A composite traditional Chinese medicine preparation for chronic osteomyelitis and its preparation method

Abstract

The invention discloses a composite Chinese medicine preparation for chronic osteomyelitis, and its raw materials include: Wuwei Xiaoduyin extractant, sodium alginate, p-toluenesulfonic acid, cyclodextrin, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, emulsifier, chitosan, sodium chloride, calcium chloride; the mass ratio of Wuwei Xiaoduyin extractant, sodium alginate, p-toluenesulfonic acid, cyclodextrin, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, emulsifier, chitosan, sodium chloride, and calcium chloride is 10-20: 1-3: 0.1-0.2: 2-8: 0.01-0.1: 0.1-0.2: 1-2: 0.1-0.2: 0.1-0.2. The invention also discloses a preparation method of the composite Chinese medicine preparation for chronic osteomyelitis. The present invention can not only solve the disadvantage that antibiotics are difficult to penetrate into the bone to form an effective antibacterial concentration, but also the method is easy to implement, simple to prepare, and convenient to use.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine preparations for chronic osteomyelitis, and in particular to a composite traditional Chinese medicine preparation for chronic osteomyelitis and a preparation method thereof. Background Art

[0002] Osteomyelitis is a serious bone disease caused by persistent infection with pathogens and is usually divided into acute or chronic osteomyelitis. Chronic osteomyelitis is characterized by the formation of septa and a series of inflammatory reactions. The common treatments for chronic osteomyelitis are surgery and antibiotics. Since antibiotics are difficult to penetrate into the affected area and form effective antibacterial concentrations locally, clinical treatment of osteomyelitis often requires systemic high-dose, long-term use of antibiotics for 4 to 6 weeks. Long-term intravenous use of excessively high doses will increase the systemic toxic side effects of antibiotics and may cause antibiotic resistance. Therefore, there is an urgent need to explore new methods for the treatment of chronic osteomyelitis.

[0003] Wuwei Xiaoduyin is a traditional Chinese medicine decoction, which is often used to treat suppurative skin infections. The decoction contains five Chinese medicinal herbs: honeysuckle, wild chrysanthemum, dandelion, Viola yedoensis, and Gynura odoratum. Previous studies have shown that Wuwei Xiaoduyin has anti-inflammatory and antibacterial effects. Chinese medicinal raw materials still contain a variety of functional ingredients, and pharmacological activity evaluation shows that its alkaloid components have significant broad-spectrum antibacterial effects, and have a strong inhibitory effect on pathogenic bacteria such as Staphylococcus aureus and Shigella dysenteriae. It also has a variety of physiological activities such as obvious anti-inflammatory and antipyretic effects, and has the characteristics of mild, long-lasting effects, and small toxic and side effects. Its mechanism of action is characterized by multi-component, multi-pathway, and multi-effect characteristics and advantages.

[0004] At present, the use of Wuwei Xiaoduyin for chronic osteomyelitis has excellent research prospects. Existing studies have shown that Wuwei Xiaoduyin can prolong the inhibition time of STAT5 phosphorylation and inhibit the activation of STAT5-Foxp3 signaling pathway by downregulating the expression of IL-2, which further inhibits the immune response of Treg cells, thereby playing a therapeutic role in chronic osteomyelitis. However, the current medicinal drink not only has a bitter taste, but is also very susceptible to environmental influences and deterioration, and eventually loses its biological activity, which greatly limits its application in the field of chronic osteomyelitis. Summary of the invention

[0005] Based on the technical problems existing in the background technology, the present invention proposes a composite Chinese medicine preparation for chronic osteomyelitis and a preparation method thereof.

[0006] The invention provides a composite Chinese medicine preparation for chronic osteomyelitis, wherein the raw materials include: a Wuwei Xiaoduyin extractant, sodium alginate, p-toluenesulfonic acid, cyclodextrin, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, an emulsifier, chitosan, sodium chloride and calcium chloride; the mass ratio of the Wuwei Xiaoduyin extractant, sodium alginate, p-toluenesulfonic acid, cyclodextrin, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, an emulsifier, chitosan, sodium chloride and calcium chloride is 10-20: 1-3: 0.1-0.2: 2-8: 0.01-0.1: 0.1-0.2: 1-2: 0.1-0.2: 0.1-0.2.

[0007] Preferably, the concentration of the Wuwei Xiaoduyin extractant is 1.5-3 g / mL.

[0008] Preferably, the Wuwei Xiaodu Yin extract is prepared by the following specific steps: adding honeysuckle, chrysanthemum, dandelion, Viola yedoensis, and Gynura odorifera to water and soaking them for 1-5 hours, then boiling and concentrating them to obtain the extract.

