Ganoderma lucidum superfine powder-loaded gradient drug release type plaster patch and preparation method thereof

Abstract

The invention relates to the technical field of pharmaceutical preparations, and discloses a gradient drug release type plaster patch loaded with ganoderma lucidum superfine powder and a preparation method, the patch comprises a double-layer hydrogel matrix, and A-type quick release microcapsules and B-type long-acting self-emulsifying permeation enhancing microcapsules are dispersed in the matrix. A type micro-capsules and B type micro-capsules are dispersed in the lower-layer matrix, and only the B type micro-capsules are dispersed in the upper-layer matrix. Ganoderma triterpenes are entrapped by the A-type microcapsules, so that quick release is realized. Ganoderma lucidum oligosaccharide modified by enzymolysis is entrapped in the B-type microcapsule, the core of the B-type microcapsule is a self-microemulsification system precursor, and long-acting release and efficient permeation enhancement are realized after the B-type microcapsule is entrapped by a hydrophobic wall material. The preparation method comprises the steps of enzymolysis modification, double-microcapsule preparation and construction of an integrated double-layer structure through a laminated coating and temperature-sensitive gel curing technology. According to the invention, the problem of cooperative delivery of active components of ganoderma lucidum with different properties is solved, programmed release is realized, and bioavailability is remarkably improved.

Description

Technical Field

[0001] The invention relates to the technical field of pharmaceutical preparations, in particular to a gradient drug-releasing plaster patch loaded with ganoderma lucidum ultrafine powder and a preparation method thereof. Background Art

[0002] Ganoderma, a traditional medicinal fungus, contains active ingredients primarily consisting of fat-soluble triterpenes and water-soluble polysaccharides, which exhibit synergistic pharmacological effects. Transdermal administration is an effective route for drug delivery, avoiding the first-pass effect of oral administration and providing sustained, consistent blood drug concentrations. Therefore, formulating Ganoderma active ingredients into patches is an important direction for their modern application.

[0003] However, integrating multiple active ingredients from Ganoderma lucidum into a single transdermal drug delivery system and achieving efficient delivery faces significant technical bottlenecks. First, Ganoderma lucidum triterpenes and Ganoderma lucidum polysaccharides differ significantly in their physicochemical properties. The former are small, fat-soluble molecules, while the latter are large, water-soluble molecules. This difference makes it difficult to achieve both stable coexistence and effective release within the same matrix. Conventional single-matrix patches often only optimize the delivery of one component, sacrificing the efficiency and stability of the other.

[0004] Secondly, the hydrophobic barrier function of the skin's stratum corneum is a major obstacle to transdermal drug delivery. High-molecular-weight, highly hydrophilic substances like Ganoderma lucidum polysaccharides inherently have extremely low transdermal permeability, making it difficult for them to penetrate the skin barrier and reach their site of action. Even for fat-soluble Ganoderma lucidum triterpenes, transdermal efficiency is suboptimal, often requiring additional permeation enhancement techniques to achieve effective tissue concentrations. However, existing permeation enhancement methods are often limited and ineffective.

[0005] In addition, existing patch products also have limitations in drug release patterns. Most patches have a single homogeneous structure and a single drug release profile, usually showing an initial rapid burst release or a simple zero-order / first-order release, which cannot achieve programmed drug release behavior based on therapeutic needs. In actual applications, a gradient release pattern that can first take effect quickly and then have a long-lasting effect often leads to better clinical results, but existing patch technology makes it difficult to achieve this complex, time-controlled release of different components. Summary of the Invention

[0006] In response to the shortcomings of the existing technology, the present invention provides a gradient-release plaster patch loaded with ultrafine Ganoderma lucidum powder and a preparation method, which solves the problem that fat-soluble triterpenes and water-soluble polysaccharides with very different physicochemical properties in Ganoderma lucidum are difficult to stably coexist, programmatically release, and synergistically and efficiently absorb through the skin in the same transdermal drug delivery system.

[0007] In order to achieve the above objectives, the present invention provides the following technical solutions.

[0008] The first aspect of the present invention provides a gradient drug-release plaster patch loaded with ultrafine Ganoderma lucidum powder.

[0009] The patch consists of a core double-layer hydrogel matrix loaded with and dispersed within it two functionally distinct microcapsules: Type A rapid-release microcapsules and Type B long-acting self-emulsifying permeation-enhancing microcapsules. Through the specific arrangement of structure and components, the patch achieves gradient release and synergistic penetration of the drug.

