Pharmaceutical composition for treating migraine and preparation method thereof

Abstract

The invention belongs to the technical field of medicines, and provides a pharmaceutical composition for treating migraine and a preparation method thereof, the pharmaceutical composition is prepared from the following raw materials by mass: 30%-60% of armillaria mellea polysaccharide, 3%-7% of low molecular weight fucoidan polysaccharide sulfate, 8%-17% of gamma-aminobutyric acid, and 15%-60% of an auxiliary material maltodextrin. Active ingredients of food are used as raw materials, and medicinal and edible substances are adopted, so that the food is safe to eat, has good prevention and treatment effects, is suitable for people with migraine to take for a long time and is convenient to carry; the monoamine transmitter level of a composition treatment group is obviously improved, and 5-HT, NE and DA of brain tissues are obviously increased, so that the composition can nourish related neurons of an organism and promote the generation of neurotransmission substances controlled by the neurons to be increased, and therefore, the migraine symptom is improved; compared with a model group, the change of the NE has an obvious improvement effect; experiments show that the composition has a preventive treatment effect on rats suffering from migraine caused by nitroglycerin.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a medicinal composition for treating migraine and a preparation method thereof. Background Art

[0002] Migraine is a common neurological disorder that affects a large number of people worldwide. Its attacks are often accompanied by severe headaches, nausea, vomiting, and sensitivity to light and sound. Although a variety of medications are currently used to treat migraine, including analgesics and preventive medications, most treatment options still have some shortcomings.

[0003] First, the side effects of many drugs are worrying. For example, some nonsteroidal anti-inflammatory drugs (NSAIDs), while relieving pain, may cause gastrointestinal discomfort, impaired kidney function, and other problems. Preventive medications, such as β-adrenergic blockers and antidepressants, although effective for some patients, may also cause a variety of side effects such as weight gain and mood changes with long-term use, making it difficult for patients to adhere to long-term use.

[0004] Secondly, existing migraine treatments often fail to target the biological mechanisms that cause migraines. These problems, such as neurotransmitter imbalance and neuronal dysfunction, are not effectively addressed by most drugs. In particular, the relationship between migraine and monoamine neurotransmitters (such as 5-HT, NE, and DA) is gaining increasing attention. Existing treatments fail to adequately regulate the levels of these neurotransmitters, thus limiting the effectiveness of treatment.

[0005] Furthermore, the convenience of treatment methods is a concern for patients. Existing medications typically require strict schedules and dosages, lacking flexibility and making them difficult for patients to adapt to in their daily lives. Therefore, there is an urgent need to develop safe, effective, and long-term migraine treatment compositions to improve patients' quality of life. Summary of the Invention

[0006] In order to solve the above technical problems, the present invention provides a pharmaceutical composition for treating migraine and a preparation method thereof, so as to solve the problems mentioned in the background technology.

[0007] According to a first aspect of the present disclosure, a pharmaceutical composition for treating migraine is provided, which is prepared from the following raw materials in percentage by mass: 30% to 60% of Armillaria polysaccharide, 3% to 7% of low molecular weight fucoidan sulfate, 8% to 17% of gamma-aminobutyric acid, and 15% to 60% of maltodextrin as an auxiliary material;

[0008] The Armillaria polysaccharide is extracted from the mycelium of Armillaria mellea which is a wild hazel mushroom or artificially fermented and cultured;

[0009] The low molecular weight fucoidan sulfate is prepared by degrading fucoidan extracted from Laminaria japonica or Fucus vesiculosus by arabinofuranosidase, and has a molecular weight of 60,000 to 130,000.

[0010] Preferably, it is made of the following raw materials in percentage by mass: 60% of Armillaria polysaccharide, 7% of low molecular weight fucoidan sulfate, 17% of γ-aminobutyric acid, and 16% of auxiliary material maltodextrin.

[0011] According to the second aspect of the present disclosure, a method for preparing a pharmaceutical composition for treating migraine is proposed. S1. The specific preparation method of the active composition granules is as follows: 1000 g of Armillaria polysaccharide, 100 g of low molecular weight fucoidan sulfate, 250 g of γ-aminobutyric acid, and 1650 g of maltodextrin are weighed respectively, mixed evenly, and made into wet granules using a 14-16 mesh nylon sieve. The granules are dried at 50°C to 60°C to a moisture content of ≤5.0%, and bagged, 3 g per bag, to make 1000 bags.