[0009] Honeysuckle, also known as honeysuckle, is the flower bud of the perennial semi-evergreen twining woody vine of the Caprifoliaceae family. It is picked in early summer when the flowers are in bud and not yet opened, and dried in the shade. It can be used raw, fried or made into a dew. It tastes sweet and is cold in nature. It enters the lung, heart and stomach meridians. It has the effects of clearing away heat and detoxifying, and evacuating wind-heat. It is mainly used to treat carbuncle, furuncle, redness, swelling, heat and pain, exogenous wind-heat, early stage of febrile disease, heat-toxic bloody diarrhea, summer heat and thirst, and sore throat. This product has a broad-spectrum antibacterial effect, and has a strong inhibitory effect on pathogenic bacteria such as Staphylococcus aureus and Shigella dysenteriae; it has obvious anti-inflammatory and antipyretic effects; the water and wine extracts of honeysuckle have obvious cytotoxic effects on sarcoma 180 and Ehrlich ascites tumor. This product has a certain effect on lowering cholesterol.

[0010] Chrysanthemum is the flower head of the chrysanthemum plant and its variants in the Asteraceae family. It tastes sweet and bitter, and is slightly cold in nature. It enters the lung and liver meridians. It can dispel wind and heat, soothe the liver and improve eyesight. It is used for wind-heat colds, headaches and dizziness, red and swollen eyes, and blurred vision. It has a sedative and antipyretic effect. It has an inhibitory effect on Staphylococcus aureus, Streptococcus B, Shigella dysenteriae, Salmonella typhi, Salmonella paratyphi, Escherichia coli, Pseudomonas aeruginosa, human tuberculosis and influenza viruses. It can significantly dilate the coronary arteries and increase blood flow.

[0011] Dandelion is a perennial herb belonging to the Asteraceae family. It is a plant used for both medicine and food. Dig it up with roots before or just after flowering in spring and summer, remove the soil, and dry it in the sun. Pick out impurities, wash off the soil, cut it into sections, and dry it in the sun. Pick out the original medicine, wash off the mud with water, take it out and spread it out to dry, cut it into 1-1.5mm sections, dry it in the sun, and sieve out the ash. It tastes sweet and bitter, and is cold in nature. It enters the stomach meridian and liver meridian. It has the effects of clearing away heat and detoxifying, reducing swelling and dispersing knots, and promoting diuresis and relieving stranguria. It is used for carbuncle swelling and poison, breast carbuncle, scrofula, red eyes, sore throat, lung carbuncle, intestinal carbuncle, damp-heat jaundice, and hot stranguria. It has a strong killing effect on Staphylococcus aureus and hemolytic streptococci; it also has a certain killing effect on Pneumococcus, meningococcus, diphtheria bacillus, Pseudomonas aeruginosa, Proteus, Shigella dysenteriae, Salmonella typhi and catarrhalis; it also has an inhibitory effect on certain viruses, fungi and leptospira. In addition, it has the effects of choleretic, diuretic, bitter stomachic, mild laxative, etc.

[0012] Viola yedoensis is the whole herb of Viola yedoensis, a plant of the Viola yedoensis family. It is harvested in spring and autumn, removed of impurities, washed and dried. It tastes bitter and pungent, and is cold in nature. It enters the heart and liver meridians. It can clear away heat and detoxify, cool blood and reduce swelling. It is mainly used to treat jaundice, dysentery, mastitis, red and swollen eyes, pharyngitis; it can be applied externally to treat injuries from falls, carbuncle, snake bites, etc.

[0013] Purple-backed Gynura is a perennial stemless herbaceous plant of the genus Begonia in the Begoniaceae family. It is endemic to China. It tastes sweet, slightly sour, and cool in nature. It has the effects of clearing away heat and detoxifying, moistening the lungs and relieving cough, dispersing blood stasis and reducing swelling, and promoting body fluid and quenching thirst. It is mainly used to treat exogenous high fever, heatstroke, cough due to lung heat, hoarseness due to cold, carbuncle, sore, bruise, swelling and pain, etc. Its tender stems and leaves are rich in calcium, iron, etc., with high nutritional value, and have medicinal value such as clearing away heat and detoxifying, and preventing the growth of malignant cells.

[0014] Preferably, the HLB value of the emulsifier is 10, and the emulsifier includes: Span-80 and Tween-60.

[0015] Preferably, the mass ratio of Span-80 to Tween-60 is 10-12:12-14.

[0016] Preferably, the degree of deacetylation of chitosan is 85-95%.

