Method for enhancing permeability of transdermal drug delivery by using biological material

Abstract

The invention relates to the technical field of biological medicine and drug delivery, and discloses a method for enhancing the permeability of transdermal drug delivery by using a biological material, and the method comprises the following steps: S1, preparing a multifunctional nano-carrier containing a high-molecular polymer core and a functional shell layer; s2, mixing the multifunctional nano-carrier with a matrix material; s3, carrying out surface modification on the nano-carrier, and adding a targeting ligand or a penetration enhancer; s4, applying an intelligent responsive material which is a temperature-sensitive material or a pH-sensitive material and is used for controlling drug release when the temperature or the pH changes; and S5, adding a reinforcing agent into the matrix material, wherein the reinforcing agent is selected from hyaluronic acid, chitosan or other natural polysaccharides. Through a multi-layer nano-carrier, targeted ligand modification, an intelligent response material and a biodegradable microneedle, transdermal absorption, targeting and controlled release of the drug are improved, and the problems of insufficient absorption and poor targeting of a traditional system are solved.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine and drug delivery, in particular to a method for enhancing the permeability of transdermal drug delivery by utilizing biological materials. Background Art

[0002] In modern medicine, transdermal drug delivery systems have become a hot area of ​​research and application due to their non-invasiveness and high patient compliance. Transdermal drug delivery refers to the absorption of drugs through the skin to achieve systemic or local therapeutic effects. However, the natural barrier function of the skin, namely the stratum corneum, limits the transdermal absorption of most drugs. This results in low bioavailability of many drugs in transdermal delivery, making it difficult to achieve effective therapeutic concentrations. In addition, uneven release of drugs in transdermal delivery often leads to poor therapeutic effects.

[0003] Current transdermal drug delivery systems face multiple challenges, including low drug permeability, insufficient targeting, and difficulty in achieving dynamically controlled release mechanisms. Although traditional transdermal patches and creams have achieved certain success in certain applications, they are unable to meet diverse clinical needs due to the difficulty in controlling drug release, low transdermal absorption efficiency, and the lack of the ability to adjust for specific pathological conditions. At the same time, due to the lack of an effective targeting mechanism, the distribution of drugs is often difficult to concentrate on the lesion site, resulting in unsatisfactory efficacy and increased side effects.

[0004] In order to improve the transdermal absorption efficiency of drugs, scientists have tried a variety of methods, including the use of chemical promoters, physical methods (such as ion introduction and ultrasound), and nanotechnology. However, these methods still have limitations, such as chemical promoters may cause skin irritation or allergies, and physical methods are complicated and unsuitable for some patients. In addition, although nanotechnology provides certain solutions, its performance in drug release control, targeting, and safety still needs to be further optimized.

[0005] In the face of these challenges, the development of transdermal drug delivery systems requires an innovative solution to overcome the limitations of existing methods. Therefore, in response to the above problems, the present invention proposes a method for enhancing the permeability of transdermal drug delivery using biomaterials. Summary of the invention

[0006] In view of the deficiencies of the prior art, the present invention provides a method for enhancing the permeability of transdermal drug delivery using biomaterials, which solves the problems of low transdermal drug absorption efficiency, insufficient targeting, and difficulty in precise release control in existing transdermal drug delivery systems.

[0007] To achieve the above objectives, the present invention is implemented by the following technical scheme: A method for enhancing the permeability of transdermal drug delivery using biomaterials comprises the following steps:

[0008] S1. preparing a multifunctional nanocarrier comprising a polymer core and a functionalized shell;

[0009] S2, mixing the multifunctional nanocarrier with the matrix material;

[0010] S3, modifying the surface of the nanocarrier and adding a targeting ligand or a penetration enhancer;

[0011] S4. Application of intelligent responsive materials, which are temperature-sensitive materials or pH-sensitive materials, to control drug release when temperature or pH changes;

[0012] S5, adding an enhancer to the matrix material, wherein the enhancer is selected from hyaluronic acid, chitosan or other natural polysaccharides;

[0013] S6. Prepare a microneedle array, wherein the microneedles are made of a biodegradable material selected from PLGA or PVP, and combine the microneedles with a carrier matrix.

[0014] Preferably, in step S1, the core is selected from one of PLGA or PLA, the shell layer is selected from one of gold nanoparticles or silica nanoparticles, and in step S2, the matrix material is selected from one of a cream, a gel or a patch.

[0015] Preferably, the preparation method of the core material in step S1 is optionally a nanoprecipitation method and a double emulsion method, and the encapsulation technology used for the multifunctional nanocarrier in step S1 is to form a multilayer structure on the surface of the nanocarrier, each layer encapsulating a different drug or protective agent.

[0016] Preferably, the targeting ligand in step S3 is selected from antibodies or peptide chains, and the penetration enhancer in step S3 is selected from chitosan derivatives, wherein the chitosan derivative is optionally selected from methylated chitosan, carboxymethyl chitosan or quaternized chitosan.

[0017] Preferably, the step S3 specifically includes the following steps:

[0018] S3.1. Targeted ligand modification

[0019] Ligand selection and connection method: First, according to the specific antigens or receptors of the target tissue or cells, appropriate targeting ligands are selected. The methods for connecting these ligands to the surface of nanocarriers include chemical bonding and physical adsorption;

[0020] Optimization and post-processing: To achieve the best effect, the concentration of the targeting ligand needs to be controlled within the range of 0.1-2 mg / ml, and the reaction time needs to be controlled within the range of 1-3 h, the temperature needs to be 4-25 °C, and the pH value needs to be controlled within the range of 7-8. After the reaction is completed, the unreacted ligand should be removed by dialysis, centrifugation or chromatography;

[0021] Testing and verification: Enzyme-linked immunosorbent assay and surface plasmon resonance technology are used to determine whether the ligand binding efficiency is between 70-90%. Cell experiments are also needed to verify the targeting ability of the nanocarrier;

[0022] S3.2, Penetration Enhancer Modification:

[0023] Enhancer selection and modification method: According to the requirements of drug transdermal absorption, select appropriate penetration enhancers. Modification methods include coating, blending or covalent bonding;

[0024] Concentration control and post-processing: During the implementation process, the concentration of the enhancer needs to be controlled within the range of 0.1-5% w / v, and the solution is prepared according to the target concentration. After the modification is completed, the unbound enhancer is removed by dialysis or ultrafiltration.

[0025] Preferably, in step S4, the temperature-sensitive material is optionally one of N-isopropylacrylamide, polyurethane or lactide-glycolide, and the pH-sensitive material is optionally one of polyacrylic acid, sodium polyacrylate or polyethyleneimine.

[0026] Preferably, the specific steps of applying the smart response material in step S4 are as follows:

[0027] S4.1. Intelligent material selection: one of the temperature-sensitive material and pH-sensitive material can be selected;

[0028] S4.2, Material integration: In the process of preparing nanocarriers, PNIPAAm is blended with polymer materials to form nanoparticles with core-shell structure. PNIPAAm can be used as an outer layer to coat the core drug or carrier. The mass ratio of PNIPAAm to PLGA is between 1:4 and 1:1.

[0029] S4.3, preparation process: PNIPAAm and PLGA are dissolved in an organic solvent to form a uniform mixed solution, and the solution is added dropwise to the aqueous phase using a nanoprecipitation method, the dropping speed is controlled at 0.5-5 ml / min, the volume of the aqueous phase is 10-100 times that of the organic phase, the stirring speed is controlled at 300-1000 rpm, and nanoparticles are formed by solvent evaporation, and PNIPAAm is evenly distributed in the outer layer of the nanoparticles;

[0030] S4.4. Solvent removal and purification: Remove the organic solvent by rotary evaporation at 40-50°C, or use ultrafiltration with a membrane with a molecular weight cutoff point of 10-100 kDa to remove free low molecular weight components. Nanoparticle purification is performed by dialysis with a dialysis bag with a molecular weight cutoff point of 10-100 kDa and a dialysis time of 4-24 h.

[0031] Preferably, the step S6 specifically includes the following steps:

[0032] S6.1. Microneedle material selection and manufacturing:

[0033] Pour the PLGA or PVP polymer solution into the prefabricated microneedle mold, dry it at room temperature or under vacuum conditions, further solidify the molded microneedles, build the microneedle structure layer by layer using high-precision 3D printing technology, and make the PLGA or PVP material into a liquid or powder form suitable for 3D printing;

[0034] S6.2. Microneedle parameter design

[0035] Length range: 150-1500μm;

[0036] Diameter range: 50-500μm;

[0037] Density range: 200-2000 pieces / cm 2 ;

[0038] S6.3, Microneedle and matrix combination:

[0039] Drug loading: During the process of manufacturing microneedles, drugs are embedded directly into the polymer;

[0040] Binding to a carrier matrix: Binding the prepared microneedle array to a carrier matrix such as a cream, gel or patch;

[0041] Binding method: The microneedle array is fixed to the substrate by physical attachment or chemical bonding.

[0042] Preferably, the high-precision 3D printing technology in step S6.1 is optionally photo-curing stereolithography or fused deposition modeling technology, and the matrix material in step S6.2 is optionally one of water-based, oil-based or mixed matrices.

[0043] The present invention provides a method for enhancing the permeability of transdermal drug delivery using biomaterials. It has the following beneficial effects:

[0044] 1. The present invention realizes multi-level drug encapsulation by combining a high molecular polymer core with a functionalized shell in a nanocarrier. This design not only improves the stability and bioavailability of drugs, but also allows for the simultaneous delivery and controlled release of multiple drugs, thereby solving the problem in the prior art that a single-layer nanocarrier structure often leads to uneven drug release, poor stability, and insufficient bioavailability.

[0045] 2. The present invention modifies the surface of the nanocarrier with targeting ligands and penetration enhancers, which can significantly improve the targeting and skin permeability of the drug. The selection and modification methods of the targeting ligands are optimized to achieve precise targeting of specific tissues or cells. The penetration enhancer further reduces the resistance of the skin barrier and improves the transdermal absorption efficiency of the drug, thus solving the problems of the lack of targeting of traditional transdermal drug delivery methods, the difficulty of drugs to accurately act on specific tissues or cells, and the serious limitation of the absorption efficiency of drugs by the skin barrier effect.

[0046] 3. The present invention can dynamically control drug release according to environmental changes by coating these materials on the outer layer of the nanocarrier. This feature allows the drug to be released in a targeted manner under specific pathological conditions, thereby improving the accuracy and safety of the therapeutic effect and solving the problem that existing drug delivery systems are often difficult to achieve precise drug release control under different pathological conditions, resulting in insufficient concentration of drug effects or unsatisfactory release time.

[0047] 4. The present invention adopts biodegradable materials to prepare microneedle arrays, combined with high-precision 3D printing technology, to achieve precise design and manufacturing of microneedles. The parameters of the microneedles can be flexibly adjusted according to treatment needs to ensure that the drug effectively penetrates the skin while reducing the patient's pain and discomfort. The combination of microneedles and carrier matrices further optimizes the sustained release and absorption of drugs, and solves the problem that traditional transdermal drug delivery systems, such as patches and creams, often cannot provide sufficient drug penetration depth, resulting in insignificant therapeutic effects and possible discomfort to patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 It is a flow chart of the present invention. DETAILED DESCRIPTION

[0049] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings 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 creative work are within the scope of protection of the present invention.

[0050] Please see attached Figure 1 The present invention provides a method for enhancing the permeability of transdermal drug delivery using biomaterials, comprising the following steps:

[0051] S1. preparing a multifunctional nanocarrier comprising a polymer core and a functionalized shell;

[0052] S2, mixing the multifunctional nanocarrier with the matrix material;

[0053] S3, modifying the surface of the nanocarrier and adding a targeting ligand or a penetration enhancer;

[0054] S4. Application of intelligent responsive materials, which are temperature-sensitive materials or pH-sensitive materials, to control drug release when temperature or pH changes;

[0055] S5, adding an enhancer to the matrix material, wherein the enhancer is selected from hyaluronic acid, chitosan or other natural polysaccharides;

[0056] S6. Prepare a microneedle array, wherein the microneedles are made of a biodegradable material selected from PLGA or PVP, and combine the microneedles with a carrier matrix.

[0057] In step S1, the core is selected from one of PLGA or PLA, the shell layer is selected from one of gold nanoparticles or silica nanoparticles, and in step S2, the matrix material is selected from one of cream, gel or patch.

[0058] The preparation method of the core material in step S1 can be any of nanoprecipitation method and double emulsion method. The encapsulation technology used for the multifunctional nanocarrier in step S1 is to form a multilayer structure on the surface of the nanocarrier, and each layer encapsulates different drugs or protective agents.

[0059] In step S3, the targeting ligand is selected from an antibody or a peptide chain, and the penetration enhancer is selected from a chitosan derivative, which can be any one of methylated chitosan, carboxymethyl chitosan or quaternized chitosan.

[0060] The S3 step specifically includes the following steps:

[0061] S3.1. Targeted ligand modification

[0062] Ligand selection and connection method: First, according to the specific antigens or receptors of the target tissue or cells, appropriate targeting ligands are selected. The methods for connecting these ligands to the surface of nanocarriers include chemical bonding and physical adsorption;

[0063] Optimization and post-processing: To achieve the best effect, the concentration of the targeting ligand needs to be controlled within the range of 0.1-2 mg / ml, and the reaction time needs to be controlled within the range of 1-3 h, the temperature needs to be 4-25 °C, and the pH value needs to be controlled within the range of 7-8. After the reaction is completed, the unreacted ligand should be removed by dialysis, centrifugation or chromatography;

[0064] Testing and verification: Enzyme-linked immunosorbent assay and surface plasmon resonance technology are used to determine whether the ligand binding efficiency is between 70-90%. Cell experiments are also needed to verify the targeting ability of the nanocarrier;

[0065] S3.2, Penetration Enhancer Modification:

[0066] Enhancer selection and modification method: According to the requirements of drug transdermal absorption, select appropriate penetration enhancers. Modification methods include coating, blending or covalent bonding;

[0067] Concentration control and post-processing: During the implementation process, the concentration of the enhancer needs to be controlled within the range of 0.1-5% w / v, and the solution is prepared according to the target concentration. After the modification is completed, the unbound enhancer is removed by dialysis or ultrafiltration.

[0068] In step S4, the temperature-sensitive material is optionally selected from N-isopropylacrylamide, polyurethane or lactide-glycolide, and the pH-sensitive material is optionally selected from polyacrylic acid, sodium polyacrylate or polyethyleneimine.

[0069] The specific steps for applying the smart response material in step S4 are as follows:

[0070] S4.1. Intelligent material selection: one of the temperature-sensitive material and pH-sensitive material can be selected;

[0071] S4.2, Material integration: In the process of preparing nanocarriers, PNIPAAm is blended with polymer materials to form nanoparticles with core-shell structure. PNIPAAm can be used as an outer layer to coat the core drug or carrier. The mass ratio of PNIPAAm to PLGA is between 1:4 and 1:1.

[0072] S4.3, preparation process: PNIPAAm and PLGA are dissolved in an organic solvent to form a uniform mixed solution, and the solution is added dropwise to the aqueous phase using a nanoprecipitation method, the dropping speed is controlled at 0.5-5 ml / min, the volume of the aqueous phase is 10-100 times that of the organic phase, the stirring speed is controlled at 300-1000 rpm, and nanoparticles are formed by solvent evaporation, and PNIPAAm is evenly distributed in the outer layer of the nanoparticles;

[0073] S4.4. Solvent removal and purification: Remove the organic solvent by rotary evaporation at 40-50°C, or use ultrafiltration with a membrane with a molecular weight cutoff point of 10-100 kDa to remove free low molecular weight components. Nanoparticle purification is performed by dialysis with a dialysis bag with a molecular weight cutoff point of 10-100 kDa and a dialysis time of 4-24 h.

[0074] Step S6 specifically includes the following steps:

[0075] S6.1. Microneedle material selection and manufacturing:

[0076] Pour the PLGA or PVP polymer solution into the prefabricated microneedle mold, dry it at room temperature or under vacuum conditions, further solidify the molded microneedles, build the microneedle structure layer by layer using high-precision 3D printing technology, and make the PLGA or PVP material into a liquid or powder form suitable for 3D printing;

[0077] S6.2. Microneedle parameter design

[0078] Length range: 150-1500μm;

[0079] Diameter range: 50-500μm;

[0080] Density range: 200-2000 pieces / cm 2 ;

[0081] S6.3, Microneedle and matrix combination:

[0082] Drug loading: During the process of manufacturing microneedles, drugs are embedded directly into the polymer;

[0083] Binding to a carrier matrix: Binding the prepared microneedle array to a carrier matrix such as a cream, gel or patch;

[0084] Binding method: The microneedle array is fixed to the substrate by physical attachment or chemical bonding.

[0085] In step S6.1, the high-precision 3D printing technology may be photo-curing stereolithography or fused deposition modeling technology, and in step S6.2, the matrix material may be water-based, oil-based or mixed matrix.

[0086] Example 1: Basic multifunctional nanocarrier transdermal drug delivery system

[0087] Materials and Methods

[0088] Preparation of multifunctional nanocarriers:

[0089] Core material: PLGA, Shell material: silica nanoparticles.

[0090] Preparation method: Nanoprecipitation method. The mass ratio of PLGA to silica is 2:1, and the total concentration is 5% (w / v).

[0091] Encapsulated drug: Aspirin, encapsulation rate 75%.

[0092] Matrix material mixing:

[0093] Matrix material: cream, concentration 10% (w / v), mixed with nanocarriers at a ratio of 1:10.

[0094] Surface modification:

[0095] Targeting ligand: Antibody EGFR, concentration 1 mg / ml.

[0096] Penetration enhancer: methylated chitosan, concentration 2% (w / v).

[0097] Smart Responsive Materials:

[0098] