A Mongolian medicine composition capsule for treating and preventing stroke and preparation method thereof

Abstract

The present invention relates to the field of medical technology, and in particular to a Mongolian medicine composition capsule for treating and preventing stroke and a preparation method thereof, wherein the Mongolian medicine composition capsule comprises a capsule shell and contents, wherein the contents comprise the following raw materials in parts by weight: 30-40 parts of Panax notoginseng, 36-44 parts of Roselle Thistle, 15-20 parts of Fructus Choerospondiatis, 20-30 parts of Clematis, 25-32 parts of Ginseng, 18-25 parts of Aconitum, 20-30 parts of synergist, and 8-12 parts of promoter. The present invention also provides a preparation method for the Mongolian medicine composition capsule, wherein the raw materials are easily available, the process is simple, a variety of Mongolian medicines are combined, synergistically enhanced, and the prepared capsule can effectively treat and prevent stroke caused by a variety of causes, and has no toxic side effects, and the synergist and promoter can enhance the therapeutic effect and improve the drug absorption utilization rate.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to a Mongolian medicine composition capsule for treating and preventing cerebral stroke and a preparation method thereof. Background Art

[0002] Stroke is a common and serious neurological disease. Its occurrence is related to the rupture or blockage of cerebral blood vessels, which leads to insufficient blood supply to the brain, and then causes brain tissue damage or even death. Risk factors for stroke (hypertension, hyperglycemia, etc.) and ischemia can lead to the activation of reactive oxygen species (ROS)-related enzyme systems, producing a large amount of ROS, which far exceeds the endogenous clearance capacity of the body's antioxidant defense system, resulting in oxidative stress. After the onset of stroke, superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT), glutathione peroxidase (GSH-Px), and total antioxidant capacity (TAC) are significantly reduced, and malondialdehyde (MDA) is significantly increased. Changes in the above indicators aggravate the condition by promoting platelet aggregation, increasing endothelial permeability, and focal lesions of endothelial cells. The high morbidity and mortality of stroke have placed a huge burden on patients. Therefore, the treatment and prevention of stroke are crucial to the life and quality of life of patients. Early acute treatment and comprehensive rehabilitation treatment can minimize brain damage, restore function, and improve the quality of life and prognosis of patients. Mongolian medicine is a kind of traditional Chinese medicine, which is considered to have the effects of harmonizing qi and blood, promoting blood circulation and removing blood stasis, and dispelling wind and unblocking collaterals in traditional Chinese medicine theory. In recent years, more and more studies have shown that the active ingredients in Mongolian medicine have biological activities such as anti-inflammatory, antioxidant, anticoagulant, and angiogenesis promotion, and may have potential effects on the treatment and prevention of stroke. At present, the drugs for treating stroke mainly include thrombolytic drugs and anticoagulant drugs. Among them, thrombolytic drugs are used to shrink thrombi and reduce the degree of vascular occlusion. Anticoagulant drugs are mainly used to improve microcirculation and restore blood flow. However, thrombolytic drugs need to be used within a few hours after the occurrence of stroke to achieve the best therapeutic effect, and cannot play a preventive effect, which limits the opportunity for some patients to receive thrombolytic therapy. In addition, these drug compositions often have a single efficacy, or poor safety, and low bioavailability. Although these drugs have achieved certain results in acute treatment, there are also some shortcomings. Therefore, how to develop a Mongolian medicine composition capsule that can have both therapeutic and preventive effects, high bioavailability, good efficacy, and good safety for the treatment and prevention of stroke is a technical problem that needs to be solved urgently. Summary of the invention

[0003] In view of the above technical problems, the present invention proposes a Mongolian medicine composition capsule for treating and preventing stroke and a preparation method thereof. The present invention selects a variety of Mongolian medicines with the effect of treating and preventing stroke and combines them to create a unique drug combination, which can synergize and effectively treat and prevent stroke. The present invention uses Spirulina, Ceratium obtusifolium and Chlorella proteoglycans to prepare a synergist, which can improve cerebrovascular function and reduce the risk of stroke, and help prevent the occurrence of stroke. The present invention prepares a promoter, which can improve the solubility of the drug, enhance the bioaffinity of the drug, and promote the absorption and utilization of the drug. The present invention uses citric acid and NaOH solution to extract Mongolian medicine, which can neutralize or degrade toxic substances in the medicinal materials and improve the safety of the drug. The present invention adds glycyrrhizic acid, gelatin and alginate, which can reduce the toxicity of the drug and enhance the efficacy of the drug. The present invention uses glycolic acid to modify Mongolian medicine particles, improve dispersibility, and enhance absorbability. A Mongolian medicine composition capsule for treating and preventing stroke prepared by the present invention has good efficacy, high safety, high bioavailability, and can effectively treat and prevent stroke.

[0004] The present invention is achieved through the following technical solutions:

[0005] A Mongolian medicine composition capsule for treating and preventing cerebral stroke comprises a capsule shell and contents, wherein the contents comprise the following raw materials in parts by weight: 30-40 parts of Panax notoginseng, 36-44 parts of Cirsium japonicum, 15-20 parts of Fructus zoidis, 20-30 parts of Clematis chinensis, 25-32 parts of Ginseng, 18-25 parts of Aconitum carmichaelii, 20-30 parts of synergist and 8-12 parts of accelerator.

