A Chinese medicine granule and its preparation method and application

Abstract

The present invention discloses a Chinese medicine granule and a preparation method and application thereof, and belongs to the technical field of Chinese medicine granule preparation. The Chinese medicine granule is composed of an extract of Chinese medicine formula raw materials, a filler, a disintegrant, a sustained-release agent and a flavoring agent, and the Chinese medicine formula raw materials are composed of Rehmannia glutinosa, Cornus officinalis, Cyathula officinalis, Rhizoma Alismatis, Poria, Rhizoma Atractylodis Macrocephalae, Rhizoma Clematidis, Rhizoma Corydalis and Radix Glycyrrhizae. The present invention obtains a Chinese medicine formula granule with good therapeutic significance for cartilage repair, which is not only easy to carry and store, but also shows a positive therapeutic significance for cartilage repair, and the innovative process prescription has a sustained-release effect. Compared with the decoction, the drug release time is extended, the number of administrations can be reduced, and the quality of life of patients is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of preparation of Chinese medicine granules, in particular to a Chinese medicine granule and a preparation method and application thereof. Background Art

[0002] Chronic diseases caused by articular cartilage degeneration and injury have clinical manifestations such as pain, functional impairment, deformity, etc., which seriously affect people's lives and work and bring huge social and economic burdens. Therefore, timely treatment of articular cartilage degeneration and injury at an early stage to prevent further development of the disease is crucial in clinical treatment.

[0003] Based on many years of clinical experience, the inventor has obtained a Chinese medicine formula with cartilage repair effect, which can effectively treat cartilage repair, such as the previous document "Treatment of knee osteoarthritis by tonifying the kidney and removing dampness combined with platelet-rich plasma to regulate serum TGF-β1 and Smad-1 levels". However, the prescription is a traditional Chinese medicine decoction, the liquid is not easy to preserve, it is inconvenient to carry, and it is taken 2-3 times a day, and the drug cannot be effectively absorbed.

[0004] At present, there is an urgent need to provide a Chinese medicine preparation that is easy to store and carry and can provide a good absorption effect. Capsule-type prescriptions are prone to mildew and softening in a hot and humid environment, so preservation is relatively difficult. Tablet-type prescriptions take a long time to disintegrate in gastric juice, which will cause mild or severe damage to the gastric mucosa, so many tablets need to be taken after meals. At the same time, the whole tablet must be swallowed and cannot be chewed. The acidity and alkalinity of the drug will cause damage to the esophagus. The oral liquid drug has a large dispersion degree and is easily affected by the dispersion medium, which can easily cause chemical degradation of the drug, reduce the efficacy or even make it ineffective; liquid preparations are large in size, and are inconvenient to carry, transport, and store; aqueous liquid preparations are prone to mildew and need to add preservatives; non-uniform liquid preparations have a large dispersion degree of the drug, and the dispersed particles have a large specific surface area, which is prone to a series of physical stability problems.

[0005] Chinese medicine granules are solid dosage forms of certain sized particles made by soaking, extracting, concentrating, drying and other processes of Chinese medicine slices and evenly mixing them with appropriate auxiliary materials, and then granulating them. Chinese medicine granules have the advantages of reliable efficacy, stable quality between batches, easy long-term storage, easy to take, and easy to carry. Therefore, Chinese medicine granules have great advantages in the modernization and development of traditional Chinese medicine technology. Summary of the invention

[0006] In view of the problems existing in the prior art, the present invention provides a Chinese medicine granule and a preparation method and application thereof, which solve the problems of the existing water decoction that is inconvenient to store and carry, the need for frequent medication, and the difficulty in storage. The Chinese medicine granule provided by the present invention has the effect of slow release, improves the drug action time, shows a good therapeutic effect on cartilage repair, and is easy to store and carry.

[0007] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0008] In one aspect, the present invention provides a Chinese medicine granule, which is composed of the following components in weight percentage: 33.1-52.7% of the extract of the Chinese medicine formula raw material, 27.4-46% of the filler, 4-8.7% of the disintegrant, 8-17% of the sustained-release agent, and 2-4.7% of the flavoring agent;

[0009] Wherein, the filler is one or more of microcrystalline cellulose, dextrin, and lactose; the disintegrant is one or more of cross-linked polyvinyl pyrrolidone, sodium carboxymethyl starch, and low-substituted hydroxypropyl cellulose; the sustained-release agent is one or more of ethyl cellulose, polyethylene, and polypropylene; the flavoring agent is sodium saccharin and / or aspartame;

[0010] The extract of the raw materials of the Chinese medicine formula is a concentrated liquid or dry paste containing active ingredients obtained by soaking, decocting, extracting, concentrating or freeze-drying the raw materials of the Chinese medicine formula.

