Traditional Chinese medicine composition for treating deep venous thrombosis

Abstract

The invention relates to the technical field of traditional Chinese medicine compositions, in particular to a traditional Chinese medicine composition for treating deep venous thrombosis. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 5-8 parts of modified traditional Chinese medicine sustained release microspheres, 2-3 parts of a traditional Chinese medicine compound extract, 15-23 parts of modified polyacrylamide, 5-7 parts of chitosan and 40-60 parts of deionized water, 10-15 parts of astragalus membranaceus, 3-5 parts of peach kernels, 8-12 parts of rhizoma atractylodis, 4-6 parts of salvia miltiorrhiza, 1-2 parts of radix ophiopogonis, 4-7 parts of pseudo-ginseng and 5-8 parts of ligusticum wallichii are taken, traditional Chinese medicine effective components are extracted through water extraction, organic solvent extraction and fermentation, and the prepared traditional Chinese medicine composition can effectively treat deep venous thrombosis.

Description

Technical Field

[0001] The present invention relates to the technical field of Chinese medicine compositions, and in particular to a Chinese medicine composition for treating deep vein thrombosis. Background Art

[0002] Deep vein thrombosis refers to a disease in which blood coagulates in deep veins, causing complete or incomplete blockage of blood vessels. It is usually caused by blood coagulation in the deep vein circulation, which is common in the lower limbs. If the thrombus formed in the lower limbs breaks off and flows to the lungs through the blood, it can cause pulmonary artery obstruction and then cause pulmonary embolism. If deep vein thrombosis is not treated in time or is not treated correctly, sequelae are likely to occur. The limbs may swell, causing inconvenience in limb movement, accompanied by soreness, heaviness, fatigue, and swelling of the limbs. Large areas of pigmentation in the lower limbs may occur, or superficial secondary venous dilatation, itching of the skin, etc. Although there are many drugs for treating deep vein thrombosis at present, there is a general problem of unsatisfactory efficacy, and most of the existing therapeutic drugs are Western medicines. Although these Western medicines have certain efficacy, they also have obvious toxic and side effects. There are few existing Chinese patent medicines for treating deep vein thrombosis of the lower limbs, and most of them are single prescriptions, such as β-escin sodium, etc., which lack the advantages of compound Chinese medicine interaction, enhanced efficacy, and reduced toxicity. Summary of the invention

[0003] In view of the deficiencies of the prior art, the present invention provides a Chinese medicine composition for treating deep vein thrombosis.

[0004] The present invention is achieved through the following technical solutions: A Chinese medicine composition for treating deep vein thrombosis, wherein the preparation raw materials include the following components in parts by weight: 5-8 parts of modified Chinese medicine sustained-release microspheres, 2-3 parts of Chinese medicine composite extracts, 15-23 parts of modified polyacrylamide, 5-7 parts of chitosan, and 40-60 parts of deionized water.

[0005] Furthermore, the raw materials for preparing the modified Chinese medicine sustained-release microspheres include the following components in parts by weight: 4-6 parts of Chinese medicine water extract, 3-4 parts of gelatin, 10-15 parts of polylactic acid-glycolic acid copolymer, 4-7 parts of sodium alginate, 10-15 parts of polyvinyl alcohol, and 3-5 parts of citric acid.

[0006] Furthermore, the preparation method of the Chinese medicine water extract comprises the following steps: washing and drying 10-15 parts of Astragalus, 3-5 parts of Peach Kernel, 8-12 parts of Atractylodes, 4-6 parts of Salvia Miltiorrhiza, 1-2 parts of Ophiopogon, 4-7 parts of Panax Notoginseng, and 5-8 parts of Chuanxiong, respectively, crushing and passing through an 80-mesh sieve to obtain a Chinese medicine mixed powder, adding it to deionized water, heating and boiling to extract for 1 h, extracting 2-3 times, filtering, combining the filtrate, and using the filter residue for later use, simmering the filtrate at 60°C, 10 -3MPa to 20% of the original volume and freeze-dried at -40°C to obtain the Chinese medicine water extract.

[0007] Furthermore, the dosage ratio of the traditional Chinese medicine mixed powder to deionized water is 1 g:15-20 mL.

[0008] Furthermore, the preparation method of the modified Chinese medicine sustained-release microspheres comprises the following steps: A1: Add the Chinese medicine water extract and gelatin into deionized water and mix well to obtain an aqueous phase. Dissolve polylactic acid-glycolic acid in a dichloromethane / ethyl acetate mixed solvent to obtain an oil phase. Continue to slowly add the aqueous phase to the oil phase while stirring at 100 rpm. After adding, continue stirring at 180 rpm for 20-30 min. Remove the organic solvent under reduced pressure, centrifuge at 5000 rpm for 20-30 min, wash the precipitate with deionized water, and vacuum dry at 60-70°C to obtain Chinese medicine microspheres. A2: Add sodium alginate and polyvinyl alcohol to deionized water, heat at 80-90°C, stir at 150-200 rpm for 20-25 min to obtain a sodium alginate / polyvinyl alcohol solution, add the Chinese medicine microspheres obtained in step A1 to the sodium alginate / polyvinyl alcohol solution, stir at 200-300 rpm for 30 min, add dropwise to the boric acid solution of calcium chloride, stir at 200-300 rpm for 40-60 min, centrifuge at 5000-6000 rpm for 15-20 min, wash the precipitate with deionized water, and freeze-dry at -20°C to obtain Chinese medicine sustained-release microspheres; A3: The Chinese medicine sustained-release microspheres obtained in step A2 are added to deionized water, and ultrasonicated at 300 W for 20-30 min to obtain a dispersion of Chinese medicine sustained-release microspheres. In a nitrogen atmosphere, citric acid is added to tetrahydrofuran, stirred at 300-400 r / min for 50-60 min, 1,3-dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinimide (NHS) are added, and stirring is continued for 3 h. Tetrahydrofuran is removed under reduced pressure at room temperature, and added to the dispersion of Chinese medicine sustained-release microspheres, stirred at 200-300 rpm for 10-12 h, filtered, the filter cake is washed with deionized water, and vacuum dried at 50-60°C to obtain modified Chinese medicine sustained-release microspheres.

[0009] Furthermore, in step A1, the mass concentration of gelatin in deionized water is 30-40 mg / mL.

[0010] Furthermore, in step A1, the mass concentration of the polylactic acid-glycolic acid copolymer in the dichloromethane / ethyl acetate mixed solvent is 0.08-0.12 g / mL.

[0011] Furthermore, in step A1, the volume ratio of dichloromethane to ethyl acetate in the dichloromethane / ethyl acetate mixed solvent is 1:1.

[0012] Furthermore, in step A2, the mass concentration of the polyvinyl alcohol in deionized water is 40-50 mg / mL.

[0013] Furthermore, in step A2, in the calcium chloride boric acid solution, the mass concentration of calcium chloride is 2-3wt%, and the mass concentration of boric acid is 1-2wt%.

[0014] Furthermore, in step A3, the dispersion concentration of the Chinese medicine sustained-release microspheres in deionized water is 40-50 mg / mL.

[0015] Furthermore, in step A3, the mass concentration of citric acid in tetrahydrofuran is 8-10 mg / mL.

[0016] Furthermore, in step A3, the mass ratio of citric acid, DCC and NHS is 2-2.5:3-4:1.5-2.

[0017] Furthermore, the preparation method of the traditional Chinese medicine composite extract comprises the following steps: B1: Take the filter residue obtained when preparing the aqueous extract of traditional Chinese medicine, add it to 10 times the weight of the mixed organic solvent, heat it to 50°C, extract it 2-3 times, filter it, combine the filtrate, and set aside the filter residue. Remove the organic solvent from the filtrate under reduced pressure, and dry it at 70-80°C to obtain an organic solvent extract; B2: The filter residue obtained in step B1 was mixed evenly with 15 times the weight of deionized water, glucose and urea were added, and ultraviolet sterilization was performed to obtain a fermentation substrate. The activated Lactobacillus plantarum was added at a rate of 10 7 cfu / mL was inoculated into the fermentation substrate and fermented at 50 r / min and 37°C for 48 h; B3: After the cultivation in step B2 is completed, centrifuge at 8000 rpm for 8-12 min, freeze-dry the supernatant at -40°C, and mix evenly with the organic solvent extract obtained in step B1 at a mass ratio of 2:1 to obtain a traditional Chinese medicine composite extract.

[0018] Furthermore, in step B1, the mixed organic solvent is ethanol, ethyl acetate and petroleum ether mixed in a mass ratio of 5:2:1.

[0019] Furthermore, in step B2, the mass ratio of the filter residue, glucose and urea is 3:5:3.

[0020] Furthermore, in step B2, the plant lactobacillus is purchased from China General Microbiological Culture Collection Center with a number of CGMCC 1.12974.

[0021] Furthermore, the raw materials for preparing the modified polyacrylamide include the following components in parts by weight: 5-7 parts of nano-hydroxyapatite, 2-3 parts of vinyltrimethoxysilane, 3-5 parts of chlorogenic acid, 1.2-2 parts of N-(hydroxymethyl)acrylamide, 15-20 parts of acrylamide, and 5-7 parts of N-(2-hydroxypropyl)methacrylamide.

[0022] C1: Add 40-50 nm nano-hydroxyapatite and vinyltrimethoxysilane into tetrahydrofuran, stir at 40-50°C and 150-200 rpm for 6-8 h, filter, wash the filter cake with deionized water, and dry at 80°C to obtain modified hydroxyapatite; C2: Add chlorogenic acid to tetrahydrofuran, ultrasonicate at 300W for 30 min, add 1-hydroxybenzotriazole (HOBt) and N,N'-dicyclohexylcarbodiimide (DCC), stir at 200-300 rpm for 5-6 h, add N-(hydroxymethyl)acrylamide, stir at 40°C and 150-200 rpm for 12-14 h under nitrogen protection, filter, wash the filter cake with deionized water, and vacuum dry to obtain modified chlorogenic acid; C3: Add 1 / 2 amount of acrylamide and N-(2-hydroxypropyl)methacrylamide to a DMSO / acetone mixed solvent, ultrasonicate at 300 W for 30 min, add azobisisobutyronitrile, react at 40°C for 40-50 min under nitrogen protection, add the remaining acrylamide and N-(2-hydroxypropyl)methacrylamide as well as modified hydroxyapatite and modified chlorogenic acid, heat to 50°C and react for 4-5 h, dialyze in deionized water for 12 h with a 3 kDa dialysis bag, and freeze-dry the dialyzed product at -40°C to obtain modified polyacrylamide.

[0023] Furthermore, in step C1, the mass concentration of the vinyltrimethoxysilane in tetrahydrofuran is 16-20 mg / mL.

[0024] Furthermore, in step C2, the mass concentration of chlorogenic acid in tetrahydrofuran is 20-25 mg / mL.

[0025] Furthermore, in step C2, the mass ratio of chlorogenic acid, HOBt and DCC is 3-5:1.2-1.8:1.7-2.9.

[0026] Furthermore, in step C3, the amount of azobisisobutyronitrile added is 3-5wt% of acrylamide.

[0027] Furthermore, in step C3, the volume ratio of DMSO to acetone in the DMSO and acetone mixed solvent is 1:1.

[0028] Furthermore, in step C3, the ratio of acrylamide to DMSO / acetone is 1 g:15-20 mL.

[0029] Furthermore, the present invention also provides a method for preparing the Chinese medicine composition for treating deep vein thrombosis, comprising the following steps: adding modified polyacrylamide to deionized water and stirring evenly, adding the Chinese medicine composite extract, stirring at 40-50°C and 200-300 rpm for 1-1.5 h, adding modified Chinese medicine sustained-release microspheres and chitosan, continuing to stir for 2 h, pouring into a mold, and placing at 40°C for 8-10 h to obtain a Chinese medicine composition for treating deep vein thrombosis.