[0006] Furthermore, the preparation method of the synergist comprises the following steps:

[0007] (1) Spirulina, Ceratium obtusifolium, Chlorella pyrenoidosa and water are mixed to obtain a mixed algae liquid, wherein the mass ratio of algae to water is 1:8, and the mixed algae liquid is uniformly stirred and homogenized at a homogenization pressure of 10 MPa and a homogenization temperature of 70°C to obtain a mixed slurry;

[0008] (2) sterilizing the mixed slurry obtained in step (1) at 85° C. for 30 min, then cooling to 37° C., adjusting the pH value to 5-6 with citric acid, adding biological enzyme, the weight ratio of biological enzyme to mixed slurry being 1:1000, stirring for 3-5 h, heating to 95° C. for 10 min, and obtaining a hydrolyzate;

[0009] (3) The hydrolyzate obtained in step (2) was centrifuged at a speed of 4000 r / min for 10 min, the supernatant was mixed with a 10 vol% ethanol solution, the volume ratio of the ethanol solution to the supernatant was 8:1, hexadecyl betaine was added, and the mass volume ratio of hexadecyl betaine to the hydrolyzate was 1 g:1 L, and the mixture was extracted at 40° C. for 1 h, filtered, and filtrate 1 was obtained. The filtrate 1 was concentrated to a solid content of 20 wt% to obtain a concentrated solution;

[0010] (4) Add galactose to the concentrated solution obtained in step (3) at a mass ratio of galactose to the concentrated solution of 1:99, react at 121° C. for 30 min, mix the product with benzyl formate at a ratio of 1 g:2 mL, filter to obtain filtrate 2, mix with activated carbon at a mass volume ratio of activated carbon to filtrate 2 of 1 g:80 mL, stir at 30° C. for 40 min, filter to obtain filtrate 3, and freeze-dry to obtain a synergist.

[0011] Furthermore, the biological enzyme in step (2) is acid cellulase, neutral cellulase, neutral protease and papain mixed in a mass ratio of 2:1:1.5:1.2.

[0012] Furthermore, the preparation method of the accelerator comprises the following steps:

[0013] I. In an ice water bath, diethanolamine and p-aminobenzoyl chloride were added to a dimethylformamide solvent, the mass ratio of diethanolamine to p-aminobenzoyl chloride was 2:3, the mass volume ratio of diethanolamine to dimethylformamide was 1 g:2 mL, and then pyridine and triethylamine were added in a mass ratio of 2:1, and the mass ratio of pyridine to diethanolamine was 5-8:18, and the mixture was stirred for 15-20 min, and then stirred at room temperature for 24 h to obtain a reaction mixture I;

[0014] II. To the reaction mixture obtained in step I, a fatty acid was added, the molar ratio of fatty acid to p-aminobenzoyl chloride was 1:1, and then dimethyl carbonate was added, the mass ratio of dimethyl carbonate and p-aminobenzoyl chloride was 9:13, and stirred at room temperature for 6-8 h at a stirring speed of 60 rpm to obtain a reaction mixture II;

[0015] III. The reaction mixture II obtained in step II was concentrated under reduced pressure to 30% of the original volume, washed with acetone three times, the product was recrystallized in tetrahydrofuran, filtered, washed with 95wt% ethanol solution and deionized water three times in sequence, and dried under vacuum at 40-50°C for 2-3 h to obtain a promoter.

[0016] Furthermore, the fatty acid is one of capric acid, undecanoic acid, myristic acid, palmitic acid, stearic acid, linoleic acid and linolenic acid.

[0017] Furthermore, the capsule is a hard capsule or a soft capsule.

[0018] Furthermore, the capsule shell is prepared from gastric-soluble capsule excipients or enteric-soluble capsule excipients.

[0019] Furthermore, the material of the capsule shell includes but is not limited to one or more of gelatin, glycerol, sodium alginate, and starch.

[0020] The present invention also provides a method for preparing the Mongolian medicine composition capsule for treating and preventing stroke, comprising the following steps:

[0021] S1: Wash and dry Panax notoginseng, Fructus jujubae, Ginseng, Clematis, Aconitum and Cirsium chinense, crush them through an 80-mesh sieve to obtain a Mongolian medicine mixture, soak them in a 1 mol / L citric acid solution for 1-2 h, filter them to obtain a filtrate A and a precipitate A, soak the precipitate A in a 1 mol / L sodium hydroxide solution for 30-40 min, filter them to obtain a filtrate B and a precipitate B, the mass volume ratio of the Mongolian medicine mixture to the citric acid solution and the sodium hydroxide solution is 1 g:8 mL:8 mL, combine the filtrate A and the filtrate B to obtain a combined filtrate, add glycyrrhizic acid, gelatin and alginic acid in a mass ratio of 2:1:1.2, the amount ratio of the combined filtrate to glycyrrhizic acid is 1 L:10 mg, heat to 50-60°C, maintain for 3-4 h, rotary evaporate at 60°C for 1-2 h, spray dry, and obtain Mongolian medicine granules;

[0022] S2: The Mongolian medicine particles obtained in step S1 are added to deionized water, the mass volume ratio of Mongolian medicine particles to deionized water is 1 mg:100 mL, after stirring at 180 rpm for 30 min, centrifuged at 10000 rpm for 30-40 min, the precipitate is mixed with deionized water again at a ratio of 1 g:100 mL, glycolic acid is added, the mass ratio of Mongolian medicine particles to glycolic acid is 5:1, stirred at room temperature for 4 h, centrifuged at 8000 rpm for 15 min, the precipitate is washed three times with deionized water, and dried at 50°C to obtain a Mongolian medicine preparation;

[0023] S3: The Mongolian medicine preparation, accelerator and synergist are mixed evenly, ground and crushed to pass through a 500-mesh sieve, sterilized by ultraviolet light for 1 hour, and filled into capsules, with each capsule containing 0.5 g, to obtain a Mongolian medicine composition capsule for treating and preventing stroke.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The present invention selects and combines a variety of Mongolian medicines with the effect of treating and preventing stroke, creates a unique drug combination, can synergize, effectively treat and prevent stroke, the drug selection and ratio are reasonable, the efficacy is good and the safety is high. Algae is a kind of plant rich in nutrients and bioactive compounds. Some algae are considered to have the potential to treat and prevent stroke. The present invention uses Spirulina, Ceratium obtusifolium and Chlorella pyrenoidosa to prepare a synergist, which has anti-thrombotic, antioxidant, anti-inflammatory, hypotensive and neuroprotective effects, can improve cerebrovascular function and reduce the risk of stroke, and helps prevent the occurrence of stroke. The present invention prepares a promoter, which can improve the solubility of the drug, enhance the bioaffinity of the drug, promote the absorption and utilization of the drug, and thus enhance the efficacy. The present invention prepares Mongolian medicine particles by spray drying, and the particles prepared by the spray drying method have a small particle size, good biocompatibility, are more easily absorbed by the human body, and have high bioavailability. The present invention uses citric acid and NaOH solution to extract Mongolian medicine, which can neutralize or degrade toxic substances in medicinal materials, reduce drug toxicity, and improve drug safety. The present invention adds glycyrrhizic acid, gelatin and alginic acid in a mass ratio of 2:1:1.2, which can reduce the toxicity of the drug and enhance the efficacy of the drug. The present invention adopts glycolic acid to modify the Mongolian medicine particles, improve the dispersibility, increase the hydrophilicity of the Mongolian medicine particles, make it easier to disperse evenly in an aqueous solution, help improve the stability of the Mongolian medicine particles, prevent them from aggregating and precipitating in the solution, and further increase the compatibility of the Mongolian medicine particles with biological tissues, and enhance the absorbability. A Mongolian medicine composition capsule for treating and preventing cerebral stroke prepared by the present invention has good efficacy, high safety, high bioavailability, and can effectively treat and prevent cerebral stroke. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings in the following description are only for the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0027] Figure 1 The chemical structure diagram of the accelerator described in Example 1 of the present invention;

[0028] Figure 2 is the nuclear magnetic resonance spectrum of the promoter described in Example 1 of the present invention, wherein a is 1 H figure, b is 13 Figure C;

[0029] Figure 3 This is a scanning electron microscope image of the Mongolian medicine preparation described in Example 1 of the present invention;

[0030] Figure 4The cytotoxicity test diagram of the contents of the Mongolian medicine composition capsules described in Examples 1-3 and Comparative Examples 1-3 of the present invention;

[0031] Figure 5 The effect diagram of the effects of the contents of the Mongolian medicine composition capsules described in Examples 1-3 and Comparative Examples 1-3 of the present invention on oxidative stress in rats;

[0032] Figure 6 This is a test chart of the dissolution rate of the contents of the Mongolian medicine composition capsule described in Examples 1-3 of the present invention and Comparative Examples 1-3. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments, but the present invention is not limited to the following embodiments.

[0034] It should be noted that, unless otherwise specified, the chemical reagents involved in the present invention were purchased through commercial channels.

[0035] Example 1: This example provides a Mongolian medicine composition capsule content for treating and preventing stroke, comprising the following raw materials in parts by weight: 40 parts of Panax notoginseng, 44 parts of Cirsium japonicum, 20 parts of Fructus jujubae, 30 parts of Clematis chinensis, 32 parts of Ginseng, 25 parts of Aconitum, 30 parts of Enhancer, and 12 parts of Accelerator.

[0036] The preparation method of the synergist described in this embodiment comprises the following steps:

[0037] (1) Spirulina, Ceratium obtusifolium, Chlorella pyrenoidosa and water are mixed to obtain a mixed algae liquid, wherein the mass ratio of algae to water is 1:8, and the mixed algae liquid is uniformly stirred and homogenized at a homogenization pressure of 10 MPa and a homogenization temperature of 70°C to obtain a mixed slurry;

[0038] (2) sterilizing the mixed slurry obtained in step (1) at 85° C. for 30 min, then cooling to 37° C., adjusting the pH value to 6 with citric acid, adding biological enzymes, wherein the biological enzymes are acid cellulase, neutral cellulase, neutral protease, and papain mixed in a mass ratio of 2:1:1.5:1.2, and the weight ratio of the biological enzyme to the mixed slurry is 1:1000, stirring for 5 h, heating to 95° C. for 10 min, and obtaining a hydrolyzate;

[0039] (3) The hydrolyzate obtained in step (2) was centrifuged at a speed of 4000 r / min for 10 min, the supernatant was mixed with a 10 vol% ethanol solution, the volume ratio of the ethanol solution to the supernatant was 8:1, hexadecyl betaine was added, and the mass volume ratio of hexadecyl betaine to the hydrolyzate was 1 g:1 L, and the mixture was extracted at 40° C. for 1 h, filtered, and filtrate 1 was obtained. The filtrate 1 was concentrated to a solid content of 20 wt% to obtain a concentrated solution;

[0040] (4) Add galactose to the concentrated solution obtained in step (3) at a mass ratio of galactose to the concentrated solution of 1:99, react at 121° C. for 30 min, mix the product with benzyl formate at a ratio of 1 g:2 mL, filter to obtain filtrate 2, mix with activated carbon at a mass volume ratio of activated carbon to filtrate 2 of 1 g:80 mL, stir at 30° C. for 40 min, filter to obtain filtrate 3, and freeze-dry to obtain a synergist.

[0041] The preparation method of the accelerator described in this embodiment comprises the following steps:

[0042] I. In an ice water bath, diethanolamine and p-aminobenzoyl chloride were added to a dimethylformamide solvent, the mass ratio of diethanolamine to p-aminobenzoyl chloride was 2:3, and the mass volume ratio of diethanolamine to dimethylformamide was 1 g:2 mL. Pyridine and triethylamine were added at a mass ratio of 2:1, and the mass ratio of pyridine to diethanolamine was 8:18. The mixture was stirred for 20 min, and then stirred at room temperature for 24 h to obtain a reaction mixture I;

[0043] II. To the reaction mixture obtained in step I, decanoic acid was added, the molar ratio of decanoic acid to p-aminobenzoyl chloride was 1:1, and then dimethyl carbonate was added, the mass ratio of dimethyl carbonate to p-aminobenzoyl chloride was 9:13, and stirred at room temperature for 8 h at a stirring speed of 60 rpm to obtain a reaction mixture II;

[0044] III. The reaction mixture II obtained in step II was concentrated under reduced pressure to 30% of the original volume, washed with acetone three times, and the product was recrystallized in tetrahydrofuran, filtered, washed with 95wt% ethanol solution and deionized water three times in sequence, and dried under vacuum at 50°C for 3 h to obtain an accelerator having the chemical structure shown in Figure 1 The NMR spectrum is shown in Figure 2 As shown, the molecular formula is C 21 H 34 O 4 N 2 .

[0045] This embodiment also provides a method for preparing the Mongolian medicine composition capsule for treating and preventing stroke, comprising the following steps:

[0046] S1: 40 g of Panax notoginseng, 20 g of Fructus Axillae, 32 g of Ginseng, 30 g of Clematis, 25 g of Aconitum and 44 g of Cirsium chinense were washed and dried, crushed and passed through an 80-mesh sieve to obtain a Mongolian medicine mixture, soaked in a 1 mol / L citric acid solution for 2 h, filtered to obtain a filtrate A and a precipitate A, and the precipitate A was soaked in a 1 mol / L sodium hydroxide solution for 40 min, filtered to obtain a filtrate B and a precipitate B, the mass volume ratio of the Mongolian medicine mixture to the citric acid solution and the sodium hydroxide solution was 1 g:8 mL:8 mL, the filtrate A and the filtrate B were combined to obtain a combined filtrate, glycyrrhizic acid, gelatin and alginic acid were added in a mass ratio of 2:1:1.2, the amount ratio of the combined filtrate to glycyrrhizic acid was 1 L:10 mg, heated to 60°C, maintained for 4 h, rotary evaporated at 60°C for 2 h, spray dried, and Mongolian medicine granules were obtained;

[0047] S2: The Mongolian medicine particles obtained in step S1 were added to deionized water, the mass volume ratio of Mongolian medicine particles to deionized water was 1 mg:100 mL, stirred at 180 rpm for 30 min, centrifuged at 10000 rpm for 40 min, the precipitate was re-mixed with deionized water at a ratio of 1 g:100 mL, hydroxyacetic acid was added, the mass ratio of Mongolian medicine particles to hydroxyacetic acid was 5:1, stirred and reacted at room temperature for 4 h, centrifuged at 8000 rpm for 15 min, the precipitate was washed three times with deionized water, and dried at 50°C to obtain a Mongolian medicine preparation. The scanning electron microscope image of the Mongolian medicine preparation is shown in the following figure: Figure 3 As shown, the particle size is small, the shape is approximately spherical, and it is in a well-dispersed state, indicating that it has good stability and is easy to absorb and utilize;

[0048] S3: The Mongolian medicine preparation prepared in step S2 is evenly mixed with 12 g of a promoter and 30 g of a synergist, and the mixture is ground and crushed to pass through a 500-mesh sieve, and sterilized by ultraviolet light for 1 h to obtain a Mongolian medicine composition capsule for treating and preventing stroke.

[0049] Example 2: This example provides a Mongolian medicine composition capsule content for treating and preventing stroke, comprising the following raw materials in parts by weight: 30 parts of Panax notoginseng, 36 parts of Cirsium japonicum, 15 parts of Ziziphus jujuba, 20 parts of Clematis chinensis, 25 parts of Ginseng, 18 parts of Aconitum, 20 parts of synergist, and 8 parts of promoter.

[0050] The preparation method of the synergist described in this embodiment comprises the following steps:

[0051] (1) Spirulina, Ceratium obtusifolium, Chlorella pyrenoidosa and water are mixed to obtain a mixed algae liquid, wherein the mass ratio of algae to water is 1:8, and the mixed algae liquid is uniformly stirred and homogenized at a homogenization pressure of 10 MPa and a homogenization temperature of 70°C to obtain a mixed slurry;

[0052] (2) sterilizing the mixed slurry obtained in step (1) at 85° C. for 30 min, then cooling to 37° C., adjusting the pH value to 5 with citric acid, adding biological enzymes, wherein the biological enzymes are acid cellulase, neutral cellulase, neutral protease, and papain mixed in a mass ratio of 2:1:1.5:1.2, and the weight ratio of the biological enzyme to the mixed slurry is 1:1000, stirring for 3 h, and heating to 95° C. for 10 min to obtain a hydrolyzate;

[0053] (3) The hydrolyzate obtained in step (2) was centrifuged at a speed of 4000 r / min for 10 min, the supernatant was mixed with a 10 vol% ethanol solution, the volume ratio of the ethanol solution to the supernatant was 8:1, hexadecyl betaine was added, and the mass volume ratio of hexadecyl betaine to the hydrolyzate was 1 g:1 L, and the mixture was extracted at 40° C. for 1 h, filtered, and filtrate 1 was obtained. The filtrate 1 was concentrated to a solid content of 20 wt% to obtain a concentrated solution;

[0054] (4) Add galactose to the concentrated solution obtained in step (3) at a mass ratio of galactose to the concentrated solution of 1:99, react at 121° C. for 30 min, mix the product with benzyl formate at a ratio of 1 g:2 mL, filter to obtain filtrate 2, mix with activated carbon at a mass volume ratio of activated carbon to filtrate 2 of 1 g:80 mL, stir at 30° C. for 40 min, filter to obtain filtrate 3, and freeze-dry to obtain a synergist.

[0055] The preparation method of the accelerator described in this embodiment comprises the following steps:

[0056] I. In an ice water bath, diethanolamine and p-aminobenzoyl chloride were added to a dimethylformamide solvent, the mass ratio of diethanolamine to p-aminobenzoyl chloride was 2:3, and the mass volume ratio of diethanolamine to dimethylformamide was 1 g:2 mL. Pyridine and triethylamine were added at a mass ratio of 2:1, and the mass ratio of pyridine to diethanolamine was 5:18. The mixture was stirred for 15 min, and then stirred at room temperature for 24 h to obtain a reaction mixture I;

[0057] II. To the reaction mixture obtained in step I, linolenic acid was added, the molar ratio of linolenic acid to p-aminobenzoyl chloride was 1:1, and then dimethyl carbonate was added, the mass ratio of dimethyl carbonate to p-aminobenzoyl chloride was 9:13, and stirred at room temperature for 6h at a stirring speed of 60 rpm to obtain a reaction mixture II;

[0058] III. The reaction mixture II obtained in step II was concentrated under reduced pressure to 30% of the original volume, washed with acetone three times, the product was recrystallized in tetrahydrofuran, filtered, washed with 95wt% ethanol solution and deionized water three times in sequence, and dried under vacuum at 40°C for 2 h to obtain a promoter.

