Eprinomectin and selamectin as inhibitors for resisting porcine epidemic diarrhea virus and application of iprinomectin and selamectin

Abstract

The invention discloses eprinomectin and selamectin as inhibitors for resisting porcine epidemic diarrhea virus and application of the eprinomectin and selamectin as inhibitors for resisting porcine epidemic diarrhea virus (PEDV). The eprinomectin and selamectin as inhibitors for resisting porcine epidemic diarrhea virus (PEDV) are found for the first time. The PEDV resisting effect of eprinomectin and selamectin is verified through the indirect immunofluorescence technology, the TCID50 technology, the qRT-PCR technology and the like. According to the invention, through a new way of using old medicines, the research and development period of the medicines is greatly shortened, the medicines can be quickly put into use, the defects of vaccines are made up, meanwhile, the epidemic of variant strains can be effectively handled, and meanwhile, a new treatment scheme is provided for infection of PEDV, even coronavirus and other viruses; the invention also provides an important scientific basis for the research and development of the application of the eprinomectin and selamectin in further inhibition of coronavirus and other viruses.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to eprimectin and selamectin as anti-porcine epidemic diarrhea virus inhibitors and applications thereof. Background Art

[0002] Porcine epidemic diarrhea (PED) caused by porcine epidemic diarrhea virus (PED virus, PEDV) is a highly contagious enteric disease of pigs, which has caused huge economic losses to the pig farming industry and is also a serious animal disease that has plagued the world's pig farming industry for many years. In recent years, researchers at home and abroad have conducted a lot of research on vaccines and antiviral drugs for porcine epidemic diarrhea. Among them, there are many achievements in the development of vaccines, which are also the main measures to prevent PEDV. The vaccines currently developed include PED inactivated vaccines, attenuated vaccines, and genetically engineered vaccines. Although vaccination can prevent PEDV infection to a certain extent, with the continuous mutation of the strain and the complexity of the disease in production practice, the immune effects of these vaccines are not ideal. There are not many achievements in drug research and development, mainly due to the long development time, high cost, side effects and other problems, resulting in no very effective specific drugs for this disease on the market, and even some new drugs developed are almost ineffective.

[0003] New uses for old drugs (also known as drug repurposing and drug repositioning) is a new drug development strategy that re-explores and develops the applications of known drugs (including those already on the market) and uses them to treat other diseases. New uses for old drugs as a drug development strategy are gaining more and more attention. Since most of the target drugs used in new uses of old drugs have been on the market for many years or have a long period of clinical use, people already have a relatively sufficient understanding of the safety and potential dangers of the drugs, so the failure rate of research and development can be reduced during the research and development process, effectively reducing research and development costs. Currently, a variety of old drugs have been proven to have new effects against coronaviruses, and some have even been approved for the treatment of some coronavirus diseases.

[0004] Eprinomectin (MK-397) and Selamectin are both avermectin anthelmintics, and both are veterinary drugs approved for use by the U.S. Food and Drug Administration (FDA). The most common derivative of avermectin is ivermectin, which has various forms and can be used not only to prevent heartworms and treat external parasites, but also as a microfilarian. The eprinomectin proposed in this application has anthelmintic, insecticidal and acaricidal activities, and has excellent broad-spectrum antiparasitic effects when applied topically to cattle; Selamectin can be used as a topical parasiticide and anthelmintic for dogs and cats, and is used to treat and prevent heartworms, fleas, ear mites, scabies and certain types of tick infections in dogs, and to prevent heartworms, fleas, ear mites, hookworms and roundworms in cats.