[0010] Specifically, the bilayer hydrogel matrix consists of a lower and upper layer structured in a stacked structure. The lower layer directly contacts the skin surface and contains both Type A immediate-release microcapsules and Type B long-acting self-emulsifying and permeation-enhancing microcapsules. The upper layer, located above the lower layer, serves as a drug reservoir and contains only Type B long-acting self-emulsifying and permeation-enhancing microcapsules. This distribution creates a concentration gradient of the long-acting active ingredient from the upper layer to the lower layer.

[0011] The structure of the Type A rapid-release microcapsule is composed of a core and a water-soluble wall material, wherein the mass ratio of the core to the wall material is 30-45:55-70. The core contains two components: One component comprises a Ganoderma triterpenoid extract obtained by supercritical CO2 extraction of ultrafine Ganoderma lucidum powder, and the other comprises a penetration enhancer. In one specific embodiment, the ratio of Ganoderma triterpenoid extract to penetration enhancer in the core is 70 to 85 parts by weight: 15 to 30 parts by weight. When the patch is applied, moisture in the underlying matrix rapidly dissolves the water-soluble wall material of the Type A microcapsules, rapidly releasing their contents (triterpenes and penetration enhancer) to the skin's stratum corneum.

[0012] The Type B long-lasting self-emulsifying permeability-enhancing microcapsules consist of a core and a hydrophobic wall material, with the core to wall material ratio being 35-50:50-65 by weight. The core is a precursor to a self-microemulsifying drug delivery system (SMEDDS). This precursor is composed, by weight, of 10-20 parts modified Ganoderma lucidum oligosaccharides, 25-40 parts phospholipids, 30-45 parts hydrophilic surfactants, and 10-20 parts lipophilic surfactants. The modified Ganoderma lucidum oligosaccharides are derived from natural Ganoderma lucidum polysaccharides through a specific enzymatic hydrolysis reaction, reducing their molecular weight to a predetermined range. Because the Type B microcapsule wall material is hydrophobic, it slowly dissolves or degrades in the hydrogel environment, releasing its contents. When the water in the matrix penetrates into the microcapsule core, the SMEDDS precursor comes into contact with water and spontaneously forms nano-scale microemulsion droplets encapsulating the modified Ganoderma lucidum oligosaccharides, thereby converting the originally water-soluble oligosaccharides into a nano-carrier form that can efficiently penetrate the skin barrier.

[0013] In a preferred embodiment, the lower layer matrix is ​​composed of the following components, in parts by mass: 1.0-2.5 parts of carbomer, 5.0-8.0 parts of gelatin, 5.0-10.0 parts of type A rapid-release microcapsules, 2.0-4.0 parts of type B long-acting self-emulsifying osmotic microcapsules, and the balance of water and moisturizer.

[0014] In another preferred embodiment, the upper matrix is ​​a thermosensitive hydrogel, which is composed of the following components, in parts by mass: 20-28 parts of poloxamer, 1.0-2.0 parts of a viscosity enhancer (such as chitosan), 8.0-15.0 parts of type B long-acting self-emulsifying osmotic microcapsules, and the balance water.

[0015] The second aspect of the present invention provides a method for preparing the gradient drug-release plaster patch loaded with the Ganoderma lucidum ultrafine powder.

[0016] The method comprises the following steps: First, the active substance is prepared and modified (step S1): the natural polysaccharide extracted from Ganoderma lucidum is placed under specific reaction conditions and subjected to molecular structure modification through enzymatic hydrolysis, so that it is degraded into modified Ganoderma lucidum oligosaccharides of a predetermined molecular weight.

[0017] In a specific embodiment, this step uses β-glucanase to enzymatically hydrolyze the Ganoderma lucidum polysaccharide until the average molecular weight of the product reaches the range of 1-5 kDa. Simultaneously, a triterpene-rich extract is obtained from the Ganoderma lucidum ultrafine powder using a supercritical CO2 extraction method.

[0018] Next, functional microcapsules are prepared (step S2): Type A rapid-release microcapsules and Type B long-lasting self-emulsifying and permeation-enhancing microcapsules are prepared. The preparation of Type B microcapsules is a key step in this method. The modified Ganoderma lucidum oligosaccharide obtained in step S1 is mixed with phospholipids, hydrophilic and lipophilic surfactants to form a uniform oily liquid, the precursor of the self-microemulsifying system, which serves as the core. This core is then encapsulated with a hydrophobic wall material.