[0059] This embodiment also provides a method for preparing the Mongolian medicine composition capsule for treating and preventing stroke, comprising the following steps:

[0060] S1: 30 g of Panax notoginseng, 15 g of Fructus jujubae, 25 g of Radix ginseng, 20 g of Clematis, 18 g of Aconitum and 36 g of Cirsium chinense were washed and dried, crushed and passed through an 80-mesh sieve to obtain a Mongolian medicine mixture, soaked in a 1 mol / L citric acid solution for 1 h, filtered to obtain a filtrate A and a precipitate A, and the precipitate A was soaked in a 1 mol / L sodium hydroxide solution for 30 min, filtered to obtain a filtrate B and a precipitate B, the mass volume ratio of the Mongolian medicine mixture to the citric acid solution and the sodium hydroxide solution was 1 g:8 mL:8 mL, the filtrate A and the filtrate B were combined to obtain a combined filtrate, glycyrrhizic acid, gelatin and alginic acid were added in a mass ratio of 2:1:1.2, the amount ratio of the combined filtrate to glycyrrhizic acid was 1 L:10 mg, heated to 50°C, maintained for 3 h, rotary evaporated at 60°C for 1 h, spray dried, and Mongolian medicine granules were obtained;

[0061] S2: The Mongolian medicine particles obtained in step S1 are added to deionized water, the mass volume ratio of Mongolian medicine particles to deionized water is 1 mg:100 mL, after stirring at 180 rpm for 30 min, centrifuged at 10000 rpm for 30 min, the precipitate is mixed with deionized water again at a ratio of 1 g:100 mL, glycolic acid is added, the mass ratio of Mongolian medicine particles to glycolic acid is 5:1, stirred for reaction at room temperature for 4 h, centrifuged at 8000 rpm for 15 min, the precipitate is washed three times with deionized water, and dried at 50°C to obtain a Mongolian medicine preparation;

[0062] S3: The Mongolian medicine preparation obtained in step S2 is mixed evenly with 8 g of a promoter and 20 g of a synergist, ground and crushed to pass through a 500-mesh sieve, and sterilized by ultraviolet light for 1 h to obtain the contents of a Mongolian medicine composition capsule for treating and preventing stroke.

[0063] Example 3: This example provides a Mongolian medicine composition capsule content for treating and preventing stroke, comprising the following raw materials in parts by weight: 35 parts of Panax notoginseng, 40 parts of Cirsium japonicum, 18 parts of Fructus zoidis, 25 parts of Clematis chinensis, 30 parts of Ginseng, 20 parts of Aconitum, 25 parts of Enhancer, and 10 parts of Accelerator.

[0064] The preparation method of the synergist described in this embodiment comprises the following steps:

[0065] (1) Spirulina, Ceratium obtusifolium, Chlorella pyrenoidosa and water are mixed to obtain a mixed algae liquid, wherein the mass ratio of algae to water is 1:8, and the mixed algae liquid is uniformly stirred and homogenized at a homogenization pressure of 10 MPa and a homogenization temperature of 70°C to obtain a mixed slurry;

[0066] (2) sterilizing the mixed slurry obtained in step (1) at 85° C. for 30 min, then cooling to 37° C., adjusting the pH value to 5.5 with citric acid, adding biological enzymes, wherein the biological enzymes are acid cellulase, neutral cellulase, neutral protease, and papain mixed in a mass ratio of 2:1:1.5:1.2, and the weight ratio of the biological enzyme to the mixed slurry is 1:1000, stirring for 4 h, heating to 95° C. for 10 min, and obtaining a hydrolyzate;

[0067] (3) The hydrolyzate obtained in step (2) was centrifuged at a speed of 4000 r / min for 10 min, the supernatant was mixed with a 10 vol% ethanol solution, the volume ratio of the ethanol solution to the supernatant was 8:1, hexadecyl betaine was added, and the mass volume ratio of hexadecyl betaine to the hydrolyzate was 1 g:1 L, and the mixture was extracted at 40° C. for 1 h, filtered, and filtrate 1 was obtained. The filtrate 1 was concentrated to a solid content of 20 wt% to obtain a concentrated solution;

[0068] (4) Add galactose to the concentrated solution obtained in step (3) at a mass ratio of galactose to the concentrated solution of 1:99, react at 121° C. for 30 min, mix the product with benzyl formate at a ratio of 1 g:2 mL, filter to obtain filtrate 2, mix with activated carbon at a mass volume ratio of activated carbon to filtrate 2 of 1 g:80 mL, stir at 30° C. for 40 min, filter to obtain filtrate 3, and freeze-dry to obtain a synergist.

[0069] The preparation method of the accelerator described in this embodiment comprises the following steps:

[0070] I. In an ice water bath, diethanolamine and p-aminobenzoyl chloride were added to a dimethylformamide solvent, the mass ratio of diethanolamine to p-aminobenzoyl chloride was 2:3, and the mass volume ratio of diethanolamine to dimethylformamide was 1 g:2 mL. Pyridine and triethylamine were added at a mass ratio of 2:1, and the mass ratio of pyridine to diethanolamine was 7:18. The mixture was stirred for 18 min, and then stirred at room temperature for 24 h to obtain a reaction mixture I;

[0071] II. To the reaction mixture obtained in step I, myristic acid was added, the molar ratio of myristic acid to p-aminobenzoyl chloride was 1:1, and then dimethyl carbonate was added, the mass ratio of dimethyl carbonate and p-aminobenzoyl chloride was 9:13, and the mixture was stirred at room temperature for 7 h at a stirring speed of 60 rpm to obtain a reaction mixture II;

[0072] III. The reaction mixture II obtained in step II was concentrated under reduced pressure to 30% of the original volume, washed with acetone three times, the product was recrystallized in tetrahydrofuran, filtered, washed with 95wt% ethanol solution and deionized water three times in sequence, and dried under vacuum at 45°C for 2.5h to obtain a promoter.