[0011] Furthermore, the Chinese medicine formula raw materials are composed of the following components by weight:

[0012] 12-15 parts by weight of Rehmannia glutinosa;

[0013] Cornus officinalis 9-12 parts by weight;

[0014] 6-9 parts by weight of Cyathula capitata;

[0015] Rhizoma Alismatis 6-9 parts by weight;

[0016] Poria 12-15 parts by weight;

[0017] Stir-fried Atractylodes macrocephala 6-9 parts by weight;

[0018] 6-9 parts by weight of Qiannianjian;

[0019] 6-9 parts by weight of Clematis chinensis;

[0020] Corydalis yanhusuo 7-10 parts by weight;

[0021] Licorice 3-6 parts by weight.

[0022] Preferably, the weight ratio of Cyathula capitata, Rhizoma Alismatis, Rhizoma Atractylodis Macrocephalae, Rhizoma Cibotii and Radix Clematis is 1:1:1:1:1:1.

[0023] Preferably, the filler is microcrystalline cellulose, the disintegrant is cross-linked polyvinyl pyrrolidone, the sustained-release agent is ethyl cellulose, and the flavoring agent is saccharin sodium.

[0024] On the other hand, the present invention also provides a method for preparing the above-mentioned Chinese medicine granules, comprising:

[0025] 1) Preparation of extracts of Chinese medicine formula raw materials: soaking, decocting, extracting, concentrating or freeze-drying the Chinese medicine formula raw materials in the prescribed amount to obtain a concentrated solution or dry paste containing active ingredients;

[0026] 2) re-dissolving the above concentrate or dry paste with purified water, and mixing with fillers, disintegrants, sustained-release agents, and flavoring agents to prepare granules;

[0027] Among them, the granulation method uses fluidized bed wet granulation, the main fan temperature is 63°C-65°C, the fan frequency is 15Hz, the infusion pump speed is 5-8mL / min, and the atomization pressure is 1.6-1.9bar;

[0028] 3) Sieve the above granules through a sieve to obtain the desired Chinese medicine granules;

[0029] The mesh number of the upper sieve of the sieve is 16-18 meshes, and the mesh number of the lower sieve is 30-35 meshes.

[0030] Furthermore, the step 1) is specifically as follows:

[0031] a. Weigh the raw materials of Chinese medicine formula according to the proportion, then mix the raw materials of Chinese medicine formula with purified water at a mass ratio of 1:7-8, and soak at room temperature for 90-150 minutes;

[0032] b. After soaking, extracting the medicinal liquid by water decoction method, the water decoction method conditions are as follows: boil the mixture in step a, then decoct for 50-70 minutes, filter and collect the decoction; decoct the residue twice, continue boiling for 35-40 minutes, filter and collect the decoction; mix the two decoctions and filter the residue to obtain the extract;

[0033] c. The extract obtained in step b is concentrated by vacuum concentration to obtain a concentrated solution or a dry paste.

[0034] Preferably, in step c, the pressure of the reduced pressure concentration method is -0.04~0.06MPa and the temperature is 60°C.

[0035] Preferably, in step c, the dry cream is obtained by freeze-drying the concentrated liquid, and the dry cream is stored for no more than 5 months;

[0036] The freeze-drying conditions are as follows: the concentrated liquid is cooled to about 5°C and maintained for 1 hour, then cooled to -40°C and maintained for 2 hours; then heated to -20°C, vacuum degree 20Pa, heating rate 0.4°C / min, and maintained for 20 hours; heated to zero again, vacuum degree unchanged, heating rate unchanged, and maintained for 5 hours; then heated to 40°C, vacuum degree 10Pa, heating rate 1°C / min, and maintained for 4 hours.

[0037] Furthermore, in the step 2), the density of the concentrated solution is 1.04-1.08 g / mL, and the density of the dry paste after reconstitution with purified water is 1.04-1.08 g / mL.

[0038] In another aspect, the present invention also provides an application of the above-mentioned Chinese medicine granules in arthritis and cartilage repair drugs.

[0039] Compared with the prior art, the present invention has the following beneficial effects:

[0040] The invention fully soaks and decocts the raw materials of the Chinese medicine formula with a specific ratio, extracts the active ingredients to the maximum extent, and then adopts the processes of reduced pressure concentration, freeze drying, etc., with mild conditions, shortening the concentration time, and being conducive to maintaining the activity of the effective ingredients. Finally, a certain density concentrated liquid is obtained and mixed with ethyl cellulose, microcrystalline cellulose, and cross-linked polyvinyl pyrrolidone, and the Chinese medicine granules are prepared by wet granulation. The obtained Chinese medicine granules have a high yield, and the prepared granules have the advantages of stable efficacy, low hygroscopicity, physical and chemical stability, reliable quality, etc.

