Use of FB23-2 in inhibiting PEDV or TGEV cell replication

Abstract

The application discloses application of FB23-2 in inhibition of PEDV or TGEV cell replication, relates to the technical field of preventive veterinary medicine, and has the technical points that the FB23-2 is used for inhibiting porcine epidemic diarrhea virus (PEDV) or porcine transmissible gastroenteritis virus (TGEV) replication in cells; experiments show that the FB23-2 can efficiently inhibit PEDV and TGEV replication in cells, the concentration of the FB23-2 is controlled to be 2.5-25 muM, the inhibition effect is best, the FB23-2 can be used for prevention and control of PEDV and TGEV viruses, and has important significance for related scientific research.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of preventive veterinary technology, and particularly relates to application of FB23-2 in inhibiting cell replication of PEDV or TGEV. BACKGROUND

[0002] Porcine epidemic diarrheavirus (PEDV) and porcine transmissible gastroenteritis virus (TGEV) are both members of the alpha coronavirus genus of the Coronaviridae family, are single-stranded RNA viruses with envelopes, are widely present in nature and artificial environment, have a diameter of about 80-150 nm, and have a total length of about 28 kb. PEDV and TGEV mainly harm newborn piglets within 7 days of age, and the clinical symptoms are very similar, mainly including diarrhea, vomiting, and dehydration. The mortality of newborn piglets infected with the above two viruses can be more than 95%, which is an important epidemic disease hindering the healthy development of the pig industry. Vaccine immunization is the main means for preventing and controlling PEDV and TGEV in the current pig breeding industry.

[0003] At present, the PEDV and TGEV vaccines on the market are all attenuated live vaccines and inactivated vaccines, and no other vaccines such as subunit vaccines have been approved for marketing. However, the above viruses mainly harm newborn piglets, and newborn piglets have no complete immune system, so vaccination cannot stimulate humoral immunity and cellular immunity, and therefore the effect is poor. In addition, the maternal antibodies stimulated by immunizing sows can only be transmitted to piglets through breast milk, and the transmission efficiency is low and the protection effect is poor.

[0004] Therefore, the present application aims to provide the application of FB23-2 in inhibiting cell replication of PEDV or TGEV to solve the above problems. SUMMARY

[0005] The purpose of the present application is to solve the above problems, provide the application of FB23-2 in inhibiting cell replication of PEDV or TGEV, and FB23-2 can efficiently inhibit the replication of PEDV or TGEV in cells.

[0006] In order to achieve the above purpose, the technical scheme of the present application is as follows:

[0007] The present application provides the application of FB23-2 in inhibiting cell replication of PEDV or TGEV, and the FB23-2 can inhibit the replication of PEDV and TGEV in cells. When the concentration of the FB23-2 is 1 μM to <50 μM, the replication of PEDV in cells is inhibited. When the concentration of the FB23-2 is 1 μM to <50 μM, the replication of TGEV in cells is inhibited.

[0008] The chemical formula of the FB23-2 is C 18 H 15 Cl2N3O3.

[0009] Compared with the prior art, the beneficial effects of the present application are:

[0010] The present application finds that FB23-2 can be used to inhibit the replication of porcine epidemic diarrhea virus (PEDV) or porcine transmissible gastroenteritis virus (TGEV) in cells, and FB23-2 can be used for the prevention and control of PEDV and TGEV viruses, which has important significance for related scientific research. BRIEF DESCRIPTION OF DRAWINGS

[0011] Figure 1 is a column chart of PEDV replication and a curve chart of cell toxicity in the embodiment of the present application, wherein A is a column chart of FB23-2 of different concentrations inhibiting PEDV replication; B is a curve chart of FB23-2 of different concentrations not having significant cell toxicity;

[0012] Figure 2 is a result chart of FB23-2 inhibiting PEDV replication in the embodiment of the present application, which shows that FB23-2 of different concentrations can inhibit PEDV replication and reduce the number of plaques;

[0013] Figure 3 is a column chart of TGEV replication and a curve chart of cell toxicity in the embodiment of the present application, wherein A is a column chart of FB23-2 of different concentrations inhibiting TGEV replication; B is a curve chart of FB23-2 of different concentrations not having significant cell toxicity;

[0014] Figure 4 is a result chart of FB23-2 inhibiting TGEV replication in the embodiment of the present application, which shows that FB23-2 of different concentrations can inhibit TGEV replication and reduce the number of plaques;

[0015] Figure 5 is a structural formula of FB23-2 in the embodiment of the present application. DETAILED DESCRIPTION

[0016] In order to enable the persons skilled in the art to better understand the present application, the technical solutions of the present application will be further described in detail below with reference to the embodiments of the present application and the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor should belong to the protection scope of the present application.

[0017] It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict. The present application will be described in detail below with reference to the embodiments.

[0018] Example 1

[0019] Experiments show that when the concentration is less than 50 μM, different concentrations of FB23-2 can inhibit the replication of PEDV, and after FB23-2 treatment, there is no significant effect on cell activity.