[0005] Existing studies have reported that ivermectin also has the effect of preventing or treating viral diseases. For example, Chinese patent application document CN118510506A discloses an aqueous composition containing avermectin, wherein at least one active pharmaceutical ingredient selected from the avermectin group is ivermectin, which is used to prevent or treat viral diseases, especially for preventing or treating SARS-Covid-2 infection or HIV infection; for another example, a US patent document US08741856B2 discloses avermectin and milbemycin compounds, especially ivermectin, for preparing antiviral drugs for treating, preventing or improving flavivirus infections, such as YFV (yellow fever virus), DENV (dengue virus), JEV (Japanese encephalitis virus), TBEV (tick-borne encephalitis virus) or MODV (Modoc virus).

[0006] However, ivermectin has not been proven to be effective in preventing and treating porcine epidemic diarrhea virus. Moreover, neither ivermectin nor selamectin proposed in this application has been reported to be useful in preventing or treating viral diseases, let alone porcine epidemic diarrhea virus. As mentioned above, the safety of these drugs is already available, but whether they have an anti-porcine epidemic diarrhea virus effect requires further study. Summary of the invention

[0007] The immune effects of existing porcine epidemic diarrhea vaccines are not ideal, and there is no very effective specific medicine for the disease on the market. The present invention provides an anti-porcine epidemic diarrhea virus inhibitor and its application. By using old drugs in new ways, the avermectin-type anthelmintics epramectin and selamectin are screened out, both of which have good anti-porcine epidemic diarrhea virus effects, greatly shortening the drug research and development cycle, and can be quickly put into use, making up for the shortcomings of vaccines, and effectively responding to the prevalence of mutant strains. The specific technical scheme is as follows:

[0008] One of the objects of the present invention is to provide a porcine epidemic diarrhea virus inhibitor, the active ingredient of which includes any one of epromectin or selamectin or a pharmaceutically acceptable salt of epromectin and selamectin.

[0009] The aforementioned porcine epidemic diarrhea virus inhibitor, the molecular structure formula of the epramectin is shown in Formula 1, and the molecular structure formula of the selamectin is shown in Formula 2;

[0010] Formula 1:

[0011]

[0012] Formula 2:

[0013]

[0014] The aforementioned porcine epidemic diarrhea virus inhibitor, the concentration of epramectin is 2.5 μmol / L to 10 μmol / L; the concentration of selamectin is 2.5 μmol / L to 9 μmol / L.

[0015] Preferably, in the aforementioned porcine epidemic diarrhea virus inhibitor, the concentration of eprimectin is 7 μmol / L, 8 μmol / L, 9 μmol / L or 10 μmol / L.

[0016] Further preferably, in the aforementioned porcine epidemic diarrhea virus inhibitor, the concentration of eprimectin is 10 μmol / L.

[0017] Preferably, in the aforementioned porcine epidemic diarrhea virus inhibitor, the concentration of selamectin is 6 μmol / L, 8 μmol / L or 9 μmol / L.

[0018] Further preferably, in the aforementioned porcine epidemic diarrhea virus inhibitor, the concentration of selamectin is 9 μmol / L.

[0019] Another object of the present invention is to provide an application of the aforementioned porcine epidemic diarrhea virus inhibitor in the preparation of a drug for treating or preventing porcine epidemic diarrhea virus.

[0020] Beneficial effects of the present invention:

[0021] 1) Through the approach of repurposing old drugs, the avermectin-type anthelmintics epramectin and selamectin were screened out, both of which have good anti-porcine epidemic diarrhea virus effects, greatly shortening the drug research and development cycle. They can be put into use quickly, making up for the shortcomings of vaccines and effectively responding to the spread of mutant strains.

[0022] 2) The present invention uses a combination of cell biology and virology techniques to discover for the first time that eprimectin and selamectin have a new effect in inhibiting PEDV infection, providing a new treatment option for PEDV and even coronavirus and other viral infections.

