299, the anti-proliferative activity against cancer cells is significantly higher than that against normal cells (Reference DOI:10.1016 / j.ejmech.2017.06.064).

[0006] (2) Signal pathway regulation: This inhibitor affects the signal transduction of STAT3, mitochondrial apoptosis pathway and other pathways by inhibiting PKM2 activity, and has the potential for application in tumor treatment research (Reference DOI:10.1038 / srep16082). Summary of the Invention

[0007] To address the aforementioned technical problems, this invention unexpectedly discovered that Compound 3k can efficiently inhibit the replication of porcine sapellovirus with low cytotoxicity. Therefore, Compound 3k can be used to prepare antiviral drugs for the prevention or treatment of porcine sapellovirus infection, showing promising application prospects. Specifically, it includes the following: In a first aspect, the present invention provides the use of Compound 3k or a pharmaceutically acceptable salt thereof in the preparation of antiviral drugs, wherein the structural formula of Compound 3k is shown in formula (I) below:

[0008] Equation (Ⅰ).

[0009] Preferably, the virus is swine sapello virus.

[0010] Preferably, the Compound 3k or its pharmaceutically acceptable salt is formulated into any pharmaceutically acceptable dosage form by adding pharmaceutically acceptable excipients.

[0011] Secondly, the present invention provides the use of Compound 3k or a pharmaceutically acceptable salt thereof in the preparation of a drug for preventing porcine sapellovirus infection, wherein the structural formula of Compound 3k is shown in formula (I) below:

[0012] Equation (Ⅰ).

[0013] Preferably, the Compound 3k or its pharmaceutically acceptable salt is formulated into any pharmaceutically acceptable dosage form by adding pharmaceutically acceptable excipients.

[0014] Thirdly, the present invention provides the use of Compound 3k or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating porcine sapellovirus infection, wherein the structural formula of Compound 3k is shown in formula (I) below:

[0015] Equation (Ⅰ).

[0016] Preferably, the Compound 3k or its pharmaceutically acceptable salt is formulated into any pharmaceutically acceptable dosage form by adding pharmaceutically acceptable excipients.

[0017] The beneficial effects of this invention are: This invention unexpectedly discovered that adding 1.0 to 2.0 μM of Compound 3k to PSV-infected cells can significantly inhibit viral replication, which is of great significance for the prevention and control of PSV and related research. Attached Figure Description

[0018] Figure 1 Cytotoxicity test results of Compound 3k.

[0019] Figure 2 A histogram of Compound 3k inhibiting PSV replication.

[0020] Figure 3 The image shows the fluorescent staining results of Compound 3k inhibiting PSV replication. Detailed Implementation

[0021] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the scope of the present invention in any way. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in this technical field. Unless otherwise specified, the reagents and materials used in the following embodiments are all commercially available.

[0022] Compound 3k (C 18 H 19 NO2S2 (CAS No.: 94164-88-2) was purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd.

[0023] The PSV strain (PSV / GN / CN / 2024) involved in the following examples was isolated from pig feces samples and preserved by the Microbiology Laboratory of Gansu Nationalities Normal University. It has about 97% homology with PSV GS01 strain (GenBank No. OM328111) and can be used to verify the activity of Compound 3k in inhibiting PSV described in this application.

[0024] Example 1: Cytotoxicity assay of Compound 3k Compound 3k was diluted with culture medium to 0 μM, 0.5 μM, 1.0 μM, 1.5 μM, 2.0 μM, and 3.0 μM, and added to well-grown PK-15 cells in 96-well plates. Each concentration was replicated in triplicate, and the cells were cultured for 12 h. Subsequently, 10 μL of CCK-8 reagent was added to each well, and the cells were incubated for 1 h. The OD of each well was then measured using a UV spectrophotometer. 450 Absorbance. The formula for calculating cell viability is: Cell viability (%) = (Average value of drug-treated group / Average value of control group) × 100%.

[0025] The results are as follows Figure 1 As shown, treatment with different concentrations of Compound 3k had no significant effect on the viability of PK-15 cells, demonstrating good safety.

[0026] Example 2: Effect of Compound 3k on PSV Virus Titer PSV was inoculated into PK-15 cells at an MOI of 0.01. One hour after infection, the virus was washed away, and then Compound 3k was added to the culture medium to achieve final concentrations of 0 μM, 0.5 μM, 1.0 μM, 1.5 μM, and 2.0 μM. The culture medium was harvested at 12 h, 36 h, and 60 h post-infection and treated with TCID50. 50 The method determines the viral content.

[0027] The results are as follows Figure 2 As shown, after 12, 36, and 60 h of treatment with 1.5 μM and 2.0 μM Compound 3k, the viral titer of PSV-infected cells was significantly lower than that of the control group without Compound 3k, with differences ranging from 100 to 10,000 times. In particular, the virus was virtually undetectable in the 2.0 μM Compound 3k treatment group at 12 and 36 h post-infection. This indicates that Compound 3k significantly inhibits PSV replication in a dose-dependent manner.

[0028] Example 3: Inhibition of PSV virus replication by Compound 3k PSV was inoculated into PK-15 cells at an MOI of 0.01. After infection for 1 h, the virus was washed away, and Compound 3k was added to the culture medium to achieve final concentrations of 0 μM, 0.5 μM, 1.0 μM, 1.5 μM, and 2.0 μM. At 60 h post-infection, cells were fixed with 4% paraformaldehyde at room temperature for 30 min, then the fixative was washed away, and cells were permeabilized with 0.5% Triton-X100 solution. After permeabilization for 15 min, cells were fixed with 5% skim milk, and then incubated with anti-PSV-VP1 monoclonal antibody (1:2000 dilution) and anti-mouse FITC secondary antibody (1:500) for 1 h each. Finally, cells were stained with DAPI for 10 min and the fluorescence signal was observed under a microscope.

[0029] The results are as follows Figure 3 As shown, after PSV infection, treatment with different concentrations of Compound 3k can significantly inhibit PSV replication, with the 2.0 μM concentration showing the most significant inhibitory effect.

[0030] In summary, the Compound 3k described in this application can significantly inhibit PSV replication and has low cytotoxicity. It can be used as a candidate drug or in combination with other inhibitors for the prevention or treatment of PSV infection.

[0031] The above embodiments are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Therefore, all equivalent changes or modifications made in accordance with the structure, features and principles of the present invention should be included in the scope of the present invention.

Claims

1. Use of Compound 3k or a pharmaceutically acceptable salt thereof in the manufacture of an antiviral medicament, characterized in that, The structural formula of the Compound 3k is shown in the following formula (I): Formula (I).

2. Use according to claim 1, wherein The virus is porcine sapelovirus.

3. Use according to claim 1 or 2, characterized in that, The Compound 3k or its pharmaceutically acceptable salt is added into pharmaceutically acceptable adjuvant to form any pharmaceutically acceptable dosage form.

4. Use of compound 3k or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for preventing porcine sapovirus infection, characterized in that, The structural formula of the Compound 3k is shown in the following formula (I): Formula (I).

5. Use according to claim 4, characterized in that, The Compound 3k or its pharmaceutically acceptable salt is added into pharmaceutically acceptable adjuvant to form any pharmaceutically acceptable dosage form.

6. Use of Compound 3k or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of porcine sapovirus infection, characterized in that, The structural formula of the Compound 3k is shown in the following formula (I): Formula (I).

7. Use according to claim 6, characterized in that, The Compound 3k or its pharmaceutically acceptable salt is added into pharmaceutically acceptable adjuvant to form any pharmaceutically acceptable dosage form.

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