Preparation method and application of traditional Chinese medicine sustained release preparation for targeting bone marrow focus

Abstract

The invention relates to the technical field of traditional Chinese medicine preparations, in particular to a preparation method and application of a bone marrow focus targeting traditional Chinese medicine sustained release preparation. The traditional Chinese medicine sustained release preparation for targeting the bone marrow focus comprises the following raw materials in parts by mass: 15-25 parts of oldenlandia diffusa, 5-15 parts of sculellaria barbata, 20-30 parts of astragalus membranaceus, 10-20 parts of dandelion, 5-12 parts of herba violae, 5-15 parts of gynura bicolor, 1-3 parts of sodium alginate, 1-3 parts of hyperbranched polylysine, 1-2 parts of poloxamer, 1-5 parts of a polylactic acid-glycolic acid copolymer and 5-15 parts of a calcium chloride solution. And 5-10 parts of polyvinyl alcohol. The sustained and controllable release of the traditional Chinese medicine components in the osteomyelitis focus part is realized, the long-acting anti-infection and bone repair promoting potential is achieved, and the preparation method is simple and suitable for large-scale popularization and application.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparation technology, and in particular to a method for preparing and applying a sustained-release traditional Chinese medicine preparation targeting bone marrow lesions. Background Technology

[0002] Osteomyelitis is a common but difficult-to-cure infectious disease in orthopedics, with bacterial infection being its direct cause. Traditional treatment relies on high-dose systemic antibiotics; however, poor local blood circulation at the site of bone infection makes it difficult for antibiotics to effectively penetrate and form sufficient antibacterial concentrations at the lesion site. Furthermore, studies have shown that excessively high local antibiotic concentrations can inhibit osteoblast activity during bone regeneration, affecting bone repair. Clinically, high-dose intravenous administration is often required for several weeks initially. This long-term medication not only increases the systemic toxicity of antibiotics but also easily leads to the development of drug-resistant bacteria, further complicating the treatment of osteomyelitis.

[0003] Traditional Chinese medicine (TCM) treatment emphasizes holistic regulation, employing formulas that invigorate Qi and nourish Yin, warm the kidneys and unblock the Du meridian, offering advantages in anti-infection and bone repair promotion with multiple targets and low drug resistance. However, existing TCM preparations also have significant shortcomings in clinical application. On the one hand, ordinary TCM preparations have poor sustained-release properties, releasing drugs too quickly and making it difficult to maintain long-term effective concentrations at the osteomyelitis lesion site. Patients need frequent administration, leading to low treatment compliance, and drastic fluctuations in blood drug concentration can also affect the sustainability of efficacy. On the other hand, TCM preparations generally have poor stability, and their active ingredients are easily degraded or inactivated by environmental factors. Many TCM components undergo chemical structural changes or even deterioration under high temperature, high humidity, or strong light conditions, resulting in decreased or lost biological activity.

[0004] These factors severely restrict the promotion and application of traditional Chinese medicine in the treatment of osteomyelitis. How to improve the sustained-release properties of traditional Chinese medicine preparations, enabling them to achieve sustained and controllable drug release at the lesion site, and enhance their resistance to external environmental influences, has become an urgent technical problem to be solved. Summary of the Invention

[0005] The purpose of this invention is to address the shortcomings of existing technologies by proposing a method for preparing and applying a sustained-release traditional Chinese medicine preparation targeting bone marrow lesions.

[0006] A sustained-release preparation of traditional Chinese medicine targeting bone marrow lesions comprises the following raw materials by weight: 15-25 parts of Hedyotis diffusa, 5-15 parts of Scutellaria barbata, 20-30 parts of Astragalus membranaceus, 10-20 parts of Taraxacum mongolicum, 5-12 parts of Viola yedoensis, 5-15 parts of Smilax china, 1-3 parts of sodium alginate, 1-3 parts of hyperbranched polylysine, 1-2 parts of poloxamer, 1-5 parts of polylactic acid-glycolic acid copolymer, 5-15 parts of calcium chloride solution, and 5-10 parts of polyvinyl alcohol.