[0030] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a Chinese medicine composition for treating deep vein thrombosis, which has a dual sustained-release pathway, increases drug loading, can release Chinese medicine ingredients slowly, improves the utilization rate of effective ingredients, and thus enhances the therapeutic effect. Modified Chinese medicine sustained-release microspheres, modified polyacrylamide, and chitosan are cross-linked with each other, enhance tensile properties, and are easier to fit the skin, and can effectively treat deep vein thrombosis. The present invention compound Chinese medicines astragalus, peach kernel, atractylodes, salvia miltiorrhiza, ophiopogon, Panax notoginseng, and chuanxiong, and all the medicines are used together to invigorate qi and activate blood circulation, reduce swelling and relieve pain. The Chinese medicine is subjected to water extraction, organic solvent extraction, and fermentation extraction, and the effective ingredients of the Chinese medicine are fully utilized, and different effective ingredients synergize to effectively treat deep vein thrombosis. The present invention adopts a solvent volatilization method to prepare a Chinese medicine water extract into a Chinese medicine microsphere coated with polylactic acid and glycolic acid, and uses a calcium chloride boric acid solution for crosslinking and curing, and coats a sodium alginate / polyvinyl alcohol layer to improve the sustained release performance, so that the surface of the Chinese medicine sustained-release microsphere is connected with a hydroxyl group, which is beneficial to the improvement of the crosslinking performance and is convenient for subsequent treatment. The carboxyl group of citric acid is covalently grafted with the hydroxyl group on the surface of the Chinese medicine sustained-release microsphere, so that the surface of the Chinese medicine sustained-release microsphere is connected with a carboxyl group, which is beneficial to improve the degree of crosslinking. The present invention grafts carbon-carbon double bonds on the surface of hydroxyapatite and chlorogenic acid, participates in the polymerization of polyacrylamide, reduces agglomeration, and makes hydroxyapatite and chlorogenic acid uniformly dispersed. Hydroxyapatite has a multilayer pore structure and a large specific surface area, which is beneficial to the loading of Chinese medicine components, can adsorb effective ingredients, increase drug loading, and enhance sustained release performance. At the same time, it can be used as a filler to improve mechanical properties. Chlorogenic acid contains multiple phenolic hydroxyl groups, has effective antioxidant and antibacterial properties, and at the same time helps to enhance the degree of crosslinking and improve the mechanical properties of the composition. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] 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.

[0032] Figure 1 It is the tensile strength of the Chinese medicine composition described in Example 1 and Comparative Examples 1-5 of the present invention; Figure 2 The antibacterial properties of the traditional Chinese medicine compositions described in Example 1 and Comparative Examples 1-5 of the present invention. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical scheme and advantages of the present invention more clear, 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. It should be noted that, unless otherwise specified, the chemical reagents involved in the present invention are purchased through commercial channels.

[0034] Example 1: A Chinese medicine composition for treating deep vein thrombosis, the preparation raw materials include the following components in parts by weight: 8 parts of modified Chinese medicine sustained-release microspheres, 3 parts of Chinese medicine composite extracts, 23 parts of modified polyacrylamide, 7 parts of chitosan, and 60 parts of deionized water.

[0035] The raw materials for preparing the modified Chinese medicine sustained-release microspheres include the following components in parts by weight: 6 parts of Chinese medicine water extract, 4 parts of gelatin, 15 parts of polylactic acid-glycolic acid copolymer, 7 parts of sodium alginate, 15 parts of polyvinyl alcohol, and 5 parts of citric acid.

[0036] The preparation method of Chinese medicine water extract comprises the following steps: washing and drying 15 g of Astragalus, 5 g of Peach Kernel, 12 g of Atractylodes, 6 g of Salvia Miltiorrhiza, 2 g of Radix Ophiopogonis, 7 g of Panax Notoginseng and 8 g of Rhizoma Chuanxiong respectively, crushing and passing through an 80-mesh sieve to obtain a mixed Chinese medicine powder, adding the mixed Chinese medicine powder to deionized water in a ratio of 1 g:20 mL, heating and boiling to extract for 1 h, extracting 3 times, filtering, combining the filtrate, and using the filter residue for later use, and simmering the filtrate at 60°C and 10 -3 MPa to 20% of the original volume and freeze-dried at -40°C to obtain the Chinese medicine water extract.

[0037] The preparation method of modified Chinese medicine sustained-release microspheres comprises the following steps: A1: Add 6 g of Chinese medicine water extract and 4 g of gelatin into 100 mL of deionized water and mix well to obtain an aqueous phase. Dissolve 15 g of polylactic acid-glycolic acid in 125 mL of a dichloromethane / ethyl acetate mixed solvent (v / v=1:1) to obtain an oil phase. Slowly add the aqueous phase to the oil phase while stirring at 100 rpm. Continue stirring at 180 rpm for 20-30 min after the addition is complete. Remove the organic solvent under reduced pressure, centrifuge at 5000 rpm for 20-30 min, wash the precipitate with deionized water, and dry in a vacuum at 60-70°C to obtain Chinese medicine microspheres. A2: Add 7 g of sodium alginate and 15 g of polyvinyl alcohol to 300 mL of deionized water, heat at 90°C, and stir at 200 rpm for 25 min to obtain a sodium alginate / polyvinyl alcohol solution. Add the Chinese medicine microspheres obtained in step A1 to the sodium alginate / polyvinyl alcohol solution, stir at 300 rpm for 30 min, add dropwise to a boric acid solution of calcium chloride (the mass concentration of calcium chloride is 3wt%, and the mass concentration of boric acid is 2wt%), stir at 300 rpm for 60 min, centrifuge at 6000 rpm for 20 min, wash the precipitate with deionized water, and freeze-dry at -20°C to obtain Chinese medicine sustained-release microspheres; A3: The Chinese medicine sustained-release microspheres obtained in step A2 were added to deionized water, and ultrasonicated at 300 W for 20-30 min to obtain a 50 mg / mL Chinese medicine sustained-release microsphere dispersion. In a nitrogen atmosphere, 5 g of citric acid was added to 500 mL of tetrahydrofuran, and stirred at 400 r / min for 60 min. 8 g of DCC and 4 g of NHS were added, and stirring was continued for 3 h. Tetrahydrofuran was removed under reduced pressure at room temperature, and added to the Chinese medicine sustained-release microsphere dispersion, and stirred at 300 rpm for 12 h. Filtered, the filter cake was washed with deionized water, and vacuum dried at 60°C to obtain modified Chinese medicine sustained-release microspheres.

