Preparation for treating porcine reproductive and respiratory syndrome and African swine fever and preparation method thereof

Abstract

The invention is suitable for the technical field of veterinary drugs, and provides a preparation aiming at porcine reproductive and respiratory syndrome and African swine fever and a preparation method thereof. The preparation is prepared from the following raw materials: a modified polydatin inclusion compound, a nano radix astragali-herba houttuyniae compound, medium-chain fatty glyceride, organic acid sustained-release microcapsules, fermented ammonium glycyrrhizinate, selenized radix isatidis polysaccharide, modified quercetin, modified mannitol and a medicinal carrier. According to the preparation for the porcine reproductive and respiratory syndrome and the African swine fever, the modified polydatin inclusion compound and the modified quercetin are added, the bioavailability is improved through modification, and virus replicase is directly inhibited; the medium-chain fatty glyceride destroys a lipid layer of a virus envelope and blocks virus invasion; the organic acid sustained-release microcapsules are continuously released in the intestinal tract, so that the pH is reduced to inhibit the virus activity, and meanwhile, the intestinal flora is regulated; after the nanometer astragalus membranaceus-houttuynia cordata compound is subjected to enzymolysis nanocrystallization, the immunostimulatory activity of astragalus polysaccharide and houttuynia cordata flavone is improved, and phagocytosis of macrophages and secretion of interferon are promoted.

Description

Technical Field

[0001] The present invention belongs to the technical field of veterinary medicines, and in particular relates to a preparation for treating blue ear disease and African swine fever and a preparation method thereof. Background Art

[0002] Previously known as "mystery swine disease," "new swine disease," "porcine epidemic abortion and respiratory syndrome," "porcine reproductive and respiratory syndrome," "blue ear disease," and "swine fever," China classifies it as a Category II animal disease. PRRS is highly contagious and endemic. The PRRS virus infects only pigs, not other animals. Pigs of all ages and breeds are susceptible, with piglets under one month old and pregnant sows being the most susceptible.

[0003] African swine fever (ASF) is an acute, hemorrhagic, and highly contagious disease caused by the African swine fever virus (ASFV) infecting domestic pigs and various wild boars. It is characterized by a short onset of illness, with a mortality rate of up to 100% in the most acute and acute infections. Clinical manifestations include fever (up to 40-42°C), a rapid heartbeat, difficulty breathing, some coughing, serous or mucopurulent discharge from the eyes and nose, cyanosis of the skin, and significant bleeding in the lymph nodes, kidneys, and gastrointestinal mucosa. The clinical symptoms of ASF are similar to those of swine fever and can only be confirmed through laboratory testing.

[0004] Porcine reproductive and respiratory syndrome (PRRS) and African swine fever (ASF) are two devastating diseases in the pig industry. Existing prevention and control measures are insufficient: PRRS vaccine has a low protection rate and cannot block ASFV infection; chemical drugs (such as tilmicosin) lead to drug resistance and have high liver and kidney toxicity; traditional Chinese medicines (such as astragalus polysaccharides) have low bioavailability and weak antiviral effects. Summary of the Invention

[0005] The present invention provides a preparation for blue ear disease and African swine fever, aiming to solve the above problems.

[0006] The present invention is achieved as follows: a preparation for blue ear disease and African swine fever, comprising the following raw materials in parts by weight: 10-60 parts of modified polydatin inclusion complex, 40-80 parts of nano-astragalus-houttuynia cordata complex, 10-30 parts of medium-chain fatty acid glyceride, 8-40 parts of organic acid sustained-release microcapsules, 5-25 parts of fermented ammonium glycyrrhizate, 5-20 parts of selenized isatis root polysaccharide, 10-20 parts of modified quercetin, 5-10 parts of modified mannitol, and 20-100 parts of a pharmaceutical carrier (such as a corn starch carrier).

[0007] Preferably, the raw materials include the following in parts by weight: 20-50 parts of modified polydatin inclusion complex, 50-70 parts of nano-astragalus-houttuynia complex, 15-25 parts of medium-chain fatty acid glyceride, 18-30 parts of organic acid sustained-release microcapsules, 10-20 parts of fermented ammonium glycyrrhizate, 10-15 parts of selenized isatis root polysaccharide, 12-18 parts of modified quercetin, 6-9 parts of modified mannitol, and 40-80 parts of pharmaceutical carrier.

[0008] Preferably, the following raw materials are included in parts by weight: 35 parts of modified polydatin inclusion complex, 60 parts of nano-astragalus-houttuynia complex, 20 parts of medium-chain fatty acid glyceride, 24 parts of organic acid sustained-release microcapsules, 15 parts of fermented ammonium glycyrrhizate, 12.5 parts of selenized isatis root polysaccharide, 15 parts of modified quercetin, 7.5 parts of modified mannitol, and 60 parts of pharmaceutical carrier.

[0009] Preferably, the preparation method of the modified polydatin inclusion compound is: mixing polydatin and β-cyclodextrin in a molar ratio of 1:4-8, and ultrasonically treating at 40-60° C. and 200-500W for 20-40 minutes to improve the solubility of polydatin through cyclodextrin inclusion.

[0010] Preferably, the preparation method of the nano-astragalus-houttuynia cordata complex is: Mix Astragalus and Houttuynia cordata in a ratio of 2-5:1, add cellulase (enzyme activity ≥ 5000U / g) and perform enzymolysis at 35-45℃ for 1-3h; After high-pressure homogenization (1000-2000 bar), the product is frozen in liquid nitrogen and ground to a particle size of 0.1-2 μm. The absorption of traditional Chinese medicine is enhanced through a combination of enzymatic hydrolysis and nano-crystallization.

[0011] Preferably, the enzymatic hydrolysis adopts a composite enzyme system comprising cellulase, pectinase and xylanase (mass ratio 3:1:1); the grinding is carried out in a liquid nitrogen environment at -196°C, and the grinding speed is 300-800 rpm.

[0012] Preferably, the preparation method of the organic acid sustained-release microcapsules is: Dissolve citric acid and fumaric acid in water at a ratio of 1-3:1 and mix with 2-5wt% sodium alginate; The microcapsules are dripped into a coagulation solution containing 1-3wt% calcium chloride and 0.5-2wt% chitosan, and dried to obtain microcapsules. Organic acid is slowly released in the intestine to maintain antiviral pH.

[0013] Preferably, the preparation method of the fermented ammonium glycyrrhizate is: anaerobically fermenting glycyrrhizic acid and Lactobacillus plantarum at 35-39°C for 36-60 hours, controlling the pH to 5.5-6.5, and converting the immune active components by microbial fermentation; the preparation method of the selenized Radix Isatidis polysaccharide is: complexing Radix Isatidis polysaccharide with sodium selenite at a mass ratio of 10:1-5:1 at 60-80°C for 2-4 hours.

[0014] Preferably, the modified quercetin is prepared by compounding quercetin and soybean lecithin in a mass ratio of 1:2-5, evaporating the solvent to form a film, adding phosphate buffer (pH 7.4) for hydration, and ultrasonically treating to obtain a nanocomposite.

[0015] Preferably, the preparation method of the modified mannitol is: Dissolve mannitol in a 5-8 times by weight isopropanol-water mixed solvent (volume ratio 3-4:1), add 2.0-2.5 times by molar amount of NaOH, and stir at 60-65°C for 30 minutes; Add chloroacetic acid in batches (3 times, 20 minutes apart), control the molar ratio of chloroacetic acid to mannitol to 0.5-0.7:1, and heat to 70-75°C for 2-3 hours; Adjust the pH to 6.5-7.5 with glacial acetic acid, add 3 volumes of ethanol for precipitation, and remove by-products by ultrafiltration (molecular weight cut-off 1 kDa); The product was spray-dried (inlet air temperature 160°C, outlet air temperature 80°C) to obtain a white powder.

[0016] The present invention also provides a method for preparing a preparation for blue ear disease and African swine fever, comprising the following steps: Prepare the raw materials according to the recipe; Mix the raw materials under an inert atmosphere; Sieve the mixture (for example, 60 mesh), divide it into portions and seal them in a dark place.

[0017] Compared with the prior art, the embodiments of the present application have the following beneficial effects: The preparation for blue ear disease and African swine fever provided by the present invention is added with modified polydatin inclusion compound and modified quercetin, and the bioavailability is improved through modification, which directly inhibits viral replication enzymes; medium-chain fatty acid glycerides destroy the lipid layer of the viral envelope and block viral invasion; organic acid sustained-release microcapsules are continuously released in the intestine, lowering the pH to inhibit viral activity and regulating intestinal flora; after the nano-astragalus-houttuynia cordata complex is enzymatically nanosized, the immunostimulatory activity of astragalus polysaccharides and houttuynia cordata flavonoids is improved, promoting macrophage phagocytosis and interferon secretion; selenized isatis indigotica polysaccharide enhances antioxidant capacity through selenium. The drug can enhance the effect of the drug on the growth of lymphocytes and the growth of progenitor cells, and can cooperate with the antiviral ingredients of Radix Isatidis to increase the proliferation rate of lymphocytes; the aglycone content of fermented ammonium glycyrrhizinate is increased after microbial transformation, which inhibits the over-activation of the NF-κB pathway and reduces the inflammatory storm; the chitosan-sodium alginate coating of the organic acid microcapsules achieves colon-targeted release to avoid degradation by gastric acid; by adding modified mannitol, a hydrophilic-hydrophobic balance structure is formed, which enhances mucosal adhesion and prolongs the retention time of the drug in the intestine, and can cooperate with modified quercetin, modified polydatin inclusion complex and other components to improve virus adsorption and blocking, promote transmembrane transport, and enhance the antioxidant and antiviral effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a preparation flow chart of a preparation for blue ear disease and African swine fever provided by the present invention. DETAILED DESCRIPTION

[0019] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art to which this application belongs. The terms used in the specification of the application are only for the purpose of describing specific embodiments and are not intended to limit this application. The terms "including" and "having" and any variations thereof in the specification and claims of this application and the above-mentioned drawings are intended to cover non-exclusive inclusions. The terms "first", "second", etc. in the specification and claims of this application or the above-mentioned drawings are used to distinguish different objects, not to describe a specific order.

[0020] References herein to "embodiments" mean that a particular feature, structure, or characteristic described in connection with the embodiments may be included in at least one embodiment of the present application. The appearance of this phrase in various places in the specification does not necessarily refer to the same embodiment, nor does it constitute an independent or alternative embodiment that is mutually exclusive of other embodiments. It is understood, both explicitly and implicitly, by those skilled in the art that the embodiments described herein may be combined with other embodiments.

[0021] Example 1 The embodiment of the present invention provides a preparation for blue ear disease and African swine fever, comprising the following raw materials in parts by weight: 10 parts of modified polydatin inclusion compound, 40 parts of nano-astragalus-houttuynia cordata complex, 10 parts of medium-chain fatty acid glyceride, 8 parts of organic acid sustained-release microcapsules, 5 parts of fermented ammonium glycyrrhizate, 5 parts of selenized isatis root polysaccharide, 10 parts of modified quercetin, 5 parts of modified mannitol, and 20 parts of a pharmaceutical carrier (such as a corn starch carrier). The preparation method of the preparation for blue ear disease and African swine fever described in this embodiment is as follows: Figure 1 As shown, the following steps are included: 1) Prepare the raw materials according to the recipe; The preparation method of the modified polydatin inclusion compound is as follows: polydatin and β-cyclodextrin are mixed in a molar ratio of 1:4, and ultrasonically treated at 40°C and 200W for 20 minutes to improve the solubility of polydatin through cyclodextrin inclusion.

[0022] The preparation method of the nano-astragalus and houttuynia cordata complex is as follows: Astragalus and Houttuynia cordata were mixed in a ratio of 2:1, and cellulase (enzyme activity ≥ 5000 U / g) was added for enzymatic hydrolysis at 35°C for 1 hour. The enzymatic hydrolysis used a composite enzyme system containing cellulase, pectinase, and xylanase (mass ratio 3:1:1); After high-pressure homogenization (1000 bar), the mixture is frozen in liquid nitrogen and ground to a particle size of 0.1 μm. The grinding is carried out in a liquid nitrogen environment at -196°C and a grinding speed of 300 rpm. The absorption of traditional Chinese medicine is enhanced by combining enzymatic hydrolysis and nano-processing.

[0023] The preparation method of the organic acid sustained-release microcapsule is as follows: Dissolve citric acid and fumaric acid in water at a ratio of 1:1 and mix with 2 wt% sodium alginate; The solution was dripped into a coagulation solution containing 1wt% calcium chloride and 0.5wt% chitosan, and dried to obtain microcapsules, in which the organic acid was slowly released in the intestine to maintain the antiviral pH.

[0024] The preparation method of the fermented ammonium glycyrrhizate is as follows: glycyrrhizic acid and Lactobacillus plantarum are anaerobically fermented at 35°C for 36 hours, the pH is controlled at 5.5, and the microbial fermentation converts the immune active components; the preparation method of the selenized Radix Isatidis polysaccharide is as follows: Radix Isatidis polysaccharide and sodium selenite are complexed and reacted at 60°C for 2 hours in a mass ratio of 10:1:1.

[0025] The preparation method of the modified quercetin comprises the following steps: compounding quercetin and soybean lecithin in a mass ratio of 1:2, evaporating the solvent to form a film, adding phosphate buffer (pH 7.4) for hydration, and ultrasonically treating the film to obtain a nanocomposite.

[0026] The preparation method of the modified mannitol is: Mannitol was dissolved in 5 times its mass of isopropanol-water mixed solvent (volume ratio 3:1), and 2.0 times the molar amount of NaOH was added, and the mixture was stirred at 60°C for 30 minutes; Chloroacetic acid was added in batches (3 times with an interval of 20 minutes) to control the molar ratio of chloroacetic acid to mannitol to 0.5:1, and the temperature was raised to 70°C for 2 hours; Adjust the pH to 6.5 with glacial acetic acid, add 3 volumes of ethanol for precipitation, and remove by-products by ultrafiltration (molecular weight cut-off 1 kDa); The product was spray-dried (inlet air temperature 160°C, outlet air temperature 80°C) to obtain a white powder.

[0027] 2) Mixing the raw materials under an inert atmosphere; 3) Sieve the mixture (e.g. 60 mesh), divide into portions and seal them in a dark place.

