Pharmaceutical composition for preventing or treating porcine epidemic diarrhea

Abstract

The invention belongs to the technical field of animal medicine, and discloses a pharmaceutical composition for preventing or treating porcine epidemic diarrhea. The paeoniflorin extract, the pawpaw total triterpenes, the mesalazine, the honeysuckle flower water extract and the mannan oligosaccharide are scientifically compounded, so that multi-target synergistic treatment is realized. Mesalazine can inhibit intestinal inflammation, and pawpaw total triterpenes can relieve oxidative damage and protect intestinal epithelium; the honeysuckle water extract is used for blocking virus infection, and the paeoniflorin extract is used for repairing intestinal villi; and the mannan oligosaccharide can regulate intestinal flora. All the components have a synergistic effect, the porcine epidemic diarrhea symptom is effectively controlled, the intestinal function is repaired, the cad pig incidence rate is reduced, and compared with a traditional scheme, pig herd health and breeding benefits can be more efficiently guaranteed.

Description

Technical Field

[0001] This invention belongs to the field of animal medicine technology, and specifically relates to a pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea. Background Technology

[0002] Porcine epidemic diarrhea (PED) is a highly contagious intestinal infectious disease caused by porcine epidemic diarrhea virus (PEDV), characterized by vomiting, diarrhea, and dehydration. In recent years, PED has become widespread in rural areas of my country, causing significant economic losses to the pig farming industry. PEDV is highly pathogenic; the mortality rate in infected newborn piglets can reach up to 100%, while the mortality rate in weaned piglets and finishing pigs is approximately 10%-30%, directly leading to a sharp decline in herd survival rates. PEDV infection primarily causes piglets to exhibit severe watery diarrhea, vomiting, severe dehydration, and hemorrhagic enteritis. These clinical symptoms appear very rapidly, often spreading quickly within hours of infection, greatly increasing the difficulty of treatment. Even after recovery, infected pigs often suffer from intestinal malabsorption due to thinned intestinal walls and atrophied villi, resulting in "runts." Data shows that runts can have a 20%-30% lower feed conversion rate and a 30-45 day longer growth cycle, significantly increasing farming costs.

[0003] While traditional Western medicines (such as antiviral drugs and antibiotics) can provide short-term symptom relief, they easily induce drug-resistant viral mutations, and long-term use may damage the liver and kidney function of pigs. Traditional Chinese medicine (TCM), with its advantages of being natural, safe, and having fewer side effects, is gradually gaining attention in the treatment of porcine epidemic diarrhea (PED). However, although TCM single-herb or compound formulas have the advantages of being naturally low in toxicity and having multi-target regulation, they also suffer from problems such as slow onset of action, unclear active ingredients, and unstable dosage forms, making it difficult to quickly control acute infections when used alone. Therefore, providing a drug composition that can quickly control symptoms and provide long-term repair is of great significance for improving the treatment effect of PED and ensuring the healthy development of grassroots pig farming. Summary of the Invention

[0004] The present invention aims to solve at least one of the technical problems existing in the prior art, and provides a pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, wherein the components work synergistically to effectively control the symptoms of porcine epidemic diarrhea, repair intestinal function, and reduce the incidence of runts.

[0005] The first aspect of the present invention is to provide a pharmaceutical composition.

[0006] The second aspect of the present invention aims to provide the application of the pharmaceutical composition of the first aspect of the present invention.

[0007] The third aspect of this invention is to provide a product.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides a pharmaceutical composition comprising paeoniflorin extract, total papaya triterpenes, mesalazine, honeysuckle extract, and mannan oligosaccharides.

[0009] In some embodiments of the present invention, the pharmaceutical composition comprises, by weight, 5-25 parts of paeoniflorin extract, 8-25 parts of total papaya triterpenes, 3-15 parts of mesalazine, 10-30 parts of honeysuckle extract, and 15-45 parts of mannan oligosaccharides.