[0041] Through the creative work of the experimenters, through the screening of auxiliary materials and process optimization, a Chinese medicine granule with good sustained-release effect was obtained, which improved the drug effect and other advantages. At the same time, through cytological tests, the granule of the present invention achieved a good cartilage repair effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 This is a graph of the cumulative release rate of rehmannia glycoside D in the traditional Chinese medicine granules prepared in Examples 1-8 of the present invention;

[0043] Figure 2 These are the pathological sections with different treatments;

[0044] Figure 3 The staining images of type II collagen (CⅡ) in articular cartilage treated differently. DETAILED DESCRIPTION

[0045] In order to make the purpose, technical solution and advantages of the present application more clearly understood, the present application is further described in detail through embodiments below. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0046] Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as understood by any technician in the field to which this application belongs. The terms used to describe this application are intended to describe specific implementations only and are not intended to limit the scope of teachings.

[0047] The technical terms appearing in this application are used according to their common sense or meaning to those skilled in the art. If specific meanings are expressed for certain terms, the definitions of the terms will be given below in the context of using these terms.

[0048] The following examples are used to further illustrate the advantages and characteristics of the present method, but are not intended to limit the present application. The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions.

[0049] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0050] Example 1

[0051] A Chinese medicine granule, composed of the following components in percentage by weight: 42.6% of an extract of a Chinese medicine formula raw material, 36.2% of microcrystalline cellulose, 6.0% of cross-linked polyvinyl pyrrolidone, 12.0% of ethyl cellulose, and 3.2% of saccharin sodium;

[0052] A method for preparing the above-mentioned Chinese medicine granules comprises the following steps:

[0053] 1) Soak, decoct, extract and concentrate the following amounts of Chinese medicine formula raw materials to obtain a concentrated solution containing active ingredients;

[0054] Among them, the Chinese medicine ingredients in the formula are: 30g of Rehmannia glutinosa, 24g of Cornus officinalis, 18g of Cyathula officinalis, 18g of Alisma orientalis, 30g of Poria cocos, 18g of Rhizoma Atractylodis Macrocephalae, 18g of Millennium Jian, 18g of Clematidis, 20g of Corydalis yanhusuo, and 12g of Licorice.

[0055] a. Soak the raw materials of Chinese medicine formula in purified water at a mass ratio of 1:8 for 120 minutes at room temperature;

[0056] b. After soaking, extracting the medicinal liquid by water decoction method, the water decoction conditions are: decoction twice, boiling the mixture in step a, and then decocting for 60 minutes, filtering the decoction, decocting the residue twice, boiling for 40 minutes, filtering the decoction, mixing the two decoctions and filtering the residue to obtain the extract;

[0057] c. The obtained extract was concentrated by vacuum concentration to obtain a concentrated solution. The pressure of the vacuum concentration method was -0.04~0.06MPa and the temperature during concentration was 60°C. The obtained concentrated solution had a drug density of 1.05g / mL. The drug density effectively prevented excessive local concentration, ensured the overall uniform distribution of the active ingredients of the drug, and was not easy to cause the particles to adhere to each other and agglomerate, which was beneficial to the subsequent drying.

[0058] 2) mixing the concentrate with other ingredients to form unsieved granules;

[0059] Among them, other ingredients include 175.1g of microcrystalline cellulose, 58g of ethyl cellulose, 29g of cross-linked polyvinyl pyrrolidone, and 15.47g of saccharin sodium;

[0060] Among them, fluidized bed wet granulation was used, with the conditions of main fan temperature of 63°C-65°C, fan frequency of 15Hz, infusion pump speed of 7mL / min, and atomization pressure of 1.8bar;

[0061] 3) The unscreened granules are sieved through a sieve to obtain the desired Chinese medicine granules;

[0062] Among them, the unscreened particles are screened through an upper sieve with a mesh size of 17 and a lower sieve with a mesh size of 33 to obtain the desired Chinese medicine granules.

[0063] Example 2

[0064] In this example, the contents of the components in the granules are shown in Table 1, and the preparation process conditions are the same as those in Example 1.

[0065] Table 1

[0066]

[0067] Example 3

[0068] In this example, the contents of the components in the granules are shown in Table 2, and the preparation process conditions are the same as those in Example 1.

[0069] Table 2

[0070]

[0071] Example 4

[0072] In this example, the contents of the components in the granules are shown in Table 3, and the preparation process conditions are the same as those in Example 1.

[0073] Table 3

[0074]

[0075] Example 5

[0076] In this example, the contents of the components in the granules are shown in Table 4, and the preparation process conditions are the same as those in Example 1.

[0077] Table 4

[0078]

[0079] Example 6

[0080] In this example, the contents of the components in the granules are shown in Table 5, and the preparation process conditions are the same as those in Example 1.