[0073] This embodiment also provides a method for preparing the Mongolian medicine composition capsule for treating and preventing stroke, comprising the following steps:

[0074] S1: 35 g of Panax notoginseng, 18 g of Fructus Axillae, 30 g of Radix Ginseng, 25 g of Clematis, 20 g of Aconitum and 40 g of Cirsium chinense were washed and dried, crushed and passed through an 80-mesh sieve to obtain a Mongolian medicine mixture, soaked in a 1 mol / L citric acid solution for 1.5 h, filtered to obtain a filtrate A and a precipitate A, and the precipitate A was soaked in a 1 mol / L sodium hydroxide solution for 35 min, filtered to obtain a filtrate B and a precipitate B, the mass volume ratio of the Mongolian medicine mixture to the citric acid solution and the sodium hydroxide solution was 1 g:8 mL:8 mL, the filtrate A and the filtrate B were combined to obtain a combined filtrate, glycyrrhizic acid, gelatin and alginic acid were added in a mass ratio of 2:1:1.2, the amount ratio of the combined filtrate to glycyrrhizic acid was 1 L:10 mg, heated to 55°C, maintained for 3.5 h, rotary evaporated at 60°C for 1.5 h, spray dried to obtain Mongolian medicine granules;

[0075] S2: The Mongolian medicine particles obtained in step S1 are added to deionized water, the mass volume ratio of Mongolian medicine particles to deionized water is 1 mg:100 mL, after stirring at 180 rpm for 30 min, centrifuged at 10000 rpm for 35 min, the precipitate is mixed with deionized water again at a ratio of 1 g:100 mL, glycolic acid is added, the mass ratio of Mongolian medicine particles to glycolic acid is 5:1, stirred at room temperature for 4 h, centrifuged at 8000 rpm for 15 min, the precipitate is washed three times with deionized water, and dried at 50°C to obtain a Mongolian medicine preparation;

[0076] S3: The Mongolian medicine preparation obtained in step S2 is evenly mixed with 10 g of a promoter and 25 g of a synergist, ground and crushed to pass through a 500-mesh sieve, and sterilized by ultraviolet light for 1 h to obtain the contents of a Mongolian medicine composition capsule for treating and preventing stroke.

[0077] The difference between Comparative Example 1 and Example 1 is that no synergist is added.

[0078] The only difference between Comparative Example 2 and Example 1 is that no accelerator is added.

[0079] The only difference between Comparative Example 3 and Example 1 is that the preparation method of the Mongolian medicine granules comprises the following steps: washing and drying Panax notoginseng, jujube, ginseng, clematis, aconite and roselle thistle, crushing them through an 80-mesh sieve to obtain a Mongolian medicine mixture, adding the Mongolian medicine mixture to deionized water with a mass volume ratio of 1 g:16 mL, heating to 60°C, soaking for 4 h, rotary evaporating the filtrate at 60°C for 2 h, spray drying, and obtaining Mongolian medicine granules.

[0080] Experimental Example 1: The cytotoxicity of the capsule contents of the Mongolian medicine composition prepared in Examples 1-3 and Comparative Examples 1-3 was determined by the MTT method. According to the ratio of m capsule contents / v complete medium = 50 mg / mL, the capsule contents sterilized by ultraviolet light for 1 h were completely immersed in complete medium, and extracted in a 37°C water bath for 24 h to obtain the extract solution. L-929 in the logarithmic growth phase was taken, and the L-929 concentration was adjusted to 2×10 4 100 μL per well was inoculated into a 96-well plate and placed at 37°C with 5% CO 2 After the cells adhered to the wall, 100 μL of extract was added to each well as the experimental group. Another well with only 200 μL of complete medium was set as the blank control group, and the well with 100 μL of complete medium + 100 μL of cells was set as the negative control. After adding the extract, the cells were cultured for 5 days. Every day, 20 μL of MTT (5 mg / mL prepared in PBS) was added to each well in the dark. On the last day, MTT was added and cultured for 4 h. After discarding all the liquid in the well, 150 μL of DMSO solution was added to each well. The cells were shaken at a low speed for 15 min in an ELISA reader, and the absorbance at λ=490 nm was measured. The toxicity of cells was determined by calculating the relative growth rate (RGR, %): RGR = (OD experimental group - OD blank group) / (OD negative control group - OD blank group). The specific toxicity grading standards are: Level 0: RGR ≥ 100%; Level 1: 99% ≥ RGR ≥ 75; Level 2: 74 ≥ RGR ≥ 50; Level 3: 49 ≥ RGR ≥ 25; Level 4: 24 ≥ RGR ≥ 1; Level 5 = 0. Levels 2 and above can be considered as cytotoxic reactions. The results are shown in Figure 4 shown.

[0081] Figure 4 The results showed that the RGR of comparative examples 1-3 was smaller than that of the embodiment 1-3 group, among which comparative example 3 was the smallest. However, the toxicity levels of the contents of the capsules prepared from the embodiments 1-3 and comparative examples 1-3 were both 0-1, indicating that the Mongolian medicine composition capsules for treating and preventing stroke of the present invention are non-cytotoxic and safe and reliable.