[0010] In some embodiments of the present invention, the weight parts of the paeoniflorin extract may be selected from any value of 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, or 25 parts, or a range formed by any two of these values.

[0011] In some embodiments of the present invention, the weight parts of the total triterpenes of papaya may be selected from any value of 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, or a range formed by any two of these values.

[0012] In some embodiments of the present invention, the weight parts of mesalazine may be selected from any value of 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, or 15 parts, or a range formed by any two of these values.

[0013] In some embodiments of the present invention, the weight parts of the honeysuckle extract may be selected from any value of 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, or 30 parts, or a range formed by any two of these values.

[0014] In some embodiments of the present invention, the weight parts of the mannan oligosaccharide may be selected from any value of 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 parts, 31 parts, 32 parts, 33 parts, 34 parts, 35 parts, 36 parts, 37 parts, 38 parts, 39 parts, or 40 parts, or a range formed by any two of these values.

[0015] In some embodiments of the present invention, the paeoniflorin extract is an alcoholic extract of peony.

[0016] Peony belongs to the Paeoniaceae family ( Paeoniaceae The dried root of the perennial herbaceous plant peony is bitter, sour, and slightly cold in nature. It enters the liver and spleen meridians and has the functions of calming the liver and relieving pain, nourishing blood and regulating menstruation, and astringing yin and stopping sweating. Paeoniflorin is the main active ingredient in peony. Modern pharmacological studies have shown that it has pharmacological effects such as inhibiting autoimmune reactions, anti-inflammation, analgesia, liver protection, increasing cardiac output, and improving cardiopulmonary function.

[0017] In some embodiments of the present invention, the paeoniflorin extract is prepared by a method comprising: mixing peony with ethanol and reflux extraction to obtain the paeoniflorin extract.

[0018] In some preferred embodiments of the present invention, before mixing, the peony is pretreated, including drying the roots of the peony and crushing them through a 50-60 mesh.

[0019] In some preferred embodiments of the present invention, the ratio (g / mL) of peony to ethanol is 1:(8-15).

[0020] In some preferred embodiments of the present invention, the ratio of peony root to ethanol (g / mL) can be selected from any value of 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15 or a range of any two.

[0021] In some preferred embodiments of the present invention, the reflux extraction conditions are: extraction at 70-80°C for 1-3 hours; and cooling and drying at 25-60°C.

[0022] In some preferred embodiments of the present invention, the concentration of ethanol is 60%-80%.

[0023] In some preferred embodiments of the present invention, the reflux extraction is repeated 2-3 times.

[0024] In some preferred embodiments of the present invention, the method further includes a purification step, comprising concentrating the reflux extract under reduced pressure and purifying it by adsorption using a macroporous resin (selected from AB-8, HPD100 or D101), with the eluent being a gradient concentration of ethanol solution (e.g., 30% ethanol, 70% ethanol elution).

[0025] In some preferred embodiments of the present invention, the method further includes a drying step.

[0026] In some embodiments of the present invention, the honeysuckle extract is an aqueous extract of honeysuckle.

[0027] Honeysuckle is rich in chlorogenic acid, trace elements and vitamins, and has broad-spectrum antiviral, antibacterial and heat-clearing and detoxifying effects. It is used to treat carbuncles, boils, sore throat, erysipelas, dysentery caused by heat and toxins, colds caused by wind and heat, and fever caused by febrile diseases.

[0028] In some embodiments of the present invention, the honeysuckle extract is prepared by a method comprising: mixing honeysuckle with water and extracting to obtain the honeysuckle extract.

[0029] In some preferred embodiments of the present invention, the honeysuckle is dried honeysuckle buds.

[0030] In some preferred embodiments of the present invention, the ratio of honeysuckle to water (g / mL) is 1:(8-15).

[0031] In some preferred embodiments of the present invention, the ratio of honeysuckle to water (g / mL) can be selected from any value of 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15 or a range of any two.