[0017] Preferably, the cyclodextrin is β-cyclodextrin.

[0018] Preferably, the viscosity-average molecular weight of the sodium alginate is 430 kDa.

[0019] The preparation method of the above-mentioned composite Chinese medicine preparation for chronic osteomyelitis comprises the following steps:

[0020] (1) dissolving sodium alginate in dimethylformamide, stirring at 50-60° C., adding dried p-toluenesulfonic acid, stirring for 1-2 hours, adding cyclodextrin, stirring for 1-3 hours, adding 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, washing, drying, adding water, stirring at 50-60° C. for 10-20 minutes, and cooling to room temperature to obtain a pretreated cyclodextrin solution;

[0021] (2) adding the Wuwei Xiaoduyin extract and the emulsifier to the pretreated cyclodextrin solution, stirring at 40-50° C. for 5-15 min, and homogenizing at 10000-15000 r / min for 1-4 min to obtain a single-layer coated emulsion;

[0022] (3) Dissolve chitosan in acetic acid solution, add sodium chloride thereto and stir evenly, adjust the stirring speed to 1000-1500 r / min, add the monolayer coating emulsion thereto at 20-40° C. and continue stirring for 10-20 min, adjust the pH value of the system to 6.0-6.4, add calcium chloride, stir at 1-5° C. for 1-2 h, centrifuge for 5-10 min, collect the precipitate, and freeze-dry to obtain a composite Chinese medicine preparation for chronic osteomyelitis.

[0023] Preferably, the drying temperature of toluenesulfonic acid is 105-120° C., the drying time is 20-60 min, and the vacuum degree is maintained at 10-20 kPa during the drying process.

[0024] The technical effects of the present invention are as follows:

[0025] The present invention adopts cyclodextrin and sodium alginate to compound and pre-treat the product, and the product has extremely high water solubility, and the larger hydrophobic cavity can complex the components of the Wuwei Xiaoduyin extract with larger molecular weight, effectively improving the stability of the Wuwei Xiaoduyin extract, and even if the cyclodextrin is pre-treated at high temperature, the Wuwei Xiaoduyin extract can be stably wrapped, thereby improving its thermal stability and effectively preventing the Wuwei Xiaoduyin extract from deteriorating or evaporating. The single-layer coated emulsion is further used as a template, and the chitosan wall material is further coated on the surface through self-assembly. The pre-treated cyclodextrin has excellent affinity with chitosan, which can effectively enhance the adsorption strength of chitosan on the surface of the single-layer coated emulsion, and compounded with the sodium alginate on the surface of the single-layer coated emulsion, and solidified by calcium chloride, not only the product structure size is uniform, but also the coating strength of the content is extremely high, and the storage stability is excellent.

[0026] The present invention can not only solve the disadvantage that antibiotics are difficult to penetrate into the local bone to form an effective antibacterial concentration, but also avoid the disadvantage that high-dose and long-term intravenous application of antibiotics are required for the clinical treatment of chronic osteomyelitis. At the same time, the method is easy to implement, the preparation is simple, the administration is convenient, the treatment effect is significant, the safety is high, the systemic toxicity is small, and the drug effect lasts for a long time. The present invention has good biocompatibility, in the in vivo drug release experiment, the "burst release" effect is not obvious, the sustained release effect is excellent, and it has an excellent treatment effect for chronic osteomyelitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 The particle size distribution diagram of the pretreated cyclodextrin solution obtained in step (1) in Example 5, the single-layer coated emulsion obtained in step (2) and the composite Chinese medicine preparation for chronic osteomyelitis obtained in step (3) is shown.

[0028] Figure 2 This is a comparison chart of the cumulative release rates of the composite Chinese medicine preparations for chronic osteomyelitis obtained in Example 5 and Comparative Example 1 at different storage temperatures.

[0029] Figure 3 The thermogravimetric analysis diagrams of the composite Chinese medicine preparation for chronic osteomyelitis obtained in Comparative Example 1 and the Wuwei Xiaodu Yin extract obtained in Comparative Example 2 are shown.

[0030] Figure 4 It is a cumulative release curve diagram of the composite Chinese medicine preparation for chronic osteomyelitis obtained in Example 5 and Comparative Example 1, and the Wuwei Xiaodu Yin extract obtained in Comparative Example 2 in the digestive system. DETAILED DESCRIPTION

[0031] The technical solution of the present invention is described in detail below through specific embodiments.