[0038] The preparation method of the Chinese medicine composite extract comprises the following steps: B1: Take the filter residue obtained when preparing the aqueous extract of traditional Chinese medicine, add it to 10 times the weight of a mixed organic solvent, wherein the mixed organic solvent is ethanol, ethyl acetate, and petroleum ether mixed in a mass ratio of 5:2:1, heat it to 50°C, extract it 3 times, filter it, combine the filtrate, and set aside the filter residue, remove the organic solvent from the filtrate under reduced pressure, and dry it at 80°C to obtain an organic solvent extract; B2: The filter residue obtained in step B1 was mixed evenly with 15 times the weight of deionized water, and glucose and urea were added, with the mass ratio of the filter residue, glucose and urea being 3:5:3, and ultraviolet sterilization was performed to obtain a fermentation substrate, and the activated Lactobacillus plantarum was added at 10 7 cfu / mL was inoculated into the fermentation substrate and fermented at 50 r / min and 37°C for 48 h; B3: After the cultivation in step B2, centrifuge at 8000 rpm for 12 min, freeze-dry the supernatant at -40°C, and mix evenly with the organic solvent extract obtained in step B1 at a mass ratio of 2:1 to obtain a traditional Chinese medicine composite extract.

[0039] The raw materials for preparing modified polyacrylamide include the following components in parts by weight: 7 parts of nano-hydroxyapatite, 3 parts of vinyltrimethoxysilane, 5 parts of chlorogenic acid, 2 parts of N-(hydroxymethyl)acrylamide, 20 parts of acrylamide, and 7 parts of N-(2-hydroxypropyl)methacrylamide.

[0040] C1: Add 7 g of 50 nm nanohydroxyapatite and 3 g of vinyltrimethoxysilane into 150 mL of tetrahydrofuran, stir at 50°C and 200 rpm for 8 h, filter, wash the filter cake with deionized water, and dry at 80°C to obtain modified hydroxyapatite; C2: Add 5 g of chlorogenic acid to 200 mL of tetrahydrofuran, ultrasonicate at 300 W for 30 min, add 1.8 g of HOBt and 2.9 g of DCC, stir at 300 rpm for 6 h, add 2 g of N-(hydroxymethyl)acrylamide, stir at 40 ° C and 200 rpm for 14 h under nitrogen protection, filter, wash the filter cake with deionized water, and vacuum dry to obtain modified chlorogenic acid; C3: 10 g acrylamide and 3.5 g N-(2-hydroxypropyl)methacrylamide were added to 200 mL DMSO / acetone mixed solvent (v / v=1:1), ultrasonicated at 300 W for 30 min, 0.5 g azobisisobutyronitrile was added, and the mixture was reacted at 40°C for 50 min under nitrogen protection. The remaining acrylamide and N-(2-hydroxypropyl)methacrylamide as well as modified hydroxyapatite and modified chlorogenic acid were added, and the mixture was heated to 50°C for 5 h. The mixture was dialyzed in deionized water for 12 h using a 3 kDa dialysis bag, and the dialyzed product was freeze-dried at -40°C to obtain modified polyacrylamide.

[0041] This embodiment also provides a method for preparing the Chinese medicine composition for treating deep vein thrombosis, comprising the following steps: adding 23 g of modified polyacrylamide to 60 g of deionized water and stirring evenly, adding 3 g of the Chinese medicine composite extract, stirring at 50°C and 300 rpm for 1.5 h, adding 5 g of modified Chinese medicine sustained-release microspheres and 7 g of chitosan, continuing to stir for 2 h, pouring into a mold, and placing at 40°C for 10 h to obtain a Chinese medicine composition for treating deep vein thrombosis.

[0042] Example 2: A Chinese medicine composition for treating deep vein thrombosis, the preparation raw materials include the following components in parts by weight: 5 parts of modified Chinese medicine sustained-release microspheres, 2 parts of Chinese medicine composite extract, 15 parts of modified polyacrylamide, 5 parts of chitosan, and 40 parts of deionized water.

[0043] The raw materials for preparing the modified Chinese medicine sustained-release microspheres include the following components in parts by weight: 4 parts of Chinese medicine water extract, 3 parts of gelatin, 10 parts of polylactic acid-glycolic acid copolymer, 4 parts of sodium alginate, 10 parts of polyvinyl alcohol, and 3 parts of citric acid.

[0044] The preparation method of Chinese medicine water extract comprises the following steps: washing and drying 10 g of Astragalus, 3 g of Peach Kernel, 8 g of Atractylodes, 4 g of Salvia Miltiorrhiza, 1 g of Radix Ophiopogonis, 4 g of Panax Notoginseng and 5 g of Rhizoma Chuanxiong respectively, crushing and passing through an 80-mesh sieve to obtain a mixed Chinese medicine powder, adding the mixed Chinese medicine powder to deionized water in a ratio of 1 g:15-20 mL, heating and boiling to extract for 1 h, extracting twice, filtering, combining the filtrate, and using the filter residue for later use, and simmering the filtrate at 60°C and 10 -3 MPa to 20% of the original volume and freeze-dried at -40°C to obtain the Chinese medicine water extract.

