Application of tannin compounds in preparation of drugs for resisting porcine reproductive and respiratory disorder virus

Abstract

The invention discloses application of tannin compounds in preparation of drugs for resisting porcine reproductive and respiratory disorder viruses. The inventor of the invention performs activity screening on natural tannin compounds, and finds that the tannin compounds as shown in the formula 1 have excellent anti-PRRSV activity, and the EC50 value of the tannin compounds to PRRSV is lower than or obviously lower than that of ribavirin; therefore, the inhibiting effect of the compounds on the PRRSV is greater than or even far greater than that of a positive control drug ribavirin. The compounds have an unexpected effect of resisting PRRSV (porcine reproductive and respiratory syndrome virus).

Description

Technical Field

[0001] The invention relates to the field of pharmaceutical chemistry, and in particular to the application of tannin compounds in the preparation of anti-porcine reproductive and respiratory syndrome virus drugs or pharmaceutical feed additives. Background Art

[0002] Porcine reproductive and respiratory syndrome virus (PRRSV) is an acute, high fever and highly contagious disease of pigs. This infectious disease can cause obvious respiratory symptoms in pigs of different breeding ages. Severe infection can cause pig death and often lead to reproductive disorders such as miscarriage, stillbirth and mummification in sows, which seriously endangers the healthy development of the global pig farming industry and causes billions of dollars in economic losses to the global pig farming industry every year.

[0003] At present, there is no effective means to prevent and control blue ear disease in pigs. The main means of prevention and control of blue ear disease in pigs is vaccination. However, due to the high gene mutation rate of PRRSV and the frequent recombination between different strains, there are many kinds of mutant strains, which leads to the inability of vaccination to effectively block the infection of the virus and the poor prevention and control effect. Therefore, the research on anti-PRRSV drugs is of great significance in the prevention and control of blue ear disease in pigs. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide an application of a tannin compound in the preparation of an anti-PRRSV drug in order to overcome the insufficient development of anti-PRRSV drugs in the prior art.

[0005] The tannin compounds described in the present invention are mainly derived from pomegranate (Punica granatum L.), Phyllanthus emblica L., Oenothera biennis L., Sanguisorba officinalis and Terminalia chebula Retz., which are a class of known compounds. After searching and analyzing, there are literature reports that these compounds have inhibitory effects on viruses such as influenza virus, human immunodeficiency virus (HIV), hepatitis B virus (HBV), but it is not reported that they have anti-PRRSV effects. In the field of pharmaceutical chemistry, the antiviral effect of drugs is less predictable. A certain compound has an inhibitory effect on a certain virus, which does not mean that it also has an inhibitory effect on other viruses. For example, the commonly used anti-influenza virus drug oseltamivir phosphate only has an inhibitory effect on influenza virus, and has no inhibitory effect or no obvious inhibitory effect on other viruses. That is to say, based on the fact that these compounds disclosed in the literature have an inhibitory effect on certain viruses, those skilled in the art cannot predict that these compounds also have an inhibitory effect on PRRSV.

[0006] The above technical problems to be solved by the present invention are achieved through the following technical solutions:

[0007] The present invention provides the use of tannin compounds in the preparation of anti-porcine reproductive and respiratory syndrome virus (PRRSV) drugs, wherein the tannin compound is any of the following compounds:

[0008] Punicalagin (1, punicalagin), corilagin (2, corilagin), punicalin (3, punicalin), 2-O-galloylpunicalin (4), chebulagic acid (5, chebulagic acid), chebulinic acid (6, chebulinic acid), oenothein B (7, oenothein B), gemin D (8), lagerstannin C (9), granatin A (10), granatin B (11), tellimagrandin I (12, tellimagrandin I), tellimagrandin II (13, tellimagrandin II), casuarin (14, casuarinin), punigluconin (15, punigluconin), pedunculagin (16, pedunculagin), terflavin A (17), terflavin D (18), castanein (19, (+)-castalin), casuarin (20, (-)-strictinin), hippomanin A (21), punicarin C (22, punicacortein C), tercatain (23), heterophylliin A (24), castanein (25, castalagin), casuarin (26, casuarictin), sanguiin H1 (27), sanguiin H4 (28), (-)-quercetin (29, (-)-vescalin), pomegranate leaf tannin (30, punicafolin), cornusin A (31, cornusiin A), cornusiin B (32, cornusiin B), cornusiin C (33, cornusiin C), woodfordin B (34, woodfordin B), woodfordin H (35, woodfordin H), casuarin (36, casuariin), 2,3-(S)-HHDP-D-glucose (37), eucalbanin A (38), eucalbanin B (39), emblicanin A (40), emblicanin B (41), mallotusinin (42), rugosin B (43) and isoterchebulin (44), the structures of which are shown in Formula I:

[0009]

[0010]

[0011]

[0012]

[0013] Preferably, the drug is a drug for treating blue ear disease in pigs.

[0014] Preferably, the tannin compound is compound 1, 5, 14, 17, 22 and / or 44.

[0015] Preferably, the dosage form of the drug is powder, oral solution or injection.

[0016] The present invention also provides that the above tannin compounds can be added to animal feed as feed additives, that is, used as feed additives.

[0017] The present invention also provides an anti-porcine reproductive and respiratory syndrome virus drug containing the above-mentioned tannin compound as an active ingredient.