[0012] S2. The specific preparation method of the active composition tablets is as follows: 3000 g of Armillaria polysaccharide, 350 g of low molecular weight fucoidan sulfate, 850 g of γ-aminobutyric acid, and 800 g of maltodextrin are weighed respectively, mixed evenly, and made into 16-mesh granules, tableted, and packaged; each tablet is 0.5 g, and 10,000 tablets are prepared.

[0013] Preferably, the preparation method of the Armillaria mellea polysaccharide is:

[0014] a. Preparation of fruiting body polysaccharide: 20 times the mass ratio of the weighed Armillaria fruiting body, washed and dried at 60 ℃, crushed into a coarse powder of about 1mm, the material-liquid ratio is 1:20, the temperature is 100 ℃, water reflux extraction 2 times, the first 3 hours, the second 2 hours; the combined extracts were vacuum concentrated to a relative density of 1.10 to 1.20 clear paste; the clear paste was precipitated with 3 to 4 times the amount of 95% ethanol for 24 hours, the precipitate was washed 3 times with anhydrous ethanol, and then dissolved with 4 times the amount of ≥80 ℃ hot water, let cool to 40 ℃ ~ 50 ℃, 1% to 3% papain was added, enzymatic hydrolysis was carried out for 4 to 5 hours, according to the polysaccharide aqueous solution: chloroform: n-butanol was added in a ratio of 25:4:1 chloroform / n-butanol mixed solution, stirred for 30 minutes, centrifuged at 3000r / min, the clarified liquid was concentrated to a thick paste, vacuum dried below 60 ℃, crushed into a fine powder, that is;

[0015] b. Preparation of mycelial polysaccharide: 10 times the mass ratio of Armillaria mellea mycelium is washed and dried at 60°C, with a solid-liquid ratio of 1:5 at a temperature of 90°C-100°C. Extract the mixture twice with water under reflux, first for 2 hours and then for 1 hour. Combine the extracts and vacuum concentrate to a clear paste with a relative density of 1.10-1.20. The paste is precipitated with 3-4 times the mass of 95% ethanol for 24 hours. The precipitate is washed three times with anhydrous ethanol and then dissolved with 4 times the mass of hot water (≥80°C). After cooling to 40-50°C, 1%-3% papain is added and enzymatic hydrolysis is carried out for 4-5 hours. A chloroform / n-butanol mixture is added in a ratio of 25:4:1 of polysaccharide aqueous solution:chloroform:n-butanol. The mixture is stirred for 30 minutes and centrifuged at 3000 rpm. The clarified liquid is concentrated to a thick paste, dried under vacuum below 60°C, and pulverized into a fine powder.

[0016] Preferably, the preparation method of the low molecular weight fucoidan sulfate is as follows: fucoidan sulfate extracted from kelp or fucus is weighed at 1.2 to 1.5 times the mass ratio, dissolved in water at a ratio of 1.5% to 3.0%, sterilized at 100°C for 20 to 30 minutes, and when the fucoidan sulfate aqueous solution is cooled to 60°C to 70°C, arabinofuranosidase is added at a ratio of 1% to 3%, the optimal pH is ≤ 7, the optimal reaction temperature is 60°C, and the reaction time is 4 to 5 hours to degrade the fucoidan sulfate; vacuum concentration is carried out, ethanol is added to the concentrate to 75%, the precipitate is collected, dried, and crushed to obtain low molecular weight fucoidan sulfate, whose molecular weight is 60,000 to 130,000.

[0017] Preferably, the γ-aminobutyric acid is derived from sodium L-glutamate as a raw material and is fermented by Lactobacillus hilgardii, and has a γ-aminobutyric acid content of ≥20%, a moisture content of ≤10%, and an ash content of ≤18%.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] 1. The present invention uses active ingredients of food as raw materials and adopts medicine-food homologous substances. It is safe to eat and has good preventive and therapeutic effects. It is suitable for long-term use by people with migraine and is easy to carry.