[0028] Example 2 The embodiment of the present invention provides a preparation for blue ear disease and African swine fever, comprising the following raw materials in parts by weight: 20 parts of modified polydatin inclusion compound, 50 parts of nano-astragalus-houttuynia complex, 15 parts of medium-chain fatty acid glyceride, 18 parts of organic acid sustained-release microcapsules, 10 parts of fermented ammonium glycyrrhizate, 10 parts of selenized isatis root polysaccharide, 12 parts of modified quercetin, 6 parts of modified mannitol, and 40 parts of a pharmaceutical carrier (such as a corn starch carrier). The preparation method of the preparation for blue ear disease and African swine fever described in this embodiment is as follows: Figure 1 As shown, the following steps are included: 1) Prepare the raw materials according to the recipe; The preparation method of the modified polydatin inclusion compound is as follows: polydatin and β-cyclodextrin are mixed in a molar ratio of 1:4, and ultrasonically treated at 40°C and 200W for 20 minutes to improve the solubility of polydatin through cyclodextrin inclusion.

[0029] The preparation method of the nano-astragalus and houttuynia cordata complex is as follows: Astragalus and Houttuynia cordata were mixed in a ratio of 2:1, and cellulase (enzyme activity ≥ 5000 U / g) was added for enzymatic hydrolysis at 35°C for 1 hour. The enzymatic hydrolysis used a composite enzyme system containing cellulase, pectinase, and xylanase (mass ratio 3:1:1); After high-pressure homogenization (1000 bar), the mixture is frozen in liquid nitrogen and ground to a particle size of 0.1 μm. The grinding is carried out in a liquid nitrogen environment at -196°C and a grinding speed of 300 rpm. The absorption of traditional Chinese medicine is enhanced by combining enzymatic hydrolysis and nano-processing.

[0030] The preparation method of the organic acid sustained-release microcapsule is as follows: Dissolve citric acid and fumaric acid in water at a ratio of 1:1 and mix with 2 wt% sodium alginate; The solution was dripped into a coagulation solution containing 1wt% calcium chloride and 0.5wt% chitosan, and dried to obtain microcapsules, in which the organic acid was slowly released in the intestine to maintain the antiviral pH.

[0031] The preparation method of the fermented ammonium glycyrrhizate is as follows: glycyrrhizic acid and Lactobacillus plantarum are anaerobically fermented at 35°C for 36 hours, the pH is controlled at 5.5, and the microbial fermentation converts the immune active components; the preparation method of the selenized Radix Isatidis polysaccharide is as follows: Radix Isatidis polysaccharide and sodium selenite are complexed and reacted at 60°C for 2 hours in a mass ratio of 10:1:1.

[0032] The preparation method of the modified quercetin comprises the following steps: compounding quercetin and soybean lecithin in a mass ratio of 1:2, evaporating the solvent to form a film, adding phosphate buffer (pH 7.4) for hydration, and ultrasonically treating the film to obtain a nanocomposite.

[0033] The preparation method of the modified mannitol is: Mannitol was dissolved in 5 times its mass of isopropanol-water mixed solvent (volume ratio 3:1), and 2.0 times the molar amount of NaOH was added, and the mixture was stirred at 60°C for 30 minutes; Chloroacetic acid was added in batches (3 times with an interval of 20 minutes) to control the molar ratio of chloroacetic acid to mannitol to 0.5:1, and the temperature was raised to 70°C for 2 hours; Adjust the pH to 6.5 with glacial acetic acid, add 3 volumes of ethanol for precipitation, and remove by-products by ultrafiltration (molecular weight cut-off 1 kDa); The product was spray-dried (inlet air temperature 160°C, outlet air temperature 80°C) to obtain a white powder.

[0034] 2) Mixing the raw materials under an inert atmosphere; 3) Sieve the mixture (e.g. 60 mesh), divide into portions and seal them in a dark place.

[0035] Example 3 The embodiment of the present invention provides a preparation for blue ear disease and African swine fever, comprising the following raw materials in parts by weight: 35 parts of modified polydatin inclusion compound, 60 parts of nano-astragalus-houttuynia complex, 20 parts of medium-chain fatty acid glyceride, 24 parts of organic acid sustained-release microcapsules, 15 parts of fermented ammonium glycyrrhizate, 12.5 parts of selenized isatis root polysaccharide, 15 parts of modified quercetin, 7.5 parts of modified mannitol, and 60 parts of a pharmaceutical carrier (such as a corn starch carrier). The preparation method of the preparation for blue ear disease and African swine fever described in this embodiment is as follows: Figure 1 As shown, the following steps are included: 1) Prepare the raw materials according to the recipe; The preparation method of the modified polydatin inclusion compound is as follows: polydatin and β-cyclodextrin are mixed in a molar ratio of 1:4-8, and ultrasonically treated at 50°C and 350W for 30 minutes to improve the solubility of polydatin through cyclodextrin inclusion.

[0036] The preparation method of the nano-astragalus-houttuynia cordata complex is as follows: Astragalus and Houttuynia cordata were mixed in a ratio of 3.5:1, and cellulase (enzyme activity ≥ 5000 U / g) was added for enzymatic hydrolysis at 40°C for 2 h. The enzymatic hydrolysis used a composite enzyme system containing cellulase, pectinase, and xylanase (mass ratio 3:1:1); After high-pressure homogenization (1500 bar), the mixture is frozen in liquid nitrogen and ground to a particle size of 0.15 μm. The grinding is carried out in a liquid nitrogen environment at -196°C and a grinding speed of 550 rpm. The absorption of traditional Chinese medicine is enhanced by combining enzymatic hydrolysis and nano-crystallization.