[0032] Example 1

[0033] A composite Chinese medicine preparation for chronic osteomyelitis, the raw materials of which include: 10kg of Wuwei Xiaoduyin extractant, 1kg of sodium alginate with a viscosity-average molecular weight of 430kDa, 0.1kg of p-toluenesulfonic acid, 2kg of beta-cyclodextrin, 0.01kg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 0.1kg of emulsifier, 1kg of chitosan with a deacetylation degree of 85%, 0.1kg of sodium chloride and 0.1kg of calcium chloride.

[0034] The extract of Wuwei Xiaoduyin is prepared by the following specific steps: 5kg of honeysuckle, 5kg of chrysanthemum, 5kg of dandelion, 5kg of Viola yedoensis, and 5kg of Gynura purpurea are added into 30kg of water and soaked for 1 hour, then boiled and concentrated to a concentration of 1.5g / mL. The HLB value of the emulsifier is 10, and the emulsifier is composed of Span-80 and Tween-60 in a mass ratio of 5:6.

[0035] The preparation method of the above-mentioned composite Chinese medicine preparation for chronic osteomyelitis comprises the following steps:

[0036] (1) Sodium alginate was dissolved in 10 kg of dimethylformamide, stirred at 50° C., dry p-toluenesulfonic acid was added, stirring was continued for 1 h, cyclodextrin was added, stirring was continued for 1 h, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride was added, stirring was continued for 1 h, washed with anhydrous ethanol, dried, added to 10 kg of water, stirred at 50° C. for 10 min, and cooled to room temperature to obtain a pretreated cyclodextrin solution;

[0037] (2) adding the Wuwei Xiaoduyin extract and the emulsifier to the pretreated cyclodextrin solution, stirring at 40°C for 5 minutes, homogenizing for 1 minute, and homogenizing at a speed of 10,000 r / min to obtain a single-layer coated emulsion;

[0038] (3) Chitosan was dissolved in 10 kg of 1% acetic acid solution, sodium chloride was added thereto and stirred evenly, a single-layer coating emulsion was added thereto at a temperature of 20°C and a stirring speed of 1000 r / min, stirring was continued for 10 min, the pH value of the system was adjusted to 6.0-6.4, calcium chloride was added, stirred at a temperature of 1°C for 1 h, centrifuged at a speed of 3000 r / min for 5 min, the precipitate was collected, and freeze-dried to obtain a composite Chinese medicine preparation for chronic osteomyelitis.

[0039] Example 2

[0040] A composite Chinese medicine preparation for chronic osteomyelitis, the raw materials of which include: 20kg of Wuwei Xiaoduyin extractant, 3kg of sodium alginate with a viscosity-average molecular weight of 430kDa, 0.2kg of p-toluenesulfonic acid, 8kg of beta-cyclodextrin, 0.1kg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 0.2kg of emulsifier, 2kg of chitosan with a deacetylation degree of 95%, 0.2kg of sodium chloride, and 0.2kg of calcium chloride.

[0041] The extract of Wuwei Xiaoduyin is prepared by the following specific steps: 10kg honeysuckle, 1kg chrysanthemum, 1kg dandelion, 1kg Viola yedoensis, and 1kg Gynura purpurea are added into 60kg water and soaked for 5h, then boiled and concentrated to a concentration of 3g / mL. The HLB value of the emulsifier is 10, and the emulsifier is composed of Span-80 and Tween-60 in a mass ratio of 6:7.

[0042] The preparation method of the above-mentioned composite Chinese medicine preparation for chronic osteomyelitis comprises the following steps:

[0043] (1) Sodium alginate is dissolved in 20 kg of dimethylformamide, stirred at 60° C., dry p-toluenesulfonic acid is added, stirring is continued for 2 h, cyclodextrin is added, stirring is continued for 3 h, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is added, stirring is continued for 3 h, washed with anhydrous ethanol, dried, added into 20 kg of water, stirred at 60° C. for 20 min, and cooled to room temperature to obtain a pretreated cyclodextrin solution;

[0044] (2) adding the Wuwei Xiaoduyin extract and the emulsifier to the pretreated cyclodextrin solution, stirring at 50°C for 15 minutes, homogenizing for 4 minutes at a homogenizing speed of 15000 r / min, and obtaining a single-layer coated emulsion;

[0045] (3) Chitosan was dissolved in 20 kg of 2% acetic acid solution, sodium chloride was added thereto and stirred evenly, a single-layer coating emulsion was added thereto at a temperature of 40°C and a stirring speed of 1500 r / min, stirring was continued for 20 min, the pH value of the system was adjusted to 6.0-6.4, calcium chloride was added, stirring was performed at a temperature of 5°C for 2 h, centrifuged at a speed of 4000 r / min for 10 min, the precipitate was collected, and freeze-dried to obtain a composite Chinese medicine preparation for chronic osteomyelitis.