[0032] In some preferred embodiments of the present invention, the extraction conditions are: extraction at 70-85°C for 1-3 hours; further, extraction at 77-85°C for 1.2-2 hours; and even further, extraction at 79-82°C for 1.4-1.6 hours. The extraction temperature ensures sufficient dissolution of chlorogenic acid while reducing its thermal degradation. Low temperatures can easily lead to incomplete extraction and loss of active ingredients, while high temperatures can cause chlorogenic acid to be destroyed or insufficiently dissolved, affecting antiviral activity.

[0033] In some preferred embodiments of the present invention, the extraction is repeated 2-3 times.

[0034] In some preferred embodiments of the present invention, the method further includes concentration (e.g., concentration under reduced pressure at 50-65°C to a relative density of 1-1.2) and drying steps.

[0035] In some embodiments of the present invention, the total triterpenes of papaya contain oleanolic acid, ursolic acid, betulinic acid, 3-O-acetylsourlic acid, 3-O-acetylsourmicic acid, hawthorn acid, cinnamyl acid and speciosaperoxide.

[0036] Papaya, known as the "fruit of a hundred benefits," is a very important ornamental and edible plant, and was listed as one of the first batch of foods recognized for both medicinal and edible uses in 2003. Traditional Chinese medicine believes that papaya has the effects of relaxing muscles and tendons, promoting blood circulation, strengthening the spleen and stomach, soothing the liver and relieving pain, and dispelling wind and dampness. It can be used to prevent and treat rheumatism, cholera, dysentery, gastroenteritis, beriberi, and vitamin C deficiency. Total triterpenes in papaya are important active ingredients, mainly including oleanolic acid, ursolic acid, betulinic acid, 3-O-acetylursolic acid, 3-O-acetylporphyrin, hawthorn acid, potassium oleate, and speciosaperoxide, which possess excellent antioxidant and immune-enhancing functions.

[0037] In some embodiments of the present invention, the total triterpenes of papaya are prepared by the following method: papaya is mixed with ethanol and refluxed for extraction to obtain total triterpenes of papaya.

[0038] In some preferred embodiments of the present invention, before mixing, the papaya is pretreated, including pulverizing the papaya and drying it (drying at 70-90℃ for 2-3 hours) and passing it through a 40-50 mesh.

[0039] In some preferred embodiments of the present invention, the ratio of papaya to ethanol (g / mL) is 1:(8-15).

[0040] In some preferred embodiments of the present invention, the ratio of papaya to ethanol (g / mL) can be selected from any value of 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15 or a range of any two.

[0041] In some preferred embodiments of the present invention, the reflux extraction conditions are: extraction at 60-80°C for 1-3 hours; and concentration and drying at 25-60°C.

[0042] In some preferred embodiments of the present invention, the concentration of ethanol is 60%-80%.

[0043] In some preferred embodiments of the present invention, the reflux extraction is repeated 3-4 times.

[0044] In some preferred embodiments of the present invention, the method further includes a purification step, comprising concentrating the reflux extract under reduced pressure, and purifying it by adsorption using a macroporous resin (selected from AB-8, HPD100, or D101), followed by elution with water and 70%-80% ethanol solution. Eluent is then used to elute until the effluent is colorless, followed by elution with ethanol solution. In some preferred embodiments of the present invention, the method further includes a drying step.

[0045] Mesalazine, also known as 5-aminosalicylic acid, is the active ingredient of sulfasalazine. It is an effective drug for treating mild to moderate active ulcerative colitis and segmental ileitis. It can specifically act on the site of intestinal inflammation, inhibit the production and release of inflammatory mediators, reduce the inflammatory response of the intestinal mucosa, and relieve symptoms such as diarrhea and abdominal pain.

