Traditional Chinese medicine preparation for osteoporosis rehabilitation and preparation method thereof

Abstract

The invention relates to the field of traditional Chinese medicine preparations, and discloses a traditional Chinese medicine preparation for osteoporosis rehabilitation and a preparation method thereof.The traditional Chinese medicine preparation comprises a traditional Chinese medicine active extract, a multi-metal synergistic complexing center, an acid-sensitive modifier and an auxiliary stabilizer. A stable complex is formed by active extracts of traditional Chinese medicines and metal ions such as calcium, zinc, magnesium and iron, and is combined with an acid-sensitive modifier to realize targeted release of bone tissues, so that the medicine utilization rate and the treatment effect are remarkably improved. The preparation method comprises the processes of traditional Chinese medicine extract extraction, multi-metal complexing, acid-sensitive modification and granulation coating, and the stability, the controlled release performance and the batch consistency of the preparation are improved. The invention overcomes the defects of poor stability of active ingredients, insufficient drug targeting and poor controlled release effect in the prior art, and can be widely applied to treatment and rehabilitation of osteoporosis and related bone diseases.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine preparations, and specifically to a traditional Chinese medicine preparation for osteoporosis rehabilitation and its preparation method. Background Art

[0002] Osteoporosis is a systemic disease characterized by increased bone fragility and fracture risk caused by reduced bone mass and degenerated bone tissue structure. With the aggravation of population aging, the incidence of osteoporosis is rising continuously, and it has become a major public health problem globally. The existing treatment methods for osteoporosis mainly include calcium supplementation, vitamin D, bisphosphonates, etc. However, these methods have problems such as relatively large side effects, limited efficacy, and insufficient compliance in long-term applications. Therefore, in recent years, the rehabilitation treatment of osteoporosis based on traditional Chinese medicine has gradually attracted attention. It exerts an overall therapeutic effect by regulating multiple targets, especially suitable for the long-term management of chronic diseases. However, there are still many technical bottlenecks in the current traditional Chinese medicine-based osteoporosis treatment preparations.

[0003] In the prior art, traditional Chinese medicine preparations are mainly directly used through single components or simple extracts in the rehabilitation application of osteoporosis, but show the following deficiencies in actual applications:

[0004] Poor stability of active ingredients and difficult to guarantee efficacy

[0005] Active ingredients of traditional Chinese medicine (such as flavonoids, glycosides, polyphenols) are highly sensitive to environmental factors such as light, heat, and oxygen, and are prone to oxidative degradation, resulting in reduced drug efficacy. Although in the prior art, certain preparation means (such as embedding or adding antioxidants) are used to improve the stability of traditional Chinese medicine components, the effect is limited. Especially under high-temperature storage or long-term storage conditions, the effectiveness of the drug still cannot be fully guaranteed. Therefore, traditional Chinese medicine preparations still need to be further optimized in terms of stability.

[0006] Lack of bone-targeted delivery mechanism and low drug utilization rate

[0007] The pathogenesis of osteoporosis is complex, and drugs need to act concentratedly on the lesion sites of bone tissue (such as active bone resorption areas) to exert the best efficacy. However, existing traditional Chinese medicine preparations are mostly administered systemically and lack a bone-targeted delivery mechanism, resulting in the distribution of drugs throughout the body, which not only reduces the effective drug concentration in bone tissue but also may cause adverse reactions in non-target tissues. Although in the prior art, some chemical modification or carrier delivery methods are tried to improve the bone targeting of drugs, their designs are complex and costly, making it difficult to be widely applied.

[0008] Drug release is uncontrolled and precise treatment cannot be achieved

[0009] The pathological environment of osteoporosis (such as the acidic microenvironment in the active area of osteoclasts) is specific. However, existing traditional Chinese medicine preparations are usually immediate-release types, and it is difficult to regulate the time and location of drug release, resulting in insufficient efficacy or even drug waste. Although some studies have attempted to improve drug release behavior through technologies such as sustained release and controlled release, these technologies are usually less applicable to specific environmental responses, especially the response release technology for the bone tissue-specific acidic environment, which limits their therapeutic effects.

[0010] The process of traditional Chinese medicine preparations is simple, and the compound synergistic effect is insufficient.

[0011] The pathogenesis of osteoporosis involves the dynamic balance of bone formation and bone resorption. In existing technologies, single components or simple extracts are usually used for treatment, and it is impossible to take into account the comprehensive regulatory effects of osteogenesis and anti-bone resorption. Some studies have attempted to introduce metal ions (such as calcium, zinc, and magnesium) combined with traditional Chinese medicine components to enhance the efficacy. However, the selection of metal ions is single, the ratio is improper, the synergistic effect is limited, and the chemical stability of the complex is poor, making it difficult to form a functional composite preparation.

[0012] Therefore, the present invention proposes a traditional Chinese medicine preparation for the rehabilitation of osteoporosis and its preparation method to solve the deficiencies of existing technologies. Summary of the Invention

[0013] Aiming at the problems of poor stability of active ingredients, lack of bone-targeted delivery mechanism, uncontrolled drug release, and insufficient compound regulatory effect in the rehabilitation treatment of osteoporosis by traditional Chinese medicine preparations in existing technologies, the present invention provides a traditional Chinese medicine preparation for the rehabilitation of osteoporosis and its preparation method. Through scientific component design and preparation process optimization, the present invention improves the existing technologies from four aspects: stability, targeting, controlled release, and comprehensive regulatory effect, significantly enhancing the therapeutic effect and application value of traditional Chinese medicine preparations.

