Use of tricin-7-o-β-d-glucopyranoside in the preparation of an antiallergic drug

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to the use of red clover extract-7-O-β-D-glucopyranoside in the preparation of anti-allergic drugs. Through in vitro and in vivo pharmacological experiments, this invention demonstrates that red clover extract-7-O-β-D-glucopyranoside can effectively inhibit IgE-mediated allergic reactions and suppress the release of type 2 inflammatory factors, exhibiting significant anti-allergic activity. It can be used to prevent or treat allergic diseases such as allergic rhinitis, allergic asthma, atopic dermatitis, and allergic gastrointestinal diseases, with high safety and low toxicity. It provides a new candidate drug for the treatment of allergic diseases and has broad clinical application prospects.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to the use of red clover extract-7-O-β-D-glucopyranoside in the preparation of anti-allergy drugs. Background Technology

[0002] Allergic diseases, also known as hypersensitivity diseases, are a collective term for a group of diseases caused by an abnormal immune response of the body's immune system to normally harmless foreign substances (allergens) such as pollen and dust mites. They encompass various types, including allergic rhinitis, allergic asthma, and atopic dermatitis. According to epidemiological survey data from 30 countries conducted by the World Allergy Organization (WAO), approximately 22% of the global population is affected. Although these diseases were rare at the beginning of the 20th century, their prevalence has risen sharply globally in recent decades, now affecting 10% to 40% of the world's population. The World Health Organization has listed them as one of the three major diseases requiring focused research and prevention in the 21st century. Focusing on China, the "2022 China Allergic Disease Epidemiological Survey Report and Current Status Analysis," as my country's first specialized report in this field, systematically reviews the disease burden, risk factors, and current treatment status. Studies show that allergic diseases not only impose a huge economic burden but also have a particularly severe impact on children. In 2016, the proportion of infants under 2 years old in my country who had experienced or were experiencing allergic symptoms was as high as 40.9%. How to effectively prevent and treat allergic diseases has become a core issue in the medical and pharmaceutical communities both at home and abroad.

[0003] Currently, the clinical treatment of allergic diseases with Western medicine mainly relies on the following three categories of drugs: first, antihistamines, such as diphenhydramine, loratadine, and cetirizine; second, glucocorticoids, such as hydrocortisone, dexamethasone, and prednisone, which aim to relieve allergy symptoms by inhibiting inflammatory mediators; and third, allergy mediator release inhibitors, such as ketotifen and sodium cromoglycate. However, existing Western medicines have significant clinical limitations: on the one hand, adverse reactions are frequent and have serious consequences. For example, astemizole can induce arrhythmias or even cardiac arrest, while montelukast may cause serious psychosomatic reactions such as stuttering and suicidal tendencies; on the other hand, these drugs can mostly only treat the symptoms and cannot fundamentally correct immune system disorders, making it difficult to block the disease process, resulting in a persistently high recurrence rate of allergies.

[0004] In recent years, natural medicinal ingredients have shown potential in the field of anti-allergy treatment, such as baicalin, compound glycyrrhizin, and curcumin. However, related research and clinical applications have confirmed that these ingredients still have significant side effects. Animal experiments have shown that high doses of baicalin can significantly reduce the percentage of lymphocytes, indicating a risk of immunosuppression, accompanied by signs of kidney damage such as elevated serum creatinine and glomerular inflammatory infiltration. Compound glycyrrhizin can easily cause multi-system adverse reactions such as hypokalemia, edema, elevated blood pressure, and muscle weakness. Although curcumin has pharmacological activity, its water solubility is extremely poor (only 16.82 μg / mL), and it is rapidly metabolized due to enterohepatic circulation. The effective therapeutic dose usually needs to exceed 8g per day, which can easily cause gastrointestinal reactions such as nausea, abdominal pain, and diarrhea. Furthermore, there are two reported cases of acute hepatitis and even liver failure induced by taking curcumin supplements.

[0005] Therefore, developing a novel anti-allergy drug with proven efficacy and improved safety is a pressing technical problem that needs to be solved in this field. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the present invention adopts the following technical solution:

[0007] A first aspect of the present invention provides the use of thymol-7-O-β-D-glucopyranoside in the preparation of an anti-allergy drug;

[0008] Furthermore, the anti-allergy drug is selected from any one or more of allergic rhinitis, allergic asthma, atopic dermatitis, and allergic gastrointestinal diseases; even further, the anti-allergy drug is selected from any one or more of type 2 inflammation-mediated allergic rhinitis, allergic asthma, atopic dermatitis, and allergic gastrointestinal diseases.