[0045] The preparation method of modified Chinese medicine sustained-release microspheres comprises the following steps: A1: Add 4 g of Chinese medicine water extract and 3 g of gelatin into 100 mL of deionized water and mix well to obtain an aqueous phase. Dissolve 10 g of polylactic acid-glycolic acid in 125 mL of a dichloromethane / ethyl acetate mixed solvent (v / v=1:1) to obtain an oil phase. Slowly add the aqueous phase to the oil phase while stirring at 100 rpm. Continue stirring at 180 rpm for 20 min after adding. Remove the organic solvent under reduced pressure, centrifuge at 5000 rpm for 20 min, wash the precipitate with deionized water, and dry in a vacuum at 60°C to obtain Chinese medicine microspheres. A2: Add 4 g of sodium alginate and 10 g of polyvinyl alcohol to 250 mL of deionized water, heat at 80°C, and stir at 150 rpm for 20 min to obtain a sodium alginate / polyvinyl alcohol solution. Add the Chinese medicine microspheres obtained in step A1 to the sodium alginate / polyvinyl alcohol solution, stir at 200 rpm for 30 min, add dropwise to a boric acid solution of calcium chloride (the mass concentration of calcium chloride is 2wt%, and the mass concentration of boric acid is 1wt%), stir at 200 rpm for 40 min, centrifuge at 5000 rpm for 15 min, wash the precipitate with deionized water, and freeze-dry at -20°C to obtain Chinese medicine sustained-release microspheres; A3: The Chinese medicine sustained-release microspheres obtained in step A2 were added to deionized water and ultrasonicated at 300 W for 20 min to obtain a 40 mg / mL Chinese medicine sustained-release microsphere dispersion. In a nitrogen atmosphere, 3 g of citric acid was added to 375 mL of tetrahydrofuran, stirred at 300 r / min for 50 min, 4.5 g of DCC and 2.25 g of NHS were added, and stirring was continued for 3 h. Tetrahydrofuran was removed under reduced pressure at room temperature, and added to the Chinese medicine sustained-release microsphere dispersion, stirred at 200 rpm for 10 h, filtered, the filter cake was washed with deionized water, and vacuum dried at 50°C to obtain modified Chinese medicine sustained-release microspheres.

[0046] The preparation method of the Chinese medicine composite extract comprises the following steps: B1: Take the filter residue obtained when preparing the aqueous extract of traditional Chinese medicine, add it to 10 times the weight of a mixed organic solvent, wherein the mixed organic solvent is ethanol, ethyl acetate, and petroleum ether mixed in a mass ratio of 5:2:1, heat it to 50°C, extract it twice, filter it, combine the filtrate, and set aside the filter residue, remove the organic solvent from the filtrate under reduced pressure, and dry it at 70°C to obtain an organic solvent extract; B2: The filter residue obtained in step B1 was mixed evenly with 15 times the weight of deionized water, and glucose and urea were added, with the mass ratio of the filter residue, glucose and urea being 3:5:3, and ultraviolet sterilization was performed to obtain a fermentation substrate, and the activated Lactobacillus plantarum was added at 10 7 cfu / mL was inoculated into the fermentation substrate and fermented at 50 r / min and 37°C for 48 h; B3: After the cultivation in step B2, centrifuge at 8000 rpm for 8 min, freeze-dry the supernatant at -40°C, and mix evenly with the organic solvent extract obtained in step B1 at a mass ratio of 2:1 to obtain a traditional Chinese medicine composite extract.

[0047] The raw materials for preparing modified polyacrylamide include the following components in parts by weight: 5 parts of nano-hydroxyapatite, 2 parts of vinyltrimethoxysilane, 3 parts of chlorogenic acid, 1.2 parts of N-(hydroxymethyl)acrylamide, 15 parts of acrylamide, and 5 parts of N-(2-hydroxypropyl)methacrylamide.

[0048] C1: 5 g of 40 nm nanohydroxyapatite and 2 g of vinyltrimethoxysilane were added to 125 mL of tetrahydrofuran, stirred at 40°C and 150 rpm for 6 h, filtered, the filter cake was washed with deionized water, and dried at 80°C to obtain modified hydroxyapatite; C2: 3 g of chlorogenic acid was added to 150 mL of tetrahydrofuran, ultrasonicated at 300 W for 30 min, 1.2 g of HOBt and 1.7 g of DCC were added, stirred at 200 rpm for 5 h, 1.2 g of N-(hydroxymethyl)acrylamide was added, stirred at 40 ° C and 150 rpm for 12 h under nitrogen protection, filtered, the filter cake was washed with deionized water, and vacuum dried to obtain modified chlorogenic acid; C3: 7.5 g acrylamide and 2.5 g N-(2-hydroxypropyl)methacrylamide were added to 112.5 mL DMSO / acetone mixed solvent (v / v=1:1), ultrasonicated at 300 W for 30 min, 0.225 g azobisisobutyronitrile was added, and the mixture was reacted at 40 °C for 40 min under nitrogen protection. The remaining acrylamide and N-(2-hydroxypropyl)methacrylamide as well as modified hydroxyapatite and modified chlorogenic acid were added, and the mixture was heated to 50 °C for 4 h. The mixture was dialyzed in deionized water for 12 h using a 3 kDa dialysis bag, and the dialyzed product was freeze-dried at -40 °C to obtain modified polyacrylamide.

[0049] This embodiment also provides a method for preparing the Chinese medicine composition for treating deep vein thrombosis, comprising the following steps: adding 15 g of modified polyacrylamide to 40 g of deionized water and stirring evenly, adding 2 g of the Chinese medicine composite extract, stirring at 40°C and 200 rpm for 1 h, adding 5 g of modified Chinese medicine sustained-release microspheres and 5 g of chitosan, continuing to stir for 2 h, pouring into a mold, and placing at 40°C for 8 h to obtain a Chinese medicine composition for treating deep vein thrombosis.

[0050] Example 3: A Chinese medicine composition for treating deep vein thrombosis, the preparation raw materials include the following components in parts by weight: 7 parts of modified Chinese medicine sustained-release microspheres, 2.5 parts of Chinese medicine composite extract, 20 parts of modified polyacrylamide, 6 parts of chitosan, and 50 parts of deionized water.

[0051] The raw materials for preparing the modified Chinese medicine sustained-release microspheres include the following components in parts by weight: 5 parts of Chinese medicine water extract, 3.5 parts of gelatin, 12 parts of polylactic acid-glycolic acid copolymer, 5 parts of sodium alginate, 12 parts of polyvinyl alcohol, and 4 parts of citric acid.

[0052] The preparation method of a Chinese medicine water extract comprises the following steps: washing and drying 12 g of Astragalus, 4 g of Peach Kernel, 10 g of Atractylodes, 5 g of Salvia Miltiorrhiza, 1.5 g of Radix Ophiopogonis, 6 g of Panax Notoginseng and 6 g of Rhizoma Chuanxiong respectively, crushing and passing through an 80-mesh sieve to obtain a Chinese medicine mixed powder, adding the Chinese medicine mixed powder to deionized water in a ratio of 1 g:15-20 mL, heating and boiling to extract for 1 h, extracting 3 times, filtering, combining the filtrate, and using the filter residue for later use, and simmering the filtrate at 60°C and 10 -3 MPa to 20% of the original volume and freeze-dried at -40°C to obtain the Chinese medicine water extract.