[0081] Table 5

[0082]

[0083] Example 7

[0084] A Chinese medicine granule, the formula of which is the same as that of Example 1;

[0085] The preparation method of the above-mentioned Chinese medicine granules is as follows, comprising:

[0086] 1) Weighing the raw materials of the Chinese medicine formula according to the proportion (same as in Example 1), soaking, decocting, extracting, concentrating and freeze-drying to obtain a dry paste containing active ingredients;

[0087] a. Weigh the raw materials of Chinese medicine formula according to the proportion, and soak them in purified water at room temperature for 90 minutes according to the mass ratio of Chinese medicine formula raw materials to purified water of 1:7 times;

[0088] b. After soaking, extracting the medicinal liquid by water decoction method, the water decoction conditions are: decoction twice, boiling the mixture in step a, and then decocting for 70 minutes, filtering the decoction, decocting the residue twice, boiling for 35 minutes, filtering the decoction, mixing the two decoctions and filtering the residue to obtain the extract;

[0089] c. The extract obtained in step b is concentrated by vacuum concentration to obtain a concentrated solution; the pressure of the vacuum concentration method is -0.04~0.06MPa, and the temperature during concentration is 60°C; the concentrated solution has a liquid density of 1.08g / mL;

[0090] The concentrated solution was freeze-dried: the concentrated solution was cooled to about 5°C, then cooled to -40°C over 1 hour, maintained for 2 hours, then heated to -20°C, vacuum degree 20Pa, heating rate 0.4°C / min, maintained for 20 hours, heated to zero again, vacuum degree unchanged, heating rate unchanged, maintained for 5 hours, then heated to 40°C, vacuum degree 10Pa, heating rate 1°C / min, maintained for 4 hours;

[0091] 2) The dry paste is reconstituted with purified water to a density of 1.05 g / mL, and mixed with other ingredients to form unsieved granules;

[0092] Among them, the granulation method uses fluidized bed wet granulation, the main fan temperature is 63°C, the fan frequency is 15Hz, the infusion pump speed is 8mL / min, and the atomization pressure is 1.6bar;

[0093] 3) The unscreened granules are sieved through a sieve to obtain the desired Chinese medicine granules;

[0094] Among them, the unscreened particles are screened through an upper sieve with a mesh size of 18 and a lower sieve with a mesh size of 30 to obtain the required Chinese medicine granules.

[0095] Example 8

[0096] A Chinese medicine granule, the formula of which is the same as that of Example 1;

[0097] The preparation method of the above-mentioned Chinese medicine granules is as follows, comprising:

[0098] 1) Weighing the raw materials of the Chinese medicine formula according to the proportion (same as in Example 1), soaking, decocting, extracting, concentrating and freeze-drying to obtain a dry paste containing active ingredients;

[0099] a. Weigh the raw materials of Chinese medicine formula according to the proportion, and soak them in purified water at room temperature for 90 minutes according to the mass ratio of Chinese medicine formula raw materials to purified water of 1:7 times;

[0100] b. After soaking, extracting the medicinal liquid by water decoction method, the water decoction conditions are: decoction twice, boiling the mixture in step a, and then decocting for 50 minutes, filtering the decoction, decocting the residue twice, boiling for 4 minutes, filtering the decoction, mixing the two decoctions and filtering the residue to obtain the extract;

[0101] c. The extract obtained in step b is concentrated by vacuum concentration to obtain a concentrated solution; the pressure of the vacuum concentration method is -0.04~0.06MPa, and the temperature during concentration is 60°C; the concentrated solution has a liquid density of 1.5g / mL;

[0102] 2) mixing the above concentrate with other ingredients to form unsieved granules;

[0103] Among them, the granulation method uses fluidized bed wet granulation, the main fan temperature is 65°C, the fan frequency is 15Hz, the infusion pump speed is 5mL / min, and the atomization pressure is 1.9bar;

[0104] 3) The unscreened granules are sieved through a sieve to obtain the desired Chinese medicine granules;

[0105] Among them, the unscreened particles are screened through an upper sieve with a mesh size of 16 and a lower sieve with a mesh size of 35 to obtain the required Chinese medicine granules.

[0106] To further illustrate the beneficial effects of the present invention, the following comparative example is constructed taking Example 1 as an example.

[0107] Comparative Example 1: Preparation of a decoction having the same composition and content as in Example 1

[0108] Preparation of water decoction: Take the raw materials of Chinese medicine formula, including 30g of Rehmannia glutinosa, 24g of Cornus officinalis, 18g of Cyathula officinalis, 18g of Rhizoma Alismatis, 30g of Poria, 18g of Rhizoma Atractylodis Macrocephalae, 18g of Rhizoma Clematidis, 20g of Rhizoma Corydalis, and 12g of Radix Glycyrrhizae. Put the above ingredients into a sand pot, add 600ml of purified water, and boil to obtain 300ml of water decoction.