[0019] In a specific embodiment, the hydrophobic wall material is poly(lactic acid-co-glycolic acid) (PLGA), and the encapsulation process adopts a solvent evaporation method.

[0020] Next, the double-layer matrix is ​​constructed and the patch is formed. This process involves dispersing the prepared Type A rapid-release microcapsules and Type B long-acting self-emulsifying and permeation-enhancing microcapsules in a predetermined ratio into the gel solution of the lower matrix layer, followed by coating and gelation to form a solidified lower layer (step S3). Then, the Type B long-acting self-emulsifying and permeation-enhancing microcapsules are dispersed into the gel solution of the upper matrix layer and evenly coated onto the formed lower layer (step S4).

[0021] Finally, the double-layer structure is solidified as an integrated whole (step S5): by regulating the ambient temperature, the liquid upper matrix gel solution undergoes a sol-gel phase transition, transforming into a semi-solid state and tightly combining with the lower matrix to form an integral double-layer structure.

[0022] In a specific embodiment, the upper matrix uses poloxamer as a thermosensitive material, and its gel phase transition is triggered by raising the ambient temperature to 32-37°C to complete the final patch formation.

[0023] The present invention provides a gradient-release plaster patch loaded with ultrafine Ganoderma lucidum powder and a preparation method thereof. It has the following beneficial effects: 1. The present invention successfully solves the technical problem of the stable coexistence and efficient delivery of two core active ingredients in Ganoderma lucidum with huge differences in physicochemical properties in the same drug delivery system by designing two microcapsule systems with different functions, namely, co-loading the fat-soluble triterpenoid components and permeation enhancers into Type A rapid-release microcapsules, and loading the enzymatically modified water-soluble oligosaccharide components into Type B long-acting self-emulsifying permeation-enhancing microcapsules. This synergistic encapsulation strategy ensures the stability of each component and lays the foundation for their subsequent programmed release and targeted absorption.

[0024] 2. The present invention constructs a double-layer gradient hydrogel matrix and differentially arranges the two types of microcapsules. The water-soluble wall material of the type A microcapsules in the lower matrix allows for rapid release of their contents, quickly establishing an effective osmotic concentration in the skin's stratum corneum, achieving "fast-acting" results. The hydrophobic wall material of the type B microcapsules and their gradient distribution in the upper and lower layers form a long-acting drug reservoir, continuously and slowly releasing active oligosaccharides, achieving "long-acting" results. This structural design enables the patch to achieve programmed gradient drug release, first rapid-acting and then long-acting, within a single dosing cycle, balancing both speed of onset and duration of action.

[0025] 3. The present invention significantly improves the transdermal absorption efficiency of active ingredients by combining the bio-enzymatic modification technology with the self-microemulsification delivery technology. First, the macromolecular Ganoderma lucidum polysaccharide is pre-enzymatically hydrolyzed into easily absorbed small-molecule oligosaccharides; second, the oligosaccharide is encapsulated in a self-microemulsification system precursor. When it is released from the microcapsule and contacts the moisture on the skin surface, it can spontaneously form nano-scale microemulsion droplets in situ. This nanocarrier greatly improves the skin permeability of the water-soluble oligosaccharide, thereby effectively overcoming the physiological barrier of the skin and improving the bioavailability of the drug. DETAILED DESCRIPTION

[0026] The following will be combined with the present invention specification to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0027] The sources and specifications of the main raw materials and reagents used in the following examples and comparative examples are as follows. Reagents not specifically specified are all commercially available products of analytical grade or higher.