[0014] To achieve the above objectives, the present invention is realized through the following technical solutions: A traditional Chinese medicine preparation for the rehabilitation of osteoporosis, the traditional Chinese medicine preparation comprising a composition of the following components:

[0015] 3%-10% of traditional Chinese medicine active extracts, including flavonoids, glycosides, and polyphenolic compounds;

[0016] Multi-metal complexation center, containing 10%-30% of calcium ions, 5%-15% of zinc ions, 5%-10% of magnesium ions, and 2%-10% of iron ions;

[0017] Auxiliary stabilizer, containing 1%-5% of chitosan and 0.5%-2% of hydroxypropyl methylcellulose;

[0018] Environment-responsive modifier, containing 0.5%-3% of acid-sensitive bond modifier.

[0019] Preferably, the flavonoid is quercetin, the glycoside is tanshinone IIA, and the polyphenol is chlorogenic acid.

[0020] Preferably, the molar ratio of calcium ions, zinc ions, magnesium ions and iron ions in the multi-metal complex center is 2:1:1:1.

[0021] Preferably, the acid-sensitive bond modifier is succinate or its derivative.

[0022] Active Chinese medicine extract: Among them, flavonoids, glycosides and polyphenol compounds have good osteogenic promotion and antioxidant activities. As a representative of flavonoids, quercetin can improve bone metabolism by promoting the proliferation and differentiation of osteoblasts; tanshinone IIA and chlorogenic acid have synergistic anti-inflammatory effects and promote microcirculation in bone tissue, providing a multi-target action basis for the comprehensive treatment of osteoporosis.

[0023] Multi-metal complex center: Through the multi-metal complexation of calcium, zinc, magnesium and iron, a complex network is constructed. Calcium ions directly participate in the mineralization process of bone tissue; zinc ions promote new bone formation by activating the activity of osteoblast alkaline phosphatase; magnesium ions stabilize the bone matrix structure and improve the toughness of bone tissue; iron ions promote angiogenesis in the bone tissue microenvironment, providing blood support for bone repair. Through coordination chemistry, different metal ions coordinate synergistically to stabilize the active ingredients of traditional Chinese medicine, and at the same time enhance the bone targeting of drugs through the bone affinity of metal ions.

[0024] Auxiliary stabilizer: Chitosan forms a protective film to enhance the stability of complex particles and prevent them from being inactivated by external factors (such as oxidation or humidity) in vivo or during storage; hydroxypropyl methylcellulose improves the molding performance of the preparation by increasing the particle viscosity and delays drug release at the same time.

[0025] Environment-responsive modifier: The acid-sensitive bond modifier (such as succinate) will break in the acidic microenvironment (pH 5.5 - 6.8) of bone tissue, triggering the decomposition of the complex and releasing the active ingredients, thus achieving targeted and controlled drug release.

[0026] Preferably, the preparation method of the traditional Chinese medicine preparation for the rehabilitation of osteoporosis includes the following steps:

[0027] S1. Preparation of Chinese medicine extract: Extract Chinese medicine raw materials with an ethanol aqueous solution, and obtain the Chinese medicine extract through concentration, purification and drying;

[0028] S2. Preparation of metal ion solutions: Prepare calcium ion, zinc ion, magnesium ion and iron ion solutions respectively, and mix them in proportion;

[0029] S3. Complexation reaction: The metal ion solution is added dropwise to the traditional Chinese medicine extract solution, controlling the pH to be 6.8 - 7.5, and stirring and reacting at 40°C - 50°C for 1 - 2 hours;

[0030] S4. Environment-responsive modification: After the complexation reaction, an acid-sensitive modifier (succinate solution) is added and the reaction continues for 30 - 60 minutes; The introduction of succinate endows the complex with acid-sensitive properties, and it combines with the surface active groups of the complex to form cleavable bonds. Under the acidic conditions (pH 5.5 - 6.8) of the bone microenvironment, the acid-sensitive bonds break, releasing the active ingredients in the complex, achieving precise drug delivery and controlled release.

[0031] S5. Separation and drying: The complex precipitate is separated by centrifugation and freeze-dried at low temperature to obtain the traditional Chinese medicine complex powder;

[0032] Centrifugal separation effectively removes unreacted metal ions and traditional Chinese medicine molecules, obtaining pure complex powder. Freeze-drying avoids damage to the complex structure through low-temperature sublimation, while maintaining its chemical activity and physical stability.

[0033] S6. Granulation: The complex powder is mixed with a stabilizer to make a granule preparation;

[0034] Auxiliary stabilizers (chitosan, HPMC) enhance the stability by coating the complex granules, while delaying the release rate of the granules in the body. The acid-sensitive coating technology endows the granule preparation with environment-responsive properties by spraying succinate, further enhancing the bone targeting and controlled release effects.

[0035] Preferably, in step S1, a 50% - 70% ethanol aqueous solution is used as the extraction solvent, the extraction temperature is 60°C - 70°C, and the extraction time is 2 - 3 hours.

[0036] During the extraction process, a 50% - 70% ethanol solution can dissolve active ingredients such as flavonoids, glycosides, and polyphenols, while maintaining the stability of their structures. The concentration and purification processes remove non-medicinal ingredients (such as polysaccharides and proteins) from the traditional Chinese medicine extract, ensuring the purity of the active ingredients. Spray drying is completed under low-temperature conditions to prevent oxidation or degradation of the active ingredients.

[0037] Preferably, in step S2, the concentration range of the metal ion solution is as follows: calcium ion 0.1 - 0.2 mol / L, zinc ion 0.05 - 0.1 mol / L, magnesium ion 0.05 - 0.1 mol / L, iron ion 0.02 - 0.05 mol / L.

[0038] Metal ions provide a coordination center in the complexation reaction, forming coordination bonds with hydroxyl, carboxyl, and keto groups in the traditional Chinese medicine extract. By optimizing the proportion of metal ions, the stability and functionality of the complex are ensured. The calcium ion content is higher than other metal ions, which helps to enhance the bone affinity and mineralization ability of the complex.

[0039] Preferably, the pH value of the complexation reaction in step S3 is adjusted by 1 mol / L NaOH solution or HCl solution.