[0109] Comparative Example 2

[0110] In this comparative example, the microcrystalline cellulose in Example 1 was replaced with sucrose, and the proportion of each auxiliary material was adjusted. The other conditions were the same as those in Example 1. The specific ingredients are shown in Table 6.

[0111] Table 6

[0112]

[0113] Comparative Example 3

[0114] In this comparative example, the microcrystalline cellulose in Example 1 was replaced with starch, and the proportion of each auxiliary material was adjusted. The other conditions were the same as those in Example 1. The specific ingredients are shown in Table 7.

[0115] Table 7

[0116]

[0117] Comparative Example 4

[0118] A Chinese medicine granule, the formula of which is the same as that of Example 1;

[0119] The preparation method of the above-mentioned Chinese medicine granules is as follows, comprising:

[0120] 1) Weighing the raw materials of the Chinese medicine formula according to the proportion (same as in Example 1), soaking, decocting, extracting, concentrating and freeze-drying to obtain a dry paste containing active ingredients;

[0121] a. Weigh the raw materials of Chinese medicine formula according to the proportion, and soak them in purified water at room temperature for 90 minutes according to the mass ratio of Chinese medicine formula raw materials to purified water of 1:7 times;

[0122] b. After soaking, extracting the medicinal liquid by water decoction method, the water decoction conditions are: decoction twice, boiling the mixture in step a, and then decocting for 50 minutes, filtering the decoction, decocting the residue twice, boiling for 4 minutes, filtering the decoction, mixing the two decoctions and filtering the residue to obtain the extract;

[0123] c. The extract obtained in step b is concentrated by vacuum concentration to obtain a concentrated solution; the pressure of the vacuum concentration method is -0.04~0.06MPa, and the temperature during concentration is 60°C; the concentrated solution has a liquid density of 1.5g / mL;

[0124] 2) mixing the above concentrate with other ingredients to form unsieved granules;

[0125] Among them, the granulation method uses fluidized bed wet granulation, the main fan temperature is 58°C, the fan frequency is 15Hz, the infusion pump speed is 8mL / min, and the atomization pressure is 1.3bar;

[0126] 3) The unscreened granules are sieved through a sieve to obtain the desired Chinese medicine granules;

[0127] Among them, the unscreened particles are screened through an upper sieve with a mesh size of 16 and a lower sieve with a mesh size of 35 to obtain the required Chinese medicine granules.

[0128] Comparative Example 5

[0129] A Chinese medicine granule, the formula of which is the same as that of Example 1;

[0130] The preparation method of the above-mentioned Chinese medicine granules is as follows, comprising:

[0131] 1) Weighing the raw materials of the Chinese medicine formula according to the proportion (same as in Example 1), soaking, decocting, extracting, concentrating and freeze-drying to obtain a dry paste containing active ingredients;

[0132] a. Weigh the raw materials of Chinese medicine formula according to the proportion, and soak them in purified water at room temperature for 90 minutes according to the mass ratio of Chinese medicine formula raw materials to purified water of 1:7 times;

[0133] b. After soaking, extracting the medicinal liquid by water decoction method, the water decoction conditions are: decoction twice, boiling the mixture in step a, and then decocting for 50 minutes, filtering the decoction, decocting the residue twice, boiling for 4 minutes, filtering the decoction, mixing the two decoctions and filtering the residue to obtain the extract;

[0134] c. The extract obtained in step b is concentrated by vacuum concentration to obtain a concentrated solution; the pressure of the vacuum concentration method is -0.04~0.06MPa, and the temperature during concentration is 60°C; the concentrated solution has a liquid density of 1.5g / mL;

[0135] 2) mixing the above concentrate with other ingredients to form unsieved granules;

[0136] Among them, the granulation method uses fluidized bed wet granulation, the main fan temperature is 60°C, the fan frequency is 15Hz, the infusion pump speed is 8mL / min, and the atomization pressure is 1.4bar;

[0137] 3) The unscreened granules are sieved through a sieve to obtain the desired Chinese medicine granules;

[0138] Among them, the unscreened particles are screened through an upper sieve with a mesh size of 16 and a lower sieve with a mesh size of 35 to obtain the required Chinese medicine granules.

[0139] Experimental results:

[0140] 1. Moisture absorption test

[0141] According to the current pharmacopoeia guidelines, the hygroscopicity test was carried out on the Chinese herbal granules prepared in the above Examples 1-10 and Comparative Examples 1-5. A dry stoppered weighing bottle was placed in a 25°C constant temperature dryer 24 hours before the experiment. A saturated solution of ammonium chloride or ammonium sulfate was placed at the bottom. The temperature was set at 25±1°C and the relative humidity was 80%±2% in an artificial climate box. The weight was accurately weighed and m1 was measured. 10 g of the test sample was spread flat in the above weighing bottle and m2 was accurately weighed. The weighing bottle was opened and placed under constant temperature and humidity conditions for 24 hours together with the bottle cap. Then the weighing bottle cap was covered and the weight was measured and m3 was measured. The weight gain percentage was (m3-m2 / m2-m1)×100%. The specific results are shown in Table 8.