[0046] Mannooligosaccharides (MOS) are formed by the synthesis of mannose units through β-transferases. Manno-oligosaccharides are oligosaccharides linked by 1,4 glycosidic bonds, belonging to the category of functional oligosaccharides. They are primarily derived from the hemicellulose components of plant cell walls, such as the hydrolysates of galactomannan widely found in the seeds of legumes. As a bioactive substance, manno-oligosaccharides possess various biological effects, including enhancing the body's immune function, promoting growth and development, and improving gut health. They can selectively promote the growth and reproduction of beneficial gut bacteria (such as Bifidobacteria and Lactobacilli), thus regulating the balance of the gut microbiota.

[0047] The pharmaceutical composition of this invention, mesalazine, reduces the release of acute inflammatory mediators by inhibiting the activity of key enzymes such as cyclooxygenase and lipoxygenase, thereby alleviating intestinal mucosal edema and inflammatory exudation. Papain triterpenes simultaneously activate the Nrf2 pathway, promoting the expression of antioxidant enzymes and promptly neutralizing reactive oxygen species generated during mesalazine metabolism, reducing oxidative stress damage to intestinal mucosal cells. The combined use of these two ingredients accelerates the resolution of intestinal mucosal congestion, promotes ulcer healing, and achieves dual anti-inflammatory and cell-protective effects. Simultaneously, papaya triterpenes upregulate the expression of tight junction proteins, effectively preventing the leakage of harmful substances from the intestine; paeoniflorin extract accelerates the differentiation and proliferation of villous basal cells, promoting rapid repair and regeneration of damaged intestinal villi. The synergistic effect of these two ingredients efficiently restores the intestinal physical barrier function and nutrient absorption function. In this invention, multiple active ingredients (such as chlorogenic acid) in the honeysuckle water extract can bind to PEDV surface proteins, blocking the process of viral adsorption and invasion of intestinal cells, effectively inhibiting viral replication in vivo, reducing viral antigen load, and, in conjunction with mesalazine, inhibiting the excessive inflammatory response induced by the virus, effectively reducing the risk of secondary intestinal mucosal damage. Mannooligosaccharides, as prebiotics, can selectively promote the growth and reproduction of beneficial intestinal bacteria such as Bifidobacteria and Lactobacillus, rapidly rebuilding the intestinal microecological balance and forming a microbial barrier. After the intestinal mucosa is repaired under the action of other components, the microbial barrier and the intestinal mucosa together constitute a "bacterial-membrane dual barrier," reducing pathogen reinfection and nutrient absorption disorders, thereby reducing the probability of runt formation.

[0048] A second aspect of the invention provides the use of the pharmaceutical composition of the first aspect of the invention in (1) or (2): (1) Preparation of products against swine epidemic diarrhea virus; (2) Prepare products for the prevention and / or treatment of diseases caused by porcine epidemic diarrhea virus infection.

[0049] In some embodiments of the present invention, the product includes reagents, reagent kits, and drugs.

[0050] In some embodiments of the present invention, the disease caused by the porcine epidemic diarrhea virus infection includes porcine epidemic diarrhea.

[0051] In some embodiments of the present invention, when the pharmaceutical composition is used to prevent and / or treat diseases caused by porcine epidemic diarrhea virus infection, the effective dose is 1-2 g / kg body weight per day.

[0052] A third aspect of the present invention provides a product comprising the pharmaceutical composition of the first aspect of the present invention.

[0053] In some embodiments of the present invention, the product includes reagents, reagent kits, and drugs.

[0054] In some embodiments of the present invention, the product further includes pharmaceutically acceptable excipients.

[0055] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one selected from fillers, disintegrants, diluents, dispersants, excipients, stabilizers, lubricants, binders, humectants, flavoring agents, solubilizers, suspending agents, solvents, sustained-release agents, emulsifiers, absorption enhancers, surfactants, preservatives, pigments, fragrances, and solvents.