[0040] The complexation reaction is based on the principle of dynamic coordination chemistry. Metal ions form complexes with active groups in the traditional Chinese medicine extract through multiple sites, generating a stable coordination network. These reaction conditions (such as temperature, pH) can prevent the precipitation of metal ions or the degradation of traditional Chinese medicine molecules, while optimizing the complexation efficiency.

[0041] Preferably, the freeze-drying temperature range in step S5 is -40°C to -20°C, and the drying time is 8 - 12 hours.

[0042] Preferably, the coating of the granule preparation in step S6 is completed by spraying a succinate solution, the spraying temperature is 40°C - 50°C, the spraying speed is 1 - 2 mL / min, and the spraying is repeated 3 - 5 times.

[0043] The present invention provides a traditional Chinese medicine preparation for the rehabilitation of osteoporosis and its preparation method. It has the following

[0044] Beneficial effects:

[0045] 1. The present invention adopts a multi-metal synergistic complexation technology. Through the complexation of calcium, zinc, magnesium, and iron multi-metal ions with the active extract of traditional Chinese medicine, a complex with relatively high chemical stability is generated, achieving the technical effect of protecting the active ingredients from oxidation and degradation during preparation, storage, and in vivo transportation. Compared with the technical solution in the prior art that directly uses traditional Chinese medicine extract and is easily affected by environmental factors (such as light, oxygen, or humidity) resulting in the loss of active ingredients, it solves the problems of poor preparation stability and serious loss of effective ingredients.

[0046] 2. The present invention adopts a technical solution that combines a metal complexation center with an environment-responsive modifier, endowing the preparation with the characteristic of triggered release in an acidic environment, achieving the technical effects of enhanced bone targeting and improved drug delivery efficiency. Compared with the problems of poor drug targeting and uneven drug efficacy distribution in traditional Chinese medicine preparations in the prior art, it solves the problems of low bone tissue absorption efficiency and insufficient drug utilization.

[0047] 3. The present invention selectively introduces multi-metal ions such as calcium, zinc, magnesium, and iron to synergistically act with the active extracts of traditional Chinese medicine. Through the optimized design of the proportion of multi-metal ions, the technical effect of comprehensive treatment through multiple functions such as bone metabolism regulation, osteoblast activity enhancement, and bone tissue angiogenesis promotion is achieved. Compared with the technical solutions in the prior art that rely only on a single component or a single action target, the problem of single treatment effect and insufficiency in dealing with the complex pathological mechanism of osteoporosis is solved.

[0048] 4. The present invention adopts preparation technical solutions such as acid-sensitive modification, granulation, and freeze-drying, significantly improving the physical stability and process repeatability of the preparation, and achieving the technical effects of facilitating industrial production, good preparation uniformity and stability. Compared with the problems of complex preparation process, poor stability, and insufficient quality consistency between batches in the prior art, the deficiencies of high preparation production difficulty and limited process scale-up are solved. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 It is a flow chart of the preparation method of the traditional Chinese medicine preparation for osteoporosis rehabilitation. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0050] Next, the technical solutions of the present invention will be clearly and completely described in conjunction with the drawings of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0051] Please refer to the attached Figure 1 :

[0052] Example:

[0053] Example 1:

[0054] Step 1: Preparation of traditional Chinese medicine extract

[0055] The raw medicinal materials (quercetin, salvia miltiorrhiza, and the plant source of chlorogenic acid) are cleaned, dried, and then pulverized to 60 mesh.

[0056] Weigh 500 g of the medicinal materials and perform reflux extraction with 50% ethanol aqueous solution (the mass ratio of the medicinal materials to the solvent is 1:8). The extraction temperature is 65 °C, the extraction time is 2 hours, and the extraction is repeated 2 times.

[0057] Concentrate the extract to 1 / 3 of the original volume and purify it through macroporous resin: the sample loading amount is the mass ratio of the medicinal liquid to the resin of 1:10, and the eluent is 80% ethanol solution.

[0058] The purified liquid is spray-dried (inlet temperature 130 °C, outlet temperature 70 °C) to obtain 65 g of the active extract of traditional Chinese medicine.

[0059] Step 2: Preparation of metal ion solutions

[0060] Prepare 0.15 mol / L CaCl₂ solution, 0.08 mol / L ZnSO₄ solution, 0.08 mol / L MgCl₂ solution and 0.03 mol / L FeCl₃ solution.

[0061] Mix the above solutions in a molar ratio of 2:1:1:1 to obtain a multi-metal ion mixture.

[0062] Step 3: Complexation reaction

[0063] Place the 5% (w / v) traditional Chinese medicine extract solution under stirring conditions (400 rpm), and gradually add the multi-metal ion mixture dropwise, controlling the liquid addition rate at 1 mL / min.

[0064] Adjust the pH to 7.0 with 1 mol / L NaOH solution, the reaction temperature is 45 °C, and the stirring time is 1.5 hours to form a complex solution.

[0065] Step 4: Environment-responsive modification

[0066] Add 3% (w / v) succinate solution after the complexation reaction and continue stirring for 45 minutes.

[0067] Step 5: Separation and drying

[0068] Separate the precipitate by centrifugation (5000 rpm, 10 minutes) and wash it twice with deionized water.

[0069] Freeze-dry (-30 °C, 10 hours) to obtain 78 g of multi-metal complex powder.

[0070] Step 6: Granulation

[0071] Mix the multi-metal complex powder with chitosan (mass ratio 1:10) and hydroxypropyl methylcellulose (0.5% w / w), add deionized water (mass ratio 1:2) to prepare wet granules, and form them by extrusion-spheronization.

[0072] The drying temperature is 55 °C and the drying time is 4 hours.

[0073] Spray-coat with 5% (w / v) succinate solution, the spraying temperature is 45 °C, the spraying speed is 1.5 mL / min, and repeat spraying 3 times to obtain the granule preparation.