[0028] Poloxamer 407: provided by BASF. CAS number: 9003-11-6; Carbomer 940: provided by Lubrizol. CAS number: 9003-01-4; Gelatin: Pharmaceutical grade, supplied by Rousselot. CAS number: 9000-70-8. Chitosan: medium viscosity, deacetylation degree greater than 90%, purchased from Sinopharm Chemical Reagent Co., Ltd. CAS number: 9012-76-4; Sodium Alginate: Food grade, purchased from Sinopharm Chemical Reagent Co., Ltd. CAS No.: 9005-38-3; Gum Arabic (Acacia Gum): purchased from Sinopharm Chemical Reagent Co., Ltd. CAS No.: 9000-01-5; Poly(lactic-co-glycolic acid) (PLGA): lactic acid to glycolic acid copolymer ratio of 75:25, intrinsic viscosity 0.6 dL / g, purchased from Jinan Daigang Bioengineering Co., Ltd. CAS number: 26780-50-7; Laurocapram / Azone: Pharmaceutical grade, purity greater than 98%, purchased from Sigma-Aldrich. CAS number: 59227-89-3; Soybean lecithin: injection grade, purchased from Avituo (Shanghai) Pharmaceutical Technology Co., Ltd. CAS No.: 8002-43-5; Tween-80 (Polysorbate 80): pharmaceutical excipient grade, purchased from Sinopharm Chemical Reagent Co., Ltd. CAS No.: 9005-65-6; Sorbitan Oleate: Pharmaceutical excipient grade, purchased from Sinopharm Chemical Reagent Co., Ltd. CAS No.: 1338-43-8; β-Glucanase: From Aspergillus niger, with an activity greater than 100,000 U / g, purchased from Novozymes. EC number: 3.2.1.6; Glutaraldehyde: 25% aqueous solution, purchased from Sinopharm Chemical Reagent Co., Ltd. CAS number: 111-30-8; Dichloromethane: analytical grade, purchased from Sinopharm Chemical Reagent Co., Ltd. CAS number: 75-09-2; Glycerin: analytical grade, purchased from Sinopharm Chemical Reagent Co., Ltd. CAS No.: 56-81-5.

[0029] Example 1: This embodiment provides a specific method for preparing a gradient-release plaster patch loaded with ultrafine Ganoderma lucidum powder.

[0030] S1. Preparation of Ganoderma lucidum triterpenoid extract: Take 1000 parts of ultrafine Ganoderma lucidum powder and place it in a supercritical CO2 extraction vessel. Dynamically extract for 2 hours at an extraction pressure of 25 MPa, an extraction temperature of 40°C, and a CO2 flow rate of 20 L / h. Collect the extract and concentrate under reduced pressure to remove residual solvent to obtain a yellow-brown Ganoderma lucidum triterpenoid extract, which is set aside.

[0031] S2. Preparation of modified Ganoderma lucidum oligosaccharides: Dissolve 100 parts of crude Ganoderma lucidum polysaccharide in 2000 parts of acetate buffer at a pH of 5.0. Heat the mixture to 50°C and add 100 units of β-glucanase per part of polysaccharide. Stir and react at 50°C for 4 hours. After the reaction is complete, heat the reaction mixture to 100°C for 15 minutes to inactivate the enzyme. After cooling, filter the mixture using an ultrafiltration membrane with a molecular weight cutoff of 1 kDa to remove impurities. Collect the filtrate and freeze-dry it to obtain a white powdered modified Ganoderma lucidum oligosaccharide with an average molecular weight of 2-3 kDa. Set aside.

[0032] Preparation of S3, Type A rapid-release microcapsules: Accurately weigh 80 parts of the Ganoderma lucidum triterpene extract prepared in step S1 and 20 parts of azone (as a penetration enhancer) and mix them evenly to form the microcapsule core. Separately, take 60 parts of gelatin and 60 parts of gum arabic as water-soluble wall materials. Dissolve 60 parts of gelatin in 1200 parts of purified water at 60°C, and dissolve 60 parts of gum arabic in 1200 parts of purified water at 60°C. Add the core material to the gelatin solution and emulsify under high shear for 30 minutes to form an O / W emulsion. Slowly add the gum arabic solution to this emulsion while stirring. Adjust the pH of the mixture to 4.0 using 10% acetic acid solution, then slowly cool to 5°C to allow the wall material to precipitate and coat the oil droplets. Add 0.5% glutaraldehyde solution for cross-linking and curing, and react for 4 hours. Finally, filter, wash, and freeze-dry the product to obtain Type A rapid-release microcapsule powder.

[0033] Preparation of S4, Type B long-acting self-emulsifying permeability-enhancing microcapsules: Accurately weigh 15 parts of the modified Ganoderma lucidum oligosaccharide prepared in Step 2, 35 parts of soy lecithin (as a phospholipid), 40 parts of Tween-80 (as a hydrophilic surfactant), and 10 parts of Span-80 (as a lipophilic surfactant). Mix and stir in a 60°C water bath until a clear, homogeneous oily liquid forms. This is the precursor of the self-microemulsifying system, which serves as the microcapsule core. Weigh 55 parts of poly(lactic-co-glycolic acid) (PLGA, 75:25) as the hydrophobic wall material and dissolve it in an appropriate amount of dichloromethane. Add the prepared core oily liquid to the PLGA solution and emulsify it using a high-speed homogenizer at 10,000 rpm for 5 minutes to form colostrum. This colostrum is rapidly injected into an aqueous solution containing 1% polyvinyl alcohol with continuous stirring. The solvent evaporates, allowing the microcapsules to solidify and form. The product is centrifuged, washed, and freeze-dried to obtain Type B long-lasting self-emulsifying, permeability-enhancing microcapsule powder.

[0034] S5. Integrated molding of double-layer gradient patch: Prepare the lower matrix gel solution: Weigh, by weight, 1.5 parts carbomer, 6.0 parts gelatin, 8.0 parts Type A immediate-release microcapsules, 3.0 parts Type B long-acting self-emulsifying osmotic microcapsules, and 15 parts glycerin. Make up the remainder with purified water. Dissolve the carbomer, gelatin, and glycerin in water, heat to swell, then cool to 40°C. Add the Type A and Type B microcapsules and stir thoroughly to obtain the lower matrix gel solution.

[0035] Prepare the upper matrix gel solution: Weigh 25 parts by mass of poloxamer 407, 1.5 parts of chitosan, and 12.0 parts of type B long-acting self-emulsifying osmotic microcapsules. Make up the remainder with purified water. Dissolve the poloxamer and chitosan in 4°C purified water. Then add the type B microcapsules, stir well, and store at 4°C until ready to use. This will yield the upper matrix gel solution.

[0036] Molding: The lower matrix gel solution is evenly coated onto a release film and allowed to gel at room temperature. Subsequently, the upper matrix gel solution, in a low-temperature liquid state, is evenly coated onto the formed lower layer. This two-layer structure is placed in an environment of 34°C. The poloxamer solution in the upper matrix undergoes a temperature-sensitive phase transition, solidifying into a gel that tightly bonds with the lower layer to form an integrated structure. Finally, a release layer is applied and the product is cut to obtain the gradient-release plaster patch described herein.

[0037] Example 2: The preparation method of this embodiment is basically the same as that of Example 1, with the only difference being the formula of the upper layer matrix gel solution in step 5.

[0038] Preparation of upper matrix gel solution: Weigh 22 parts by mass of poloxamer 407, 1.0 part of sodium alginate (as a thickener), and 15.0 parts of type B long-acting self-emulsifying osmotic microcapsules. Make up the remainder with purified water. All other preparation steps and conditions are the same as in Example 1.

[0039] Example 3: The preparation method of this embodiment is basically the same as that of Example 1, with the only difference being the component composition of the inner core of the type A rapid-release microcapsule in step 3.

[0040] Preparation of Type A Rapid-Release Microcapsules: Accurately weigh 75 parts of the Ganoderma lucidum triterpene extract prepared in Step 1 and 25 parts of laurocaprol (as a penetration enhancer) and mix them evenly to form the microcapsule core. All other preparation steps, components, and conditions are identical to those in Example 1.

[0041] Comparative Example 1: Compared with Example 1, the difference is that the microencapsulation preparation in steps 3 and 4 is not carried out, and the polysaccharide enzymatic modification in step 2 is not carried out. The Ganoderma triterpenoid extract prepared in step 1 and the unmodified Ganoderma crude polysaccharide are directly mixed with the lower layer matrix component (without microcapsules) in step 5(1) of Example 1 to prepare a single-layer hydrogel patch.

[0042] Comparative Example 2: Compared with Example 1, the difference is that the type A rapid-release microcapsules prepared in step 3 and the type B long-acting self-emulsifying permeation-enhancing microcapsules prepared in step 4 are mixed in the total number of portions used for the two layers in step 5 of Example 1 and added to a single hydrogel matrix to prepare a single-layer patch instead of forming a double-layer gradient structure.

[0043] Comparative Example 3: Compared to Example 1, the difference lies in the preparation of Type B long-lasting self-emulsifying and permeation-enhancing microcapsules in Step 4: the core of the microcapsules consists solely of the modified Ganoderma lucidum oligosaccharide prepared in Step 2, without the addition of soy lecithin, Tween-80, and Span-80. All other steps, components, and conditions are identical to those in Example 1.

[0044] Comparative Example 4: Compared to Example 1, the enzymatic hydrolysis and modification in step 2 were omitted. In step 4, when preparing Type B long-lasting self-emulsifying and permeation-enhancing microcapsules, the active ingredient in the core was directly unhydrolyzed crude Ganoderma lucidum polysaccharide, replacing the modified Ganoderma lucidum oligosaccharide. All other steps, components, and conditions were identical to those in Example 1.