[0018] Beneficial effects:

[0019] The inventors of the present invention screened the activity of tannin compounds from natural sources and found that the tannin compounds shown in Formula 1 all have excellent anti-PRRSV activity. 50 The value should be lower or significantly lower than the EC of ribavirin 50 This indicates that the inhibitory effect of these compounds on PRRSV is greater than or even much greater than the positive control drug ribavirin. These compounds have unexpected anti-PRRSV effects. DETAILED DESCRIPTION

[0020] The present invention is further explained below in conjunction with specific examples, but the examples do not limit the invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0021] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0022] The structure of the tannin compound is shown in Formula 1, specifically punicalagin (1), corilagin (2), punicalin (3), 2-O-galloylpunicalin (4), chebulagic acid (5), chebulinic acid (6), oenothein B (7), gemin D (8), lagerstannin C (9), granatin A (10), granatin B (11), tellimagrandin I (12), tellimagrandin II (13), casuarinin (14), punigluconin (15), pedunculagin (16), terflavin A (17), terflavin D (18), castanein (19, (+)-castalin), casuarin (20, (-)-strictinin), hippomanin A (21), punicarin C (22, punicacortein C), tercatain (23), heterophylliin A (24), castanein (25, castalagin), casuarin (26, casuarictin), sanguiin H1 (27), sanguiin H4 (28), (-)-quercetin (29, (-)-vescalin), pomegranate leaf tannin (30, punicafolin), cornusin A (31, cornusiin A), cornusiin B (32, cornusiin B), cornusiin C (33, cornusiin C), woodfordin B (34, woodfordin B), woodfordin H (35, woodfordin H), casuarin (36, casuariin), 2,3-(S)-HHDP-D-glucose (37), eucalbanin A (38), eucalbanin B (39), emblicanin A (40), emblicanin B (41), mallotusinin (42), rugosin B (43) and isoterchebulin (44), the structures of which are shown in Formula I:

[0023]

[0024]

[0025]

[0026]

[0027] Example 1: In vitro anti-PRRSV activity test

[0028] Cytotoxicity assay: 6×10 4 Marc-145 cells / mL were plated in a 96-well plate and placed in a cell culture incubator at 5% CO2 and 37°C overnight. Then, the test compound (a total of 6 concentrations) diluted 2-fold in DMEM (cell maintenance medium) containing 2% FBS and 1% double antibody (penicillin-streptomycin) was added. At the same time, a cell control group (maintenance medium without compound) was set up. Three replicate wells were set up for each concentration of the compound and the cell control group. The plates were cultured in a cell culture incubator for 72 hours. The old culture medium was discarded, and DMEM containing 10% MTS was added to continue culturing for 0.5 to 2 hours. Then, the OD value was measured, and the half-lethal concentration (CC) of the compound was calculated using Graphpad Prime6.0 software. 50 ). The positive control drug was ribavirin.

[0029] Cytopathic effect (CPE) assay anti-PRRSV activity: 6×10 4 Marc-145 cells / mL were plated in a 96-well plate and cultured overnight in a 5% CO2, 37°C cell culture incubator. The old solution was discarded and 100 μL of 100 TCID 50 The PRRSV virus solution was diluted with maintenance solution, and the test compound (a total of 6 concentrations) diluted 2-fold with cell maintenance solution was added as the compound treatment group. In the virus control group, 100 μL of DMEM culture solution containing 2% FBS and 100 μL of virus solution were added to each well, and only 200 μL of maintenance solution was added to the cell control group. Three replicate wells were set up in each group and cultured in a 37°C, 5% CO2 cell culture incubator for 2-3 days. The cell pathological changes and fluorescence were observed every day, and the cell pathological changes of each well were recorded and photographed. 0-25%: +; 25%-50%: ++; 50-75%: +++; 75-100%: ++++, and EC was calculated by Reed and Muench method 50 The results are shown in Table 1. The positive control drug was ribavirin.

[0030] The CC of compounds 1 to 44 in formula 1 and the positive control drug Ribavirin on host cells Marc-145 cells and PRRSV CHR6 / Li11 virus strain were tested by the above method. 50 IC50 The specific results are shown in Table 1.

[0031] Table 1. Test results of tannin compounds against porcine reproductive and respiratory syndrome virus

[0032]

[0033]

[0034] From the results of the in vitro anti-PRRSV activity test in Table 1, it can be seen that the tannin compounds shown in Formula 1 have inhibitory effects on PRRSV CHR6 and Li11, and their antiviral activities are equivalent to or better than the positive control drug ribavirin. It is worth noting that the EC values ​​of compounds 1, 5, 14, 17, 22 and 44 against PRRSV 50 Less than 10 μM, its activity is dozens of times better than ribavirin.

Claims

1. Use of tannin compounds in the preparation of anti-porcine reproductive and respiratory syndrome virus drugs, wherein the tannin compound is any of the following compounds:

2. The use according to claim 1, characterized in that: The medicine is a medicine for treating blue ear disease in pigs.

3. The use according to claim 1 or 2, characterized in that: The tannin compound is compound 1, 5, 14, 17, 22 and / or 44.

4. The use according to claim 1, characterized in that: The dosage form of the medicine is powder, oral solution or injection.

5. Use of tannin compounds in the preparation of feed additives, wherein the tannin compound is any of the following compounds:

6. An anti-porcine reproductive and respiratory syndrome virus drug, characterized in that: Contains tannin compounds as active ingredients; The tannin compound is any of the following compounds:

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