[0007] Hedyotis diffusa, bitter and sweet in taste, cold in nature; enters the heart, liver, spleen, and large intestine meridians. It can clear heat and detoxify; promote diuresis. It is used to treat lung heat cough with wheezing; sore throat; intestinal abscess; boils and sores; snake bites; painful urination due to heat; edema; dysentery; enteritis; damp-heat jaundice; and cancer.

[0008] Scutellaria barbata, pungent and bitter in taste, cold in nature. It enters the lung, liver, and kidney meridians. It can clear heat and detoxify, resolve blood stasis and promote diuresis. It is used for boils and carbuncles, sore throat, snake bites, traumatic injuries, edema, and jaundice.

[0009] Astragalus membranaceus (Huang Qi) is sweet in taste and slightly warm in nature. It enters the lung, spleen, liver, and kidney meridians. It has the effects of tonifying qi and strengthening the exterior, astringing sweat and preventing prolapse, promoting tissue regeneration and relieving sores, and promoting diuresis and reducing swelling. It is used to treat qi deficiency and fatigue, sinking of middle qi, chronic diarrhea and rectal prolapse, hematochezia and metrorrhagia, spontaneous sweating due to exterior deficiency, carbuncles that are difficult to heal, chronic ulcers that do not heal, anemia and chlorosis, internal heat and thirst, chronic nephritis, proteinuria, and diabetes. Prepared astragalus membranaceus tonifies qi and strengthens the middle jiao, while raw astragalus membranaceus strengthens the exterior and promotes tissue regeneration.

[0010] Dandelion, sweet and slightly bitter in taste, and cold in nature. It clears heat and detoxifies, reduces swelling and dissipates nodules. It is mainly used to treat upper respiratory tract infections, conjunctivitis, mumps, mastitis, gastritis, dysentery, hepatitis, cholecystitis, acute appendicitis, urinary tract infections, pelvic inflammatory disease, boils and carbuncles, pharyngitis, acute mastitis, lymphadenitis, scrofula, boils and carbuncles, acute conjunctivitis, fever from colds, acute tonsillitis, acute bronchitis, and urinary tract infections.

[0011] Viola yedoensis, dried whole herb. Harvested in spring and autumn, impurities removed, washed, and sun-dried. Bitter and pungent in taste, cold in nature; enters the heart and liver meridians. Clears heat and detoxifies, cools the blood and reduces swelling. Mainly used to treat jaundice, dysentery, mastitis, red and swollen eyes, and pharyngitis; externally applied to treat traumatic injuries, carbuncles, and snake bites.

[0012] Purple-backed amaranth has a pungent, bitter, and sweet taste, and is neutral or cold in nature. It can reduce swelling, detoxify, and promote urination. It is mainly used to treat scrofula, hernia, and difficulty urinating.

[0013] Preferably, the molecular weight of the polylactic acid-glycolic acid copolymer is 10-13 kDa.

[0014] Preferably, the calcium chloride solution is obtained by mixing calcium chloride and water in a mass ratio of 1-2:5-15.

[0015] Preferably, the molecular weight of polyvinyl alcohol is 20-35 kDa.

[0016] The preparation method of the above-mentioned sustained-release traditional Chinese medicine preparation targeting bone marrow lesions includes the following steps: S1. Mix and grind Hedyotis diffusa, Scutellaria barbata, Astragalus membranaceus, Taraxacum mongolicum, Viola yedoensis, and Smilax china, add to an ethanol aqueous solution, and extract once at 60-70℃ for 1-2 hours, then filter; add the residue to an ethanol aqueous solution, and extract a second time at 60-70℃ for 1-2 hours, then filter; combine the two extracts and concentrate at 50-60℃ to obtain an extract; add the extract, sodium alginate, hyperbranched polylysine, and poloxamer 407 to water and stir evenly to obtain pre-mixed material a; S2. Add polylactic acid-glycolic acid copolymer to dichloromethane and stir until homogeneous. While stirring, add preform a and calcium chloride solution dropwise in sequence. Shear at high speed at 1-5℃ for 5-10 minutes to obtain preform b. S3. Add polyvinyl alcohol to water and stir until homogeneous. While stirring, add pre-mixed material b dropwise. Rotary evaporate until dichloromethane is completely volatilized. Collect by centrifugation, wash, and freeze-dry.