[0020] Vero cells were inoculated with PEDV at a dose of MOI = 0.01, and after 1 hour of infection, the unabsorbed virus was washed away, and then FB23 or DMSO was added in the culture medium, so that the final concentration of FB23 was 0 μM, 1 μM, 2.5 μM, 10 μM and 50 μM. At 12 hours, 36 hours and 60 hours after infection, the culture medium was collected, and the virus content was determined by the TCID 50 Method. At the same time, the control cells without PEDV infection but with FB23-2 were set up for cell activity determination.

[0021] The results show that at 12 hours and 36 hours after PEDV infection, the virus titers of the 2.5 μM, 10 μM and 50 μM FB23-2 treatment groups are significantly lower than those of the 0 μM FB23-2 control group, indicating that FB23-2 can inhibit the replication of PEDV, with a difference of 5-10 times Figure 1 A). It is shown that FB23-2 inhibits the replication of PEDV Figure 1 A). The cell activity detection of FB23-2 treated cells without PEDV infection shows that after the cells are treated with 0.1-10 μM concentration of FB23-2, there is no significant difference in cell activity, but after the cells are treated with FB23-2 at a concentration of 50 μM or more, the cell activity is significantly decreased Figure 1 B).

[0022] Vero cells were infected with PEDV, and after 1 hour of infection, the unabsorbed virus was washed away, and then 1% methyl cellulose containing different concentrations of FB23-2 was added, and the culture was continued at 37°C for 48 hours. After removing the methyl cellulose, 1% crystal violet was used for staining, and the size and number of plaques were observed.

[0023] The results show that the cells infected with PEDV without FB23-2 concentration group appear a small amount of pinhead-like plaques; the number of plaques of the PEDV infected cells treated with 2.5-10 μM FB23-2 is reduced Figure 2 ). The above results fully show that FB23-2 significantly inhibits the infection and replication of PEDV, and the effect increases with the increase of the concentration of FB23-2, showing a dose-dependent manner.

[0024] Example 2

[0025] The experiment shows that when the concentration is less than 100 μM, different concentrations of FB23-2 can inhibit the replication of TGEV, and after FB23-2 treatment, there is no significant effect on cell activity.

[0026] PK-15 cells were inoculated with TGEV at an MOI of 0.01, and after 1 hour of infection, the virus was washed away, and then FB23-2 or DMSO was added to the culture medium, so that the final concentration of FB23-2 was 1 μM, 2.5 μM, 10 μM and 50 μM FB23-2. The culture medium was collected at 12 hours, 36 hours and 60 hours after infection, and the TCID50 of the virus was determined. 50 The method was used to determine the virus content. At the same time, TGEV-uninfected FB23-2-added control cells were set up for cell activity determination.

[0027] The results show that at 12, 36 and 60 hours after TGEV infection, the virus titers of the 01 μM, 2.5 μM, 10 μM and 50 μM FB23-2 treatment groups are significantly lower than those of the 0 μM FB23-2 control group, with a difference of 5-10 times, indicating that FB23-2 can inhibit the replication of TGEV Figure 3 A). The CCK-8 experiment results of FB23-2-treated cells without TGEV infection show that after being treated with 1-50 μM concentration of FB23-2, the cell activity has no significant difference, but after being treated with 100 μM concentration of FB23-2, the cell activity decreases significantly Figure 3 B)

[0028] PK-15 cells were infected with TGEV, and after 1 hour of infection, the unabsorbed virus was washed away, and then 1% methyl cellulose containing different concentrations of FB23-2 was added, and the culture was continued at 37°C for 48 hours. After removing the methyl cellulose, 1% crystal violet was used for staining, and the size and number of plaques were observed.

[0029] The results show that TGEV-infected cells without FB23-2 concentration group appear a large number of needle-shaped plaques; the number of plaques of TGEV-infected cells treated with 2.5-10 μM FB23-2 is reduced Figure 4 The above results fully demonstrate that FB23-2 significantly inhibits the infection and replication of TGEV, and the effect increases with the increase of the concentration of FB23-2, showing a dose-dependent manner.

[0030] In summary, FB23-2 has a significant effect on inhibiting the replication of PEDV and TGEV in cells, and can be used as an antiviral drug.

[0031] The above specific embodiments are only an explanation of the present application, which is not a limitation of the present application, and the person skilled in the art can make a modification of the present embodiment without a creative contribution according to the need after reading the present specification, but as long as it is within the scope of the claims of the present application, it is protected by the patent law.

Claims

1. Use of FB23-2 in the preparation of a drug for inhibiting PEDV or TGEV replication, characterized by: The FB23-2 can inhibit the replication of PEDV and TGEV in cells.

2. Use of FB23-2 according to claim 1 in the preparation of a drug for inhibiting PEDV or TGEV replication, characterized in that: The chemical formula of FB23-2 is C 18 H 15 Cl2N3O3.

3. Use of FB23-2 according to claim 1 in the preparation of a drug for inhibiting PEDV or TGEV replication, characterized in that: The FB23-2 inhibits the replication of PEDV in cells at a concentration of 1 μM to <50 μM.

4. Use of FB23-2 according to claim 1 in the preparation of a drug for inhibiting PEDV or TGEV replication, characterized in that: The FB23-2 inhibits the replication of TGEV in cells at a concentration of 1 μM to <50 μM.

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