[0023] 3) The present invention verifies for the first time the effective and safe concentrations of epramectin and selamectin for inhibiting PEDV infection, providing an important scientific basis for their further development of drug applications for inhibiting coronaviruses and other viruses. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 The results of indirect immunofluorescence experiments of epromectin and selamectin inhibiting PEDV;

[0025] Figure 2 The results of the toxicity test of epramectin and selamectin on Vero cells;

[0026] Figure 3 The RNA load of PEDV inhibited by different concentrations of epramectin;

[0027] Figure 4 The RNA load of PEDV inhibited by different concentrations of selamectin;

[0028] Figure 5 is the inhibitory effect of epromectin on PEDV;

[0029] Figure 6 This is the inhibitory effect of selamectin on PEDV. DETAILED DESCRIPTION

[0030] The specific implementation methods of the present invention are further described in detail below in conjunction with the examples. The reagents not provided with the source in the following experiments are all commercially available reagents, and the methods not described in detail are all conventional well-known experimental methods. The old drugs involved in the examples of the present invention are all derived from FDA-approved veterinary drugs. The molecular structure formula of the compound eprimectin involved in each example is as follows:

[0031]

[0032] The molecular structure of selamectin is as follows:

[0033]

[0034] The present invention uses two old drugs, ipromectin and selamectin, as examples to illustrate the inhibitory effect on porcine viruses. Pharmaceutically acceptable salts of these two old drugs also have inhibitory effects on PEDV. Specific examples are as follows:

[0035] Example 1: Identification of the preliminary inhibitory effects of epremectin and selamectin on Vero cells infected with PEDV

[0036] In order to verify whether eprimectin and selamectin have inhibitory effects on Vero cells infected with PEDV, indirect immunofluorescence assay (IFA) was used for identification.

[0037] Vero cells were cultured at 2.5 × 10 5 24-well cell culture plates were laid with 10% (v / v) heat-inactivated fetal bovine serum, 100U / mL penicillin, and 100μg / mL streptomycin in each well, and 1mL of DMEM medium containing 10% (v / v) heat-inactivated fetal bovine serum, 100U / mL penicillin, and 100μg / mL streptomycin was added to each well, and cultured in a 37°C, 5% CO2 incubator for 24h; the original cell culture medium was removed, and the cells were washed three times with phosphate buffered saline (PBS), and then 1mL of culture medium containing different concentrations (2.5μmol / L, 5μmol / L, 10μmol / L) of epromectin or selamectin was added to each well and incubated for 2h; new DMEM medium was replaced, and PEDV with a virus infection multiplicity (MOI) of 0.1 was used. The AJ1102 strain infected Vero cells at 37°C for 2 hours; discarded the cell supernatant, washed the cells three times with phosphate buffered saline (PBS) to wash away the virus particles that did not invade the cells; added 1 mL of DMEM culture medium containing different concentrations (2.5 μmol / L, 5 μmol / L, 10 μmol / L) of epromectin or selamectin to the wells of the cell culture plate, and continued to culture at 37°C for 18 hours. The cells were fixed with 4% paraformaldehyde, incubated with PEDV N antibody at room temperature for 2 hours, incubated with fluorescent secondary antibody in the dark for 1 hour, incubated with DAPI for 15 minutes, and observed the results with an Olympus BX63 fully electric upright fluorescence microscope.

[0038] The results are as follows Figure 1 As shown in the figure, at a concentration of 10 μmol / L, both compounds had a significant inhibitory effect on PEDV. Compared with the cell activity of the group without compound treatment (DMSO group), at a concentration of 2.5 μmol / L, the green fluorescence of the epromectin group and the selamectin group did not decrease significantly; at a concentration of 5 μmol / L, the green fluorescence of the epromectin group and the selamectin group decreased to a certain extent, but the decrease was not very obvious; at a concentration of 10 μmol / L, the green fluorescence of the epromectin group and the selamectin group decreased significantly, and both compounds showed a certain concentration dependence, indicating that epromectin and selamectin have an inhibitory effect on Vero cells infected with PEDV.