[0017] Preferably, in S1, the material-to-liquid ratio is 1:5-8 during a single extraction process.

[0018] Preferably, in S1, the material-to-liquid ratio during the secondary extraction process is 1:8-10.

[0019] Preferably, in S1, the relative density of the extract is 1.1-1.2.

[0020] Preferably, in S2, the shearing rate is 5000-8000 r / min.

[0021] The application of the above-mentioned sustained-release traditional Chinese medicine preparations targeting bone marrow lesions in the preparation of drugs for treating osteomyelitis.

[0022] Compared with existing technologies, the present invention has the following advantages: This invention uses a combination of Oldenlandia diffusa, Scutellaria barbata, Astragalus membranaceus, Taraxacum mongolicum, Viola yedoensis, and Smilax china, which not only helps to inhibit the formation of bacterial biofilms, but also regulates the body's overall immunity, improves local microcirculation, and promotes bone repair while fighting infection. It has a good sustained-release effect and maximizes the therapeutic advantages of traditional Chinese medicine.

[0023] This invention utilizes the cross-linking of calcium chloride and sodium alginate ions, combined with hyperbranched polylysine, to form a stable and dense three-dimensional network that effectively encapsulates the drug, effectively restricts the free migration of drug molecules, and enhances the physical and chemical stability of the entire system. This helps to cope with the effects of storage and complex environments. After the microspheres enter the liquid environment of the lesion, they can quickly form a hydrophilic gel network, effectively delaying the initial burst release of the drug. This is combined with the slow hydrolysis of the polylactic acid-glycolic acid copolymer backbone. Moreover, the polylactic acid-glycolic acid copolymer itself is hydrophobic, which can reduce water penetration and protect the drug from hydrolysis. Combined with the thermosensitive properties of poloxamer 407, it can change from a sol to a gel state at body temperature, increasing the adhesion of the formulation to the lesion site, thereby prolonging the duration of action, enabling slow and controlled drug release, and promoting the maintenance of drug concentration at the lesion site for several weeks, significantly reducing the frequency of drug administration.

[0024] This invention enables the continuous and controllable release of traditional Chinese medicine components at the site of osteomyelitis lesions, possessing both long-lasting anti-infection and bone repair promotion potential. Moreover, the preparation method is simple and suitable for large-scale promotion and application. Attached Figure Description

[0025] Figure 1 The graph shows the cumulative release rate of the sustained-release preparations of traditional Chinese medicine obtained in Example 5 and Comparative Examples 1-2 during 60 days of constant temperature storage at 25°C.

[0026] Figure 2 The cumulative release curves of the sustained-release preparations of traditional Chinese medicine obtained in Example 5 and Comparative Examples 1-2 in the digestive system are shown.

[0027] Figure 3 Comparison of bone volume fraction of tibia in mice from the normal group, Example 5 group, Comparative Example 1 group, and Comparative Example 2 group.

[0028] Figure 4 This is a comparison of bacterial colony counts in the tibial bone marrow tissues of mice in the model group, Example 5 group, Comparative Example 1 group, and Comparative Example 2 group. Detailed Implementation

[0029] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0030] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0031] The hyperbranched polylysine (D3-BOC) used below16 The following materials were purchased from Weihai Mouyuan Molecular New Materials Co., Ltd. The polylactic acid-glycolic acid copolymer (50 / 50) used below was purchased from Shenzhen Moufeng Biotechnology Co., Ltd. The following polyvinyl alcohol (food grade PVA2499) used below was sourced from Wanwei Group. The following calcium chloride solution was obtained by mixing calcium chloride and water at a mass ratio of 1.5:10.5.

[0032] Example 1: A sustained-release preparation of traditional Chinese medicine targeting bone marrow lesions, the raw materials of which include: 15g of Hedyotis diffusa, 5g of Scutellaria barbata, 20g of Astragalus membranaceus, 10g of Taraxacum mongolicum, 5g of Viola yedoensis, 5g of Smilax china, 1g of sodium alginate, 1g of hyperbranched polylysine, 1g of poloxamer 407, 1g of polylactic acid-glycolic acid copolymer, 5g of calcium chloride solution, and 5g of polyvinyl alcohol.

[0033] The preparation method of the above-mentioned sustained-release traditional Chinese medicine preparation targeting bone marrow lesions includes the following steps: S1. Mix and grind Hedyotis diffusa, Scutellaria barbata, Astragalus membranaceus, Taraxacum mongolicum, Viola yedoensis, and Smilax china, pass through a 100-mesh sieve, add to a 50% ethanol aqueous solution at a material-to-liquid ratio of 1:5, extract once at 60℃ for 1 hour, and filter. Add the residue to a 50% ethanol aqueous solution at a material-to-liquid ratio of 1:8, extract a second time at 60℃ for 1 hour, and filter. Combine the two extracts and concentrate at 50℃ to a relative density of 1.1 to obtain an extract. Add the extract, sodium alginate, hyperbranched polylysine, and poloxamer 407 to 40g of deionized water and stir evenly to obtain pre-mixed material a. S2. Add polylactic acid-glycolic acid copolymer to 100g dichloromethane and stir until homogeneous. While stirring, add preform a and calcium chloride solution dropwise in sequence. Shear at high speed for 5 minutes at 1℃ with a shearing speed of 5000r / min to obtain preform b. S3. Add polyvinyl alcohol to 200g of water and stir evenly. While stirring, add pre-made material b dropwise. Rotary evaporate until dichloromethane is completely volatilized. Collect by centrifugation, wash with deionized water, and freeze dry.

[0034] Example 2: A sustained-release preparation of traditional Chinese medicine targeting bone marrow lesions, the raw materials of which include: 25g of Hedyotis diffusa, 15g of Scutellaria barbata, 30g of Astragalus membranaceus, 20g of Taraxacum mongolicum, 12g of Viola yedoensis, 15g of Smilax china, 3g of sodium alginate, 3g of hyperbranched polylysine, 2g of poloxamer 407, 5g of polylactic acid-glycolic acid copolymer, 15g of calcium chloride solution, and 10g of polyvinyl alcohol.

[0035] The preparation method of the above-mentioned sustained-release traditional Chinese medicine preparation targeting bone marrow lesions includes the following steps: S1. Mix and grind Hedyotis diffusa, Scutellaria barbata, Astragalus membranaceus, Taraxacum mongolicum, Viola yedoensis, and Smilax china, pass through a 100-mesh sieve, and add to a 70% ethanol aqueous solution at a material-to-liquid ratio of 1:8. Extract once at 70°C for 2 hours, and filter. Add the residue to a 70% ethanol aqueous solution at a material-to-liquid ratio of 1:10, and extract a second time at 70°C for 2 hours, and filter. Combine the two extracts and concentrate at 60°C to a relative density of 1.2 to obtain an extract. Add the extract, sodium alginate, hyperbranched polylysine, and poloxamer 407 to 60g of deionized water and stir evenly to obtain pre-mixed material a. S2. Add polylactic acid-glycolic acid copolymer to 200g dichloromethane and stir until homogeneous. While stirring, add preform a and calcium chloride solution dropwise in sequence. Shear at high speed for 10min at 5℃ with a shearing speed of 8000r / min to obtain preform b. S3. Add polyvinyl alcohol to 400g of water and stir evenly. While stirring, add pre-mixed material b dropwise. Rotary evaporate until dichloromethane is completely volatilized. Collect by centrifugation, wash with deionized water, and freeze dry.

[0036] Example 3: A sustained-release preparation of traditional Chinese medicine targeting bone marrow lesions comprises the following raw materials: 18g of Hedyotis diffusa, 12g of Scutellaria barbata, 22g of Astragalus membranaceus, 17g of Taraxacum mongolicum, 6g of Viola yedoensis, 12g of Smilax china, 1.5g of sodium alginate, 2.5g of hyperbranched polylysine, 1.2g of poloxamer 407, 4g of polylactic acid-glycolic acid copolymer, 8g of calcium chloride solution, and 9g of polyvinyl alcohol.