[0009] Furthermore, the anti-allergy drug is an oral or non-oral formulation;

[0010] Furthermore, the oral formulation is made from tsaponin-7-O-β-D-glucopyranoside and a pharmaceutically acceptable carrier;

[0011] Furthermore, the oral preparation is selected from any one or more of tablets, capsules, powders, pellets, micro-pellets, and granules;

[0012] Furthermore, the tablets are selected from any one or more of uncoated tablets, coated tablets, and sustained-release tablets;

[0013] Furthermore, the capsule is selected from any one or more of hard capsules, soft capsules, and sustained-release capsules;

[0014] Furthermore, the pharmaceutically acceptable carrier is selected from any one or more of the following: fillers, diluents, binders, disintegrants, wetting agents, lubricants, colorants, and sweeteners;

[0015] Furthermore, the filler or diluent is selected from any one or more of starch, pregelatinized starch, lactose, glucose, mannitol, sorbitol, cellulose, xylitol, dextrose, fructose, calcium sulfate, light magnesium oxide, and sucrose;

[0016] Furthermore, the adhesive is selected from any one or more of the following: polyvinylpyrrolidone, microcrystalline cellulose, starch paste, syrup, maltose, microcrystalline glucose, sodium carboxymethyl cellulose, honey, sodium carboxymethyl dextran, and dextrin.

[0017] Furthermore, the disintegrant is selected from any one or more of starch, sodium carboxymethyl starch, croscarmellose sodium, hydroxypropyl starch, low-substituted hydroxypropyl cellulose, and microcrystalline cellulose;

[0018] Furthermore, the wetting agent is selected from distilled water or ethanol;

[0019] Furthermore, the lubricant is selected from any one or more of the following: talc, magnesium stearate, liquid paraffin, magnesium dodecyl sulfate, sodium dodecyl sulfate, long-chain fat emulsion, stearic acid, polyethylene glycol, micronized silica gel, and hydrogenated vegetable oil.

[0020] Furthermore, the non-oral formulation is selected from injections, nasal sprays, or topical gels;

[0021] Furthermore, the injection is made from red cloverin-7-O-β-D-glucopyranoside and a pharmaceutically acceptable carrier; further still, the injection is a fat emulsion intravenous injection formulation or a lyophilized powder injection.

[0022] Furthermore, the injectable acceptable carrier is selected from any one or more of the following: pH adjuster, antioxidant, osmotic pressure adjuster, medium or long chain fat emulsion, lyophilization protectant, solubilizer, cosolvent, preservative, and chelating agent;

[0023] Furthermore, the acid-base regulator is selected from acids, bases, or buffer salts; further still, the acid-base regulator is selected from one or more of the following: sodium hydroxide aqueous solution, ammonia, hydrochloric acid, dilute sulfuric acid, citric acid, acetic acid, maleic acid, lactic acid, phosphoric acid, tartaric acid, tris(hydroxymethyl)aminomethane, sodium carbonate, sodium bicarbonate, sodium acetate, trisodium phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, ethanolamine, triethanolamine, ethylenediamine, monosodium glutamate, and boric acid; further still, the acid-base regulator is preferably selected from ammonia, sodium hydroxide solution, or hydrochloric acid.

[0024] Furthermore, the pH value of the injection is 3 to 11; further still, the pH value of the injection is preferably 6 to 11; further still, the pH value of the injection is preferably 6.5 to 8.

[0025] Furthermore, the antioxidant is selected from any one or more of sodium bisulfite, sodium metabisulfite, potassium bisulfite, sodium thiosulfate, vitamin C, enediol palmitate, tocopherol, hydroquinone, propyl gallate, dihydroguaiacol, tert-butyl-p-hydroxyanisole, dibutylcresol, L-cysteine, L-methionine, L-lysine, L-valine, proline, histidine, cystine, disodium edetate, and calcium sodium edetate.

[0026] Furthermore, the osmotic pressure regulator is selected from any one or more of glucose, sodium chloride, potassium chloride, glycerol, mannitol, sorbitol, xylitol, and sodium lactate Ringer's solution; further still, the osmotic pressure regulator is preferably sodium chloride;

[0027] Furthermore, the freeze-drying protectant is selected from any one or more of sucrose, lactose, and mannitol; more specifically, lactose is preferred as the freeze-drying protectant.