[0045] Test Example 1: In vitro release performance evaluation This test example is intended to evaluate the in vitro drug release characteristics of different patch preparations, in order to illustrate the role of the technical solution of the present invention in achieving programmed gradient drug release.

[0046] Experimental materials and equipment: Experimental samples: plaster patches prepared in Example 1, Comparative Example 1, and Comparative Example 2.

[0047] Main equipment: Franz vertical diffusion cell, constant temperature magnetic stirrer, high performance liquid chromatography (HPLC).

[0048] Experimental medium: phosphate buffered saline (PBS, pH 7.4).

[0049] Separation membrane: cellulose acetate dialysis membrane (molecular weight cut-off 12000Da).

[0050] Experimental steps: (1) Fix the dialysis membrane between the donor chamber and the receiver chamber of the Franz diffusion cell to ensure that there are no bubbles.

[0051] (2) The receiving chamber was filled with PBS solution preheated to 37 ± 0.5 °C and stirred continuously at 300 rpm using a magnetic stirrer to simulate body fluid circulation.

[0052] (3) Each experimental sample (Example 1, Comparative Example 1, Comparative Example 2) was cut into a circular sample of a specified area, and the drug-administered surface was tightly applied to the dialysis membrane.

[0053] (4) At the preset time points (0.5, 1, 2, 4, 8, 12, and 24 hours), 1.0 mL of receiving solution was drawn from the sampling port of the receiving chamber and immediately supplemented with an equal amount of isothermal fresh PBS solution.

[0054] (5) After the samples were appropriately diluted, the concentrations of Ganoderma triterpenes and modified Ganoderma oligosaccharides were determined by HPLC system.

[0055] (6) Calculate the cumulative release percentage of the active ingredient at each time point based on the measured concentration.

[0056] Experimental data: The cumulative release rate of the active ingredient of each sample at different time points is recorded in the table below.

[0057] Table 1: Cumulative release rate of active ingredients from different patch samples (%)

[0058] Result analysis: Experimental data showed that the patch prepared according to Example 1 exhibited a clear biphasic release profile. Ganoderma triterpenes were released rapidly in the initial phase (first four hours), with a cumulative release rate exceeding 80%. This was attributed to the structure of the Type A rapid-release microcapsules in the underlying matrix. The water-soluble wall material of these microcapsules rapidly dissolved upon contact with the water in the hydrogel matrix, resulting in the rapid release of the triterpenes and permeation enhancer from the inner core. Simultaneously, the modified Ganoderma oligosaccharide exhibited a sustained, nearly constant release rate, slowly releasing over 24 hours. This corresponds to the mechanism by which the hydrophobic wall material (PLGA) of the Type B long-acting microcapsules releases its contents through slow erosion and degradation.

[0059] In contrast, the sample in Comparative Example 1, which lacked any microencapsulation technology, released both active ingredients rapidly and disorderly within a very short period of time, reaching a release plateau within 2 hours and failing to achieve any effective timing control. This comparison directly demonstrates the necessity of a microencapsulation structure for achieving controlled drug release.

[0060] Furthermore, although the sample in Comparative Example 2 employed two microcapsules, its oligosaccharide release rate was significantly faster than that of Example 1 due to the lack of a double-layer gradient structure. This is because all Type B microcapsules were located within a single layer and directly participated in the release process, without the concentration gradient and physical retardation provided by the upper matrix as a drug reservoir. Therefore, the data confirm that the dual microcapsule system and the double-layer gradient matrix structure in this technical solution, acting together, are the key to achieving the programmed release of the two active ingredients, ensuring the rapid onset of one ingredient and the steady, long-lasting effect of the other.

[0061] Test Example 2: In vitro transdermal permeation performance evaluation This test example is intended to evaluate the in vitro transdermal permeability of different patch preparations to illustrate the role of the technical solution of the present invention in improving the efficiency of active ingredient delivery across the skin barrier.

[0062] Experimental materials and equipment: Experimental samples: plaster patches prepared in Example 1, Comparative Example 1, Comparative Example 3, and Comparative Example 4.

[0063] Main equipment: Franz vertical diffusion cell, constant temperature magnetic stirrer, high performance liquid chromatography (HPLC).

[0064] Experimental barrier: full-thickness abdominal skin of isolated Kunming mice.

[0065] Receiving medium: phosphate buffered saline (PBS, pH 7.4).