[0039] Example 2: Identification of the toxicity of epremectin and selamectin to Vero cells

[0040] In order to further verify the cytotoxicity of epramectin and selamectin on Vero cells and determine the effective concentrations of the two compounds in inhibiting Vero cell infection with PEDV, CCK-8 cytotoxicity assay was used for identification.

[0041] The cells were cultured as in Example 1. After the cells were fully grown, the Vero cells were grown in a 37°C, 5% CO2 incubator. After the cells in the culture flask were fully grown, the Vero cells were plated at 2.5×10 5 96-well cell culture plates were plated with 100 μL of DMEM medium containing 10% (v / v) heat-inactivated fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin, and each well was added with 100 μL of DMEM medium containing 10% (v / v) heat-inactivated fetal bovine serum, 100 U / mL penicillin, and 100 μg / mL streptomycin, and cultured in a 37°C, 5% CO2 incubator for 24 h; the original cell culture medium was removed and replaced with 100 μL of DMEM culture medium containing different concentrations (0 μmol / L, 2.5 μmol / L, 5 μmol / L, 7.5 μmol / L, 10 μmol / L, 12.5 μmol / L, 15 μmol / L, 20 μmol / L, 25 μmol / L) of epramectin or selamectin, and incubated at 37°C for 24 h, then 10 μL of CCK-8 reagent was added and incubated in a 37°C, 5% CO2 incubator for 2 h; cytotoxicity identification was performed according to the operating instructions.

[0042] The results are as follows Figure 2 As shown, Figure A is Eprimectin CC 50 Fitting curve, reflecting the CC of eprimectin 50 Value, Figure B shows CC of selamectin 50 Fitting curve, reflecting the CC of selamectin 50 The data showed that compared with the cell activity of the group without compound treatment (DMSO group), below the concentration of 10μmol / L, after ipramectin treatment, the cells can maintain a survival rate of about 80%, and after selamectin treatment, the cell survival rate is about 75%, indicating that the maximum non-toxic concentration of ipramectin is 10μmol / L, while the maximum non-toxic concentration of selamectin is lower than 10μmol / L. Therefore, in the subsequent verification of the virus effectiveness, the maximum concentration of ipramectin was set to 10μmol / L, and the maximum concentration of selamectin was set to 9μmol / L.

[0043] Example 3: Effect of eprimectin on PEDV RNA content in Vero cells infected with

[0044] To verify whether eprimectin can reduce the PEDV RNA content in Vero cells infected with PEDV in a concentration-dependent manner, real-time fluorescence quantitative PCR (qRT-PCR) was used for identification.

[0045] Vero cells were cultured at 2.5 × 10 524-well cell culture plates were laid with 10% (v / v) heat-inactivated fetal bovine serum, 100U / mL penicillin, and 100μg / mL streptomycin in 1mL DMEM medium to each well, and cultured in a 37°C, 5% CO2 incubator for 24h; the original cell culture medium was removed, and the cells were washed three times with phosphate buffered saline (PBS), and then 1mL DMEM medium containing different concentrations of epremectin (7μmol / L, 8μmol / L, 9μmol / L, 10μmol / L) was added to each well and incubated for 2h; new DMEM medium was replaced with PEDV with a virus infection multiplicity (MOI) of 0.1 The AJ1102 strain infected Vero cells at 37°C for 2 hours; discarded the cell supernatant, washed the cells three times with phosphate buffered saline (PBS) to wash away the virus particles that did not invade the cells; added 1 mL of DMEM culture medium containing different concentrations of epremectin (7 μmol / L, 8 μmol / L, 9 μmol / L, 10 μmol / L) to the wells of the cell culture plate, and continued to culture at 37°C for 16 hours. The untreated Vero cells infected with PEDV were used as the control.