[0074] In this example, through multi-metal complexation and acid-sensitive modification technologies, the stability of traditional Chinese medicine active ingredients was significantly improved, while bone targeting and controlled release functions were achieved, effectively solving the problems of uneven distribution of drug efficacy and easy degradation of ingredients in the prior art.

[0075] Example 2:

[0076] Step 1: Preparation of traditional Chinese medicine extract

[0077] The same as in Example 1.

[0078] Step 2: Preparation of metal ion solution

[0079] Prepare 0.2 mol / L CaCl2 solution, 0.05 mol / L ZnSO4 solution, 0.05 mol / L MgCl2 solution and 0.02 mol / L FeCl3 solution.

[0080] Mix them in a molar ratio of 3:1:1:0.5 to obtain a multi-metal ion mixture.

[0081] Step 3: Complexation reaction

[0082] Using the same method as in Example 1, slowly add the metal ion solution drop by drop, but the reaction conditions are 40 °C, the pH is adjusted to 6.8, and the stirring time is 2 hours.

[0083] Step 4: Environment-responsive modification

[0084] Add 2.5% (w / v) succinate solution and react for 30 minutes.

[0085] Step 5: Separation and drying

[0086] Centrifuge the precipitate at 6000 rpm, wash it 3 times, and obtain the complex powder by freeze-drying (-40 °C, 12 hours).

[0087] Step 6: Granulation

[0088] The conditions for preparing granules and spraying coating are the same as in Example 1.

[0089] By optimizing the calcium ion content, the bone affinity of the complex was enhanced, further improving the bone tissue adsorption effect and solving the problem of insufficient drug adsorption efficiency in the prior art.

[0090] Example 3: Traditional Chinese medicine preparation with increased proportion of magnesium ions

[0091] Step 1: Preparation of traditional Chinese medicine extract

[0092] The same as in Example 1.

[0093] Step 2: Preparation of metal ion solution

[0094] Prepare 0.1 mol / L CaCl₂ solution, 0.05 mol / L ZnSO₄ solution, 0.1 mol / L MgCl₂ solution and 0.03 mol / L FeCl₃ solution.

[0095] Mix them in a molar ratio of 2:1:2:1 to obtain a multi-metal ion solution.

[0096] Step 3: Complexation reaction

[0097] In the same way as in Example 1, dropwise add the metal ion solution. The reaction conditions are 45 °C, adjust the pH to 7.5, and stir for 1.5 hours.

[0098] Step 4: Environment-responsive modification

[0099] Add 3% (w / v) succinate solution and continue the reaction for 60 minutes.

[0100] Step 5: Separation and drying

[0101] Separate by centrifugation (5000 rpm, 10 minutes). The freeze-drying conditions are -35 °C for 10 hours.

[0102] Step 6: Granulation

[0103] The conditions for preparing the granules and spray coating are the same as in Example 1.

[0104] Increasing the proportion of magnesium ions improves the stabilizing effect of the complex on the bone matrix. At the same time, through the strengthening effect of magnesium ions on the toughness of the bone matrix, the problem that bone brittleness cannot be effectively improved in the prior art is solved.

[0105] Example 4: Traditional Chinese medicine preparation using different acid-sensitive modifiers

[0106] Step 1: Preparation of traditional Chinese medicine extract

[0107] The same as in Example 1.

[0108] Step 2: Preparation of metal ion solution

[0109] The same as in Example 1.

[0110] Step 3: Complexation reaction

[0111] The same as in Example 1.

[0112] Step 4: Environment-responsive modification

[0113] Use 4% (w / v) malonate solution as the modifier. The reaction conditions are 40 minutes and the stirring speed is 400 rpm.

[0114] Step 5: Separation and Drying

[0115] Same as Example 1

[0116] Step 6: Granulation

[0117] Same as Example 1

[0118] Using malonate as a modifier endows the preparation with a stronger acid-responsive ability, achieving more precise release in the bone microenvironment and solving the problem of insufficient drug release efficiency in the prior art.

[0119] Example 5: Traditional Chinese Medicine Preparation with Improved Granulation Process Stability

[0120] Steps 1-5:

[0121] Same as Example 1

[0122] Step 6: Granulation

[0123] Adjust the chitosan content to 3%, the hydroxypropyl methylcellulose content to 1%, and the proportion of purified water used in wet granulation to 1:3.

[0124] Set the drying temperature at 50 °C and extend the drying time to 5 hours.

[0125] During the coating process, reduce the spraying speed to 1 mL / min and repeat spraying 4 times.

[0126] Adjusting the granulation process stabilizes the morphology of the preparation and improves the uniformity of the granule preparation, solving the problem of poor batch-to-batch consistency of granulated preparations in the prior art.

[0127] Comparative Example:

[0128] Comparative Example 1: Without using multi-metal synergistic complexation (corresponding to Example 1)

[0129] Preparation process:

[0130] Step 1: Preparation of Traditional Chinese Medicine Extract

[0131] Completely extract according to Step 1 of Example 1 to obtain the traditional Chinese medicine extract.

[0132] Step 2: Preparation of Metal Ion Solution

[0133] Only use single calcium ion to prepare a 0.15 mol / L CaCl2 solution without introducing zinc ion, magnesium ion and iron ion.

[0134] Step 3: Complexation Reaction

[0135] Under the same conditions (pH 7.0, stirred at 45 °C for 1.5 hours), the calcium ion solution was added dropwise to the 5% (w / v) traditional Chinese medicine extract solution to complete the single-metal complexation.

[0136] Step 4: Environment-responsive modification

[0137] Add 3% (w / v) succinate solution and stir for 45 minutes.

[0138] Step 5: Separation and drying

[0139] Centrifuge the precipitate (5000 rpm, 10 minutes) and freeze-dry (-30 °C, 10 hours) to obtain the complex powder.