[0037] The preparation method of the above-mentioned sustained-release traditional Chinese medicine preparation targeting bone marrow lesions includes the following steps: S1. Mix and grind Hedyotis diffusa, Scutellaria barbata, Astragalus membranaceus, Taraxacum mongolicum, Viola yedoensis, and Smilax china, pass through a 100-mesh sieve, add to a 55% ethanol aqueous solution at a material-to-liquid ratio of 1:7, extract once at 62℃ for 100 min, and filter. Add the residue to a 55% ethanol aqueous solution at a material-to-liquid ratio of 1:9.5, extract a second time at 62℃ for 100 min, and filter. Combine the two extracts and concentrate at 52℃ to a relative density of 1.15 to obtain an extract. Add the extract, sodium alginate, hyperbranched polylysine, and poloxamer 407 to 55g of deionized water and stir evenly to obtain pre-mixed material a. S2. Add polylactic acid-glycolic acid copolymer to 120g dichloromethane and stir until uniform. While stirring, add preform a and calcium chloride solution dropwise in sequence. Shear at high speed for 7 minutes at 4℃ with a shearing speed of 7000r / min to obtain preform b. S3. Add polyvinyl alcohol to 260g of water and stir evenly. While stirring, add pre-made material b dropwise. Evaporate until dichloromethane is completely volatilized. Collect by centrifugation, wash with deionized water, and freeze dry.

[0038] Example 4: A sustained-release preparation of traditional Chinese medicine targeting bone marrow lesions comprises the following raw materials: 22g of Hedyotis diffusa, 8g of Scutellaria barbata, 28g of Astragalus membranaceus, 13g of Taraxacum mongolicum, 10g of Viola yedoensis, 8g of Smilax china, 2.5g of sodium alginate, 1.5g of hyperbranched polylysine, 1.8g of poloxamer 407, 2g of polylactic acid-glycolic acid copolymer, 12g of calcium chloride solution, and 7g of polyvinyl alcohol.

[0039] The preparation method of the above-mentioned sustained-release traditional Chinese medicine preparation targeting bone marrow lesions includes the following steps: S1. Mix and grind Hedyotis diffusa, Scutellaria barbata, Astragalus membranaceus, Taraxacum mongolicum, Viola yedoensis, and Smilax china, pass through a 100-mesh sieve, add to a 65% ethanol aqueous solution at a material-to-liquid ratio of 1:6, extract once at 68℃ for 80 minutes, and filter. Add the residue to a 65% ethanol aqueous solution at a material-to-liquid ratio of 1:8.5, extract a second time at 68℃ for 80 minutes, and filter. Combine the two extracts and concentrate at 58℃ to a relative density of 1.15 to obtain an extract. Add the extract, sodium alginate, hyperbranched polylysine, and poloxamer 407 to 45g of deionized water and stir evenly to obtain pre-mixed material a. S2. Add polylactic acid-glycolic acid copolymer to 180g dichloromethane and stir until homogeneous. While stirring, add preform a and calcium chloride solution dropwise in sequence. Shear at high speed for 9min at 2℃ with a shearing speed of 6000r / min to obtain preform b. S3. Add polyvinyl alcohol to 340g of water and stir until homogeneous. While stirring, add pre-mixed material b dropwise. Evaporate until dichloromethane is completely volatilized. Collect by centrifugation, wash with deionized water, and freeze-dry.

[0040] Example 5: A sustained-release preparation of traditional Chinese medicine targeting bone marrow lesions, the raw materials of which include: 20g of Hedyotis diffusa, 10g of Scutellaria barbata, 25g of Astragalus membranaceus, 15g of Taraxacum mongolicum, 8g of Viola yedoensis, 10g of Smilax china, 2g of sodium alginate, 2g of hyperbranched polylysine, 1.5g of poloxamer 407, 3g of polylactic acid-glycolic acid copolymer, 10g of calcium chloride solution, and 8g of polyvinyl alcohol.

[0041] The preparation method of the above-mentioned sustained-release traditional Chinese medicine preparation targeting bone marrow lesions includes the following steps: S1. Mix and grind Hedyotis diffusa, Scutellaria barbata, Astragalus membranaceus, Taraxacum mongolicum, Viola yedoensis, and Smilax china, pass through a 100-mesh sieve, and add to a 60% ethanol aqueous solution at a material-to-liquid ratio of 1:6.5. Extract once at 65℃ for 90 minutes, and filter. Add the residue to a 60% ethanol aqueous solution at a material-to-liquid ratio of 1:9, and extract a second time at 65℃ for 90 minutes, and filter. Combine the two extracts and concentrate at 55℃ to a relative density of 1.15 to obtain an extract. Add the extract, sodium alginate, hyperbranched polylysine, and poloxamer 407 to 50g of deionized water and stir evenly to obtain pre-mixed material a. S2. Add polylactic acid-glycolic acid copolymer to 150g dichloromethane and stir until homogeneous. While stirring, add preform a and calcium chloride solution dropwise in sequence. Shear at high speed for 8 minutes at 3℃ with a shearing speed of 6500r / min to obtain preform b. S3. Add polyvinyl alcohol to 300g of water and stir evenly. While stirring, add pre-made material b dropwise. Rotary evaporate until dichloromethane is completely volatilized. Collect by centrifugation, wash with deionized water, and freeze dry.

[0042] Comparative Example 1: A sustained-release traditional Chinese medicine preparation targeting bone marrow lesions comprises the following ingredients: Astragalus membranaceus 34g, Taraxacum mongolicum 24g, Viola yedoensis 14g, Achyranthes bidentata 16g, sodium alginate 2g, hyperbranched polylysine 2g, poloxamer 407 1.5g, polylactic acid-glycolic acid copolymer 3g, calcium chloride solution 10g, and polyvinyl alcohol 8g.

[0043] The preparation method of the above-mentioned sustained-release traditional Chinese medicine preparation targeting bone marrow lesions includes the following steps: S1. Grind Astragalus membranaceus, Taraxacum mongolicum, Viola yedoensis, and Smilax china together, pass through a 100-mesh sieve, and add to a 60% ethanol aqueous solution at a material-to-liquid ratio of 1:6.5. Extract once at 65℃ for 90 minutes, and filter. Add the residue to a 60% ethanol aqueous solution at a material-to-liquid ratio of 1:9, and extract a second time at 65℃ for 90 minutes, and filter. Combine the two extracts and concentrate at 55℃ to a relative density of 1.15 to obtain an extract. Add the extract, sodium alginate, hyperbranched polylysine, and poloxamer 407 to 50g of deionized water and stir evenly to obtain pre-mixed material a. S2. Add polylactic acid-glycolic acid copolymer to 150g dichloromethane and stir until homogeneous. While stirring, add preform a and calcium chloride solution dropwise in sequence. Shear at high speed for 8 minutes at 3℃ with a shearing speed of 6500r / min to obtain preform b. S3. Add polyvinyl alcohol to 300g of water and stir evenly. While stirring, add pre-made material b dropwise. Rotary evaporate until dichloromethane is completely volatilized. Collect by centrifugation, wash with deionized water, and freeze dry.

[0044] Comparative Example 2: A sustained-release traditional Chinese medicine preparation targeting bone marrow lesions comprises the following ingredients: 20g of Hedyotis diffusa, 10g of Scutellaria barbata, 25g of Astragalus membranaceus, 15g of Taraxacum mongolicum, 8g of Viola yedoensis, 10g of Smilax china, 3g of sodium alginate, 2.5g of poloxamer 407, 3g of polylactic acid-glycolic acid copolymer, 10g of calcium chloride solution, and 8g of polyvinyl alcohol.