[0046] Example 3

[0047] A composite Chinese medicine preparation for chronic osteomyelitis, the raw materials of which include: 13kg of Wuwei Xiaoduyin extractant, 2.5kg of sodium alginate with a viscosity-average molecular weight of 430kDa, 0.13kg of p-toluenesulfonic acid, 6kg of beta-cyclodextrin, 0.02kg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 0.18kg of emulsifier, 1.7kg of chitosan with a deacetylation degree of 88%, 0.12kg of sodium chloride, and 0.18kg of calcium chloride.

[0048] The extract of Wuwei Xiaoduyin was prepared by the following specific steps: 6kg honeysuckle, 4kg chrysanthemum, 2kg dandelion, 4kg Viola yedoensis, and 2kg Gynura purpurea were added into 50kg water and soaked for 2h, then boiled and concentrated to a concentration of 2.6g / mL. The HLB value of the emulsifier was 10, and the emulsifier was composed of Span-80 and Tween-60 in a mass ratio of 7:9.

[0049] The preparation method of the above-mentioned composite Chinese medicine preparation for chronic osteomyelitis comprises the following steps:

[0050] (1) Sodium alginate was dissolved in 18 kg of dimethylformamide, stirred at 52° C., dry p-toluenesulfonic acid was added, stirring was continued for 100 min, cyclodextrin was added and stirred, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride was added and stirring was continued for 1.5 h, washed with anhydrous ethanol, dried, added to 17 kg of water, stirred at 52° C. for 17 min, and cooled to room temperature to obtain a pretreated cyclodextrin solution;

[0051] (2) adding the Wuwei Xiaoduyin extract and the emulsifier to the pretreated cyclodextrin solution, stirring at 42°C for 12 minutes, homogenizing for 2 minutes at a homogenizing speed of 13000 r / min, and obtaining a single-layer coated emulsion;

[0052] (3) Chitosan was dissolved in 12 kg of 1.5% acetic acid solution, sodium chloride was added thereto and stirred evenly, the monolayer coating emulsion was added thereto at a temperature of 25°C and a stirring speed of 1400 r / min, stirring was continued for 13 min, the pH value of the system was adjusted to 6.0-6.4, calcium chloride was added, stirred at a temperature of 4°C for 1.3 h, centrifuged at a speed of 3700 r / min for 7 min, the precipitate was collected, and freeze-dried to obtain a composite Chinese medicine preparation for chronic osteomyelitis.

[0053] Example 4

[0054] A composite Chinese medicine preparation for chronic osteomyelitis, the raw materials of which include: 17kg of Wuwei Xiaoduyin extractant, 1.5kg of sodium alginate with a viscosity-average molecular weight of 430kDa, 0.17kg of p-toluenesulfonic acid, 4kg of beta-cyclodextrin, 0.08kg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 0.12kg of emulsifier, 1.3kg of chitosan with a deacetylation degree of 92%, 0.16kg of sodium chloride, and 0.14kg of calcium chloride.

[0055] The extract of Wuwei Xiaoduyin is prepared by the following specific steps: 8kg honeysuckle, 2kg chrysanthemum, 4kg dandelion, 2kg Viola yedoensis, and 4kg Gynura purpurea are added into 40kg water and soaked for 4h, then boiled and concentrated to a concentration of 2g / mL. The HLB value of the emulsifier is 10, and the emulsifier is composed of Span-80 and Tween-60 in a mass ratio of 1:1.

[0056] The preparation method of the above-mentioned composite Chinese medicine preparation for chronic osteomyelitis comprises the following steps:

[0057] (1) Sodium alginate was dissolved in 18 kg of dimethylformamide, stirred at 52° C., dry p-toluenesulfonic acid was added, stirring was continued for 100 min, cyclodextrin was added and stirred, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride was added and stirring was continued for 1.5 h, washed with anhydrous ethanol, dried, added to 17 kg of water, stirred at 52° C. for 17 min, and cooled to room temperature to obtain a pretreated cyclodextrin solution;

[0058] (2) adding the Wuwei Xiaoduyin extract and the emulsifier to the pretreated cyclodextrin solution, stirring at 42°C for 12 minutes, homogenizing for 2 minutes at a homogenizing speed of 13000 r / min, and obtaining a single-layer coated emulsion;

[0059] (3) Chitosan was dissolved in 12 kg of 1.5% acetic acid solution, sodium chloride was added thereto and stirred evenly, the monolayer coating emulsion was added thereto at a temperature of 25°C and a stirring speed of 1400 r / min, stirring was continued for 13 min, the pH value of the system was adjusted to 6.0-6.4, calcium chloride was added, stirred at a temperature of 4°C for 1.3 h, centrifuged at a speed of 3700 r / min for 7 min, the precipitate was collected, and freeze-dried to obtain a composite Chinese medicine preparation for chronic osteomyelitis.

