Application of galloyl telichroflavane in preparation of drugs for resisting porcine reproductive and respiratory syndrome virus

Abstract

The invention discloses an application of galloyl telichroflavane in preparation of a drug for resisting porcine reproductive and respiratory syndrome virus (PRRSV). When antiviral activity screening is carried out on the galloyl terichromone compound, it is found for the first time that the galloyl terichromone compound can remarkably inhibit proliferation of porcine reproductive and respiratory syndrome virus (PRRSV), the antiviral activity of the galloyl terichromone compound is higher than that of positive control medicine ribavirin, and the galloyl terichromone compound can be used for preventing and treating infection of porcine reproductive and respiratory syndrome virus.

Description

Technical Field

[0001] The present invention relates to medicinal chemistry, in particular to a galloyl tericeroflavanoid compound and its use in resisting porcine reproductive and respiratory syndrome virus (PRRSV). Background Art

[0002] Blue ear disease (Porcine reproductive and respiratory syndrome) is a highly contagious and high-mortality swine infectious disease caused by the porcine reproductive and respiratory syndrome virus (PRRSV), characterized by respiratory symptoms in piglets and reproductive disorders in sows. The difficulties in preventing and controlling PRRSV include rapid virus mutation, co-infection of multiple pathogens in the host, and the inability of antibody detection kits to correctly indicate the immune status of the pig herd, which has caused huge economic losses to the global pig industry.

[0003] At present, the prevention and control strategy of PRRSV is mainly based on purification. Only some pig farms can maintain a double negative state of pathogens and antibodies for a long time, and most are still in a positive stable or unstable state. Recombination is one of the important ways of viral evolution, which occurs not only between wild viruses, but also between wild viruses and attenuated live vaccines. Faced with the difficulties in the prevention and control of blue ear disease, there is no specific drug for the prevention and treatment of PRRSV, and only antibiotics can be used to treat secondary bacterial infections caused by the virus.

[0004] Galloyltericeroflavanoids are flavonoids that are widely present in nature. Their molecular characteristics include the basic structural skeleton of flavonoids and galloyl groups. They have multiple physiological and pharmacological activities, such as anti-inflammatory, antioxidant, anti-tumor, and cardiovascular protection. Summary of the invention

[0005] The purpose of the present invention is to develop the application of galloyl teresin in drugs for preventing and treating blue ear disease. The general formula of this type of compound is shown in Formula 2, and a single or multiple galloyl groups (II) are connected to the benzene ring A of the skeleton (Ⅰ) or the benzene ring B substituted by C2. After retrieval and analysis, this type of compound is mainly isolated from plants and is a class of known compounds; it can also be obtained by a synthetic method. However, the compound with the structure shown in the general formula has not been reported to have an anti-PRRSV effect.

[0006] 《Antiviral flavans from the leaves of Pithecellobium clypearia》、《Studies on the bioactive flavonoids isolated from Pithecellobium clypeariaBenth》、《The potential of Paritaprevir and Emetine as inhibitors of SARS-CoV-2RdRp》、《Network pharmacology-based screening of the active ingredients andpotential targets of the genus of Pithecellobium marthae(Britton&Killip)Niezgoda&Nevl for application to Alzheimer's disease》、《Polyphenols fromAcacia nilotica leaves and evaluation of antihyperglycemic effect of aqueousextract》、《Chemical constituents from Oncocalyx glabratus and their biologicalactivities》、《In-silico investigation of antitrypanosomal phytochemicals fromNigerian medicinal plants》、《Chemical constituents ofPithecellobiumclypearia》、《HPLC fingerprint spectrum of Archidendron clypearialeaves》、《Prosopis juliflora(Sw.)DC phytochemicals induce apoptosis andinhibit cell proliferation signaling pathways,EMT,migration,invasion,angiogenesis and stem cell markers in melanoma cell lines》、《Antiplasmodialphenolic compounds from Piptadenia pervillei》、《Tannins and relatedcompounds.111.Flavan-3-ol gallates and proanthocyanidins from Pithecellobiumlobatum》、《Chemical constituents of Bergenia crassifolia roots and their growthinhibitory activity against Babesia bovis and B.bigemina》、《Separation ofAntioxidant-Rich Alternanthera Sessilis Red Extracts by Sephadex LH-20andIdentification of Polyphenols Using HPLC-QToF-MS / MS》、《Two new flavanes andbioactive compounds from Daphne tangutica Maxim》、《A new epicatechin gallate andcalpain inhibitory activity from Orostachys japonicus》 and other literatures have reported such compounds.