[0140] Step 6: Granulation

[0141] Prepare granules according to Step 6 of Example 1.

[0142] This comparative example only uses calcium ions as the complexation center, lacking the synergistic effect of multi-metal ions and unable to provide multi-functional targeting (such as zinc promoting osteogenesis and magnesium enhancing bone toughness, etc.) for comparing the advantages of multi-metal synergistic complexation technology.

[0143] Comparative Example 2:

[0144] Preparation process:

[0145] Step 1: Preparation of traditional Chinese medicine extract

[0146] Extract according to Step 1 of Example 1 to obtain the traditional Chinese medicine extract.

[0147] Step 2: Preparation of metal ion solution

[0148] Prepare the multi-metal ion solution according to Step 2 of Example 1.

[0149] Step 3: Complexation reaction

[0150] Carry out the complexation reaction exactly according to Step 3 of Example 1.

[0151] Step 4: Environment-responsive modification

[0152] Skip the acid-sensitive modification step and do not add the succinate solution.

[0153] Step 5: Separation and drying

[0154] Separate and dry according to Step 5 of Example 1.

[0155] Step 6: Granulation

[0156] Prepare granules according to Step 6 of Example 1.

[0157] This comparative example did not perform acid-sensitive modification, only forming simple complex particles and not having the ability of specific release in the bone microenvironment, which was used to compare the contribution of environmental response modification to drug controlled release and bone targeting.

[0158] Comparative Example 3:

[0159] Preparation process:

[0160] Step 1: Preparation of traditional Chinese medicine extract

[0161] Extraction was carried out according to Step 1 of Example 1 to obtain the traditional Chinese medicine extract.

[0162] Step 2: Preparation of metal ion solution

[0163] Prepare 0.15 mol / L CaCl2 solution, 0.1 mol / L ZnSO4 solution, 0.1 mol / L MgCl2 solution and 0.03 mol / L FeCl3 solution, but adjust the molar ratio of calcium ions to other metal ions to 1:1:1:1.

[0164] Step 3: Complexation reaction

[0165] The complexation reaction was carried out according to Step 3 of Example 1.

[0166] Step 4: Environmental response modification

[0167] Acid-sensitive modification was carried out according to Step 4 of Example 1.

[0168] Step 5: Separation and drying

[0169] Separation and drying were carried out according to Step 5 of Example 1.

[0170] Step 6: Granulation

[0171] Granules were prepared according to Step 6 of Example 1.

[0172] In this comparative example, the ratio of calcium ions to other metal ions was adjusted to weaken the dominant role of calcium ions, resulting in a weakened promotion effect on bone tissue mineralization, which was used to compare the influence of optimized ratio design on bone targeting and therapeutic effect.

[0173] Comparative Example 4:

[0174] Preparation process:

[0175] Step 1: Preparation of traditional Chinese medicine extract

[0176] Extraction was carried out according to Step 1 of Example 1 to obtain the traditional Chinese medicine extract.

[0177] Step 2: Preparation of metal ion solution

[0178] Prepare a multi-metal ion solution according to Step 2 of Example 1.

[0179] Step 3: Complexation reaction

[0180] Carry out the complexation reaction according to Step 3 of Example 1.

[0181] Step 4: Environment-responsive modification

[0182] Carry out acid-sensitive modification according to Step 4 of Example 1.

[0183] Step 5: Separation and drying

[0184] Separate and dry according to Step 5 of Example 1.

[0185] Step 6: Granulation

[0186] In the preparation of granules, no chitosan is added, and only hydroxypropyl methylcellulose (0.5% w / w) is added to prepare wet granules. The coating spraying conditions are the same as those in Example 1.

[0187] In this comparative example, chitosan is not used as an auxiliary stabilizer, and the surface protection performance and drug stability of the granule preparation will be significantly reduced, which is used to compare the role of chitosan in enhancing the stability of the preparation.

[0188] Comparative Example 5:

[0189] Preparation process:

[0190] Step 1: Preparation of traditional Chinese medicine extract

[0191] Carry out extraction according to Step 1 of Example 1 to obtain the traditional Chinese medicine extract.

[0192] Step 2: Preparation of metal ion solution

[0193] Prepare a multi-metal ion solution according to Step 2 of Example 1.

[0194] Step 3: Complexation reaction

[0195] Carry out the complexation reaction according to Step 3 of Example 1.

[0196] Step 4: Environment-responsive modification

[0197] Carry out acid-sensitive modification according to Step 4 of Example 1.

[0198] Step 5: Separation and drying

[0199] Use hot air drying instead of freeze drying. The drying temperature is set at 80 °C and the drying time is 6 hours.

[0200] Step 6: Granulation

[0201] The granule preparation is the same as that in Example 1.

[0202] This comparative example uses a hot air drying process, which may cause partial degradation of the active ingredients of the complex or instability of the physical structure, and is used to compare the role of freeze drying in maintaining the stability of active ingredients and the quality of the preparation.

[0203] Test experiment:

[0204] Experiment 1: Stability test of traditional Chinese medicine active ingredients

[0205] Experiment description

[0206] Experiment purpose

[0207] By comparing the stability of the traditional Chinese medicine active extracts in the examples and the comparative examples, verify the role of multi-metal synergistic complexation and chitosan stabilizer in improving the stability of the preparation.

[0208] Experiment steps

[0209] Sample preparation

[0210] Weigh the granular preparations of Example 1, Example 5, Comparative Example 1 and Comparative Example 4, dissolve them in deionized water respectively, and prepare an aqueous solution of 1 mg / mL to ensure that the sample concentrations are the same.

[0211] Storage condition setting

[0212] The samples are stored under the following two conditions respectively:

[0213] Normal temperature and dark condition: 25 °C, humidity 40%-50%.

[0214] Accelerated aging condition: 40 °C, humidity 75%.

[0215] Samples are taken for detection at 0 day, 7 days, 14 days and 30 days.