[0056] In some embodiments of the present invention, the filler is selected from starch, sucrose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, or glucose, etc.; the binder is selected from cellulose derivatives, alginate, starch, water, dextrin, gelatin, or polyvinylpyrrolidone, etc.; the disintegrant is selected from microcrystalline cellulose, sodium carboxymethyl starch, croscarmellose, low-substituted hydroxypropyl cellulose, or croscarmellose sodium; the lubricant is selected from stearic acid, polyethylene glycol, calcium carbonate, sodium bicarbonate, micronized silica gel, talc, or magnesium stearate; and the suspending agent is selected from micronized silica gel, beeswax, cellulose, or solid polyethylene glycol. The wetting agent is selected from glycerin, Tween-80, hydrogenated castor oil, or lecithin; the solvent is selected from ethanol, liquid polyethylene glycol, isopropanol, Tween-80, glycerin, propylene glycol, or vegetable oil, wherein the vegetable oil is selected from soybean oil, castor oil, peanut oil, blended oil, etc.; the surfactant is selected from sodium dodecylbenzenesulfonate, stearic acid, polyoxyethylene-polyoxypropylene copolymer, fatty acid sorbitan, or polysorbate (Tween), etc.; the flavoring agent is selected from aspartame, sucralose, flavoring, stevia, acesulfame potassium, citric acid, or sodium saccharin; the preservative is selected from at least one of methylparaben or propylparaben.

[0057] In some embodiments of the present invention, the dosage form of the drug includes a gastrointestinal dosage form or a non-gastrointestinal dosage form.

[0058] In some embodiments of the present invention, the gastrointestinal dosage form includes at least one of pills, powders, tablets, granules, capsules, sustained-release preparations, solutions, dry suspensions, effervescent tablets, emulsions, suspensions, syrups, drops, and chewable tablets.

[0059] In some embodiments of the present invention, the gastrointestinal dosage forms include, but are not limited to, enteric-coated tablets, coated tablets, film-coated tablets, sugar-coated tablets, dispersible tablets, sucking tablets, chewable tablets, effervescent tablets, scratch tablets, sustained-release and controlled-release dosage forms, sustained-release tablets, sustained-release coated tablets, controlled-release tablets, orally disintegrating tablets, lozenges, and oral patches.

[0060] In some embodiments of the present invention, the non-gastrointestinal drug delivery dosage form includes at least one of injection dosage form, respiratory dosage form, skin dosage form, mucosal dosage form, and cavity dosage form.

[0061] In some embodiments of the present invention, the injectable dosage forms include, but are not limited to, injection solutions, solutions for injection, injection solutions for intravenous infusion, suspensions for injection, sterile powders for injection, intravenous injections, water injections, emulsions for injection, powder injections, injections, sterile powder injections, lyophilized powder injections, etc.

[0062] The beneficial effects of this invention are: This invention scientifically combines paeoniflorin extract, total papaya triterpenes, mesalazine, honeysuckle water extract, and mannan oligosaccharides to achieve multi-target synergistic treatment. Mesalazine inhibits intestinal inflammation, while total papaya triterpenes reduce oxidative damage and protect the intestinal epithelium; honeysuckle water extract blocks viral infection, and paeoniflorin extract repairs intestinal villi; mannan oligosaccharides regulate the intestinal flora. The synergistic effect of these components effectively controls the symptoms of porcine epidemic diarrhea, repairs intestinal function, and reduces the incidence of runts. Compared to traditional methods, this approach more efficiently protects the health of the pig herd and improves farming efficiency. Detailed Implementation

[0063] The following will describe the concept and technical effects of the present invention clearly and completely with reference to embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are all within the scope of protection of the present invention.

[0064] Unless otherwise specified in the examples, the procedures should be performed under standard conditions or conditions recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0065] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0066] Example 1 This embodiment provides a method for preparing paeoniflorin extract, total triterpenes of papaya, and honeysuckle water extract, as detailed below: Paeoniflorin extract was prepared by the following method: dried peony root was pulverized to 60 mesh and extracted twice with 70% ethanol at a material-to-liquid ratio of 1:10 (extraction temperature was 75℃), 2 hours each time; the extracts were combined and concentrated under reduced pressure to a relative density of 1.15; the concentrate was adsorbed onto AB-8 macroporous adsorption resin and eluted successively with 30% ethanol and 70% ethanol, the eluent was collected, concentrated under reduced pressure until alcohol-free, and then cooled and dried at 60℃ to obtain paeoniflorin extract.