[0053] The preparation method of modified Chinese medicine sustained-release microspheres comprises the following steps: A1: Add 5 g of Chinese medicine water extract and 3.5 g of gelatin into 100 mL of deionized water and mix well to obtain an aqueous phase. Dissolve 12 g of polylactic acid-glycolic acid in 120 mL of a dichloromethane / ethyl acetate mixed solvent (v / v=1:1) to obtain an oil phase. Slowly add the aqueous phase to the oil phase while stirring at 100 rpm. Continue stirring at 180 rpm for 25 min after adding. Remove the organic solvent under reduced pressure, centrifuge at 5000 rpm for 25 min, wash the precipitate with deionized water, and dry in a vacuum at 65°C to obtain Chinese medicine microspheres. A2: Add 5 g of sodium alginate and 12 g of polyvinyl alcohol to 270 mL of deionized water, heat at 85°C, and stir at 180 rpm for 22 min to obtain a sodium alginate / polyvinyl alcohol solution. Add the Chinese medicine microspheres obtained in step A1 to the sodium alginate / polyvinyl alcohol solution, stir at 250 rpm for 30 min, add dropwise to a calcium chloride boric acid solution (the mass concentration of calcium chloride is 2.5wt%, and the mass concentration of boric acid is 1.5wt%), stir at 250 rpm for 50 min, centrifuge at 5500 rpm for 18 min, wash the precipitate with deionized water, and freeze-dry at -20°C to obtain Chinese medicine sustained-release microspheres; A3: The Chinese medicine sustained-release microspheres obtained in step A2 were added to deionized water, and ultrasonicated at 300 W for 25 min to obtain a 45 mg / mL Chinese medicine sustained-release microsphere dispersion. In a nitrogen atmosphere, 4 g of citric acid was added to 450 mL of tetrahydrofuran, stirred at 350 r / min for 55 min, 6.5 g of DCC and 3.5 g of NHS were added, and stirring was continued for 3 h. Tetrahydrofuran was removed under reduced pressure at room temperature, and added to the Chinese medicine sustained-release microsphere dispersion, stirred at 250 rpm for 11 h, filtered, the filter cake was washed with deionized water, and vacuum dried at 55°C to obtain modified Chinese medicine sustained-release microspheres.

[0054] The preparation method of the Chinese medicine composite extract comprises the following steps: B1: Take the filter residue obtained when preparing the aqueous extract of traditional Chinese medicine, add it to 10 times the weight of a mixed organic solvent, wherein the mixed organic solvent is ethanol, ethyl acetate, and petroleum ether mixed in a mass ratio of 5:2:1, heat it to 50°C, extract it 3 times, filter it, combine the filtrate, and set aside the filter residue, remove the organic solvent from the filtrate under reduced pressure, and dry it at 75°C to obtain an organic solvent extract; B2: The filter residue obtained in step B1 was mixed evenly with 15 times the weight of deionized water, and glucose and urea were added, with the mass ratio of the filter residue, glucose and urea being 3:5:3, and ultraviolet sterilization was performed to obtain a fermentation substrate, and the activated Lactobacillus plantarum was added at 10 7 cfu / mL was inoculated into the fermentation substrate and fermented at 50 r / min and 37°C for 48 h; B3: After the cultivation in step B2, centrifuge at 8000 rpm for 10 min, freeze-dry the supernatant at -40°C, and mix evenly with the organic solvent extract obtained in step B1 at a mass ratio of 2:1 to obtain a traditional Chinese medicine composite extract.

[0055] The raw materials for preparing modified polyacrylamide include the following components in parts by weight: 6 parts of nano-hydroxyapatite, 2.5 parts of vinyltrimethoxysilane, 4 parts of chlorogenic acid, 1.5 parts of N-(hydroxymethyl)acrylamide, 18 parts of acrylamide, and 6 parts of N-(2-hydroxypropyl)methacrylamide.

[0056] C1: 6 g of 45 nm nanohydroxyapatite and 2.5 g of vinyltrimethoxysilane were added to 150 mL of tetrahydrofuran, stirred at 45°C and 180 rpm for 7 h, filtered, the filter cake was washed with deionized water, and dried at 80°C to obtain modified hydroxyapatite; C2: 4 g of chlorogenic acid was added to 180 mL of tetrahydrofuran, ultrasonicated at 300 W for 30 min, 1.5 g of HOBt and 2 g of DCC were added, stirred at 250 rpm for 5.5 h, 1.5 g of N-(hydroxymethyl)acrylamide was added, stirred at 40 ° C and 180 rpm for 13 h under nitrogen protection, filtered, the filter cake was washed with deionized water, and vacuum dried to obtain modified chlorogenic acid; C3: Add 9 g acrylamide and 3 g N-(2-hydroxypropyl)methacrylamide to 162 mL DMSO / acetone mixed solvent (v / v=1:1), ultrasonicate at 300 W for 30 min, add 0.36 g azobisisobutyronitrile, react at 40°C for 40-50 min under nitrogen protection, add the remaining acrylamide and N-(2-hydroxypropyl)methacrylamide as well as modified hydroxyapatite and modified chlorogenic acid, heat to 50°C and react for 4-5 h, dialyze in deionized water for 12 h using a 3 kDa dialysis bag, and freeze-dry the dialyzed product at -40°C to obtain modified polyacrylamide.

[0057] This embodiment also provides a method for preparing the Chinese medicine composition for treating deep vein thrombosis, comprising the following steps: adding 20 g of modified polyacrylamide to 50 g of deionized water and stirring evenly, adding 2.5 g of the Chinese medicine composite extract, stirring at 45° C. and 250 rpm for 1.2 h, adding 7 g of modified Chinese medicine sustained-release microspheres and 6 g of chitosan, continuing to stir for 2 h, pouring into a mold, and placing at 40° C. for 9 h to obtain a Chinese medicine composition for treating deep vein thrombosis.