[0216] Detection of active ingredient content

[0217] Take 10 μL of the solution of each sample at the corresponding time point, and use high performance liquid chromatography (HPLC) to detect the contents of quercetin, tanshinone IIA and chlorogenic acid.

[0218] The detection wavelength of quercetin is 365 nm, tanshinone IIA is 270 nm, and chlorogenic acid is 320 nm.

[0219] The retention rate of the active ingredient at each time point is calculated according to the following formula:

[0220]

[0221] Data recording

[0222] Compare the retention rates of different samples under two conditions and statistically analyze their variation patterns.

[0223] Experimental data

[0224] Comparison table of the retention rates of traditional Chinese medicine active ingredients (under different storage conditions)

[0225]

[0226]

[0227] In this experiment, by comparing the stability of the active ingredients of traditional Chinese medicine in the examples and comparative examples, the action mechanisms of the multi-metal synergistic complexation technology and chitosan stabilizer were verified. First, in Examples 1 and 5, stable coordination bonds were formed through multi-metal complexation (calcium, zinc, magnesium, iron) with the active groups of traditional Chinese medicine (such as hydroxyl and carboxyl groups), significantly inhibiting the oxidative decomposition of the active ingredients; while in Comparative Example 1, only single calcium ions were used, and the chemical stability of the complex was poor, resulting in a significant decrease in the retention rate. This proves the necessity of the multi-metal synergistic effect.

[0228] Secondly, as an auxiliary stabilizer, chitosan forms a protective film by coating the particle surface, further slowing down the destructive effects of environmental humidity and oxygen on the preparation. The stability of Example 5 under normal temperature and dark conditions is better than that of Comparative Example 4, fully verifying the value of adding chitosan in the granulation process.

[0229] Finally, under accelerated aging conditions, the retention rates of the active ingredients in the examples are still significantly higher than those in the comparative examples. Especially for oxidation-sensitive components such as quercetin and chlorogenic acid, the retention rates are increased by more than 20% respectively. This is closely related to the dynamic coordination chemistry properties of the multi-metal complexation centers. The dynamic reorganization ability of the coordination bonds can effectively buffer the influence of the external environment and further improve the stability of the active ingredients. These results fully demonstrate the significant advantages of the present invention in the stability of traditional Chinese medicine preparations.

[0230] Experiment 2: Bone targeting test

[0231] Experiment description

[0232] Experiment purpose

[0233] By comparing the bone targeting properties of the examples and comparative examples, verify the promoting effects of multi-metal synergistic complexation and acid-sensitive modification on the drug distribution in bone tissue.

[0234] Experimental procedure

[0235] Sample preparation

[0236] Select the granule preparations of Example 1, Example 2, Example 4, Comparative Example 1, and Comparative Example 2.

[0237] Prepare a 1 mg / mL sample solution and label each preparation with a fluorescent labeling reagent (FITC, fluorescein isothiocyanate). After stirring for 2 hours for sufficient reaction, remove the unbound fluorescent reagent using a dialysis bag to obtain the labeled sample.

[0238] Animal experiment

[0239] Use healthy adult female SD rats (body weight 200 - 220 g), divided into the following 5 groups, with 6 rats in each group:

[0240] Example 1 group

[0241] Example 2 group

[0242] Example 4 group

[0243] Control group 1

[0244] Control group 2

[0245] Inject the fluorescently labeled sample solution into each rat via the tail vein at a dose of 10 mg / kg body weight.

[0246] Sampling and detection

[0247] At 2 hours, 4 hours, 8 hours, and 12 hours after administration, randomly sacrifice 1 rat from each group, and take the femur (bone tissue) and tissues of the main organs (liver, kidney, heart).

[0248] Use a fluorescence microscopy imaging instrument (excitation wavelength 495 nm, emission wavelength 520 nm) to detect the fluorescence intensity in the tissues.

[0249] Normalize the detection results and calculate the fluorescence intensity ratio of the drug distributed in the bone tissue.

[0250] Experimental data

[0251] Fluorescent distribution intensity of different preparations in bone tissue (after normalization)

[0252]

[0253]

[0254] In this experiment, by comparing the fluorescence intensities of different preparations in bone tissue, the mechanism of action of multi-metal synergistic complexation and acid-sensitive modification on bone targeting was verified. Example 1 showed the highest fluorescence distribution ratio in bone tissue (still as high as 88% after 12 hours), which was due to the bone affinity conferred by the multi-metal complexation of calcium, zinc, magnesium, and iron. Calcium ions directly participated in the mineralization process of bone tissue, zinc and magnesium ions further promoted the activity of osteoblasts, and iron ions increased the blood supply of bone tissue, significantly enhancing the selective distribution of the preparation in bone tissue.

[0255] The introduction of acid-sensitive modifiers also played an important role. In Example 1 and Example 2, succinate modification enabled the drug to be rapidly released in the acidic microenvironment of bone tissue (such as the active area of osteoclasts), thus further increasing bone targeting. In Comparative Example 2, due to the lack of acid-sensitive modification, the distribution of the fluorescence signal in bone tissue was significantly lower than that in Example 1.

[0256] In addition, in Example 4, a different acid-sensitive modifier (malonate) was used, which had weaker acid sensitivity, resulting in slightly lower bone targeting than Example 1. This result indicates that the selection of acid-sensitive modifiers is crucial for the precise release of the formulation at specific targets. In Comparative Example 1, the single calcium complex lacked the synergistic effect of multiple metals, resulting in the worst targeting distribution in bone tissue. This experimental result fully demonstrates the synergistic effect of multi-metal synergistic complexation and acid-sensitive modification in the technical solution of the present invention, which has significant advantages in improving bone targeting.