[0020] 2. The treatment group with the composition of the present invention had a significant improvement in the levels of monoamine neurotransmitters, and significantly increased 5-HT, NE, and DA in brain tissue, indicating that the composition of the present invention can nourish the relevant neurons in the body, promote the increased production of neurotransmitters controlled by them, and thus improve migraine symptoms; the change in NE showed a significant improvement compared with the model group.

[0021] 3. The present invention has demonstrated through experiments that the composition has a preventive therapeutic effect on rats with migraine induced by nitroglycerin. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The present invention is a flow chart of the preparation method of the pharmaceutical composition. DETAILED DESCRIPTION

[0023] The following embodiments of the present invention are described in further detail with reference to the accompanying drawings and examples. The following examples are used to illustrate the present invention but are not intended to limit the scope of the present invention.

[0024] As attached Figure 1 As shown:

[0025] Example 1: The present invention provides a pharmaceutical composition for treating migraine, which is prepared from the following raw materials in percentage by weight: 60% of Armillaria polysaccharide, 7% of low molecular weight fucoidan sulfate, 17% of γ-aminobutyric acid, and 16% of maltodextrin as an auxiliary material;

[0026] Armillaria polysaccharide is extracted from the mycelium of Armillaria mellea which is a wild hazel mushroom or artificially fermented and cultured.

[0027] Low molecular weight fucoidan sulfate is a fucoidan sulfate extracted from kelp of the Laminaria family or Fucus vesiculosus, which is degraded by arabinofuranosidase and has a molecular weight of 60,000 to 130,000.

[0028] Example 2: The present invention also provides a method for preparing a pharmaceutical composition for treating migraine, including S1, an active composition granule. The specific preparation method is: weigh 1000g of Armillaria polysaccharide, 100g of low molecular weight fucoidan sulfate, 250g of γ-aminobutyric acid, and 1650g of maltodextrin respectively, mix them evenly, and use a 14-16 mesh nylon sieve to form wet granules, dry them at 50℃~60℃ to a moisture content of ≤5.0%, and bag them, 3g per bag, to make 1000 bags.

[0029] S2. The specific preparation method of the active composition tablets is as follows: 3000 g of Armillaria polysaccharide, 350 g of low molecular weight fucoidan sulfate, 850 g of γ-aminobutyric acid, and 800 g of maltodextrin are weighed respectively, mixed evenly, and made into 16-mesh granules, tableted, and packaged; each tablet is 0.5 g, and 10,000 tablets are prepared.

[0030] Preparation of Armillaria polysaccharide: a. Preparation of Armillaria fruiting body polysaccharide: weigh Armillaria fruiting body according to 20 times of mass ratio, wash and dry at 60℃, crush into coarse powder of about 1mm, with solid-liquid ratio of 1:20, temperature of 100℃, add water and reflux extract twice, the first time for 3 hours and the second time for 2 hours; combine the extracts, vacuum concentrate into a clear paste with a relative density of 1.10-1.20 (room temperature); precipitate the clear paste with 3-4 times of 95% (mass fraction) ethanol for 24 hours; take the precipitate , washed 3 times with anhydrous ethanol, then dissolved with 4 times the amount (mass fraction) of hot water >80℃, cooled to 40℃~50℃, added 1%~3% (mass ratio) of papain, and kept warm for 4~5 hours: added chloroform / n-butanol mixed solution according to the ratio of polysaccharide aqueous solution: chloroform: n-butanol at 25:4:1, stirred for 30 minutes, and centrifuged at 3000r / min; took the clarified liquid, concentrated it into a thick paste, dried it in a vacuum below 60℃, and crushed it into fine powder to obtain Armillaria fruiting body polysaccharide;

[0031] b. Preparation of Armillaria mycelium polysaccharide: Take Armillaria mycelium at 10 times the above mass ratio, wash and dry at 60°C, with a solid-liquid ratio of 1:5 and a temperature of 90°C to 100°C, add water and reflux extract twice, the first time for 2 hours and the second time for 1 hour; combine the extracts and vacuum concentrate to a clear paste with a relative density of 1.10 to 1.20 (room temperature); precipitate the clear paste with 3 to 4 times the amount of 95% (mass fraction) ethanol for 24 hours; take the precipitate and wash it with anhydrous ethanol for 3 hours times, then dissolve it with 4 times the amount (mass fraction) of hot water >80℃, cool it to 40℃~50℃, add 1%~3% (mass ratio) of papain, and keep it warm for enzymatic hydrolysis for 4~5 hours; add chloroform / n-butanol mixed solution in a ratio of polysaccharide aqueous solution: chloroform: n-butanol of 25:4:1, stir for 30 minutes, and centrifuge at 3000r / min; take the clarified liquid, concentrate it into a thick paste, dry it in a vacuum below 60℃, and grind it into fine powder to obtain Armillaria mycelium polysaccharide.