[0045] The preparation method of the above-mentioned sustained-release traditional Chinese medicine preparation targeting bone marrow lesions includes the following steps: S1. Mix and grind Hedyotis diffusa, Scutellaria barbata, Astragalus membranaceus, Taraxacum mongolicum, Viola yedoensis, and Smilax china, pass through a 100-mesh sieve, and add to a 60% ethanol aqueous solution at a material-to-liquid ratio of 1:6.5. Extract once at 65℃ for 90 minutes, and filter. Add the residue to a 60% ethanol aqueous solution at a material-to-liquid ratio of 1:9, and extract a second time at 65℃ for 90 minutes, and filter. Combine the two extracts and concentrate at 55℃ to a relative density of 1.15 to obtain an extract. Add the extract, sodium alginate, and poloxamer 407 to 50g of deionized water and stir evenly to obtain pre-mixed material a. S2. Add polylactic acid-glycolic acid copolymer to 150g dichloromethane and stir until homogeneous. While stirring, add preform a and calcium chloride solution dropwise in sequence. Shear at high speed for 8 minutes at 3℃ with a shearing speed of 6500r / min to obtain preform b. S3. Add polyvinyl alcohol to 300g of water and stir evenly. While stirring, add pre-made material b dropwise. Rotary evaporate until dichloromethane is completely volatilized. Collect by centrifugation, wash with deionized water, and freeze dry.

[0046] Accurately weigh 1.0 g of each of the sustained-release traditional Chinese medicine preparations obtained in Example 5 and Comparative Example 1, and then store them at a constant temperature of 25°C. They were taken out on the 5th, 10th, 15th, 20th, 25th, 30th and 60th days, respectively, and measured by UPLC. The peak area was recorded and the cumulative release rate was calculated.

[0047] like Figure 1 As shown, with the extension of storage time, the cumulative release rate of each group gradually increases and the release rate gradually slows down; while the cumulative release rate of Example 5 is always lower than that of the comparative example under the same conditions.

[0048] The sustained-release formulations of traditional Chinese medicine obtained in Example 5 and Comparative Examples 1-2 were first placed in simulated gastric fluid and allowed to stand at 36°C for 2 hours. After filtration, the filter cake was placed in simulated intestinal fluid and allowed to stand at 36°C for 4 hours. During the experiment, 2 mL of supernatant was taken every hour, and 2 mL of fresh simulated gastric fluid or fresh simulated intestinal fluid was added. The supernatant was filtered through a 0.22 μm filter membrane, and UPLC was performed to determine the peak area and calculate the cumulative release rate. Figure 2 As shown, Example 5 exhibits the best release effect, significantly superior to the comparative example.

[0049] A comparative experiment was conducted using the sustained-release traditional Chinese medicine preparations obtained in Example 5 and Comparative Examples 1-2, as follows: Sixty male SD rats (210±10g) were randomly divided into 5 groups (normal group, model group, Example 5 group, Comparative Example 1 group, and Comparative Example 2 group). Rats in the model group, Example 5 group, Comparative Example 1 group, and Comparative Example 2 group underwent osteomyelitis modeling treatment (SD rats were fed for 1 week and fasted for 12 hours before the procedure). After anesthesia with sodium pentobarbital, the skin was prepared and disinfected, the knee joint was flexed, and an 8-gauge needle was used to drill into the medullary cavity of the tibia from the anterior middle 1 / 3 of the tibial plateau, slightly posterior to the vertical line of the tibia. A breakthrough or loss of momentum was felt during drilling. The needle (8-gauge) was then inserted into the medullary cavity of the tibia through the anterior hole, and 100μL of Staphylococcus aureus suspension (1×10⁻⁶) was injected. 8 cfu / mL). Withdraw the needle and apply slight pressure to the puncture site.

[0050] After modeling, local reactions were observed, including redness and swelling of the affected limb, reduced movement, and flexion. Four weeks later, local redness, sinus tract formation, and purulent discharge were observed. Imaging examination revealed periosteal hyperplasia, thickening and increased density of cortical bone, and the presence of sequestra and osteolytic cavities, indicating successful experimental modeling.

[0051] After the model was successfully constructed, rats in Example 5, Comparative Example 1, and Comparative Example 2 were administered the Chinese medicine sustained-release preparations obtained in Example 5, Comparative Example 1, and Comparative Example 2 by gavage for 4 weeks (once a day), at a dose of 0.87 g / kg body weight. At the same time, the normal group and the model group were given the same dose of physiological saline by gavage.

[0052] After sacrificing rats, the bone volume fraction (BV / TV) of the tibia was measured in the normal group, Example 5 group, Comparative Example 1 group, and Comparative Example 2 group to characterize the microstructure of bone trabeculae. Bone marrow tissue from the tibia of the model group, Example 5 group, Comparative Example 1 group, and Comparative Example 2 group was weighed, homogenized, plated, and the colony count was calculated.