[0142] Table 8

[0143]

[0144] As can be seen from the table above, when microcrystalline cellulose is used as a filler, the sample has lower hygroscopicity, which can prevent the drug from absorbing moisture and becoming damp. When starch, sucrose and dextrin are used as fillers, the particles gain more weight, are hygroscopic, and the preparations are prone to agglomeration.

[0145] 2. Investigation of physical properties of granules under different preparation processes

[0146] Take 10 g of each of the Chinese medicine granules prepared in Example 1, Examples 7-8 and Comparative Examples 2-5, add 200 ml of hot water, stir for 5 minutes, and observe immediately. The results are shown in Table 9.

[0147] Table 9

[0148]

[0149] It can be seen from Table 9 that, by using the process parameters defined in Examples 1, 7, 8 and Comparative Examples 2 and 3 of the present invention, granulation can be relatively easy to obtain granules, and the granules have good solubility and are not easy to agglomerate. However, under the processes of Comparative Examples 4 and 5, when the temperature of the main fan is lowered, the water volatilization rate is too slow, resulting in the particles becoming larger and being easy to agglomerate. If the atomization pressure becomes low, the liquid medicine is not sprayed evenly, and it is also easy to agglomerate and difficult to granulate.

[0150] 3. Sustained release test

[0151] The release rate was determined by the slurry method specified in Part IV of the 2015 edition of the Chinese Pharmacopoeia, and the granules obtained in Examples 1-8 and Comparative Examples 2-5 were subjected to in vitro dissolution experiments in a pH = 6.8 potassium dihydrogen phosphate buffer. In the present invention, Rehmannia glutinosa and Cornus officinalis are the main drugs of the formula, and the rehmannia glutinosa glycoside D in Rehmannia glutinosa is used as the main test index. In the dissolution experiment, samples were taken at predetermined time points (0.5, 2, 4, 6, 8, and 10 hours), and the samples were detected by C18 column HPLC method to calculate the cumulative release rate (%). The cumulative release rate results are shown in FIG. Figure 1 As shown in Tables 10-11.

[0152] The dissolution of rehmannia glycoside D was tested at 0.5 h, 2 h, 4 h, 6 h, 8 h, and 10 h respectively to determine the release of the functional substances of the drug.

[0153] Table 10

[0154]

[0155] As can be seen from Table 10, Examples 1, 4, 5, 6, 7, and 8 have relatively ideal in vitro dissolution under the simulated gastrointestinal environment (pH 6.8), and the drug is released evenly and slowly within 0-8 hours. Examples 2 and 3 have a faster release rate within 0-2 hours, and the sustained release effect is not ideal.

[0156] Table 11

[0157]

[0158] It can be seen from Table 11 above that after adjusting the proportion of excipient components, the release rate of Comparative Examples 2 and 3 within 0-2h is faster, the drug sustained release performance is not ideal, and the drug release is incomplete. Comparative Examples 3 and 4 have a sustained release effect, but the drug release is incomplete, which is not conducive to the effective use of the drug.

[0159] 4. Pharmacokinetics

[0160] Test drugs: Chinese medicine granules of Example 1, decoction of Comparative Example 1;

[0161] Test animals: New Zealand white rabbits were made into knee arthritis models and randomly divided into 2 groups, 12 in the granule group and 12 in the decoction group;

[0162] Dosage: For the decoction group, 8 ml (kg*d) was administered orally 3 times a day; for the granule group, 10 mg was administered sublingually every day;

[0163] Determination parameters: Cmax--maximum plasma drug concentration; Tmax--time to reach maximum plasma drug concentration; AUC0-24--area under the 0-24 hour drug concentration-time curve, T1 / 2--half-life;

[0164] The measurement results are shown in Table 12.

[0165] Table 12

[0166]

[0167] According to Table 12 above, it can be seen that the granules of the present invention reach the maximum drug concentration in 5 hours, which is 4 hours later than the decoction, and the maximum drug plasma concentrations of the two are basically equal; the AUC0-24 of the granules of Example 1 is basically equivalent to that of the decoction, indicating that the granules can achieve substantially the same drug absorption degree as the decoction and have good bioavailability; the half-life of the granules is significantly longer than that of the decoction, and it can be seen that the granules of the present invention have a longer drug onset time.