[0028] Furthermore, the solubilizer or co-solvent is selected from any one or more of the following: amides or urea derivatives, amines, amino acids, alcohols, inorganic acids and their salts, sugars, glycosides, saponins, hexamethylenetetramine, morpholine, sodium cholate, monovalent soap, aloxo OT, sodium lauryl sulfate, cyclodextrin, polyoxyethylene monostearate, higher fatty acids of sucrose, Tween 80, polyoxyethylene castor oil, and polyoxyethylene hydrogenated castor oil.

[0029] Furthermore, the preservative is selected from any one or more of benzyl alcohol, hydroxypropyl butyl ester, hydroxypropyl methyl ester, phenol, chlorobutanol, and thimerosal; further still, benzyl alcohol is preferred as the preservative.

[0030] Furthermore, the chelating agent is selected from EDTA salts; preferably disodium EDTA salt;

[0031] The present invention has the following beneficial effects:

[0032] This invention is the first to demonstrate that red cloverin-7-O-β-D-glucopyranoside has significant therapeutic or ameliorative effects on a variety of allergic diseases, including but not limited to respiratory allergic diseases such as allergic rhinitis and allergic asthma, skin allergic diseases such as atopic dermatitis, and allergic gastrointestinal diseases. This broad-spectrum activity indicates that red cloverin-7-O-β-D-glucopyranoside acts on the common core links of allergic reactions, rather than providing local relief of single symptoms, and has the clinical development potential of "one drug for multiple treatments." Its mechanism of action involves synergistic regulation of multiple targets and links: on the one hand, it can inhibit IgE levels and alleviate hypersensitivity reactions; on the other hand, it can inhibit type 2 inflammatory responses, regulate Th1 / Th2 immune balance, and reduce tissue infiltration of inflammatory cells. In models of allergic rhinitis and asthma, red clover extract-7-O-β-D-glucopyranoside significantly improved airway hyperresponsiveness and nasal inflammation, and also alleviated airway wall thickening. In a model of atopic dermatitis, it had a dual effect of inhibiting skin inflammation and repairing skin barrier function. In allergic gastrointestinal diseases, it could reduce intestinal allergic reactions by maintaining intestinal mucosal immune homeostasis and barrier integrity.

[0033] Compared with existing glucocorticoids and antihistamines, red cloverin-7-O-β-D-glucopyranoside offers equal or better efficacy while exhibiting lower toxicity and higher safety. Furthermore, it can be flexibly administered via multiple routes, including oral, inhalation, nasal, and topical application, facilitating the development of appropriate formulations for different target organs. Red cloverin-7-O-β-D-glucopyranoside provides a novel drug candidate for the treatment of allergic diseases, possessing broad clinical application prospects and significant industrial value. Attached Figure Description

[0034] Figure 1 The present invention describes the effect of red clover extract-7-O-β-D-glucopyranoside on the levels of type 2 cytokines IL-5 and IL-13 in lung tissue of a mouse model of allergic asthma.

[0035] Figure 2 The effects of red cloverin-7-O-β-D-glucopyranoside on the number of peripheral blood eosinophils (EOS) and serum IgE levels in a mouse model of allergic asthma;

[0036] Figure 3 The present invention describes the effect of red cloverin-7-O-β-D-glucopyranoside on pathological changes in lung tissue in a mouse model of allergic asthma.

[0037] Figure 4 This invention, red cloverin-7-O-β-D-glucopyranoside, inhibits the activity of ILC2 cells in the lung tissue of mice in the remission phase of allergic inflammation, accounting for 55% of CD45. + The effect of cell percentage;

[0038] Figure 5 The effect of red clover extract-7-O-β-D-glucopyranoside on middle ear swelling in a mouse model of atopic dermatitis;

[0039] Figure 6 The effect of red clover extract-7-O-β-D-glucopyranoside on ear weight in a mouse model of atopic dermatitis;

[0040] Figure 7 The present invention describes the effect of red clover extract-7-O-β-D-glucopyranoside on the levels of type 2 cytokines IL-5 and IL-13 in the ear tissue of a mouse model of atopic dermatitis.

[0041] Figure 8 The present invention describes the effect of red clover extract-7-O-β-D-glucopyranoside on the pathological changes of the middle ear tissue in a mouse model of atopic dermatitis.