[0032] Preparation of low molecular weight fucoidan sulfate: fucoidan sulfate extracted from kelp or fucus is dissolved in water at a ratio of 1.5% to 3.0%, and sterilized at 100°C for 20 to 30 minutes; when the fucoidan sulfate aqueous solution is cooled to 60°C to 70°C, arabinofuranosidase is added at a ratio of 1% to 3% with an optimum pH of ≤7, an optimum reaction temperature of 60°C, and a reaction time of 4 to 5 hours to degrade the fucoidan sulfate; vacuum concentration is performed, ethanol is added to the concentrate to 75%, the precipitate is collected, dried, and pulverized to obtain low molecular weight fucoidan sulfate with a molecular weight of 60,000 to 130,000.

[0033] Experimental Example 1: The active ingredient Armillaria polysaccharide prepared according to Example 2 was subjected to spectrophotometric determination of the polysaccharide content: Reagents and reference substances: Unless otherwise specified, only reagents of analytical grade and distilled water in accordance with GB / T 6682 were used in the analysis; sulfuric acid (H2SO4), ρ = 1.84 g / mL; anhydrous ethanol (C2H6O); phenol (C6H6O), redistilled; glucose (C6H 12 O6), and dry at 105°C to constant weight before use; 80% phenol solution: weigh 80g phenol in a 100mL beaker, dissolve it with water, dilute to 100mL, transfer to a brown bottle, and store in a 4°C refrigerator away from light; 5% phenol: draw 5mL of 80% phenol solution, dissolve it in 75mL of water, mix well, and prepare it before use; 100mg / L standard glucose solution: accurately weigh 0.1000g glucose in a 100mL beaker, dissolve it with water, dilute to 1000mL volumetric flask, shake well, and store in a 4°C refrigerator with a sealed stopper.

[0034] Sample preparation: Weigh 0.5g-1.0g of sample to the nearest 0.001g and place it in a 50mL stoppered centrifuge tube. Soak the sample with 5mL of water and slowly add 20mL of anhydrous ethanol while shaking on a vortex oscillator to mix thoroughly. Place the sample in an ultrasonic extractor and extract for 30 minutes. After extraction, centrifuge at 4000r / min for 10 minutes and discard the supernatant. Wash the insoluble matter with 10mL of 80% ethanol solution (mass ratio) and centrifuge. Transfer the insoluble matter to a round-bottom flask with water, add 50mL of distilled water, install a ground-mouth air condenser, and extract in a boiling water bath for 2 hours. Cool to room temperature, filter, and transfer the supernatant to a 100mL volumetric flask. Wash the residue two to three times, transfer the washing solution to the volumetric flask, and add water to the volume. This solution is the sample determination solution.

[0035] Standard curve: Pipette 0, 0.2 mL, 0.4 mL, 0.6 mL, and 1.0 mL of standard glucose solution into 20 mL stoppered test tubes, make up to 1.0 mL with distilled water, add 1.0 mL of 80% phenol solution to the test solution, and then quickly add 5.0 mL of sulfuric acid (add vertically at the liquid surface, do not let it touch the wall of the test tube, so as to fully mix with the reaction solution), let it stand for 10 minutes, use a vortex oscillator to fully mix the reaction solution, and then place the test tube in a 30°C water bath to react for 20 minutes. Measure the absorbance at 490 nm, and develop a standard curve with the glucose mass concentration as the horizontal axis and the absorbance value as the vertical axis.

[0036] Determination: Pipette 1.00 mL of sample solution into a 20 mL stoppered test tube and measure the absorbance according to the sample preparation and standard curve procedures. Simultaneously perform a blank test.