[0058] The difference between Comparative Example 1 and Example 1 is that no modified Chinese medicine sustained-release microspheres are added; The difference between Comparative Example 2 and Example 1 is that no Chinese herbal compound extract is added; The difference between Comparative Example 3 and Example 1 is that the modified Chinese medicine sustained-release microspheres are replaced by Chinese medicine aqueous extracts, and the amount of Chinese medicine aqueous extracts added is the same as that in Example 1; The difference between Comparative Example 4 and Example 1 is that no modified hydroxyapatite is added; The only difference between Comparative Example 5 and Example 1 is that no modified chlorogenic acid is added.

[0059] Experimental Example 1: Patients clinically diagnosed with deep vein thrombosis of the lower extremities were selected, including 118 male patients and 92 female patients aged 52-57 years old, and were randomly divided into 7 groups, 30 cases in each group. The Chinese medicine compositions prepared in Examples 1-3 and Comparative Examples 1-5 were applied to the skin corresponding to the thrombus position of the lower extremities of each group of patients, fixed with non-woven gauze, and replaced every two days. The efficacy criteria are: cured: the swelling of the affected limb completely disappeared, and the circumference difference of the same horizontal position of the two lower limbs was less than 0.05 cm; markedly effective: the swelling of the affected limb was significantly reduced, and the circumference difference of the same horizontal position of the two lower limbs was 0.5-1.0 cm; effective: the swelling of the affected limb was reduced, and the circumference difference of the same horizontal position of the two lower limbs was 1.0-2.0 cm; ineffective: the swelling of the affected limb did not improve, and the circumference difference of the same horizontal position of the two lower limbs was greater than 2.0 cm. The results are shown in Table 1.

[0060] Table 1:

[0061] The results in Table 1 show that the therapeutic effects of Examples 1-3 on lower extremity venous thrombosis are better than those of Comparative Examples 1-5. Comparative Example 1 does not add modified Chinese medicine sustained-release microspheres, Comparative Example 2 does not add Chinese medicine composite extracts, the content of effective ingredients is reduced, and the therapeutic effect is reduced. Comparative Example 3 does not use sustained-release microspheres, the sustained-release function is reduced, the drug loading capacity is reduced, and the therapeutic effect is reduced. Comparative Example 4 does not add modified hydroxyapatite, the drug loading capacity is reduced, and the therapeutic effect is reduced. Comparative Example 5 does not add modified chlorogenic acid, and the therapeutic effect is reduced. In addition, no adverse reactions occurred during the treatment of Examples 1-3 and Comparative Examples 1-5. The Chinese medicine composition for treating deep vein thrombosis prepared by the present invention has good therapeutic effect and high safety.

[0062] Experimental Example 2: The tensile performance test was conducted using a universal testing machine. The sample was cut into a cylinder with a diameter of 3 mm and a height of 45 mm. The initial length gauge length was 15 mm and the loading speed was 100 mm / min. The tensile strength test results are shown in Figure 2. Figure 1 shown.

[0063] Figure 1 The results showed that the tensile strength of Example 1 was significantly better than that of Comparative Example 1 and Comparative Examples 3-5. Comparative Example 1 did not add modified Chinese medicine sustained-release microspheres, Comparative Example 3 did not use sustained-release microspheres, and the degree of cross-linking decreased. Comparative Example 4 did not add modified hydroxyapatite, the degree of cross-linking decreased, and the mechanical properties decreased. Comparative Example 5 did not add modified chlorogenic acid, the degree of cross-linking decreased, and the tensile strength decreased.

[0064] Experimental Example 3: The antibacterial properties of the Chinese medicine compositions obtained in Example 1 and Comparative Examples 1-5 were quantitatively characterized using the film pasting method according to GB / T 21866-2008 standard, and the antibacterial rate against Escherichia coli was tested. The results are as follows: Figure 2 shown.

[0065] Figure 2The results showed that the antibacterial rate of Example 1 was over 99%. Comparative Examples 1 and 2 did not add Chinese medicine sustained-release microspheres and Chinese medicine composite extracts, the content of effective ingredients of Chinese medicine decreased, and the antibacterial activity decreased. Comparative Example 3 did not use sustained-release microspheres, and the drug loading amount decreased. Comparative Example 4 did not add hydroxyapatite, the drug loading amount decreased, and the antibacterial activity decreased. Comparative Example 5 did not add modified chlorogenic acid, and the antibacterial activity decreased.

[0066] 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 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.