[0082] Experimental Example 2: 160 male SD rats (weight 240 ± 20 g) were taken. After adaptive feeding for 1 week, they were randomly divided into 8 groups, each with 20 rats, namely sham operation group, model group and Example 1-3, Comparative Example 1-3 group. The capsule contents prepared in each example and comparative example were suspended with sodium carboxymethyl cellulose solution to prepare suspensions. Shake well before administration, and gavage the corresponding drugs to each group of mice every day. The Example 1-3 and Comparative Example 1-3 groups were respectively administered at 50 mg content weight / kg body weight, and the sham operation group and the model group were given the same amount of sodium carboxymethyl cellulose solution, and both were administered for 4 weeks. On the second day after the end of administration, a rat cerebral ischemia-reperfusion model was prepared, and the rats were fasted for 12 h before the operation and had free drinking water. Each group of rats was anesthetized by intraperitoneal injection of 10% chloral hydrate solution (0.3 mL / 100 g), fixed on the operating table in a lateral position, and the rat rectal temperature was maintained at 37 ± 1 ° C during the experiment. The hair was shaved, the skin was disinfected, and the length of the midline incision of the neck was about 20 mm. The common carotid artery was exposed, the internal carotid artery was bluntly separated, the common carotid artery and the internal carotid artery were clamped, and the nylon thread was slowly and gently advanced to the direction of the internal carotid artery entering the skull through the incision of the main trunk of the external carotid artery. The common carotid artery bifurcation was used as a mark, and the thread was advanced 18±0.5mm and encountered slight resistance, that is, it reached the thinner middle cerebral artery and stopped. The middle cerebral artery was ligated and blocked to fully block the local cerebral blood flow. The wound was sutured, and a nylon thread of about 3 cm was left outside the body. The surgical area was disinfected with iodine. After 1 h of ischemia, the thread was gently pulled out for 10 min to achieve reperfusion. Sham operation group: no thread ligation was inserted, and the rest of the treatment was the same as the model group, the embodiment and the comparative group. During the operation, a transcranial Doppler blood flow analyzer was used to monitor cerebral blood flow.

[0083] The condition of the animals in each group was closely observed within 24 hours of ischemia-reperfusion, and the behavioral defects were evaluated according to the ZeaLonga scoring standard, where 0 was normal and no symptoms of nerve damage; 1 was inability to fully extend the contralateral forelimb; 2 was turning in circles to the contralateral side when crawling; 3 was the body leaning to the contralateral side when walking; 4 was inability to walk spontaneously and loss of consciousness. The results are shown in Table 1. ** indicates extremely significant difference from the model group, and * indicates significant difference from the model group.

[0084] Table 1 Behavioral scores of rats in each group

[0085]

[0086] The results in Table 1 show that compared with the model group, the behavioral scores of rats in the embodiment and control groups were significantly reduced, and the scores of the embodiment 1-3 groups were lower than those of the control groups 1-3, indicating that the Mongolian medicine composition capsule for treating and preventing stroke of the present invention can improve neurological deficits.

[0087] After 24 h of reperfusion after drug administration, the animals in each group were killed. The whole brains of 10 rats in each group were randomly taken, and the wet weight of brain tissue was weighed, sliced, stained in 2% triphenyltetrazolium chloride (TTC) solution for 30 min, fixed, and the pale area (infarction area, not stained) and non-pale area (normal area) were separated. The volume of cerebral infarction (%) = (wet weight of infarction area / wet weight of brain tissue) × 100%. The stained brain tissue was placed in an oven to dry to constant weight, weighed, and the brain water content was calculated according to the following formula: brain water content (%) = [(wet weight of brain tissue-dry weight of brain tissue) / wet weight of brain tissue] × 100%. The results are shown in Table 2. ** indicates a very significant difference from the model group, and * indicates a significant difference from the model group.

[0088] Table 2 Cerebral infarction volume and brain water content of rats in each group

[0089]

[0090] The results in Table 2 show that compared with the model group, the cerebral infarction volume and brain water content of rats in the embodiment and comparative example groups were significantly reduced, indicating that the Mongolian medicine composition capsule for treating and preventing cerebral stroke of the present invention can improve and reduce the degree of cerebral edema and reduce the area of ​​cerebral infarction.

[0091] After 24 h of reperfusion after administration, the remaining 10 rats in each group were killed, and the brains were quickly removed and weighed. A 10 wt% brain tissue homogenate was prepared with normal saline. After low-temperature centrifugation, the supernatant was taken and the malondialdehyde (MDA), reduced glutathione (GSH) content and superoxide dismutase (SOD) activity in the brain tissue were detected according to the method in the kit instructions. Figure 5 shown.

[0092] Figure 5 The results showed that compared with the model group, the MDA content in the brain tissue of rats in the embodiment and comparative example groups was significantly reduced, and the GSH content and SOD activity were significantly increased, and the embodiment group had the best effect, indicating that the Mongolian medicine composition capsule for treating and preventing cerebral stroke of the present invention can reduce oxidative stress, protect brain tissue from damage by oxygen free radicals, and protect brain tissue, and is suitable for the prevention and treatment of cerebral stroke.

[0093] Experimental Example 3: Prepare 100 mL of pH 1.2 hydrochloric acid solution and 100 mL of pH 6.8 phosphate buffer solution, add the contents of the Mongolian medicine composition capsule prepared in Examples 1-3 and Comparative Examples 1-3, the added amount is 0.3 g / 100 mL, and the weight is recorded as m 1 After stirring at 200 rpm for 3 h at room temperature, the mixture was filtered with a 0.22 μm filter paper and the precipitate was dried to a constant weight. The weight was recorded as m 2 Calculate the dissolution rate of each group of contents in hydrochloric acid solution and phosphate buffer solution, dissolution rate (%) = [(m 1 -m2 ) / m 1 ]×100%, the result is as follows Figure 6 shown.

[0094] Figure 6 The results showed that the dissolution rates of the Example 1-3 groups were higher than those of the Comparative Examples 1-3, and especially higher than those of the Comparative Example 2, indicating that the Mongolian medicine composition capsules for treating and preventing stroke of the present invention have good biocompatibility, high absorption rate, are more easily absorbed and utilized by the human body, have high bioavailability, and can improve the effect of treating and preventing stroke.