[0037] Calculation: The polysaccharide content in the sample is expressed as mass fraction w, expressed in grams per hundred grams (g / 100g), and is calculated using the following formula:

[0038]

[0039] Where m1 represents the sugar content of the sample solution as found on the standard curve, in micrograms (μg); V1 represents the sample volume, in milliliters (mL); V2 represents the volume of the sample solution pipetted for colorimetric determination, in milliliters (mL); m2 represents the sample mass, in grams (g); and 0.9 represents the correction factor for converting glucose to glucan. The calculated result should be rounded to two decimal places. The Armillaria polysaccharide content (calculated as glucose) in this product must not be less than 30g / 100g.

[0040] Experimental Example 2: The active ingredient, low molecular weight fucoidan sulfate, prepared according to Example 2 was analyzed by spectrophotometry, high performance liquid chromatography, and size exclusion chromatography to determine the fucoidan content, sulfate content, molecular weight, and molecular weight distribution.

[0041] Fucoidose content determination:

[0042] Preparation of reference solution: Take about 10 mg of fucoidan dried under reduced pressure over phosphorus pentoxide, accurately weigh it, dissolve it in a small amount of water, transfer the solution to a 100 ml volumetric flask, dilute to the mark with water, and shake well.

[0043] Preparation of the standard curve: Accurately pipette 0.15, 0.30, 0.45, 0.60, 0.75, and 0.90 ml of the reference solution into colorimetric tubes. Add water to 1.0 ml, respectively. Add 4.5 ml of 87% sulfuric acid solution to an ice-water bath and shake well. After 1 minute, heat in a boiling water bath for 10 minutes. Rapidly cool to room temperature, add 0.1 ml of 3% cysteine ​​hydrochloride solution, shake well, and let stand for 90 minutes. Determine the absorbance at wavelengths of 427 nm and 396 nm, respectively, using the spectrophotometric method (Chinese Pharmacopoeia 2015 Edition, Part IV, General Chapter 0401). Plot a standard curve using the difference in absorbance as the ordinate and the amount of reference solution as the abscissa. This product, based on the dry substance, must contain at least 25% fucoidan (mass ratio).

[0044] Sulfate content determination: Determine by high performance liquid chromatography (Chinese Pharmacopoeia 2015 Edition Part IV General Chapter 0512). This product, calculated on a dry basis, shall contain no less than 22% (mass ratio) of sulfate groups.

[0045] Molecular weight and molecular weight distribution are determined by size exclusion chromatography (Chinese Pharmacopoeia 2015 Edition, Part IV, General Chapter 0514). The weight average molecular weight (MW) of this product should be 60,000 to 130,000, and the weight average molecular weight of the 10% macromolecular fraction should not exceed 750,000.

[0046] Experimental Example 3: Active composition for treating migraine, determination of Armillaria polysaccharide content by spectrophotometry:

[0047] 1) Sample processing:

[0048] a. Sample extraction: Weigh 2.0 g of sample into a 100 mL volumetric flask, add 80 mL of water, and heat in a boiling water bath for 2 hours. After cooling to room temperature, add water to the mark, mix well, and filter. Discard the initial filtrate and collect the remaining filtrate for polysaccharide precipitation.

[0049] b. Precipitation of Crude Polysaccharide: Accurately pipette 5.0 mL of the final filtrate from step (a) into a 50 mL centrifuge tube. Add 20 mL of anhydrous ethanol and mix thoroughly for 5 minutes. Centrifuge at 3000 rpm for 5 minutes and discard the supernatant. Wash the residue with several mL of 80% (volume fraction) ethanol solution, centrifuge, and discard the supernatant. Repeat this process 3-4 times. Dissolve the residue in water and dilute to 5.0 mL. Mix thoroughly to precipitate the glucan.

[0050] c. Precipitate Dextran: Accurately pipette 2.0 mL of the final solution from step (b) into a 20 mL centrifuge tube. Add 2.0 mL of 100 g / L sodium hydroxide solution and 2.0 mL of copper reagent solution. Boil in a boiling water bath for 2 minutes. Cool, centrifuge at 3000 rpm for 5 minutes, and discard the supernatant. Wash the residue with several mL of washing solution, centrifuge, and discard the supernatant. Repeat this process three times. Dissolve the residue in 2.0 mL of 10% (volume fraction) sulfuric acid solution and transfer to a 50 mL volumetric flask. Dilute to the mark with water and mix thoroughly. This solution is the sample test solution.