[0046] The cells were collected, the cell supernatant was discarded, and the cells were washed three times with PBS; the total RNA of PEDV-infected cells was extracted using the RNA extraction reagent Trizol, and the complementary DNA (cDNA) was generated using the PrimeScript RT Master Mix reverse transcription kit as a qRT-PCR template; the PEDV nucleocapsid protein (N) RNA content and the PEDV non-structural protein 12 (Nsp12) content were measured by relative qRT-PCR to represent the viral RNA content of infected cells, respectively; the viral RNA was standardized with actin β-actin mRNA, and the expression of cDNA was detected by Methods: Relative quantification was performed; primers and reaction systems are shown in Tables 1 and 2; the staining method fluorescence quantitative premix was AceQ Universal SYBR qPCR master Mix; the reaction program used the Fast program provided with the ABI fluorescence quantitative PCR instrument.

[0047] Table 1. RT-qPCR primers are as follows

[0048]

[0049] Table 2. Reaction system and procedure

[0050]

[0051] The experiment was performed three times independently with three replicates each time. The experimental data are expressed as the group mean and standard deviation (SD). Statistical analysis was performed using GraphPad Prism 8.0 software with unpaired, two-tailed Student t-test. ***p<0.001 indicated extremely significant statistical differences.

[0052] The results are as follows Figure 3 As shown, compared with the group without epramectin (0 μmol / L), after treatment with 7 μmol / L, 8 μmol / L, 9 μmol / L, and 10 μmol / L epramectin, the PEDV RNA content decreased in a concentration-dependent manner with the increase in concentration, and the RNA content in the treatment group containing 10 μmol / L epramectin decreased most significantly (reduction>99%), indicating that epramectin can significantly reduce the PEDV RNA content in Vero cells infected with epramectin.

[0053] Example 4: Effect of selamectin on PEDV RNA content in Vero cells infected with

[0054] To verify whether selamectin can reduce the PEDV RNA content in Vero cells infected with PEDV in a concentration-dependent manner, real-time fluorescence quantitative PCR (qRT-PCR) was used for identification.

[0055] Vero cells were cultured at 2.5 × 10 5 24-well cell culture plates were laid with 10% (v / v) heat-inactivated fetal bovine serum, 100U / mL penicillin, and 100μg / mL streptomycin in each well, and 1mL of DMEM medium containing 10% (v / v) heat-inactivated fetal bovine serum, 100U / mL penicillin, and 100μg / mL streptomycin was added to each well, and cultured in a 37°C, 5% CO2 incubator for 24h; the original cell culture medium was removed, and the cells were washed three times with phosphate buffered saline (PBS), and then 1mL of DMEM medium containing different concentrations of selamectin (6μmol / L, 8μmol / L, 9μmol / L) was added to each well and incubated for 2h; new DMEM medium was replaced, and Vero cells were infected with PEDV AJ1102 strain at a virus infection multiplicity (MOI) of 0.1 at 37°C for 2h; the cell supernatant was discarded, and the cells were washed three times with phosphate buffered saline (PBS) to wash away the virus particles that did not invade the cells; 1mL of DMEM medium containing different concentrations of selamectin (6μmol / L, 8μmol / L, 9μmol / L) was added to the wells of the cell culture plate The cells were cultured in DMEM medium at 37°C for 16 h, and the untreated PEDV-infected Vero cells were used as a control.

[0056] The cells were collected, the cell supernatant was discarded, and the cells were washed three times with PBS. The total RNA of PEDV-infected cells was extracted using the RNA extraction reagent Trizol, and the complementary DNA (cDNA) was generated using the PrimeScript RT Master Mix reverse transcription kit as a qRT-PCR template. The PEDV nucleocapsid protein (N) RNA content and the PEDV non-structural protein 12 (Nsp12) content were measured by relative qRT-PCR to represent the viral RNA content of infected cells, respectively. The viral RNA was standardized with actin β-actin mRNA and the expression of cDNA was measured by 2 -△△CT The method was to perform relative quantification; the primers and reaction system were consistent with those in Tables 1 and 2; the staining method fluorescence quantitative premix was AceQ Universal SYBR qPCR master Mix; and the reaction program used the Fast program provided with the ABI fluorescence quantitative PCR instrument.