[0066] Experimental steps: (1) The abdominal skin of the sacrificed mice was shaved of hair and subcutaneous fat to prepare full-thickness skin, which was then fixed between the donor and receiver chambers of a Franz diffusion cell, with the stratum corneum facing the donor chamber.

[0067] (2) The receiving chamber was filled with PBS solution preheated to 37 ± 0.5 °C and stirred continuously at 300 rpm using a magnetic stirrer.

[0068] (3) Each experimental sample (Example 1, Comparative Example 1, Comparative Example 3, Comparative Example 4) was cut into a circular sample of a specified area, and the drug-administered surface thereof was closely applied to the stratum corneum surface of the skin.

[0069] (4) After 24 hours of the experiment, samples were taken from the receiving chamber, and the concentrations of Ganoderma triterpenes and Ganoderma oligosaccharides (or polysaccharides) in the receiving solution were determined by HPLC system.

[0070] (5) Calculate the total cumulative amount of active ingredients that penetrate the skin of each sample within 24 hours based on the measured concentration.

[0071] Experimental data: The results of the cumulative amount of active ingredient permeated through the skin of each sample after 24 hours are recorded in the table below.

[0072] Table 2: Cumulative transdermal dose of different patch samples over 24 hours (μg / cm 2 )

[0073] Result analysis: As shown in Table 2, the two active ingredients in Example 1, Ganoderma triterpenes and modified Ganoderma oligosaccharides, achieved significantly higher 24-hour cumulative transdermal delivery than all the comparative examples. Comparative Example 1, which lacked any technical treatment of the active ingredients, had the lowest transdermal delivery, demonstrating the strong barrier effect of the skin's stratum corneum on these two native substances. The superior transdermal performance of Example 1 directly reflects the overall enhanced transdermal delivery of the active ingredients by its internal composite microcapsule system.

[0074] Comparing the data of Example 1 and Comparative Example 3, it can be found that the transdermal amount of Ganoderma lucidum oligosaccharides in the former is more than 4 times that of the latter. The root of this difference lies in the composition of the core of the type B microcapsule. The core of Example 1 is a precursor of a self-microemulsifying system. After contacting the moisture on the skin surface, it can spontaneously form nano-scale microemulsion droplets encapsulating oligosaccharides in situ. This nanocarrier effectively delivers water-soluble oligosaccharides through the hydrophobic skin barrier by changing the drug's delivery pathway and increasing its distribution in the lipid bilayer of the stratum corneum. Comparative Example 3 lacks self-emulsifying components such as phospholipids and surfactants, and is therefore unable to form this efficient delivery carrier, and its penetration efficiency is therefore greatly limited.

[0075] Further comparison of the data of Example 1 and Comparative Example 4 reveals the role of bio-enzymatic modification. Comparative Example 4 uses a macromolecular Ganoderma lucidum polysaccharide that has not been enzymatically hydrolyzed. Even though it is encapsulated in a self-emulsifying system, its cumulative transdermal amount is still extremely low. This shows that excessive molecular weight is the fundamental physical barrier that limits the percutaneous absorption of polysaccharides. Example 1 first removes this obstacle by pre-using β-glucanase to degrade polysaccharides into small molecule oligosaccharides of 1-5 kDa. This step is combined with the subsequent self-microemulsification delivery technology, and the two work synergistically, ultimately enabling the water-soluble macromolecular active substances that were originally difficult to penetrate the skin to achieve efficient cross-barrier delivery.

[0076] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A gradient-release plaster patch loaded with superfine Ganoderma lucidum powder, characterized in that: include: A double-layer hydrogel matrix, wherein two functional microcapsules are dispersed: type A rapid-release microcapsules and type B long-acting self-emulsifying permeation-enhancing microcapsules; The double-layer hydrogel matrix comprises a lower layer matrix in contact with the skin and an upper layer matrix thereon; The lower matrix contains Type A quick-release microcapsules and Type B long-acting self-emulsifying and permeation-enhancing microcapsules, while the upper matrix contains only Type B long-acting self-emulsifying and permeation-enhancing microcapsules. The type A rapid-release microcapsule comprises, by weight, 30-45 parts of an inner core and 55-70 parts of a water-soluble wall material; The inner core of the type A rapid-release microcapsule contains Ganoderma lucidum triterpenoid extract and a penetration enhancer; The type B long-lasting self-emulsifying osmotic microcapsule comprises, by weight, 35-50 parts of an inner core and 50-65 parts of a hydrophobic wall material; The inner core of the type B long-acting self-emulsifying permeation-enhancing microcapsule is a precursor of a self-microemulsifying drug delivery system, which comprises enzymatically hydrolyzed modified Ganoderma lucidum oligosaccharide, phospholipid, hydrophilic surfactant and lipophilic surfactant.