[0051] 2) Standard Curve Preparation: Accurately weigh 0, 0.10, 0.20, 0.40, 0.60, 0.80, and 1.00 mL of dextran standard solution (equivalent to 0, 0.01, 0.02, 0.04, 0.06, 0.08, and 0.10 mg of dextran) into 25 mL colorimetric tubes, accurately fill to 2.0 mL with water, add 1.0 mL of 50 g / L phenol solution, and mix thoroughly on a rotary mixer. Carefully add 10.0 mL of concentrated sulfuric acid and mix thoroughly on a rotary mixer. Boil in a boiling water bath for 2 min. After cooling, measure the absorbance at 485 nm using a spectrophotometer with a 1 cm cuvette, using the reagent blank solution as the reference. Plot a standard curve with the dextran concentration as the abscissa and the absorbance as the ordinate.

[0052] 3) Sample Assay: Accurately pipette 2.0 mL of the sample solution into a 25 mL colorimetric tube. Add 1.0 mL of 50 g / L phenol solution and mix thoroughly on a rotary mixer. Carefully add 10.0 mL of concentrated sulfuric acid and mix thoroughly on a rotary mixer. Boil in a boiling water bath for 2 minutes. Cool to room temperature and measure the absorbance using a spectrophotometer at 485 nm using a reagent blank as a reference and a 1 cm cuvette. Determine the glucan content from the standard curve and calculate the crude polysaccharide content in the sample. Perform a sample blank test simultaneously.

[0053] The active composition granules of the present invention contain 200 mg or more of Armillaria polysaccharide (calculated as glucan) in each bag; the active composition tablets of the present invention contain 60 mg or more of Armillaria polysaccharide (calculated as glucan) in each tablet.

[0054] Experimental Example 4: Animal testing of the active composition for treating migraine:

[0055] Animals: Male guinea pigs, weighing 230-260 g, were provided by the Experimental Animal Center of Jilin University and housed in single cages under natural light and ventilation with free access to food and water.

[0056] Drugs: Granules of the active composition of the present invention, 3g / bag; Gastrodia elata capsules (manufactured by Kunming Pharmaceutical Group Co., Ltd.), approval number H20013044, 50mg / capsule; Nitroglycerin injection (manufactured by Sihuan Pharmaceutical Co., Ltd.), batch number 20150501, 5mg / mL.

[0057] Instruments: electrochemical detector (HPLC, 5600A, Esa, USA); Sigma 3K15 universal desktop refrigerated centrifuge; PHSJ-3F digital pH meter (Shanghai Leici Instrument Factory).

[0058] Experimental Methods: Experimental animals were randomly divided into a blank group, a model group, a group treated with the active composition of the present invention, and a group treated with Gastrodia elata extract capsules, with 10 animals in each group. Each group received prophylactic medication for 5 days prior to modeling, all administered via oral gavage. The active composition of the present invention was administered at a therapeutic dose of 0.5 g / kg body weight, and the Gastrodia elata extract capsule was administered at a dose of 14 mg / kg body weight. The model and sham-operated groups were treated identically, except for the administration of equal volumes of normal saline.

[0059] All animals in the blank control group received a subcutaneous injection of 10 mg / kg nitroglycerin on day 5 of treatment to establish an experimental migraine model. The presence of symptoms indicating head discomfort, such as redness of both ears, frequent scratching with the forelimbs, and increased cage climbing, was considered a successful model.

[0060] The time of appearance and disappearance of ear redness in rats after modeling was observed; the number of times the rats scratched their heads in each time period after modeling was observed from the time of modeling; the appearance time of head scratching was marked by the rats scratching their heads more than 5 times in a row, and the disappearance time was marked by the rats scratching their heads less than 5 times in a time period and showing signs of fatigue and exhaustion.

[0061] After behavioral assessment, the rats were decapitated and their brains removed. The brainstems were quickly separated, weighed, and placed into pre-coded cryovials. These tubes were then quickly placed in liquid helium for storage until further use. High-performance liquid chromatography (HPLC) electrochemical analysis was used to measure the levels of monoamine neurotransmitters, including 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), dopamine (DA), and norepinephrine (NE), in the rat brain. The results are shown in Tables 1–3.