[0067] Total triterpenes from papaya were prepared by the following method: Papaya was pulverized to 40 mesh and dried at 80℃ for 2 hours; the dried papaya was extracted three times by reflux with 70% ethanol at a material-to-liquid ratio of 1:10 (extraction temperature was 70℃), 2 hours each time; the extracts were combined and concentrated until no alcohol odor was detected; the concentrated extract was adsorbed onto HPD100 macroporous adsorption resin, eluted with water until the effluent was colorless, and then eluted with 80% ethanol. The eluent was collected, concentrated, and cooled and dried at 60℃ to obtain total triterpenes from papaya.

[0068] Honeysuckle water extract was prepared by the following method: dried honeysuckle buds were soaked in 10 times the amount of water twice at 80°C for 1.5 hours each time; the extracts were combined and concentrated under reduced pressure at 60°C to a relative density of 1.15, and then dried at 60°C to obtain honeysuckle water extract.

[0069] Example 2 A pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, comprising, by weight, the following ingredients: 13 parts paeoniflorin extract, 16 parts total papaya triterpenes, 7 parts mesalazine, 18 parts honeysuckle water extract, and 30 parts mannan oligosaccharides.

[0070] The preparation method of the above-mentioned pharmaceutical composition is as follows: each raw material is passed through an 80-mesh sieve and then mixed.

[0071] Example 3 A pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, comprising, by weight, the following ingredients: 10 parts paeoniflorin extract, 12 parts total papaya triterpenes, 5 parts mesalazine, 15 parts honeysuckle water extract, and 25 parts mannan oligosaccharides.

[0072] The preparation method of the above-mentioned pharmaceutical composition is as follows: each raw material is passed through an 80-mesh sieve and then mixed.

[0073] Example 4 A pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, comprising, by weight, the following ingredients: 15 parts of paeoniflorin extract, 18 parts of total papaya triterpenes, 8 parts of mesalazine, 20 parts of honeysuckle extract, and 35 parts of mannan oligosaccharides.

[0074] The preparation method of the above-mentioned pharmaceutical composition is as follows: each raw material is passed through an 80-mesh sieve and then mixed.

[0075] Comparative Example 1 A pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, comprising, by weight, the following ingredients: 13 parts paeoniflorin extract, 7 parts mesalazine, 18 parts honeysuckle water extract, and 30 parts mannan oligosaccharide.

[0076] The preparation method of the above-mentioned pharmaceutical composition is as follows: each raw material is passed through an 80-mesh sieve and then mixed.

[0077] Comparative Example 2 A pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, comprising, by weight, the following ingredients: 13 parts paeoniflorin extract, 16 parts total triterpenes of papaya, 18 parts honeysuckle water extract, and 30 parts mannan oligosaccharides.

[0078] The preparation method of the above-mentioned pharmaceutical composition is as follows: each raw material is passed through an 80-mesh sieve and then mixed.

[0079] Comparative Example 3 A pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, comprising, by weight, the following ingredients: 13 parts paeoniflorin extract, 16 parts grape seed extract (purchased from Shaanxi Yuwangtang Biotechnology Development Co., Ltd.), 7 parts mesalazine, 18 parts honeysuckle water extract, and 30 parts mannan oligosaccharide.

[0080] The preparation method of the above-mentioned pharmaceutical composition is as follows: each raw material is passed through an 80-mesh sieve and then mixed.

[0081] Comparative Example 4 A pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, comprising, by weight, the following ingredients: 13 parts paeoniflorin extract, 16 parts total triterpenes of papaya, 7 parts aspirin, 18 parts honeysuckle water extract, and 30 parts mannan oligosaccharides.