[0060] Example 5

[0061] A composite Chinese medicine preparation for chronic osteomyelitis, the raw materials of which include: 15kg of Wuwei Xiaoduyin extractant, 2kg of sodium alginate with a viscosity-average molecular weight of 430kDa, 0.15kg of p-toluenesulfonic acid, 5kg of beta-cyclodextrin, 0.05kg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 0.15kg of emulsifier, 1.5kg of chitosan with a deacetylation degree of 90%, 0.14kg of sodium chloride, and 0.16kg of calcium chloride.

[0062] The extract of Wuwei Xiaoduyin was prepared by the following specific steps: 6kg honeysuckle, 3kg chrysanthemum, 3kg dandelion, 3kg Viola yedoensis, and 3kg Gynura purpurea were added into 45kg water and soaked for 3h, then boiled and concentrated to a concentration of 2.2g / mL. The HLB value of the emulsifier was 10, and the emulsifier was composed of Span-80 and Tween-60 in a mass ratio of 11:12.5.

[0063] The preparation method of the above-mentioned composite Chinese medicine preparation for chronic osteomyelitis comprises the following steps:

[0064] (1) Sodium alginate was dissolved in 15 kg of dimethylformamide, stirred at 55° C., dry p-toluenesulfonic acid was added, stirring was continued for 90 min, cyclodextrin was added, stirring was continued for 2 h, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride was added, and stirring was continued for 2 h, washed with anhydrous ethanol, dried, added to 15 kg of water, stirred at 55° C. for 15 min, and cooled to room temperature to obtain a pretreated cyclodextrin solution;

[0065] (2) adding the Wuwei Xiaoduyin extract and the emulsifier to the pretreated cyclodextrin solution, stirring at 45°C for 10 minutes, homogenizing for 2.5 minutes at a homogenizing speed of 12500 r / min, and obtaining a single-layer coated emulsion;

[0066] (3) Chitosan was dissolved in 15 kg of 1.3% acetic acid solution, sodium chloride was added thereto and stirred evenly, a single-layer coating emulsion was added thereto at a temperature of 30° C. and a stirring speed of 1300 r / min, stirring was continued for 15 min, the pH value of the system was adjusted to 6.0-6.4, calcium chloride was added, stirred at a temperature of 3° C. for 1.5 h, centrifuged at a speed of 3500 r / min for 8 min, the precipitate was collected, and freeze-dried to obtain a composite Chinese medicine preparation for chronic osteomyelitis.

[0067] 1 mL of the pretreated cyclodextrin solution obtained in step (1), the single-layer coated emulsion obtained in step (2), and the composite Chinese medicine preparation for chronic osteomyelitis obtained in step (3) were respectively taken with a pipette and transferred into a test tube. Distilled water was added to dilute the mixture 10 times. After shaking, 1 mL was added into a polystyrene sample pool. The particle size and distribution of each sample were detected by a Malvern laser particle size analyzer. The detection temperature was 25°C, the dispersed phase was water, and each sample was measured three times to obtain an average value.

[0068] like Figure 1 As shown, from left to right are the pre-treated cyclodextrin solution obtained in step (1), the single-layer coated emulsion obtained in step (2), and the composite Chinese medicine preparation for chronic osteomyelitis obtained in step (3). The single-layer coated emulsion obtained in step (2) is filled with the extract of Wuwei Xiaoduyin in the hydrophobic cavity of the pre-treated cyclodextrin obtained in step (1), so that the particle size of the single-layer coated emulsion obtained in step (2) is larger than that of the pre-treated cyclodextrin obtained in step (1), but the difference is not large. The composite Chinese medicine preparation for chronic osteomyelitis obtained in step (3) is obtained by further embedding the single-layer coated emulsion obtained in step (2), so that its particle size is significantly increased.

[0069] The three curves are all similar to normal distribution, while the particle size range of the composite Chinese medicine preparation for chronic osteomyelitis obtained in step (3) is more concentrated, with an average particle size of 137 nm, and 85% distributed between 50 nm and 260 nm.

[0070] Comparative Example 1

[0071] A composite Chinese medicine preparation for chronic osteomyelitis, the raw materials of which include: 15 kg of Wuwei Xiaoduyin extractant, 2 kg of sodium alginate with a viscosity-average molecular weight of 430 kDa, 0.15 kg of p-toluenesulfonic acid, 5 kg of beta-cyclodextrin, 0.05 kg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 0.15 kg of emulsifier.