[0168] 5. Biological experiments

[0169] In order to compare the effects of Chinese medicine granules in animal experiments, four groups were divided into a blank group (New Zealand white rabbits), a joint model group, a Chinese medicine granule group (Example 1) and a water decoction group. The experimental animals were healthy adult New Zealand white rabbits. The water decoction group was given the water decoction prepared in Comparative Example 1 after modeling, and gavage was given three times a day, 8 ml each time, and one week was a course of treatment; the Chinese medicine granule group was given the Chinese medicine granules prepared in Example 1 after modeling, mixed in animal feed, fed once a day, 10 mg each time, and one week was a course of treatment.

[0170] (1) Detection of bone marrow stem cell-related factors

[0171] Studies have confirmed that mobilizing autologous bone marrow stem cells (MSCs) can treat articular cartilage damage and degeneration. Studies have also shown that adenosine triphosphate binding transporter G2 (ABCG2), transforming growth factor (TGF) and bone morphogenetic protein (BMP) are widely present in stem cells. When stem cells are transformed into chondrocytes, the levels of these factors are downregulated. By detecting the expression of the ABCG2 gene and the levels of BMP and TGF, it can be seen that the drug regulates the expression of the above genes. The expression of the above factors was detected in the blank group, model group, decoction group and Chinese medicine granule group, as shown in Table 13, in ng / ml.

[0172] Table 13

[0173]

[0174] It can be seen from the above table that the expression level of the Chinese medicine granule group of the present invention is significantly lower than that of the model group, and it has an advantage in converting stem cells into chondrocytes.

[0175] (2) Matrix metalloproteinase detection

[0176] Studies have found that matrix metalloproteinases play an important role in cartilage degeneration in osteoarthritis. Normal chondrocytes can synthesize a large amount of type II collagen and proteoglycans and secrete them into the extracellular matrix, forming an extracellular fiber lattice structure to provide support and protection for chondrocytes. Under the action of matrix metalloproteinases, these fiber lattice structures will continue to degrade and eventually lead to articular cartilage degeneration. Overexpression of matrix metalloproteinases in cartilage tissue causes degradation of articular cartilage matrix and is the main cause of osteoarthritis in animals.

[0177] Table 14 MMP-1, MMP-3, MMP-13 test results (χ±s) Unit ng / ml

[0178]

[0179] The detection of matrix metalloproteinases MMP-1, MMP-3 and MMP-13 confirmed that the expression levels of MMP-3 and MMP-13 in the Chinese medicine granule group were lower than those in the water decoction group, and the expression levels were significantly lower than those in the model group; compared with the blank control group, P < 0.05, it can be seen that the granule group of the present invention effectively protects the matrix collagen of the bone joints.

[0180] (3) Pathological tissue observation

[0181] After killing rabbits in different experimental groups in batches, articular cartilage specimens were collected from the proximal tibia and distal femur on both sides, fixed with formaldehyde solution, decalcified with 10% neutral EDTA buffer, dehydrated routinely, and embedded in paraffin to make 4-5 micron pathological sections.

[0182] After conventional HE staining, it can be observed that Figure 2 shown.

[0183] Arthritis model group: cartilage surface was rough, chondrocytes were hyperplastic, hypertrophic, slightly disordered, and fuzzy tide lines were observed under the microscope. After the drug was applied, the TCM granule group was comparable to the blank control group, and the inflammatory symptoms were improved.

[0184] (4) Staining of type II collagen (CⅡ) in articular cartilage

[0185] CⅡ is an important component of articular cartilage and plays an important role in maintaining the elasticity and hardness of articular cartilage. When OA occurs, the matrix structure is destroyed and the content is significantly reduced, among which the destruction of CⅡ is the most obvious. Therefore, the change of CⅡ in the matrix is ​​an important indicator of the severity of arthritis. Immunohistochemistry of CⅡ in normal articular cartilage shows that extracellular CⅡ is evenly distributed and light brown. Immunohistochemistry was used to detect the expression of CⅡ in articular cartilage. According to Figure 3It can be seen that the entire layer of cartilage in the blank control group was uniformly stained blue, the surface and middle layers of the model group were not stained, and the calcification layer was stained darker; the Chinese medicine granule group was stained more evenly, with the surface layer slightly not stained; the decoction group had a surface layer not stained, and the remaining layers were unevenly colored.

[0186] In summary, the granules made from the Chinese medicine formula raw materials with a specific formula of the present invention have a therapeutic and protective effect on articular cartilage degeneration and damage. Since the granules have a certain sustained-release effect, the number of administrations is reduced, clinical administration will be more convenient, and the quality of life of patients will be improved.

[0187] The above is a preferred embodiment of the present invention. For ordinary technicians in this technical field, without departing from the principles of the present invention, making several improvements and modifications should also be regarded as within the protection scope of the present invention.