[0042] Figure 9 This invention investigates the effects of red cloverin-7-O-β-D-glucopyranoside and other flavonoids on ear weight in a mouse model of atopic dermatitis.

[0043] Figure 10 This invention investigates the effects of red cloverin-7-O-β-D-glucopyranoside and other flavonoids on middle ear swelling in a mouse model of atopic dermatitis.

[0044] Figure 11 This invention investigates the effects of red cloverin-7-O-β-D-glucopyranoside and other flavonoids on the pathological changes in the middle ear tissue of a mouse model of atopic dermatitis.

[0045] Figure 12 This invention investigates the effects of red cloverin-7-O-β-D-glucopyranoside and other flavonoids on the thymus index and spleen index in a mouse model of atopic dermatitis. Detailed Implementation

[0046] The following specific embodiments are used to illustrate the technical solutions of the present invention, but the scope of protection of the present invention is not limited to the scope of the embodiments described.

[0047] The structure of the red cloverin-7-O-β-D-glucopyranoside described in this invention is shown in Formula I below:

[0048]

[0049] Formula I;

[0050] In particular, the application of red cloverin-7-O-β-D-glucopyranoside provided by the present invention can be made available for clinical use in the form of a pharmaceutical composition.

[0051] (I) Implementation Examples

[0052] Example 1: Red clover extract-7-O-β-D-glucopyranoside tablets

[0053] Red clover extract-7-O-β-D-glucopyranoside 200g

[0054] 800g of starch

[0055] 500g of dextrin

[0056] 450g of powdered sugar

[0057] 50g magnesium stearate

[0058] Red cloverin-7-O-β-D-glucopyranoside was thoroughly mixed with a portion of starch and dextrin in a mixer to form a powder. The remaining starch, dextrin, and powdered sugar were mixed with an appropriate amount of water and heated in a water bath to form a paste. The paste was then mixed with the powder in a mixer to form a soft mass, which was then granulated, dried, and pressed into 10,000 tablets with magnesium stearate. Each tablet contains 20 mg of red cloverin-7-O-β-D-glucopyranoside.

[0059] Example 2: Red clover extract-7-O-β-D-glucopyranoside capsules

[0060] Red clover extract-7-O-β-D-glucopyranoside: 20g;

[0061] Starch: 280g; Magnesium stearate: 12g

[0062] The above components were poured into a mixer and thoroughly mixed. The mixture was then filled into capsules using a capsule filling machine to obtain 1000 capsules of red cloverin-7-O-β-D-glucopyranoside, each capsule containing 20 mg of red cloverin-7-O-β-D-glucopyranoside.

[0063] Example 3: Red clover extract-7-O-β-D-glucopyranoside fat emulsion intravenous injection formulation

[0064] Weigh 1000 mg of dispersed red cloverin-7-O-β-D-glucopyranoside and put it into a container; add 200 g of tea oil for injection and put it into the container; place the container in a water bath and heat it to about 80°C, stirring until the drug is dispersed; cool it down to 60°C, add 25 g of synthetic phospholipids and 100 mg of vitamin E, and stir until the phospholipids dissolve to form a uniform mixed phase. 700 ml of water for injection was placed in another container. 40 g of hydroxypropyl β-cyclodextrin, 25 g of glycerol, and 15 g of synthetic phospholipids were dissolved at 60°C by stirring to form an aqueous phase. A mixture containing cloverin-7-O-β-D-glucopyranoside, soybean oil, and lecithin was mixed with the aqueous phase at 60°C under mechanical stirring, and the pH was adjusted to 6.5-7.0 with 1 wt% sodium hydroxide. Water for injection was then added to a final volume of 1000 ml, and mechanical stirring was continued at 60°C for 1 hour to prepare the colostrum. The prepared colostrum was dispersed using a high-speed disperser and then homogenized in a high-pressure homogenizer until the droplet size met the requirements. The colostrum was then filtered through a microporous membrane, filled into containers, nitrogen-purged, and capped. Finally, it was sterilized in a rotary autoclave at 100°C. Sterilize at 20°C, and after sterilization, gradually cool and store at room temperature to prepare a 100mg / 100ml red clover extract-7-O-β-D-glucopyranoside fat emulsion for intravenous injection.