[0062] Table 1 Effect of the composition of the present invention on the appearance and time of ear redness in each group of experimental rats ( min)

[0063]

[0064]

[0065] Note: Compared with the model group, *P<0.05.

[0066] Table 2 Effect of the composition of the present invention on the number of head scratching and head shaking of each group of experimental rats in 3 hours

[0067] Group n Number of head scratches Number of head shakes Normal group 10 13.47±4.05** 1.40±1.64** Model Group 10 35.47±8.21 8.00±1.70 Composition of the present invention 10 16.47±3.20** 2.80±0.48* Gastrodia elata capsule group 10 23.00±6.52* 3.20±0.86*

[0068] Note: Compared with the model group, *P<0.05, **P<0.01.

[0069] Table 3 Effects of the composition of the present invention on 5-HT, 5-HIAA, NE and DA in brain tissue of experimental rats

[0070]

[0071] Note: Compared with the model group, *P<0.05, **P<0.01.

[0072] Experiments have shown that both the treatment groups of the present invention's composition and the treatment groups of Gastrodia elata capsules have an interventional effect on behaviors such as ear redness and head scratching in migraine rats, and can significantly increase the levels of serotonin, dopamine, and norepinephrine in the rat brain tissue. The model group showed a significant decrease in the levels of neurotransmitters NE and 5-HT in the brain, while the DA level remained unchanged, indicating that migraine attacks are related to a decrease in the secretory function of certain nerve cells in the brainstem. The treatment group of the present invention's composition showed a significant improvement in the levels of monoamine neurotransmitters, with a significant increase in 5-HT, NE, and DA in brain tissue, indicating that the present invention's composition can nourish the relevant neurons in the body, promoting the increased production of neurotransmitters controlled by them, thereby improving migraine symptoms. The change in NE showed a significant improvement compared to the model group. The present invention's composition can prevent migraine attacks, and this effect may be related to the restoration of the mechanism regulating the release and metabolism of monoamine neurotransmitters in the body, thereby improving brain and vascular dysfunction.

[0073] In summary, the composition of the present invention has a preventive therapeutic effect on rats with migraine induced by nitroglycerin, and its mechanism of action is closely related to the neurotransmitters 5-HT, DA, and NE in the brain.

[0074] Additionally, in order to provide a concise description of exemplary embodiments, all features of an actual embodiment (i.e., those features that are not relevant to the best mode presently contemplated for carrying out the invention or those that are not relevant to implementing the invention) may not be described.

[0075] It will be understood that in the development of any actual embodiment, as in any engineering or design project, numerous implementation-specific decisions may be made. Such a development effort may be complex and time-consuming, but for those of ordinary skill having the benefit of this disclosure, the development effort will be a routine task of design, fabrication, and production without undue experimentation.

[0076] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A pharmaceutical composition for treating migraine, characterized in that: The invention is prepared from the following raw materials in percentage by weight: 30% to 60% of Armillaria mellea polysaccharide, 3% to 7% of low molecular weight brown algae polysaccharide sulfate, 8% to 17% of gamma-aminobutyric acid, and 15% to 60% of auxiliary material maltodextrin; The Armillaria polysaccharide is extracted from the mycelium of wild Armillaria or artificially fermented Armillaria; The low molecular weight brown algae polysaccharide is prepared by degrading brown algae polysaccharide extracted from Laminaria japonica or Fucus vesiculosus of the Laminariaceae family through arabinofuranosidase, and has a molecular weight of 60,000-130,000.

2. A pharmaceutical composition for treating migraine according to claim 1, characterized in that: The invention is prepared from the following raw materials in percentage by weight: 60% of Armillaria mellea polysaccharide, 7% of low molecular weight brown algae polysaccharide sulfate, 17% of γ-aminobutyric acid and 16% of auxiliary material maltodextrin.