Claims

1. A method for preparing a Chinese medicine granule, characterized in that: include: 1) Weigh the raw materials of the Chinese medicine formula in proportion, and then soak, decoct, extract, concentrate or freeze-dry to obtain a concentrated solution or dry paste containing active ingredients; the dry matter mass of the obtained concentrated solution or dry paste is the extract of the raw materials of the Chinese medicine formula; 2) re-dissolving the above concentrate or dry paste with purified water, and mixing with fillers, disintegrants, sustained-release agents, and flavoring agents to prepare granules; Among them, the granulation method uses fluidized bed wet granulation, the main fan temperature is 63°C-65°C, the fan frequency is 15Hz, the infusion pump speed is 5-8mL / min, and the atomization pressure is 1.6-1.9bar; 3) Sieve the above granules through a sieve to obtain the desired Chinese medicine granules; The mesh number of the upper sieve of the sieve is 16-18 meshes, and the mesh number of the lower sieve is 30-35 meshes; The Chinese medicine granules are composed of the following components in weight percentage: 33.1-52.7% of Chinese medicine formula raw material extract, 27.4-46% of filler, 4-8.7% of disintegrant, 8-17% of sustained-release agent, and 2-4.7% of flavoring agent; Wherein, the filler is one or more of microcrystalline cellulose, dextrin, and lactose; the disintegrant is one or more of cross-linked polyvinyl pyrrolidone, sodium carboxymethyl starch, and low-substituted hydroxypropyl cellulose; the sustained-release agent is one or more of ethyl cellulose, polyethylene, and polypropylene; the flavoring agent is sodium saccharin and / or aspartame; The extract of the raw material of the Chinese medicine formula is a concentrated liquid or dry paste containing active ingredients obtained by soaking, decocting, extracting, concentrating or freeze-drying the raw material of the Chinese medicine formula; The Chinese medicine formula raw materials are composed of the following components by weight: 12-15 parts by weight of Rehmannia glutinosa; Cornus officinalis 9-12 parts by weight; 6-9 parts by weight of Cyathula capitata; Rhizoma Alismatis 6-9 parts by weight; Poria 12-15 parts by weight; Stir-fried Atractylodes macrocephala 6-9 parts by weight; 6-9 parts by weight of Qiannianjian; 6-9 parts by weight of Clematis chinensis; Corydalis yanhusuo 7-10 parts by weight; Licorice 3-6 parts by weight.

2. The method for preparing the Chinese medicine granules according to claim 1, characterized in that: The raw material extracts of the Chinese medicine formula account for 42.6% by weight, the filler accounts for 36.2%, the disintegrant accounts for 6%, the sustained-release agent accounts for 12%, and the flavoring agent accounts for 3.2%.

3. The method for preparing the Chinese medicine granules according to any one of claims 1-2, characterized in that: The filler is microcrystalline cellulose, the disintegrant is cross-linked polyvinyl pyrrolidone, the sustained-release agent is ethyl cellulose, and the flavoring agent is saccharin sodium.

4. The method for preparing the Chinese medicine granules according to claim 1, characterized in that: The step 1) is specifically as follows: a. Weigh the raw materials of Chinese medicine formula according to the proportion, then mix the raw materials of Chinese medicine formula with purified water at a mass ratio of 1:7-8, and soak at room temperature for 90-150 minutes; b. After soaking, extracting the medicinal liquid by water decoction method, the water decoction method conditions are as follows: boil the mixture in step a, then decoct for 50-70 minutes, filter and collect the decoction liquid; place the filter residue in the same amount of water for a second decoction, boil for 35-40 minutes, filter and collect the decoction liquid; mix the two decoctions and filter the residue to obtain the extract; c. The extract obtained in step b is concentrated by vacuum concentration to obtain a concentrated solution or a dry paste.

5. The method for preparing the Chinese medicine granules according to claim 4, characterized in that: In the step c, the pressure of the reduced pressure concentration method is -0.04~0.06MPa and the temperature is 60°C.

6. The method for preparing the Chinese medicine granules according to claim 5, characterized in that: In step c, the dry paste is obtained by freeze-drying the concentrated liquid; The freeze-drying conditions are as follows: the concentrated liquid is cooled to about 5°C and maintained for 1 hour, then cooled to -40°C and maintained for 2 hours; then heated to -20°C, vacuum degree 20Pa, heating rate 0.4°C / min, and maintained for 20 hours; heated to zero again, vacuum degree unchanged, heating rate unchanged, and maintained for 5 hours; then heated to 40°C, vacuum degree 10Pa, heating rate 1°C / min, and maintained for 4 hours.

7. The method for preparing the Chinese medicine granules according to claim 1, characterized in that: In the step 1), the density of the concentrated solution is 1.04-1.08 g / mL, and the density of the dry paste after being reconstituted with purified water is 1.04-1.08 g / mL.

Images