[0065] Stability test method: Freshly prepared fat emulsion was placed in a stoppered centrifuge tube and centrifuged at 4000 rpm for 15 min. No stratification or drug precipitation was observed. After one year of storage at room temperature in a cool place, the appearance, particle size, zeta potential, and other physicochemical properties of the emulsion, as well as its content, showed no significant changes, indicating that the emulsion was stable. Test results: pH value of the intravenous fat emulsion: 7.0; particle size range: 0.05–1.0 μm.

[0066] Example 4: Lyophilized powder for injection of red clover extract-7-O-β-D-glucopyranoside

[0067] Red clover extract-7-O-β-D-glucopyranoside is dissolved in a medical organic solvent (methanol, acetone or one or two of these), then a surface treatment dispersant is added and stirred. The mixture is heated to 35℃~55℃ while polylactic acid is added with stirring. After the reaction is complete, an excipient is added at 35℃~55℃ and stirred until the reaction is complete. The mixture is then filtered through a microporous membrane, and the filtrate is lyophilized to produce a lyophilized powder injection.

[0068] Dissolve 40 mg of red cloverin-7-O-β-D-glucopyranoside in 10 ml of methanol at room temperature. Add 400 mg of poloxamer and stir until homogeneous at room temperature. Heat the solution to 35°C–55°C and add 3000 mg of polylactic acid (MW=3000) at a rate of 3–8 ml / min while stirring at 800–1800 rpm for 25–40 minutes. Then, at 35°C–55°C and 800–1800 rpm, add 2000 mg of lactose and continue stirring for 2–10 minutes. Filter the solution through a 0.8 μm microporous membrane. Dispense the filtrate into 2 ml vials and freeze at -45°C–-55°C for 2–4 hours to obtain the final product.

[0069] Conclusion: The prepared red clover extract-7-O-β-D-glucopyranoside-polylactic acid nanoparticle powder injection has an encapsulation efficiency of 92.9%, a drug loading of 23.9%, an average particle size of 216.5 nm, a Zate potential of -22.67 mV, and an aggregation index of 0.191.

[0070] Example 5: Red Clover Extract-7-O-β-D-Gluranoside Nasal Spray

[0071] Take 800 ml of water for injection at 80℃, add hydroxypropyl-β-cyclodextrin, stir to dissolve, add sodium chloride, stir until completely dissolved, and cool to room temperature. Pre-dissolve red cloverin-7-O-β-D-glucopyranoside powder in a small amount of ethanol, and slowly add it dropwise to the excipient solution, stirring continuously for 30 minutes. Fine-tune by adding benzalkonium chloride, stirring to dissolve, and adjust the pH to 5.5-6.5 with phosphate buffer. Add water for injection to the total volume. Filter through a 0.22 μm microporous membrane for sterilization, aseptically dispense into brown glass vials, attach a nasal spray pump head, and dispense 50-100 μL per spray. Check the active ingredient content, droplet size, and sterility of each spray.

[0072] Example 6: Red clover extract-7-O-β-D-glucopyranoside topical gel

[0073] Evenly sprinkle Carbomer 940 into 400ml of purified water at room temperature, let it stand for 4 hours or overnight and stir until a uniform gel is formed. Dissolve red cloverin-7-O-β-D-glucopyranoside in propylene glycol by sonication for 30 minutes, add glycerol and azone, and mix well. Dissolve ethylparaben in a small amount of hot water first and then add it. Slowly add the drug solution to the Carbomer gel while stirring. Add triethanolamine dropwise and stir until the pH reaches 6.0~7.0, forming a transparent gel. The amount of triethanolamine used is about 1.2 times the weight of Carbomer. The consistency needs to be observed while adding. Degas under a vacuum of -0.08MPa for 30 minutes to remove air bubbles, dispense into aluminum-plastic composite tubes or glass bottles, seal, and store in a cool, dark place.

[0074] (I) Experimental Example

[0075] Experimental Example 1: Efficacy of red cloverin-7-O-β-D-glucopyranoside in an allergic asthma model

[0076] 2.1 Number of animals and grouping: 25 male BALB / c mice, 12-15g (3 weeks old), were randomly divided into 5 groups: blank control group, allergic asthma group, red clover extract-7-O-β-D-glucopyranoside treatment group (two doses of 0.5 mg / kg and 2 mg / kg, respectively), and dexamethasone group (dose of 0.2 mg / kg), with 5 mice in each group.