3. A method for preparing a pharmaceutical composition for treating migraine, applied to a pharmaceutical composition for treating migraine as claimed in any one of claims 1 to 2, characterized in that: S1. The specific preparation method of the active composition granules is as follows: 1000 g of Armillaria polysaccharide, 100 g of low molecular weight brown algae polysaccharide sulfate, 250 g of γ-aminobutyric acid, and 1650 g of maltodextrin are weighed respectively, mixed evenly, made into wet granules through a 14-16 mesh nylon sieve, dried at 50°C-60°C to a moisture content of ≤5.0%, and packed into bags, 3 g per bag, and 1000 bags are made. S2. The specific preparation method of the active composition tablet is as follows: weigh 3000 g of Armillaria polysaccharide, 350 g of low molecular weight brown algae polysaccharide sulfate, 850 g of γ-aminobutyric acid, and 800 g of maltodextrin respectively, mix them evenly, make them into 16-mesh granules, press them into tablets, and package them; each tablet weighs 0.5 g, and 10,000 tablets are made.

4. A method for preparing a pharmaceutical composition for treating migraine according to claim 3, characterized in that: The preparation method of the Armillaria polysaccharide is as follows: a. Preparation of fruiting body polysaccharide: Weigh the fruiting body of Armillaria mellea at 20 times the mass ratio, dry it at 60°C after washing, crush it into coarse powder of about 1mm, the solid-liquid ratio is 1:20, the temperature is 100°C, add water and reflux to extract twice, the first time for 3 hours and the second time for 2 hours; combine the extracts and vacuum concentrate them into a clear paste with a relative density of 1.10 to 1.20; the clear paste is precipitated with 3 to 4 times the amount of 95% ethanol for 24 hours, the precipitate is washed with anhydrous ethanol 3 times, and then dissolved with 4 times the amount of hot water ≥80°C, and when it is cooled to 40°C to 50°C, 1% to 3% papain is added, and the enzymatic hydrolysis is kept warm for 4 to 5 hours, and a chloroform / n-butanol mixed solution is added according to the ratio of polysaccharide aqueous solution: chloroform: n-butanol of 25:4:1, stirred for 30 minutes, centrifuged at 3000r / min, and the clarified liquid is concentrated into a thick paste, vacuum dried below 60°C, and crushed into fine powder, that is; b. Preparation of mycelium polysaccharide: Take 10 times the mass ratio of honey fungus mycelium, wash and dry at 60℃, with a solid-liquid ratio of 1:5, temperature of 90℃~100℃, add water and reflux to extract twice, the first time for 2 hours, the second time for 1 hour, combine the extracts, and vacuum concentrate to a clear paste with a relative density of 1.10~1.

20. The clear paste is precipitated with 3~4 times the amount of 95% ethanol for 24 hours, the precipitate is washed with anhydrous ethanol 3 times, and then dissolved with 4 times the amount of hot water ≥80℃, cooled to 40℃~50℃, 1%~3% papain is added, and enzymolysis is kept warm for 4~5 hours; chloroform / n-butanol mixed solution is added in a ratio of polysaccharide aqueous solution: chloroform: n-butanol of 25:4:1, stirred for 30 minutes, centrifuged at 3000r / min, the clarified liquid is concentrated into a thick paste, vacuum dried below 60℃, and crushed into fine powder.

5. A method for preparing a pharmaceutical composition for treating migraine according to claim 3, characterized in that: The preparation method of the low molecular weight brown algae polysaccharide sulfate is as follows: weigh the brown algae polysaccharide sulfate extracted from kelp or fucus at a mass ratio of 1.2 to 1.5 times, dissolve it in water at a ratio of 1.5% to 3.0%, sterilize it at 100° C. for 20 to 30 minutes, add arabinofuranosidase at a ratio of 1% to 3% when the brown algae polysaccharide aqueous solution is cooled to 60° C. to 70° C., and the optimal pH is ≤7, the optimal reaction temperature is 60° C., and the reaction time is 4 to 5 hours to degrade the brown algae polysaccharide sulfate; vacuum concentrate, add ethanol to the concentrated solution to 75%, collect the precipitate, dry it, and crush it to obtain the low molecular weight brown algae polysaccharide sulfate, and the molecular weight of the low molecular weight brown algae polysaccharide is 60,000 to 130,000.

6. A method for preparing a pharmaceutical composition for treating migraine according to claim 3, characterized in that: The gamma-aminobutyric acid is derived from sodium L-glutamate as a raw material and is fermented by Lactobacillus hilarii. The gamma-aminobutyric acid content is ≥20%, the moisture content is ≤10%, and the ash content is ≤18%.

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