[0082] The preparation method of the above-mentioned pharmaceutical composition is as follows: each raw material is passed through an 80-mesh sieve and then mixed.

[0083] Comparative Example 5 A pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, comprising, by weight, the following ingredients: 13 parts paeoniflorin extract, 16 parts total papaya triterpenes, 7 parts mesalazine, and 30 parts mannan oligosaccharides.

[0084] The preparation method of the above-mentioned pharmaceutical composition is as follows: each raw material is passed through an 80-mesh sieve and then mixed.

[0085] Comparative Example 6 A pharmaceutical composition for the prevention or treatment of porcine epidemic diarrhea, comprising, by weight, the following ingredients: 13 parts of paeoniflorin extract, 16 parts of total papaya triterpenes, 7 parts of mesalazine, and 18 parts of honeysuckle water extract.

[0086] The preparation method of the above-mentioned pharmaceutical composition is as follows: each raw material is passed through an 80-mesh sieve and then mixed.

[0087] Effect Example 1. Cell-level antiviral assay Using a porcine small intestinal epithelial cell (IPEC-J2) model, cells were pretreated with different drug combinations (Examples 2-4, Comparative Examples 1-6, and individual components) for 2 hours. The pretreatment solution was then discarded, and porcine epidemic diarrhea virus (PEDV) suspension (MOI=1) was added to each well. Cells were incubated at 37°C in a 5% CO2 incubator for 1 hour. The virus solution was then discarded, and the cells were washed three times with PBS. DMEM medium containing 2% fetal bovine serum was added, and the cells were cultured for another 24 hours. A virus control group (infected with PEDV only, without any drugs) was also included in the experiment.

[0088] Virus titer determination: After culture, cell supernatants from each group were collected, and the median tissue culture infectious dose (TCID) of PEDV was determined using the Reed-Muench method. 50 The result is in LogTCID. 50 / mL represents the volume of water.

[0089] Cell viability assay: Cell viability was assessed using the CCK-8 assay. 10 μL of CCK-8 reagent was added to each well, and after incubation for 2 hours, the absorbance (OD value) at 450 nm was measured using a microplate reader. Cell viability was calculated as follows: Cell viability = (OD value of experimental group - OD value of virus control group) / (OD value of normal cell control group - OD value of virus control group) × 100%.

[0090] The test results are shown in Table 1. It can be seen that the viral titer after treatment in Examples 2-4 was significantly lower than that in the viral control group, each comparative ratio, and single components, while the cell viability was significantly higher than that in other groups. This result indicates that the pharmaceutical composition of the present invention, through the synergistic effect of multiple components, is significantly more effective than single components or combinations lacking key components in inhibiting PEDV replication and protecting intestinal epithelial cells, fully demonstrating the multi-target synergistic mechanism of "antiviral-anti-inflammatory-mucosal repair-microbiota regulation".

[0091] Table 1 Results of antiviral assays at the cellular level

[0092] Note: This indicates that compared to the virus control group, P <0.01.

[0093] 2. Clinical efficacy trial One hundred and fifty piglets, aged 7-30 days, clinically diagnosed with viral porcine epidemic diarrhea (PED) were selected. Their clinical manifestations included: normal or low body temperature; initial abdominal distension, vomiting containing a small amount of fluid, followed by watery diarrhea. Larger pigs exhibited poor mental state, abdominal distension, vomiting, and diarrhea. Based on clinical observation and laboratory tests, these pigs were diagnosed with viral PED. According to baseline indicators such as age, onset time, symptom severity, weight, and sex, the pigs were randomly divided into 10 groups (15 pigs per group) on a parallel basis, including three example groups (Examples 2-4), six comparative groups (Comparative Examples 1-6), and one blank control group.