[0077] 2.2 Drug administration: Except for the control group, mice were given 20 μL of freshly prepared HDM solution via nasal drops for a total of 5 times to induce allergic asthma. The control group was given 20 μL of normal saline via nasal drops, the red clover extract-7-O-β-D-glucopyranoside treatment group was given red clover extract-7-O-β-D-glucopyranoside, the dexamethasone group was given dexamethasone, and the control group and the allergic asthma group were given normal saline.

[0078] 2.3 Experimental Results

[0079] Note: PG in the attached image represents thymol-7-O-β-D-glucopyranoside.

[0080] (1) See Figure 1 IL-5 and IL-13 levels in mouse lung tissue: Compared with the control group, the levels of IL-5 and IL-13 in the lung tissue of mice in the allergic asthma group were significantly increased, while the levels of IL-5 and IL-13 in the lung tissue of mice treated with red clover extract-7-O-β-D-glucopyranoside were significantly decreased.

[0081] (2) See Figure 2 The number of EOS and the level of IgE in the peripheral blood of mice: Compared with the blank group, the number of EOS and the level of IgE in the peripheral blood of mice in the allergic asthma group were significantly increased, while the number of EOS and the level of IgE in the serum of mice treated with red clover extract-7-O-β-D-glucopyranoside were significantly decreased.

[0082] (3) See Figure 3 Thickening of the airway wall and peri-airway inflammatory cell infiltration in mice: Compared with the control group, the airway wall of mice in the allergic asthma group was significantly thickened and peri-airway inflammatory cell infiltration was increased. No significant thickening of the airway wall was observed in mice treated with red clover extract-7-O-β-D-glucopyranoside, and the peri-airway inflammatory cell infiltration was mild.

[0083] (4) See Figure 4The number of type 2 immune cells in the lung tissue of mice in the remission phase of allergic inflammation: Compared with the control group, the percentage of type 2 immune cells (ILC2) in the lung tissue of mice in the model remission phase was still significantly increased. Treatment with red clover extract-7-O-β-D-glucopyranoside (0.5 mg / kg) significantly reduced the proportion of ILC2 in CD45 in the lung tissue of mice. + Percentage of cells.

[0084] Example 2: Efficacy of red cloverin-7-O-β-D-glucopyranoside in an atopic dermatitis model

[0085] 2.1 Number of animals and grouping: 24 BALB / c mice, half male and half female, 23-30 g (8 weeks old), were randomly divided into 4 groups of 6 mice each, half male and half female: blank group, model group, red clover extract-7-O-β-D-glucopyranoside treatment group (1 mg / kg), and dexamethasone group (0.2 mg / kg).

[0086] 2.2 Administration: From day 1 to day 7, except for the control group, 20 μL of freshly prepared 1 nmol MC903 solution was applied to both ears of mice (15 μL for the inner ear and 5 μL for the outer ear) to induce an atopic dermatitis model. The control group and the model group were administered physiological saline by gavage. The red clover extract-7-O-β-D-glucopyranoside group was administered red clover extract-7-O-β-D-glucopyranoside 1 mg / kg by gavage, and the dexamethasone group was administered dexamethasone 0.2 mg / kg by gavage.

[0087] 2.3 Experimental Results

[0088] Note: PG in the attached image represents thymol-7-O-β-D-glucopyranoside.

[0089] (1) See Figure 5 Changes in ear swelling in mice: Compared with the blank group, the ear swelling of mice in the model group was significantly increased, while the ear swelling of mice treated with red clover extract-7-O-β-D-glucopyranoside was significantly reduced.

[0090] (2) See Figure 6 Changes in mouse ear tissue weight: Compared with the blank group, the ear tissue weight of mice in the model group was significantly increased, while the ear tissue weight of mice treated with red clover extract-7-O-β-D-glucopyranoside was significantly reduced.

[0091] (3) See Figure 7 Levels of IL-5 and IL-13 in mouse ear tissue: Compared with the blank group, the levels of IL-5 and IL-13 in the ear tissue of mice in the model group were significantly increased, while the levels of IL-5 and IL-13 in the ear tissue of mice treated with red clover extract-7-O-β-D-glucopyranoside were decreased.

[0092] (4) See Figure 8 Pathological changes in mouse ear tissue: Compared with the blank group, the epidermis of mouse ear tissue in the model group was thickened and the infiltration of inflammatory cells was increased. The epidermis of mouse ear tissue treated with red clover extract-7-O-β-D-glucopyranoside was not significantly thickened and the infiltration of inflammatory cells was mild.