[0057] The experiment was performed three times independently with three replicates each time. The experimental data are expressed as the group mean and standard deviation (SD). Statistical analysis was performed using GraphPad Prism 8.0 software with unpaired, two-tailed Student t-test. ***p<0.001 indicated extremely significant statistical differences.

[0058] The results are as follows Figure 4 As shown, compared with the group without selamectin (0 μM) treatment, after treatment with 6 μmol / L, 8 μmol / L, and 9 μmol / L selamectin, the PEDV RNA content decreased in a concentration-dependent manner with increasing concentrations, and the RNA content in the treatment group containing 9 μmol / L selamectin decreased most significantly (reduction>99%), indicating that selamectin can significantly reduce the PEDV RNA content in Vero cells infected with selamectin.

[0059] Example 5: Effect of Eprimectin on the Inhibition of PEDV Virus Infection in Vero Cells

[0060] Vero cells were cultured according to Example 1; new DMEM culture medium was replaced, and 1 mL of DMEM culture medium containing different concentrations (7 μmol / L, 8 μmol / L, 9 μmol / L, 10 μmol / L) of epremectin was added to the wells of the cell culture plate, and culture was continued at 37°C for 2 h; the PEDV AJ1102 strain was infected with Vero cells at a virus infection multiplicity (MOI) of 0.1 at 37°C for 2 h; the cell supernatant was discarded, and the cells were washed three times with PBS to wash away the virus particles that did not invade the cells; 1 mL of DMEM culture medium containing different concentrations (7 μmol / L, 8 μmol / L, 9 μmol / L, 10 μmol / L) of epremectin was added to the wells of the cell culture plate, and culture was continued at 37°C for 18 h for a virus titer experiment (TCID50), and Vero cells infected with PEDV without epremectin treatment were used as a control.

[0061] Uninfected Vero cells were diluted to 2.5×105 / mL and laid on 96-well cell culture plates, 100 μL of DMEM medium was added to each well, and the plates were cultured overnight at 37°C and 5% CO2 incubator. The supernatant of the Vero cells infected for 18 hours was diluted 10-fold and inoculated on Vero cells laid on 96-well cell culture plates, 100 μL was inoculated in each well, and 8 replicates were made for each dilution. The supernatant of Vero cells infected with PEDV without treatment was used as a control. The 96-well cell culture plates were cultured in a 37°C and 5% CO2 incubator. The growth status of the cells was observed every 24 hours, and the number of cytopathic wells was recorded. The observation was performed for 2 days until the number of cytopathic wells no longer increased.

[0062] The TCID50 of the virus was calculated using the Reed-Muench method; the experimental data were expressed as group mean and standard deviation (SD), and statistical analysis was performed using GraphPad Prism 8.0 software with unpaired, two-tailed Student t-test; ***p<0.001 indicated a highly significant statistical difference.

[0063] The results are as follows Figure 5 As shown, compared with the group without epramectin (0 μM) treatment, after treatment with 7 μmol / L, 8 μmol / L, 9 μmol / L, and 10 μmol / L epramectin, the titer of PEDV decreased in a concentration-dependent manner with the increase in concentration, among which the virus titer of the 8 μmol / L, 9 μmol / L, and 10 μmol / L epramectin treatment groups was significantly reduced, indicating that epramectin can significantly reduce the infection amount of PEDV in Vero cells.