[0257] Experiment 3: Controlled release performance test

[0258] Experimental description

[0259] Experimental purpose

[0260] By comparing the drug release behaviors of different formulations in simulated bone microenvironment and neutral environment, verify the role and effect of acid-sensitive modifiers in drug controlled release, and further evaluate the influence of multi-metal synergistic complexation on controlled release performance.

[0261] Experimental procedure

[0262] Sample preparation

[0263] Select the particulate preparations of Example 1, Example 4, and Comparative Example 2 and Comparative Example 3.

[0264] Weigh 10 mg of the particulate preparation and place it in buffer solutions containing 10 mL of simulated bone microenvironment (pH 5.5) and neutral environment (pH 7.4) respectively.

[0265] Release experiment setup

[0266] Place the sample solution in a constant temperature oscillator at 37 °C (oscillation speed is 100 rpm).

[0267] Sampling is carried out at the following time points: 0 h, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h. Each time, 1 mL of the solution is taken out, and an equal amount of fresh buffer is added at the same time.

[0268] Detection of active ingredient content

[0269] The concentrations of quercetin, tanshinone IIA, and chlorogenic acid in the buffer solution were determined using an ultraviolet spectrophotometer at wavelengths of 365 nm, 270 nm, and 320 nm, respectively.

[0270] The cumulative release rate was calculated using the following formula:

[0271]

[0272] The release behaviors of different samples were compared in acidic (pH 5.5) and neutral (pH 7.4) environments.

[0273] Experimental data

[0274] Cumulative release rates of different formulations in simulated bone microenvironment and neutral environment

[0275]

[0276]

[0277] By comparing the release behaviors of different formulations in acidic and neutral environments, it can be clearly seen that the examples of the present invention have significant advantages in controlled release performance. The acid-sensitive modifiers (succinate and malonate) in Example 1 and Example 4 endow the formulations with rapid release characteristics under acidic conditions of pH 5.5. This is because the acid-sensitive bonds break in a low pH environment, triggering the rapid release of traditional Chinese medicine active ingredients; in a neutral environment (pH 7.4), the acid-sensitive bonds remain stable, significantly slowing down the drug release rate. This property is crucial for targeted drug release in the bone microenvironment, as the acidic microenvironment in the active area of osteoclasts in osteoporosis can precisely trigger drug release and reduce drug distribution in non-target tissues.

[0278] In Comparative Example 2 and Comparative Example 3, due to the absence of acid-sensitive modifiers or the lack of optimization of the metal complexation ratio, their release behaviors showed weak environmental responsiveness, slower drug release rates, and no significant differences. This indicates that the acid-sensitive modifiers in the present invention play a decisive role in improving the controlled release performance.

[0279] In addition, the dynamic coordination effect of multi-metal synergistic complexation in Example 1 and Example 4 further stabilized the drug release behavior. At pH 7.4, the release rate of Example 1 was significantly lower than that of Comparative Example 3, indicating that the multi-metal complexation structure can delay non-targeted drug release while still maintaining a high response efficiency under acidic conditions. This result shows that the combined design of multi-metal complexation and acid-sensitive modification not only enhances the controlled release performance but also achieves a balance between environmental responsiveness and release delay, significantly improving the potential application value of the formulations of the present invention in the treatment of osteoporosis.

[0280] Experiment 4: Comprehensive treatment effect test

[0281] Experimental Instructions

[0282] Experimental Objectives

[0283] By comparing the treatment effects of osteoporosis in the examples and comparative examples, verify the enhancing effects of the multi-metal synergistic complexation and acid-sensitive modification technologies on bone density, bone biomarkers, and bone mechanical properties, and further prove the overall advantages of the present invention.

[0284] Experimental Procedures

[0285] Establishment of Animal Model

[0286] Select healthy female SD rats, weighing 200 - 220 g, and randomly divide them into an osteoporosis model group and a normal control group.

[0287] The osteoporosis model was established by bilateral ovariectomy (OVX), and the success of the osteoporosis model (significant decrease in bone density) was confirmed 6 weeks after the operation.

[0288] Experimental Grouping and Administration

[0289] Randomly divide the model rats into 6 groups (10 rats in each group):

[0290] Normal control group: No treatment;

[0291] Model control group: Administer an equal volume of normal saline;

[0292] Example 1 group: Intragastrically administer the granule preparation of Example 1 (dose: 100 mg / kg) daily;

[0293] Example 2 group: Intragastrically administer the granule preparation of Example 2 (dose: 100 mg / kg) daily;

[0294] Comparative Example 1 group: Intragastrically administer the granule preparation of Comparative Example 1 (dose: 100 mg / kg) daily;

[0295] Comparative Example 2 group: Intragastrically administer the granule preparation of Comparative Example 2 (dose: 100 mg / kg) daily.

[0296] Continuously intragastrically administer for 6 weeks, once a day.

[0297] Detection Indexes and Methods

[0298] Bone Density Detection

[0299] After 6 weeks, anesthetize the rats and use dual-energy X-ray absorptiometry (DEXA) to detect the bone mineral density (BMD) of the rat femur, with the unit of g / cm 2 .

[0300] Bone Biomarker Detection

[0301] Collect serum and detect the following bone metabolism markers:

[0302] Bone resorption markers: CTX-I (C-terminal collagen degradation product);

[0303] Bone formation markers: ALP (alkaline phosphatase);

[0304] Use enzyme-linked immunosorbent assay (ELISA) for detection and record their concentrations (ng / mL and U / L) respectively.

[0305] Bone mechanical property test

[0306] Take the femurs of rats and use three-point bending test to measure fracture toughness (N / mm) and maximum fracture load (N).