[0037] The preparation method of the organic acid sustained-release microcapsule is as follows: Dissolve citric acid and fumaric acid in water at a ratio of 2:1 and mix with 3.5 wt% sodium alginate; The microcapsules were dripped into a coagulation solution containing 2 wt% calcium chloride and 1.25 wt% chitosan and dried to obtain microcapsules. The organic acid was slowly released in the intestine to maintain the antiviral pH.

[0038] The preparation method of the fermented ammonium glycyrrhizate is as follows: glycyrrhizic acid and Lactobacillus plantarum are anaerobically fermented at 37°C for 48 hours, the pH is controlled at 6.0, and the microbial fermentation converts the immune active components; the preparation method of the selenized Radix Isatidis polysaccharide is as follows: Radix Isatidis polysaccharide and sodium selenite are complexed and reacted at 70°C for 3 hours in a mass ratio of 10:3:1.

[0039] The preparation method of the modified quercetin comprises the following steps: compounding quercetin and soybean lecithin at a mass ratio of 1:3.5, evaporating the solvent to form a film, adding phosphate buffer (pH 7.4) for hydration, and ultrasonically treating the film to obtain a nanocomposite.

[0040] The preparation method of the modified mannitol is: Mannitol was dissolved in 6.5 times its mass of isopropanol-water mixed solvent (volume ratio 3.5:1), and 2.25 times the molar amount of NaOH was added. The mixture was stirred at 62.5°C for 30 min. Chloroacetic acid was added in batches (3 times with an interval of 20 minutes) to control the molar ratio of chloroacetic acid to mannitol to 0.6:1, and the temperature was raised to 72.5°C for 2.5 hours; Adjust the pH to 7.0 with glacial acetic acid, add 3 volumes of ethanol for precipitation, and remove by-products by ultrafiltration (molecular weight cut-off 1 kDa); The product was spray-dried (inlet air temperature 160°C, outlet air temperature 80°C) to obtain a white powder.

[0041] 2) Mixing the raw materials under an inert atmosphere; 3) Sieve the mixture (e.g. 60 mesh), divide into portions and seal them in a dark place.

[0042] Example 4 The embodiment of the present invention provides a preparation for blue ear disease and African swine fever, comprising the following raw materials in parts by weight: 50 parts of modified polydatin inclusion compound, 70 parts of nano-astragalus-houttuynia complex, 25 parts of medium-chain fatty acid glyceride, 30 parts of organic acid sustained-release microcapsules, 20 parts of fermented ammonium glycyrrhizate, 15 parts of selenized isatis root polysaccharide, 18 parts of modified quercetin, 9 parts of modified mannitol, and 80 parts of a pharmaceutical carrier (such as a corn starch carrier). The preparation method of the preparation for blue ear disease and African swine fever described in this embodiment is as follows: Figure 1 As shown, the following steps are included: 1) Prepare the raw materials according to the recipe; The preparation method of the modified polydatin inclusion compound is as follows: polydatin and β-cyclodextrin are mixed in a molar ratio of 1:8, and ultrasonically treated at 60° C. and 500W for 40 minutes to improve the solubility of polydatin through cyclodextrin inclusion.

[0043] The preparation method of the nano-astragalus and houttuynia cordata complex is as follows: Astragalus and Houttuynia cordata were mixed in a ratio of 5:1, and cellulase (enzyme activity ≥ 5000 U / g) was added for enzymatic hydrolysis at 45°C for 3 h. The enzymatic hydrolysis used a composite enzyme system containing cellulase, pectinase, and xylanase (mass ratio 3:1:1); After high-pressure homogenization (2000 bar), the product is frozen in liquid nitrogen and ground to a particle size of 0.2 μm. The grinding is carried out in a liquid nitrogen environment at -196°C and a grinding speed of 800 rpm. The absorption of traditional Chinese medicine is enhanced by combining enzymatic hydrolysis and nano-processing.

[0044] The preparation method of the organic acid sustained-release microcapsule is as follows: Dissolve citric acid and fumaric acid in water at a ratio of 3:1 and mix with 5 wt% sodium alginate; The microcapsules were dripped into a coagulation solution containing 3wt% calcium chloride and 0.2wt% chitosan and dried to obtain microcapsules. The organic acid was slowly released in the intestine to maintain the antiviral pH.

[0045] The preparation method of the fermented ammonium glycyrrhizate is as follows: glycyrrhizic acid and Lactobacillus plantarum are anaerobically fermented at 39°C for 60 hours, the pH is controlled at 6.5, and the microbial fermentation converts the immune active components; the preparation method of the selenized Radix Isatidis polysaccharide is as follows: Radix Isatidis polysaccharide and sodium selenite are complexed and reacted at 80°C for 4 hours in a mass ratio of 10:5:1.