[0064] Example 6: Effect of selamectin on the inhibitory effect of PEDV infection on Vero cells

[0065] Vero cells were cultured according to Example 1; new DMEM culture medium was replaced, and 1 mL of DMEM culture medium containing different concentrations (6 μmol / L, 8 μmol / L, 9 μmol / L) of selamectin was added to the wells of the cell culture plate, and the culture was continued at 37°C for 2 h; the PEDV AJ1102 strain was infected with Vero cells at a virus infection multiplicity (MOI) of 0.1 at 37°C for 2 h; the cell supernatant was discarded, and the cells were washed three times with PBS to wash away the virus particles that did not invade the cells; 1 mL of DMEM culture medium containing different concentrations (6 μmol / L, 8 μmol / L, 9 μmol / L) of selamectin was added to the wells of the cell culture plate, and the culture was continued at 37°C for 18 h to perform a virus titer experiment (TCID50), and Vero cells infected with PEDV without epromectin treatment were used as a control.

[0066] Uninfected Vero cells were diluted to 2.5×105 / mL and laid on 96-well cell culture plates, 100 μL of DMEM medium was added to each well, and the plates were cultured overnight at 37°C and 5% CO2 incubator. The supernatant of the Vero cells infected for 18 hours was diluted 10-fold and inoculated on Vero cells laid on 96-well cell culture plates, 100 μL was inoculated in each well, and 8 replicates were made for each dilution. The supernatant of Vero cells infected with PEDV without treatment was used as a control. The 96-well cell culture plates were placed in a 37°C and 5% CO2 incubator for culture. The growth status of the cells was observed every 24 hours, and the number of cytopathic wells was recorded. The observation was performed for 2 days until the number of cytopathic wells no longer increased.

[0067] The TCID50 of the virus was calculated using the Reed-Muench method; the experimental data were expressed as group mean and standard deviation (SD), and statistical analysis was performed using GraphPad Prism 8.0 software with unpaired, two-tailed Student t-test; ***p<0.001 indicated a highly significant statistical difference.

[0068] The results are as follows Figure 6 As shown, compared with the group without selamectin (0 μM) treatment, the titer of PEDV decreased in a concentration-dependent manner after treatment with 6 μmol / L, 8 μmol / L, and 9 μmol / L selamectin, respectively, with the increase of concentration, and the virus titer was significantly reduced under these concentrations, indicating that selamectin can significantly reduce the infection amount of PEDV in Vero cells.

[0069] It is obvious to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be regarded as exemplary and non-restrictive from any point of view. In addition, it should be understood that although this specification is described in accordance with the implementation mode, it does not include only one technical solution. This narrative mode of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole, and the technical solutions in the embodiments can also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A porcine epidemic diarrhea virus inhibitor, characterized in that: The active ingredient includes any one of eprimectin or selamectin or pharmaceutically acceptable salts of eprimectin and selamectin.

2. The porcine epidemic diarrhea virus inhibitor according to claim 1, characterized in that: The molecular structure formula of the epramectin is shown in Formula 1, and the molecular structure formula of the selamectin is shown in Formula 2; Formula 1: Formula 2:

3. The porcine epidemic diarrhea virus inhibitor according to claim 1, characterized in that: The concentration of the epramectin is 2.5 μmol / L to 10 μmol / L; the concentration of the selamectin is 2.5 μmol / L to 9 μmol / L.

4. The porcine epidemic diarrhea virus inhibitor according to claim 3, characterized in that: The concentration of eprimectin is 7 μmol / L, 8 μmol / L, 9 μmol / L or 10 μmol / L.

5. The porcine epidemic diarrhea virus inhibitor according to claim 4, characterized in that: The concentration of eprimectin is 10 μmol / L.

6. The porcine epidemic diarrhea virus inhibitor according to claim 3, characterized in that: The concentration of selamectin is 6 μmol / L, 8 μmol / L or 9 μmol / L.

7. The porcine epidemic diarrhea virus inhibitor according to claim 6, characterized in that: The concentration of selamectin is 9 μmol / L.

8. Use of the porcine epidemic diarrhea virus inhibitor according to any one of claims 1 to 7, characterized in that: Application in the preparation of medicines for treating or preventing epidemic diarrhea virus.

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