[0093] Example 3: Comparison of the efficacy of daidzein, genistein, red cloverin-7-O-β-D-glucopyranoside daidzein on an atopic dermatitis model.

[0094] Daidzein and genistein are two flavonoids with aglycone structures similar to those of trifolin-7-O-β-D-glucopyranoside, and their corresponding glycosides have been reported for use in anti-allergic drug treatment. To investigate the differences in the effects of these two flavonoids and trifolin-7-O-β-D-glucopyranoside in anti-allergic treatment, genistein, daidzein, trifolin, and trifolin-7-O-β-D-glucopyranoside (all at a dose of 1 mg / kg) were administered to a mouse model of atopic dermatitis.

[0095] 3.1 Number of animals and grouping: 42 male BALB / c mice, 23-30 g (8 weeks old), were randomly divided into 7 groups of 6 mice each: blank group, model group, daidzein group (1 mg / kg), genistein group (1 mg / kg), red cloverin group (1 mg / kg), red cloverin-7-O-β-D-glucopyranoside group (1 mg / kg), and dexamethasone group (0.62 mg / kg).

[0096] 3.2 Drug administration: From day 1 to day 7, except for the control group, 20 μL of freshly prepared 1 nmol MC903 solution was applied to both ears of mice (15 μL for the inner ear and 5 μL for the outer ear) to induce an atopic dermatitis model. The control group and the model group were administered physiological saline by gavage. The daidzein group, genistein group, red cloverin group, and red cloverin-7-O-β-D-glucopyranoside group were administered the corresponding drug at 1 mg / kg by gavage. The dexamethasone group was administered dexamethasone at 0.62 mg / kg by gavage.

[0097] 3.3 Experimental Results:

[0098] Note: PG represents the red cloverin-7-O-β-D-glucopyranoside group, and P represents the red cloverin group; # Compared with the blank group, * compared with the model group, † compared with the PG group.

[0099] (1) See Figure 9Changes in mouse ear tissue weight: Compared with the blank group, the weight of the mouse ear in the model group was significantly increased; all drug administration groups could reduce the weight of mouse ear, but the effect of red clover extract-7-O-β-D-glucopyranoside was better than that of daidzein group, genistein group and red clover extract group.

[0100] (2) See Figure 10 Changes in mouse ear swelling: Compared with the blank group, the ear swelling of mice in the model group was significantly increased; except for the genistein group, all other treatment groups were able to reduce mouse ear swelling, but the red clover extract-7-O-β-D-glucopyranoside group was significantly more effective than the daidzein group.

[0101] (3) See Figure 11 Pathological changes in mouse ear tissue: Compared with the blank group, the epidermal layer of the ear tissue of the model group mice was significantly thickened, and the degree of inflammatory cell infiltration was significantly increased. Except for the genistein group, all other treatment groups could alleviate the pathological changes of epidermal thickening in the ear tissue of the model mice. However, the thickness of the epidermal layer of the ear tissue of mice in the red clover extract-7-O-β-D-glucopyranoside group was lower than that in the red clover extract group and the daidzein group, and the alleviating effect on inflammatory cell infiltration was also more significant.

[0102] (4) See Figure 12 Immunosuppressive effect: Except for the dexamethasone group, no significant immunosuppression was observed in any of the treatment groups.

Claims

1. Use of red cloverin-7-O-β-D-glucopyranoside in the preparation of anti-allergy drugs.

2. The use according to claim 1, characterized in that, The anti-allergy drug is selected from any one or more of allergic rhinitis, allergic asthma, atopic dermatitis, and allergic gastrointestinal diseases; further, the anti-allergy drug is selected from any one or more of type 2 inflammation-mediated allergic rhinitis, allergic asthma, atopic dermatitis, and allergic gastrointestinal diseases.

3. The use according to claim 1 or 2, characterized in that, The anti-allergy medication is an oral or non-oral formulation.