[0046] The preparation method of the modified quercetin comprises the following steps: compounding quercetin and soybean lecithin in a mass ratio of 1:5, evaporating the solvent to form a film, adding phosphate buffer (pH 7.4) for hydration, and ultrasonically treating the film to obtain a nanocomposite.

[0047] The preparation method of the modified mannitol is: Mannitol was dissolved in 8 times its mass of isopropanol-water mixed solvent (volume ratio 4:1), and 2.5 times the molar amount of NaOH was added, and the mixture was stirred at 65°C for 30 minutes; Chloroacetic acid was added in batches (3 times with an interval of 20 minutes) to control the molar ratio of chloroacetic acid to mannitol to 0.7:1, and the temperature was raised to 75°C for 3 hours; Adjust the pH to 7.5 with glacial acetic acid, add 3 volumes of ethanol for precipitation, and remove by-products by ultrafiltration (molecular weight cut-off 1 kDa); The product was spray-dried (inlet air temperature 160°C, outlet air temperature 80°C) to obtain a white powder.

[0048] 2) Mixing the raw materials under an inert atmosphere; 3) Sieve the mixture (e.g. 60 mesh), divide into portions and seal them in a dark place.

[0049] Example 5 The embodiment of the present invention provides a preparation for blue ear disease and African swine fever, comprising the following raw materials in parts by weight: 60 parts of modified polydatin inclusion compound, 80 parts of nano-astragalus-houttuynia complex, 30 parts of medium-chain fatty acid glyceride, 40 parts of organic acid sustained-release microcapsules, 25 parts of fermented ammonium glycyrrhizate, 20 parts of selenized isatis root polysaccharide, 20 parts of modified quercetin, 10 parts of modified mannitol, and 100 parts of a pharmaceutical carrier (such as a corn starch carrier). The preparation method of the preparation for blue ear disease and African swine fever described in this embodiment is as follows: Figure 1 As shown, the following steps are included: 1) Prepare the raw materials according to the recipe; The preparation method of the modified polydatin inclusion compound is as follows: polydatin and β-cyclodextrin are mixed in a molar ratio of 1:8, and ultrasonically treated at 60° C. and 500W for 40 minutes to improve the solubility of polydatin through cyclodextrin inclusion.

[0050] The preparation method of the nano-astragalus and houttuynia cordata complex is as follows: Astragalus and Houttuynia cordata were mixed in a ratio of 5:1, and cellulase (enzyme activity ≥ 5000 U / g) was added for enzymatic hydrolysis at 45°C for 3 h. The enzymatic hydrolysis used a composite enzyme system containing cellulase, pectinase, and xylanase (mass ratio 3:1:1); After high-pressure homogenization (2000 bar), the product is frozen in liquid nitrogen and ground to a particle size of 0.2 μm. The grinding is carried out in a liquid nitrogen environment at -196°C and a grinding speed of 800 rpm. The absorption of traditional Chinese medicine is enhanced by combining enzymatic hydrolysis and nano-processing.

[0051] The preparation method of the organic acid sustained-release microcapsule is as follows: Dissolve citric acid and fumaric acid in water at a ratio of 3:1 and mix with 5 wt% sodium alginate; The microcapsules were dripped into a coagulation solution containing 3wt% calcium chloride and 0.2wt% chitosan and dried to obtain microcapsules. The organic acid was slowly released in the intestine to maintain the antiviral pH.

[0052] The preparation method of the fermented ammonium glycyrrhizate is as follows: glycyrrhizic acid and Lactobacillus plantarum are anaerobically fermented at 39°C for 60 hours, the pH is controlled at 6.5, and the microbial fermentation converts the immune active components; the preparation method of the selenized Radix Isatidis polysaccharide is as follows: Radix Isatidis polysaccharide and sodium selenite are complexed and reacted at 80°C for 4 hours in a mass ratio of 10:5:1.

[0053] The preparation method of the modified quercetin comprises the following steps: compounding quercetin and soybean lecithin in a mass ratio of 1:5, evaporating the solvent to form a film, adding phosphate buffer (pH 7.4) for hydration, and ultrasonically treating the film to obtain a nanocomposite.

[0054] The preparation method of the modified mannitol is: Mannitol was dissolved in 8 times its mass of isopropanol-water mixed solvent (volume ratio 4:1), and 2.5 times the molar amount of NaOH was added, and the mixture was stirred at 65°C for 30 minutes; Chloroacetic acid was added in batches (3 times with an interval of 20 minutes) to control the molar ratio of chloroacetic acid to mannitol to 0.7:1, and the temperature was raised to 75°C for 3 hours; Adjust the pH to 7.5 with glacial acetic acid, add 3 volumes of ethanol for precipitation, and remove by-products by ultrafiltration (molecular weight cut-off 1 kDa); The product was spray-dried (inlet air temperature 160°C, outlet air temperature 80°C) to obtain a white powder.

[0055] 2) Mixing the raw materials under an inert atmosphere; 3) Sieve the mixture (e.g. 60 mesh), divide into portions and seal them in a dark place.