[0095] Experimental Example 4: 80 male SD rats (weight 240±20 g) were randomly divided into 8 groups, each with 10 rats, including sham operation group, model group, and Example 1-3 and Comparative Example 1-3 groups after adaptive feeding for 1 week. After adaptive feeding for 1 week, a rat cerebral ischemia-reperfusion model was prepared according to the method of Experimental Example 2. After surgery, the capsule contents prepared in each Example and Comparative Example were suspended with sodium carboxymethyl cellulose solution to prepare a suspension, and the suspension was shaken before administration. The corresponding drugs were gavaged to each group of mice every day. The Example 1-3 and Comparative Example 1-3 groups were respectively administered at 50 mg content weight / kg body weight, and the sham operation group and the model group were administered with an equal amount of sodium carboxymethyl cellulose solution. After 2 weeks of administration, behavioral scores were performed according to the method of Experimental Example 2, and the cerebral infarction volume and brain water content were calculated. The results are shown in Table 3.

[0096] Table 3 The therapeutic effect of the contents of Mongolian medicine composition capsules on stroke

[0097]

[0098] The results in Table 3 show that compared with the model group, the behavioral scores, cerebral infarction volume and brain water content of rats in the embodiment and comparative example groups were significantly reduced, and the Example 1-3 groups were lower than the comparative example 1-3 groups, indicating that the Mongolian medicine composition capsule for treating and preventing cerebral stroke of the present invention can improve neurological deficits, reduce the degree of cerebral edema, reduce the area of ​​cerebral infarction, and has the effect of treating cerebral stroke.

[0099] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present invention (including the claims) is limited to these examples. Under the concept of the present invention, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of the different aspects of the present invention as described above, which are not provided in detail for the sake of simplicity.

[0100] The present invention is intended to cover all such substitutions, modifications and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A Mongolian medicine composition capsule for treating and preventing stroke, characterized in that: The capsule comprises a capsule shell and contents, wherein the contents are composed of the following raw materials in parts by weight: 40 parts of Panax notoginseng, 44 parts of Roselle Thistle, 20 parts of Fructus Axillaris, 30 parts of Clematis, 32 parts of Ginseng, 25 parts of Aconitum, and 12 parts of an accelerator; The method for preparing the Mongolian medicine composition capsule for treating and preventing stroke is characterized by comprising the following steps: S1: Wash and dry Panax notoginseng, Fructus jujubae, Ginseng, Clematis, Aconitum and Cirsium chinense, crush them through an 80-mesh sieve to obtain a Mongolian medicine mixture, soak them in a 1 mol / L citric acid solution for 2 h, filter them to obtain filtrate A and precipitate A, soak precipitate A in a 1 mol / L sodium hydroxide solution for 40 min, filter them to obtain filtrate B and precipitate B, the mass volume ratio of the Mongolian medicine mixture to the citric acid solution and the sodium hydroxide solution is 1 g:8 mL:8 mL, combine filtrate A and filtrate B to obtain a combined filtrate, add glycyrrhizic acid, gelatin and alginic acid in a mass ratio of 2:1:1.2, the dosage ratio of the combined filtrate to glycyrrhizic acid is 1 L:10 mg, heat to 60°C, maintain for 4 h, rotary evaporate at 60°C for 2 h, spray dry, and obtain Mongolian medicine granules; S2: The Mongolian medicine particles obtained in step S1 are added to deionized water, the mass volume ratio of Mongolian medicine particles to deionized water is 1 mg: 100 mL, after stirring at 180 rpm for 30 min, centrifuged at 10000 rpm for 40 min, the precipitate is mixed with deionized water again at a ratio of 1 g: 100 mL, glycolic acid is added, the mass ratio of Mongolian medicine particles to glycolic acid is 5:1, stirred at room temperature for 4 h, centrifuged at 8000 rpm for 15 min, the precipitate is washed three times with deionized water, and dried at 50°C to obtain a Mongolian medicine preparation; S3: The Mongolian medicine preparation and the accelerator are mixed evenly, ground and crushed to pass through a 500-mesh sieve, sterilized by ultraviolet light for 1 hour, and filled into capsules, with each capsule containing 0.5 g, to obtain a Mongolian medicine composition capsule for treating and preventing stroke; The structure of the promoter is 。 2. A Mongolian medicine composition capsule for treating and preventing stroke according to claim 1, characterized in that: The preparation method of the accelerator comprises the following steps: I. In an ice water bath, diethanolamine and p-aminobenzoyl chloride were added to a dimethylformamide solvent, the mass ratio of diethanolamine to p-aminobenzoyl chloride was 2:3, and the mass volume ratio of diethanolamine to dimethylformamide was 1 g:2 mL. Pyridine and triethylamine were added at a mass ratio of 2:1, and the mass ratio of pyridine to diethanolamine was 8:

18. The mixture was stirred for 20 min, and then stirred at room temperature for 24 h to obtain a reaction mixture I; II. To the reaction mixture I obtained in step I, decanoic acid was added, the molar ratio of decanoic acid to p-aminobenzoyl chloride was 1:1, and then dimethyl carbonate was added, the mass ratio of dimethyl carbonate to p-aminobenzoyl chloride was 9:13, and stirred at room temperature for 8 h at a stirring speed of 60 rpm to obtain a reaction mixture II; III. The reaction mixture II obtained in step II was concentrated under reduced pressure to 30% of the original volume, washed with acetone three times, the product was recrystallized in tetrahydrofuran, filtered, washed with 95wt% ethanol solution and deionized water three times in sequence, and dried under vacuum at 50°C for 3 h to obtain a promoter.

3. A Mongolian medicine composition capsule for treating and preventing stroke according to claim 2, characterized in that: The capsule is a hard capsule or a soft capsule.

4. A Mongolian medicine composition capsule for treating and preventing stroke according to claim 3, characterized in that: The capsule shell is prepared from gastric-soluble capsule excipients or enteric-soluble capsule excipients.

5. A Mongolian medicine composition capsule for treating and preventing stroke according to claim 4, characterized in that: The material of the capsule shell includes but is not limited to one or more of gelatin, glycerol, sodium alginate and starch.

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