4. The use according to claim 3, characterized in that, The oral formulation is made from red cloverin-7-O-β-D-glucopyranoside and a pharmaceutically acceptable carrier; Furthermore, the oral preparation is selected from any one or more of tablets, capsules, powders, pellets, micro-pellets, and granules; Furthermore, the tablets are selected from any one or more of plain tablets, coated tablets, and sustained-release tablets; Furthermore, the capsule is selected from any one or more of hard capsules, soft capsules, and sustained-release capsules; Furthermore, the pharmaceutically acceptable carrier is selected from any one or more of the following: fillers, diluents, binders, disintegrants, wetting agents, lubricants, colorants, and sweeteners; Furthermore, the filler or diluent is selected from any one or more of starch, pregelatinized starch, lactose, glucose, mannitol, sorbitol, cellulose, xylitol, dextrose, fructose, calcium sulfate, light magnesium oxide, and sucrose; Furthermore, the adhesive is selected from any one or more of the following: polyvinylpyrrolidone, microcrystalline cellulose, starch paste, syrup, maltose, microcrystalline glucose, sodium carboxymethyl cellulose, honey, sodium carboxymethyl dextran, and dextrin. Furthermore, the disintegrant is selected from any one or more of starch, sodium carboxymethyl starch, croscarmellose sodium, hydroxypropyl starch, low-substituted hydroxypropyl cellulose, and microcrystalline cellulose; Furthermore, the wetting agent is selected from distilled water or ethanol; Furthermore, the lubricant is selected from any one or more of the following: talc, magnesium stearate, liquid paraffin, magnesium dodecyl sulfate, sodium dodecyl sulfate, long-chain fat emulsion, stearic acid, polyethylene glycol, micronized silica gel, and hydrogenated vegetable oil.

5. The use according to claim 3, characterized in that, The non-oral preparation is selected from injections, nasal sprays, or topical gels.

6. The use according to claim 5, characterized in that, The injection is made from red cloverin-7-O-β-D-glucopyranoside and a pharmaceutically acceptable carrier; furthermore, the injection is a fat emulsion intravenous injection or a lyophilized powder injection. Furthermore, the injectable acceptable carrier is selected from any one or more of the following: pH adjuster, antioxidant, osmotic pressure adjuster, medium or long chain fat emulsion, lyophilization protectant, solubilizer, cosolvent, preservative, and chelating agent; Furthermore, the acid-base regulator is selected from acids, bases, or buffer salts; further still, the acid-base regulator is selected from one or more of the following: sodium hydroxide aqueous solution, ammonia, hydrochloric acid, dilute sulfuric acid, citric acid, acetic acid, maleic acid, lactic acid, phosphoric acid, tartaric acid, tris(hydroxymethyl)aminomethane, sodium carbonate, sodium bicarbonate, sodium acetate, trisodium phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, ethanolamine, triethanolamine, ethylenediamine, monosodium glutamate, and boric acid; further still, the acid-base regulator is preferably selected from ammonia, sodium hydroxide solution, or hydrochloric acid. Furthermore, the pH value of the injection is 3 to 11; further still, the pH value of the injection is preferably 6 to 11; further still, the pH value of the injection is preferably 6.5 to 8. Furthermore, the antioxidant is selected from any one or more of sodium bisulfite, sodium metabisulfite, potassium bisulfite, sodium thiosulfate, vitamin C, enediol palmitate, tocopherol, hydroquinone, propyl gallate, dihydroguaiacol, tert-butyl-p-hydroxyanisole, dibutylcresol, L-cysteine, L-methionine, L-lysine, L-valine, proline, histidine, cystine, disodium edetate, and calcium sodium edetate. Furthermore, the osmotic pressure regulator is selected from any one or more of glucose, sodium chloride, potassium chloride, glycerol, mannitol, sorbitol, xylitol, and sodium lactate Ringer's solution; further still, the osmotic pressure regulator is preferably sodium chloride; Furthermore, the freeze-drying protectant is selected from any one or more of sucrose, lactose, and mannitol; more specifically, lactose is preferred as the freeze-drying protectant. Furthermore, the solubilizer or co-solvent is selected from any one or more of the following: amides or urea derivatives, amines, amino acids, alcohols, inorganic acids and their salts, sugars, glycosides, saponins, hexamethylenetetramine, morpholine, sodium cholate, monovalent soap, aloxo OT, sodium lauryl sulfate, cyclodextrin, polyoxyethylene monostearate, higher fatty acids of sucrose, Tween 80, polyoxyethylene castor oil, and polyoxyethylene hydrogenated castor oil. Furthermore, the preservative is selected from any one or more of benzyl alcohol, hydroxypropyl butyl ester, hydroxypropyl methyl ester, phenol, chlorobutanol, and thimerosal; further still, benzyl alcohol is preferred as the preservative. Furthermore, the chelating agent is selected from EDTA salts; preferably disodium EDTA salt.

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