2. The gradient-release plaster patch loaded with superfine Ganoderma lucidum powder according to claim 1, characterized in that: The lower layer matrix comprises, by weight, 1.0-2.5 parts of carbomer, 5.0-8.0 parts of gelatin, 5.0-10.0 parts of type A rapid-release microcapsules, 2.0-4.0 parts of type B long-acting self-emulsifying osmotic microcapsules, and the balance of water and moisturizer.

3. The gradient-release plaster patch loaded with superfine Ganoderma lucidum powder according to claim 1, characterized in that: The upper matrix is ​​a thermosensitive hydrogel, which comprises, by weight, 20-28 parts of poloxamer, 1.0-2.0 parts of a viscosity enhancer, 8.0-15.0 parts of type B long-acting self-emulsifying osmotic microcapsules, and the balance water.

4. The gradient-release plaster patch loaded with superfine Ganoderma lucidum powder according to claim 3, characterized in that: In the inner core of the type A rapid-release microcapsule, the mass ratio of the Ganoderma lucidum triterpenoid extract to the penetration enhancer is 70 to 85:15 to 30.

5. The gradient-release plaster patch loaded with superfine Ganoderma lucidum powder according to claim 1, characterized in that: The inner core of the type B long-acting self-emulsifying osmotic microcapsule comprises, by weight, 10-20 parts of modified Ganoderma lucidum oligosaccharide, 25-40 parts of phospholipid, 30-45 parts of hydrophilic surfactant and 10-20 parts of lipophilic surfactant.

6. The gradient-release plaster patch loaded with superfine Ganoderma lucidum powder according to claim 1, characterized in that: The Ganoderma lucidum triterpenoid extract in the A-type rapid-release microcapsule is obtained by subjecting Ganoderma lucidum ultrafine powder to supercritical CO2 extraction.

7. A method for preparing a gradient-release plaster patch loaded with superfine Ganoderma lucidum powder, characterized in that: The method for preparing the gradient-release plaster patch loaded with Ganoderma lucidum ultrafine powder according to any one of claims 1 to 6 comprises the following steps: S1, modifying Ganoderma lucidum polysaccharide by enzymatic hydrolysis to obtain modified Ganoderma lucidum oligosaccharide with a predetermined molecular weight; S2. Preparing type A rapid-release microcapsules and type B long-acting self-emulsifying and permeation-enhancing microcapsules respectively, wherein the preparation of the type B long-acting self-emulsifying and permeation-enhancing microcapsules comprises mixing the modified Ganoderma lucidum oligosaccharide obtained in step S1 with phospholipids and a surfactant to form a self-microemulsifying system precursor as the core, and then encapsulating with a hydrophobic wall material; S3, type A rapid-release microcapsules and type B long-acting self-emulsifying osmotic microcapsules are dispersed in the lower layer matrix gel solution, coated and gelled to form the lower layer; S4, dispersing the type B long-acting self-emulsifying permeation-enhancing microcapsules in the upper layer of matrix gel solution and coating the solution on the lower layer; S5. The upper layer matrix gel solution undergoes phase change and solidification by regulating the temperature to form an integrated double-layer structure with the lower layer.

8. The method for preparing a gradient-release plaster patch loaded with superfine Ganoderma lucidum powder according to claim 7, characterized in that: In step S1, β-glucanase is used to enzymatically hydrolyze Ganoderma lucidum polysaccharide to obtain modified Ganoderma lucidum oligosaccharides with an average molecular weight of 1-5 kDa.

9. The method for preparing a gradient-release plaster patch loaded with superfine Ganoderma lucidum powder according to claim 7, characterized in that: In step S2, the hydrophobic wall material of the type B long-lasting self-emulsifying permeation-enhancing microcapsules is polylactic acid-glycolic acid copolymer, and the encapsulation is performed by a solvent evaporation method.

10. The method for preparing a gradient-release plaster patch loaded with superfine Ganoderma lucidum powder according to claim 7, characterized in that: In step S5, the upper matrix is ​​a poloxamer thermosensitive hydrogel, which undergoes a gel phase transition by heating to 32-37°C.

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