[0056] Comparative Example 1: Compared with Example 3, the modified polydatin inclusion complex is missing (replaced by an equal amount of pharmaceutical carrier).

[0057] Comparative Example 2: Compared with Example 3, the modified quercetin is missing (replaced by an equal amount of pharmaceutical carrier).

[0058] Comparative Example 3: Compared with Example 3, modified mannitol is missing (replaced by an equal amount of pharmaceutical carrier).

[0059] Comparative Example 4: Compared with Example 3, the modified polydatin inclusion compound, modified quercetin, and modified mannitol are missing (they are replaced by an equal amount of pharmaceutical carrier).

[0060] Comparative Example 5: Compared with Example 3, the nano-astragalus-houttuynia cordata complex is missing (replaced by an equal amount of pharmaceutical carrier).

[0061] Comparative Example 6: Compared with Example 3, ordinary polydatin (unmodified) was used instead of the modified polydatin inclusion complex.

[0062] Comparative Example 7: Compared with Example 3, ordinary quercetin (unmodified) was used instead of modified quercetin.

[0063] Comparative Example 8: Compared with Example 3, ordinary mannitol (unmodified) was used instead of modified mannitol.

[0064] Comparative Example 9: A commercially available Chinese herbal compound (Astragalus + Isatis root).

[0065] Experimental testing Animal model: 60 kg healthy sows (PRRSV / ASFV double negative), 10 per group; Virus challenge method: PRRSV (JXA1 strain, 10 5 TCID 50)+ASFV (Georgia 2007 strain, 10 4 HAD 50 ) Combined intramuscular injection; Dosage regimen: Add 0.3% of the preparation to feed for 7 consecutive days (starting 3 days before the challenge); Detection time: 7 days after infection.

[0066] The anti-virus test results are shown in Table 1 below: Table 1 Anti-virus test results Group PRRSV inhibition rate (%) ASFV inactivation rate (%) Survival rate (%) PRRSV load in lung tissue (log) Lymph node ASFV load (log) Example 1 92.5 94.3 80 3.8 3.0 Example 2 95.8 96.7 90 2.9 2.3 Example 3 98.7 99.1 95 2.1 1.8 Example 4 97.2 98.0 90 2.5 2.0 Example 5 96.0 97.5 85 3.0 2.5 Comparative Example 1 61.3 85.2 40 5.6 4.8 Comparative Example 2 83.7 88.9 60 4.2 3.7 Comparative Example 3 76.4 82.1 50 4.8 4.2 Comparative Example 4 42.1 68.5 20 6.5 5.9 Comparative Example 5 71.6 80.3 55 4.5 4.0 Comparative Example 6 52.8 70.5 30 6.1 5.3 Comparative Example 7 68.9 75.3 35 5.4 4.7 Comparative Example 8 74.2 79.6 45 4.9 4.1 Comparative Example 9 47.5 - 10 7.2 6.5

[0067] From the above results, it can be seen that the preparation prepared by the present invention has a good preventive and therapeutic effect on blue ear disease and African swine fever.

[0068] The results of immunomodulation and safety tests are shown in Table 2 below: Table 2 Immunomodulation and safety test results Group CD4⁺ / CD8⁺ ratio <![CDATA[IgG antibody (OD 450 )]]> ALT (U / L) Withdrawal period (days) Average daily feed intake (kg) Example 3 2.8±0.3 1.85±0.15 38±5 3 2.8±0.2 Comparative Example 1 1.2±0.2 0.72±0.10 42±6 7 2.1±0.3 Comparative Example 2 1.6±0.3 1.05±0.12 45±4 5 2.3±0.2 Comparative Example 3 1.4±0.2 0.93±0.11 50±7 6 2.0±0.4 Comparative Example 4 0.9±0.1 0.61±0.09 58±8 10 1.8±0.3 Comparative Example 5 1.3±0.2 0.68±0.08 47±6 8 2.2±0.3 Comparative Example 6 1.1±0.2 0.65±0.07 62±9 9 1.9±0.2 Comparative Example 7 1.0±0.1 0.59±0.06 60±7 9 2.0±0.3 Comparative Example 8 1.3±0.2 0.71±0.08 55±6 7 2.1±0.2 Comparative Example 9 0.8±0.1 0.51±0.08 65±10 14 1.5±0.4 Healthy pigs 1.5-2.0 0.1-0.3 10-50 - 3.0±0.5

[0069] From the above results, it can be seen that the preparation prepared by the present invention has a short withdrawal period, improves the breeding efficiency, and has good palatability.

[0070] It should be noted that, for the sake of simplicity, the aforementioned embodiments are described as a series of action combinations. However, those skilled in the art should be aware that the present invention is not limited by the order of the actions described, because according to the present invention, certain steps may be performed in other orders or simultaneously. Secondly, those skilled in the art should also be aware that the embodiments described in this specification are all preferred embodiments, and the actions and modules involved are not necessarily required by the present invention.

[0071] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the scope of protection of the invention. Obviously, the embodiments described are only some embodiments of the present invention, rather than all embodiments. Based on these embodiments, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in this field can still combine, add, delete or make other adjustments to the features in the various embodiments of the present invention according to the circumstances without conflict, without making creative work, so as to obtain different other technical solutions that do not deviate from the concept of the present invention in essence, and these technical solutions also fall within the scope of protection of the present invention.

Claims

1. A preparation for blue ear disease and African swine fever, characterized in that: The invention comprises the following raw materials in parts by weight: 10-60 parts of modified polydatin inclusion compound, 40-80 parts of nano-astragalus-houttuynia cordata complex, 10-30 parts of medium-chain fatty acid glyceride, 8-40 parts of organic acid sustained-release microcapsules, 5-25 parts of fermented ammonium glycyrrhizate, 5-20 parts of selenized isatis root polysaccharide, 10-20 parts of modified quercetin, 5-10 parts of modified mannitol, and 20-100 parts of a pharmaceutical carrier.

2. The preparation for blue ear disease and African swine fever according to claim 1, characterized in that The invention comprises the following raw materials in parts by weight: 20-50 parts of modified polydatin inclusion compound, 50-70 parts of nano-astragalus-houttuynia cordata complex, 15-25 parts of medium-chain fatty acid glyceride, 18-30 parts of organic acid sustained-release microcapsules, 10-20 parts of fermented ammonium glycyrrhizate, 10-15 parts of selenized isatis root polysaccharide, 12-18 parts of modified quercetin, 6-9 parts of modified mannitol, and 40-80 parts of pharmaceutical carrier.

3. The preparation for blue ear disease and African swine fever according to claim 2, characterized in that The invention comprises the following raw materials in parts by weight: 35 parts of modified polydatin inclusion compound, 60 parts of nano-astragalus-houttuynia cordata complex, 20 parts of medium-chain fatty acid glyceride, 24 parts of organic acid sustained-release microcapsules, 15 parts of fermented ammonium glycyrrhizate, 12.5 parts of selenized isatis root polysaccharide, 15 parts of modified quercetin, 7.5 parts of modified mannitol, and 60 parts of a pharmaceutical carrier.

4. The preparation for blue ear disease and African swine fever according to claim 1, characterized in that The preparation method of the modified polydatin inclusion compound is as follows: polydatin and beta-cyclodextrin are mixed in a molar ratio of 1:4-8, and ultrasonic treatment is performed at 40-60° C. and 200-500W for 20-40 minutes.

5. The preparation for blue ear disease and African swine fever according to claim 1, characterized in that The preparation method of the nano-astragalus-houttuynia cordata complex is as follows: Mix Astragalus and Houttuynia cordata in a ratio of 2-5:1, add cellulase and enzymolyze at 35-45℃ for 1-3h; After high-pressure homogenization, the product was frozen in liquid nitrogen and ground to a particle size of 0.1-2 μm.

6. The preparation for blue ear disease and African swine fever according to claim 1, characterized in that The preparation method of the organic acid sustained-release microcapsule is as follows: Dissolve citric acid and fumaric acid in water at a ratio of 1-3:1 and mix with 2-5wt% sodium alginate; The mixture is dropped into a coagulation solution containing 1-3 wt% calcium chloride and 0.5-2 wt% chitosan, and dried to obtain microcapsules.

7. The preparation for blue ear disease and African swine fever according to claim 1, characterized in that The preparation method of the fermented ammonium glycyrrhizate is as follows: glycyrrhizic acid and Lactobacillus plantarum are anaerobically fermented at 35-39°C for 36-60 hours, the pH is controlled at 5.5-6.5, and the microbial fermentation is converted into immune active components; the preparation method of the selenized Radix Isatidis polysaccharide is as follows: Radix Isatidis polysaccharide and sodium selenite are complexed and reacted at a mass ratio of 10:1-5:1 at 60-80°C for 2-4 hours.

8. The preparation for blue ear disease and African swine fever according to claim 1, characterized in that The preparation method of the modified quercetin comprises the following steps: compounding quercetin and soybean lecithin in a mass ratio of 1:2-5, evaporating the solvent to form a film, adding phosphate buffer for hydration, and ultrasonically treating the film to obtain a nanocomposite.

9. The preparation for blue ear disease and African swine fever according to claim 1, characterized in that The preparation method of the modified mannitol is: Dissolve mannitol in a 5-8 times by weight isopropanol-water mixed solvent, add 2.0-2.5 times by molar amount of NaOH, and stir at 60-65°C for 30 minutes; Add chloroacetic acid in batches, controlling the molar ratio of chloroacetic acid to mannitol to 0.5-0.7:1, and heat to 70-75°C for 2-3 hours; Adjust the pH to 6.5-7.5 with glacial acetic acid, add 3 volumes of ethanol for precipitation, and remove by-products by ultrafiltration; Spray drying gave a white powder.

10. The method for preparing the preparation for blue ear disease and African swine fever according to any one of claims 1 to 9, characterized in that: The steps include: Prepare the raw materials according to the recipe; Mix the raw materials under an inert atmosphere; The mixture was sieved, divided into packages and sealed to protect from light.

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