[0307] Experimental data

[0308] Comparison of the treatment effects of osteoporosis in rats of different groups

[0309]

[0310]

[0311] In terms of bone mineral density (BMD), the bone mineral density of the femurs of the rats in Example 1 and Example 2 groups was significantly higher than that of the model control group, Comparative Example 1 group and Comparative Example 2 group, approaching the level of the normal control group, indicating that the multi-metal synergistic complexation technology in the present invention significantly promoted bone mineralization. Calcium ions were directly deposited in the bone matrix, and zinc and magnesium ions enhanced bone formation by promoting the proliferation and differentiation of osteoblasts, while iron ions further increased bone density by improving bone microcirculation. In contrast, due to the lack of multi-metal synergy in Comparative Example 1, only single calcium ion complexation was used, and the mineralization effect was limited, and the increase in bone density was not obvious.

[0312] In the detection of bone metabolism markers, CTX-I (bone resorption marker) in the Example group was significantly lower than that in the Comparative Example group, indicating that it could effectively inhibit bone resorption. At the same time, the ALP level (bone formation marker) in Example 1 was significantly higher than that in the Comparative Example, indicating that the synergistic effect of multi-metal complexation and acid-sensitive modifier promoted bone formation. This was closely related to the mechanism of the acid-sensitive modifier: succinate triggered drug release in the acidic microenvironment of bone tissue, enabling the drug to act efficiently on the active area of osteoclasts, inhibiting bone resorption while promoting the activity of osteoblasts. Due to the lack of acid sensitivity modification in Comparative Example 2, the drug release was not concentrated enough, and the treatment effect was limited.

[0313] The bone mechanical property tests further verified the advantages of the example group in bone treatment. The fracture toughness and maximum load of Example 1 were significantly higher than those of the comparative examples and were close to those of the normal control group. This indicates that the multi-metal synergistic complexation technology not only improved bone density but also enhanced the structural strength of the bone matrix. In Comparative Example 1 and Comparative Example 2, due to uneven drug distribution or low release efficiency, the repair effect of the bone matrix was poor and the mechanical property recovery was insufficient.

[0314] In summary, through the combined design of the multi-metal synergistic complexation technology and the acid-sensitive modifier, the present invention achieved an all-round effect of improving bone density, balancing bone metabolism, and improving bone mechanical properties in the treatment of osteoporosis, which was significantly superior to the single-metal complexation or uncontrolled release design in the prior art, reflecting strong technological innovation and practical value.

[0315] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A traditional Chinese medicine preparation for osteoporosis rehabilitation, characterized in that, The traditional Chinese medicine preparation comprises a composition of the following components: 3%-10% of traditional Chinese medicine active extract, which includes flavonoids, glycosides and polyphenolic compounds; Multi-metal complex center, containing 10%-30% of calcium ions, 5%-15% of zinc ions, 5%-10% of magnesium ions and 2%-10% of iron ions; Auxiliary stabilizer, containing 1%-5% of chitosan and 0.5%-2% of hydroxypropyl methylcellulose; Environment-responsive modifier, containing 0.5%-3% of acid-sensitive bond modifier.

2. The traditional Chinese medicine preparation for osteoporosis rehabilitation according to claim 1, wherein, The flavonoids are quercetin, the glycosides are tanshinone IIA, and the polyphenols are chlorogenic acid.

3. The traditional Chinese medicine preparation for osteoporosis rehabilitation according to claim 1, characterized in that, The molar ratio of calcium ions to zinc ions, magnesium ions and iron ions in the multi-metal complex center is 2:1:1:

1.

4. The traditional Chinese medicine preparation for osteoporosis rehabilitation according to claim 1, wherein, The acid-sensitive bond modifier is succinate or its derivative.

5. The preparation method of the traditional Chinese medicine preparation for osteoporosis rehabilitation according to any one of claims 1-4, characterized in that, It includes the following steps: S1. Preparation of traditional Chinese medicine extract: Extract traditional Chinese medicine raw materials with an ethanol aqueous solution, and obtain the traditional Chinese medicine extract through concentration, purification and drying; S2. Preparation of metal ion solutions: Prepare calcium ion solution, zinc ion solution, magnesium ion solution and iron ion solution respectively, and mix them in proportion; S3. Complexation reaction: Dropwise add the metal ion solution into the traditional Chinese medicine extract solution, control the pH to be 6.8-7.5, and stir and react at 40°C-50°C for 1-2 hours; S4. Environment-responsive modification: Add the acid-sensitive modifier after the complexation reaction, and continue to react for 30-60 minutes; S5. Separation and drying: Centrifuge to separate the complex precipitate, and freeze-dry at low temperature to obtain the traditional Chinese medicine complex powder; S6. Granulation: Mix the complex powder with the stabilizer to make a granule preparation.

6. The preparation method of the traditional Chinese medicine preparation for osteoporosis rehabilitation according to claim 5, characterized in that, In the step S1, 50%-70% ethanol aqueous solution is used as the extraction solvent, the extraction temperature is 60°C-70°C, and the extraction time is 2-3 hours.

7. The preparation method of the traditional Chinese medicine preparation for osteoporosis rehabilitation according to claim 5, characterized in that, In the step S2, the concentration range of the metal ion solutions is: 0.1-0.2 mol / L for calcium ions, 0.05-0.1 mol / L for zinc ions, 0.05-0.1 mol / L for magnesium ions, and 0.02-0.05 mol / L for iron ions.

8. The preparation method of the traditional Chinese medicine preparation for osteoporosis rehabilitation according to claim 5, characterized in that, In the step S3, the pH value of the complexation reaction is adjusted by 1 mol / L NaOH solution or HCl solution.

9. The preparation method of the traditional Chinese medicine preparation for osteoporosis rehabilitation according to claim 5, characterized in that, In the step S5, the freeze-drying temperature range is -40°C to -20°C, and the drying time is 8-12 hours.

10. The preparation method of the traditional Chinese medicine preparation for osteoporosis rehabilitation according to claim 5, characterized in that, In the step S6, the coating of the granule preparation is completed by spraying succinate solution, the spraying temperature is 40°C-50°C, the spraying speed is 1-2 mL / min, and the spraying is repeated 3-5 times.

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