Claims

1. A Chinese medicine composition for treating deep vein thrombosis, characterized in that: The preparation raw materials include the following components in parts by weight: 5-8 parts of modified Chinese medicine sustained-release microspheres, 2-3 parts of Chinese medicine composite extracts, 15-23 parts of modified polyacrylamide, 5-7 parts of chitosan, and 40-60 parts of deionized water; The raw materials for preparing the modified Chinese medicine sustained-release microspheres include the following components in parts by weight: 4-6 parts of Chinese medicine water extract, 3-4 parts of gelatin, 10-15 parts of polylactic acid-glycolic acid copolymer, 4-7 parts of sodium alginate, 10-15 parts of polyvinyl alcohol, and 3-5 parts of citric acid; The preparation method of the Chinese medicine water extract comprises the following steps: washing and drying 10-15 parts of astragalus, 3-5 parts of peach kernel, 8-12 parts of atractylodes, 4-6 parts of salvia miltiorrhiza, 1-2 parts of ophiopogon, 4-7 parts of panax notoginseng and 5-8 parts of ligusticum chuanxiong respectively, crushing to obtain a Chinese medicine mixed powder, adding it into deionized water, heating and boiling to extract, filtering, combining the filtrate, reserving the filter residue for later use, concentrating the filtrate, and freeze-drying to obtain the Chinese medicine water extract; The preparation method of the modified Chinese medicine sustained-release microspheres comprises the following steps: A1: Add the Chinese medicine water extract and gelatin to deionized water and mix well to obtain an aqueous phase, dissolve polylactic acid-glycolic acid in a dichloromethane / ethyl acetate mixed solvent to obtain an oil phase, and slowly add the aqueous phase to the oil phase while stirring. Continue stirring after adding, remove the organic solvent under reduced pressure, centrifuge, wash the precipitate with deionized water, and vacuum dry to obtain Chinese medicine microspheres; A2: Sodium alginate and polyvinyl alcohol are added to deionized water, heated, and stirred to obtain a sodium alginate / polyvinyl alcohol solution, and the Chinese medicine microspheres obtained in step A1 are added to the sodium alginate / polyvinyl alcohol solution, stirred, and added dropwise to a boric acid solution of calcium chloride, stirred, centrifuged, and the precipitate is washed with deionized water, and freeze-dried at -20°C to obtain Chinese medicine sustained-release microspheres; A3: Add the Chinese medicine sustained-release microspheres obtained in step A2 into deionized water, perform ultrasound treatment to obtain a dispersion of Chinese medicine sustained-release microspheres, add citric acid into tetrahydrofuran in a nitrogen atmosphere, stir, add DCC and NHS, continue stirring, remove tetrahydrofuran under reduced pressure, add to the dispersion of Chinese medicine sustained-release microspheres, stir, filter, wash the filter cake with deionized water, and vacuum dry to obtain modified Chinese medicine sustained-release microspheres; The preparation method of the traditional Chinese medicine composite extract comprises the following steps: B1: Take the filter residue obtained when preparing the aqueous extract of traditional Chinese medicine, add it into 10 times the weight of the mixed organic solvent, heat and extract, filter, combine the filtrate, set the filter residue aside, reduce the pressure of the filtrate to remove the organic solvent, and dry to obtain an organic solvent extract; B2: uniformly mixing the filter residue obtained in step B1 with 15 times the weight of deionized water, adding glucose and urea, sterilizing to obtain a fermentation substrate, inoculating the activated Lactobacillus plantarum into the fermentation substrate, and fermenting and culturing for 5 minutes; B3: After the cultivation in step B2 is completed, centrifugation is performed, the supernatant is freeze-dried, and mixed evenly with the organic solvent extract obtained in step B1 at a mass ratio of 2:1 to obtain a traditional Chinese medicine composite extract; The raw materials for preparing the modified polyacrylamide include the following components in parts by weight: 5-7 parts of nano-hydroxyapatite, 2-3 parts of vinyltrimethoxysilane, 3-5 parts of chlorogenic acid, 1.2-2 parts of N-(hydroxymethyl)acrylamide, 15-20 parts of acrylamide, and 5-7 parts of N-(2-hydroxypropyl)methacrylamide; C1: Add nano-hydroxyapatite and vinyltrimethoxysilane into tetrahydrofuran, stir, filter, wash the filter cake with deionized water, and dry at 80°C to obtain modified hydroxyapatite; C2: Add chlorogenic acid to tetrahydrofuran, sonicate, add HOBt and DCC, stir, add N-(hydroxymethyl)acrylamide, stir under nitrogen protection, filter, wash the filter cake with deionized water, and vacuum dry to obtain modified chlorogenic acid; C3: Add 1 / 2 amount of acrylamide and N-(2-hydroxypropyl)methacrylamide to a DMSO / acetone mixed solvent, sonicate, add azobisisobutyronitrile, react under nitrogen protection, add the remaining acrylamide and N-(2-hydroxypropyl)methacrylamide as well as modified hydroxyapatite and modified chlorogenic acid, heat the reaction, dialyze, and freeze-dry to obtain modified polyacrylamide.

2. The Chinese medicine composition for treating deep vein thrombosis according to claim 1, characterized in that: The dosage ratio of the traditional Chinese medicine mixed powder to deionized water is 1 g:15-20 mL.

3. The Chinese medicine composition for treating deep vein thrombosis according to claim 2, characterized in that: In step A1, the mass concentration of gelatin in deionized water is 30-40 mg / mL; the mass concentration of polylactic acid-glycolic acid copolymer in dichloromethane / ethyl acetate mixed solvent is 0.08-0.12 g / mL; and the volume ratio of dichloromethane to ethyl acetate in the dichloromethane / ethyl acetate mixed solvent is 1:

1.

4. The Chinese medicine composition for treating deep vein thrombosis according to claim 3, characterized in that: In step A2, the mass concentration of the polyvinyl alcohol in deionized water is 40-50 mg / mL; in the boric acid solution of calcium chloride, the mass concentration of calcium chloride is 2-3wt%, and the mass concentration of boric acid is 1-2wt%.

5. The Chinese medicine composition for treating deep vein thrombosis according to claim 4, characterized in that: In step A3, the dispersion concentration of the Chinese medicine sustained-release microspheres in deionized water is 40-50 mg / mL; the mass concentration of citric acid in tetrahydrofuran is 8-10 mg / mL; and the mass ratio of citric acid, DCC and NHS is 2-2.5:3-4:1.5-2.

6. The Chinese medicine composition for treating deep vein thrombosis according to claim 5, characterized in that: In step B1, the mixed organic solvent is ethanol, ethyl acetate and petroleum ether mixed in a mass ratio of 5:2:1; in step B2, the mass ratio of the filter residue, glucose and urea is 3:5:

3.

7. The Chinese medicine composition for treating deep vein thrombosis according to claim 6, characterized in that: In step C1, the mass concentration of the vinyltrimethoxysilane in tetrahydrofuran is 16-20 mg / mL.

8. The Chinese medicine composition for treating deep vein thrombosis according to claim 7, characterized in that: In step C2, the mass concentration of chlorogenic acid in tetrahydrofuran is 20-25 mg / mL; the mass ratio of chlorogenic acid, HOBt and DCC is 3-5:1.2-1.8:1.7-2.

9.

9. The Chinese medicine composition for treating deep vein thrombosis according to claim 8, characterized in that: In step C3, the amount of azobisisobutyronitrile added is 3-5wt% of acrylamide; the volume ratio of DMSO to acetone in the mixed solvent of DMSO and acetone is 1:1; and the amount ratio of acrylamide to DMSO / acetone is 1 g:15-20 mL.

10. A method for preparing a Chinese medicine composition for treating deep vein thrombosis according to any one of claims 1 to 9, characterized in that: The following steps are involved: Add modified polyacrylamide into deionized water and stir evenly, add the Chinese medicine composite extract, stir, add modified Chinese medicine sustained-release microspheres and chitosan, continue stirring, pour into a mold, place at 40°C for 8-10 hours, and obtain a Chinese medicine composition for treating deep vein thrombosis.

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