[0053] like Figure 3 and Figure 4As shown, the tibia of mice in Example 5 group had the highest bone volume fraction, significantly better than the control group; while the bone marrow of mice in Example 5 group had the lowest bacterial colony count, significantly better than the control group. This confirms that the sustained-release traditional Chinese medicine preparation obtained in Example 5 can effectively promote the recovery of bone trabeculae in patients with osteomyelitis and can penetrate deep into the bone marrow tissue to exert a bactericidal effect.

[0054] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein.

Claims

1. A traditional Chinese medicine sustained-release preparation targeting bone marrow lesions, characterized in that, The raw materials include, by mass fraction: 15-25 parts of Orthosiphon aristatus, 5-15 parts of Herba Dispori, 20-30 parts of Astragalus, 10-20 parts of Taraxacum mongolicum, 5-12 parts of Viola yedoensis, 5-15 parts of Malvastrum bicolor, 1-3 parts of sodium alginate, 1-3 parts of hyperbranched polylysine, 1-2 parts of poloxamer, 1-5 parts of polylactic acid-glycolic acid copolymer, 5-15 parts of calcium chloride solution, and 5-10 parts of polyvinyl alcohol.

2. The traditional Chinese medicine sustained-release preparation for targeting bone marrow lesions according to claim 1, characterized in that, The polylactic acid-glycolic acid copolymer has a molecular weight of 10-13 kDa.

3. The traditional Chinese medicine sustained-release preparation for targeting bone marrow lesions according to claim 1, characterized in that, The calcium chloride solution is obtained by mixing calcium chloride and water in a mass ratio of 1-2:5-15.

4. The traditional Chinese medicine sustained-release preparation for targeting bone marrow lesions according to claim 1, characterized in that, The polyvinyl alcohol has a molecular weight of 20-35 kDa.

5. A method for preparing the traditional Chinese medicine sustained-release preparation for targeting bone marrow lesions according to any one of claims 1-4, characterized in that, The method comprises the following steps: S1, mixing and grinding Orthosiphon aristatus, Herba Dispori, Astragalus, Taraxacum mongolicum, Viola yedoensis, and Malvastrum bicolor, adding them to an ethanol aqueous solution, extracting once at 60-70 ℃ for 1-2 h, filtering; adding the residues to an ethanol aqueous solution, extracting twice at 60-70 ℃ for 1-2 h, filtering; combining the two extraction solutions, concentrating at 50-60 ℃ to obtain an extract; adding the extract, sodium alginate, hyperbranched polylysine, and poloxamer 407 to water, stirring to obtain a pre-preparation a; S2, adding polylactic acid-glycolic acid copolymer to dichloromethane, stirring to obtain a pre-preparation b; S3, adding polyvinyl alcohol to water, stirring to obtain a pre-preparation b; adding the pre-preparation b to the pre-preparation b under stirring, rotary evaporation until dichloromethane is completely volatilized, centrifuging, washing, and freeze-drying. In S1, the solid-liquid ratio in the first extraction is 1:5-8.

6. The method of claim 5, wherein the preparation of the traditional Chinese medicine sustained-release preparation for targeting bone marrow lesions is characterized by, In S1, the solid-liquid ratio in the second extraction is 1:8-10.

7. The preparation method of claim 5, wherein the preparation method of the traditional Chinese medicine sustained-release preparation for targeting bone marrow lesions is characterized by, In S1, the relative density of the extract is 1.1-1.

2.

8. The method of claim 5, wherein the preparation of the Chinese medicine sustained release preparation for targeting bone marrow lesions is characterized by, In S2, the shearing speed is 5000-8000 r / min.

9. The method of claim 5, wherein the preparation of the Chinese medicine sustained release preparation for targeting bone marrow lesions is characterized by, 10. Use of the traditional Chinese medicine sustained-release preparation for targeting bone marrow lesions according to any one of claims 1-4 in the preparation of a drug for treating osteomyelitis. ​

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