[0007] The above-mentioned document also publicly reported that the compound has good binding ability with the RNA-dependent RNA polymerase (RdRp) active pocket of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and has antiviral activity against respiratory syncytial virus (RSV) and influenza virus A (H1N1), but it did not disclose that the compound has anti-PRRSV effect. In the field of medicinal chemistry, the anti-disease effect of drugs is less predictable. A certain compound has an inhibitory effect on one virus, which does not mean that it also has an inhibitory effect on other viruses. In other words, based on the above-mentioned document that the compound has an anti-influenza virus effect, the technicians in this field cannot predict that the compound also has an inhibitory effect on other viruses, let alone which virus the compound has a significant inhibitory effect on.

[0008]

[0009] Therefore, the present invention provides the use of galloyl tericorin in the preparation of an anti-porcine reproductive and respiratory syndrome virus drug, wherein the structure of the galloyl tericorin is shown in any one of the following:

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[0019] Preferably, the drug is a drug for treating blue ear disease in pigs.

[0020] Preferably, the dosage form of the drug is powder, oral solution or injection.

[0021] Preferably, the galloyl teriseflavan is compound (3), (5), (7), (21), (23) or (29).

[0022] The present invention also provides that the galloyl tericorflavane can be added to animal feed as a feed additive, that is, used as a feed additive.

[0023] The present invention also provides an anti-porcine reproductive and respiratory syndrome virus drug, which contains the galloyl terisenoflavan as an active ingredient.

[0024] Preferably, the virus is porcine reproductive and respiratory syndrome virus (PRRSV).

[0025] Beneficial effects: The inventors of the present invention first discovered that galloyl tericornicotinoids in plants have a very significant inhibitory effect on PRRSV. The results of in vitro anti-PRRSV experiments showed that the structure shown in Formula 1 has a half effective concentration (EC 50 ) is less than the half effective concentration (EC 50 ), and the selection index (SI) is greater than the selection index of ribavirin. Therefore, galloyl tericorin flavanoids can be used to prepare drugs against PRRSV.

[0026] Selectivity index (SI) = half inhibitory concentration (CC 50 ) / half effective concentration (EC 50 ) DETAILED DESCRIPTION

[0027] 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.

[0028] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0029] Example 1: In vitro anti-PRRSV activity test

[0030] Cytotoxicity assay: 6×10 4 Marc-145 cells / mL were plated in a 96-well plate and placed in a 5% CO2, 37°C cell culture incubator overnight. Then, the test compound (a total of 6 concentrations) diluted 2-fold in a cell maintenance medium (DMEM containing 2% FBS and 1% penicillin-streptomycin) was added. At the same time, a cell control group (cell 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. Fresh culture medium containing 10% MTS was added and cultured for 1 to 1.5 hours. Then, the OD value was measured and the half inhibitory concentration (CC) of the compound was calculated using Graphpad Prime 6.0 software. 50 ).

[0031] The anti-PRRSV activity of the compounds was detected based on cytopathic effect (CPE): 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 cell maintenance medium, and the test compound (a total of 6 concentrations) diluted 2-fold with the cell maintenance medium was added as the compound treatment group. In the virus control group, 100 μL of DMEM culture medium containing 2% FBS and 100 μL of virus solution were added to each well, and only 200 μL of DMEM culture medium containing 2% FBS was added to the cell control group. Three replicate wells were set up for each group and cultured in a 37°C, 5% CO2 cell culture incubator for 2-3 days. The cell pathological changes were observed every day, and the cell pathological changes in each well were recorded and photographed. 0-25%: +; 25%-50%: ++; 50-75%: +++; 75-100%: ++++, and the EC was calculated by the Reed and Muench method. 50 , the results are shown in Table 1.

[0032] Table 1. Test results of anti-porcine reproductive and respiratory syndrome virus activity of galloyl terichromanol

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Claims

1. Use of galloyl tericorin in the preparation of an anti-porcine reproductive and respiratory syndrome virus drug, wherein the structure of the galloyl tericorin has any one of the following structures:

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 dosage form of the medicine is powder, oral solution or injection.

4. The use according to claim 1, characterized in that: The galloyl teriseflavan is compound (3), (5), (7), (21), (23) or (29).

5. Use of the galloyl tericeroflavane according to claim 1 in the preparation of feed additives.

6. An anti-porcine reproductive and respiratory syndrome virus drug, characterized in that: Contains the galloyl teriseflavan according to claim 1 as an active ingredient.

7. The drug according to claim 6, characterized in that The medicine is a medicine for treating blue ear disease in pigs.

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