[0072] The extract of Wuwei Xiaoduyin was prepared by the following specific steps: 6kg honeysuckle, 3kg chrysanthemum, 3kg dandelion, 3kg Viola yedoensis, and 3kg Gynura purpurea were added into 45kg water and soaked for 3h, then boiled and concentrated to a concentration of 2.2g / mL. The HLB value of the emulsifier was 10, and the emulsifier was composed of Span-80 and Tween-60 in a mass ratio of 11:12.5.

[0073] The preparation method of the above-mentioned composite Chinese medicine preparation for chronic osteomyelitis comprises the following steps:

[0074] (1) Sodium alginate was dissolved in 15 kg of dimethylformamide, stirred at 55° C., dry p-toluenesulfonic acid was added, stirring was continued for 90 min, cyclodextrin was added, stirring was continued for 2 h, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride was added, and stirring was continued for 2 h, washed with anhydrous ethanol, dried, added to 15 kg of water, stirred at 55° C. for 15 min, and cooled to room temperature to obtain a pretreated cyclodextrin solution;

[0075] (2) adding the Wuwei Xiaoduyin extract and the emulsifier to the pretreated cyclodextrin solution, stirring at 45°C for 10 minutes, homogenizing for 2.5 minutes at a homogenizing speed of 12500 r / min, and obtaining a single-layer coated emulsion;

[0076] (3) The single-layer coated emulsion is washed with anhydrous ethanol and freeze-dried to obtain a composite Chinese medicine preparation for chronic osteomyelitis.

[0077] Accurately weigh 1.0 g of each of the composite Chinese medicine preparations for chronic osteomyelitis obtained in Example 5 and Comparative Example 1, and then place them at 4°C (refrigerated storage temperature) and 25°C (normal temperature storage temperature) for constant temperature storage, respectively. Take them out on the 5th, 10th, 15th, 20th, 25th, 30th, and 60th days, measure them by UPLC, record the peak area, and calculate the cumulative release rate.

[0078] like Figure 2As shown, with the extension of storage time, the cumulative release rate of each group gradually increased, and the release rate gradually slowed down; while the cumulative release rate of Example 5 was always lower than that of Comparative Example 1 under the same conditions. This is because the present invention uses a single-layer coated latex as a template, and further coats the chitosan wall material on the surface through self-assembly, and the pretreated cyclodextrin has excellent affinity with chitosan, which can effectively enhance the adsorption strength of chitosan on the surface of the single-layer coated latex, and is compounded with sodium alginate on the surface of the single-layer coated latex, and solidified by calcium chloride, and has excellent storage stability.

[0079] Comparative Example 2

[0080] The Wuwei Xiaoduyin extract was used as the product of this comparative example.

[0081] The Wuwei Xiaoduyin extract is prepared by the following specific steps: 6kg of honeysuckle, 3kg of chrysanthemum, 3kg of dandelion, 3kg of Viola yedoensis, and 3kg of Gynura purpurea are added into 45kg of water and soaked for 3h, then boiled and concentrated to a concentration of 2.2g / mL.

[0082] The thermodynamic properties of the composite Chinese medicine preparation for chronic osteomyelitis obtained in Comparative Example 1 and the Wuwei Xiaodu Yin extract obtained in Comparative Example 2 were analyzed using a Netzsch comprehensive thermal analyzer. Under a nitrogen flow of 50 mL / min, the temperature was increased from 50°C to 580°C at a rate of 15°C / min.

[0083] like Figure 3 As shown, the Wuwei Xiaodu Drink extract decomposes at 40-150°C, while the decomposition temperature of the product of Comparative Example 1 is 300-400°C. By comparing the two curves, it can be found that the product of the present invention, after compounding cyclodextrin and sodium alginate and pre-treating, has extremely high water solubility, and its larger hydrophobic cavity can complex the Wuwei Xiaodu Drink extract components with larger molecular weight, effectively improving the stability of the Wuwei Xiaodu Drink extract. Even if the cyclodextrin is pre-treated at high temperature, the Wuwei Xiaodu Drink extract can be stably encapsulated, thereby improving its thermal stability and effectively preventing the evaporation loss of the Wuwei Xiaodu Drink extract.

[0084] The composite Chinese medicine preparation for chronic osteomyelitis obtained in Example 5 and Comparative Example 1 and the Wuwei Xiaoduyin extract obtained in Comparative Example 2 were weighed (ensuring that the content of the Wuwei Xiaoduyin extract in each group was 1 g), first placed in artificial gastric juice, and allowed to stand at 36°C for 2 hours. After filtering, the filter cake was placed in artificial intestinal juice and allowed to stand at 36°C for 4 hours. During the test, 2 mL of supernatant was taken out every 1 hour, and 2 mL of fresh artificial gastric juice or fresh artificial intestinal juice was added. The supernatant was filtered through a 0.22 μm filter membrane, and UPLC was used for determination. The peak area was recorded and the cumulative release rate was calculated.

[0085] like Figure 4 As shown, the release effect of Example 5 is the best.

[0086] The Chinese herbal medicine used in the Wuwei Xiaoduyin extract is bitter and cold, and its active ingredients are easily neutralized in an acidic environment, resulting in a significant loss of drug activity. Comparative Example 1 uses cyclodextrin as a carrier raw material to embed and fill the Wuwei Xiaoduyin extract ingredients in its hydrophobic cavity, but cyclodextrin is hydrolyzed in the acidic environment of artificial gastric juice, resulting in the Wuwei Xiaoduyin extract being completely released in the gastric juice, and the drug efficacy is destroyed.

[0087] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A composite Chinese medicine preparation for chronic osteomyelitis, characterized in that: The raw materials include: Wuwei Xiaoduyin extract, sodium alginate, p-toluenesulfonic acid, cyclodextrin, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, emulsifier, chitosan, sodium chloride, and calcium chloride; the mass ratio of Wuwei Xiaoduyin extract, sodium alginate, p-toluenesulfonic acid, cyclodextrin, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, emulsifier, chitosan, sodium chloride, and calcium chloride is 10-20: 1-3: 0.1-0.2: 2-8: 0.01-0.1: 0.1-0.2: 1-2: 0.1-0.2: 0.1-0.2; The concentration of Wuwei Xiaoduyin extract is 1.5-3g / mL; The extract of Wuwei Xiaoduyin is prepared by the following specific steps: adding honeysuckle, chrysanthemum, dandelion, Viola yedoensis and Gynura odorifera into water and soaking them for 1-5 hours, then boiling and concentrating them to obtain the extract.

2. The composite Chinese medicine preparation for chronic osteomyelitis according to claim 1, characterized in that: The HLB value of the emulsifier is 10, and the emulsifiers include: Span-80 and Tween-60.

3. The composite Chinese medicine preparation for chronic osteomyelitis according to claim 2, characterized in that: The mass ratio of Span-80 to Tween-60 is 10-12:12-14.

4. The composite Chinese medicine preparation for chronic osteomyelitis according to claim 1, characterized in that: The degree of deacetylation of chitosan is 85-95%.

5. The composite Chinese medicine preparation for chronic osteomyelitis according to claim 1, characterized in that: The cyclodextrin is β-cyclodextrin.

6. The composite Chinese medicine preparation for chronic osteomyelitis according to claim 1, characterized in that: The viscosity average molecular weight of the sodium alginate is 430 kDa.

7. A method for preparing a composite Chinese medicine preparation for chronic osteomyelitis as claimed in any one of claims 1 to 6, characterized in that: The steps include: (1) Dissolve sodium alginate in dimethylformamide, stir evenly at 50-60°C, add dried p-toluenesulfonic acid, continue stirring for 1-2 hours, add cyclodextrin and stir evenly, add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and continue stirring for 1-3 hours, wash, dry, add to water, stir at 50-60°C for 10-20 minutes, and cool to room temperature to obtain a pretreated cyclodextrin solution; (2) Add the Wuwei Xiaoduyin extract and emulsifier to the pretreated cyclodextrin solution, stir at 40-50°C for 5-15 minutes, and homogenize at 10000-15000 r / min for 1-4 minutes to obtain a single-layer coated emulsion; (3) Dissolve chitosan in acetic acid solution, add sodium chloride thereto and stir evenly, adjust the stirring speed to 1000-1500 r / min, add the monolayer coated emulsion thereto at 20-40°C and continue stirring for 10-20 min, adjust the pH value of the system to 6.0-6.4, add calcium chloride, stir at 1-5°C for 1-2 h, centrifuge for 5-10 min, collect the precipitate, and freeze-dry to obtain a composite Chinese medicine preparation for chronic osteomyelitis.

8. The method for preparing the composite Chinese medicine preparation for chronic osteomyelitis according to claim 7, characterized in that: The drying temperature of p-toluenesulfonic acid is 105-120° C., the drying time is 20-60 min, and the vacuum degree is maintained at 10-20 kPa during the drying process.

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