[0094] Administration: The drug compositions from each example and comparative example were uniformly mixed into pig feed (Beijing Kang Enhe Animal Health Products Co., Ltd. 8125 pig feed) at a ratio of 1.5 g / kg body weight per day, and fed to the corresponding groups in sequence. The blank control group was fed pig feed without the added drug compositions. Pigs were allowed free access to feed, and the effect of treatment on viral swine diarrhea was observed after 7 days. The experimental results are expressed as the mean of three parallel trials, as detailed in Table 2.

[0095] Therapeutic effect assessment: mainly based on whether the symptoms of diarrhea in piglets have disappeared, and the specific therapeutic effect is divided into three levels: Cured: All diarrhea symptoms in the piglets disappeared, and they returned to normal. Improvement: Diarrhea symptoms in piglets partially disappear, and feces become soft or porridge-like; Ineffective: Piglets' diarrhea symptoms do not improve significantly, or worsen further, leading to death.

[0096] The results are shown in Table 2. The cure rate (80.0%-86.7%) and total effective rate (93.3%-100%) of the example group were significantly higher than those of all control groups (cure rate 40.0%-60.0%, total effective rate 66.7%-80.0%), while the total effective rate of the blank control group was only 26.7%. This demonstrates that the drug composition provided by this invention, through the synergistic effect of its components, can effectively control the symptoms of porcine epidemic diarrhea, repair intestinal function, and reduce the incidence of runts. Furthermore, observations showed that no other significant discomfort was observed in the piglets fed the drug, indicating its reliable safety and lack of obvious toxic side effects.

[0097] Table 2. Statistical analysis of treatment efficacy in each treatment group

[0098] The embodiments of the present invention have been described in detail above. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention. Furthermore, the embodiments of the present invention and the features thereof can be combined with each other unless otherwise specified.

Claims

1. A pharmaceutical composition comprising paeonol extract, total triterpenes of Chaenomelis Fructus, mesalazine, honeysuckle extract and mannan oligosaccharide.

2. The pharmaceutical composition of claim 1, wherein, The pharmaceutical composition comprises, by weight, 5-25 parts of paeonol extract, 8-25 parts of total triterpenes of Chaenomelis Fructus, 3-15 parts of mesalazine, 10-30 parts of honeysuckle extract and 15-45 parts of mannan oligosaccharide.

3. The pharmaceutical composition according to claim 1 or 2, characterized in that, The paeonol extract is an alcohol extract of Paeonia lactiflora Pall.; and / or, the honeysuckle extract is a water extract of Lonicera japonica Thunb.

4. The pharmaceutical composition of claim 3, wherein, The paeonol extract is prepared by a method comprising mixing Paeonia lactiflora Pall. with ethanol and refluxing to extract, thereby obtaining the paeonol extract.

5. The pharmaceutical composition of claim 3, wherein, The honeysuckle extract is prepared by a method comprising mixing Lonicera japonica Thunb. with water and extracting, thereby obtaining the honeysuckle extract.

6. The pharmaceutical composition according to claim 1 or 2, characterized by, The total triterpenes of Chaenomelis Fructus contain oleanolic acid, ursolic acid, betulinic acid, 3-O-acetyl ursolic acid, 3-O-acetyl pomolic acid, maslinic acid, potentilla acid and Speciosaperoxide.

7. The pharmaceutical composition of claim 6, wherein, The total triterpenes of Chaenomelis Fructus are prepared by a method comprising mixing Chaenomelis Fructus with ethanol and refluxing to extract, thereby obtaining the total triterpenes of Chaenomelis Fructus. 8.Use of the pharmaceutical composition of any one of claims 1-7 in (1) or (2): (1) preparation of a product against porcine epidemic diarrhea virus; or (2) preparation of a product for preventing and / or treating a disease caused by infection with porcine epidemic diarrhea virus. 9.A product comprising the pharmaceutical composition of any one of claims 1-7. 10.The product of claim 9, further comprising a pharmaceutically acceptable excipient. ​

Images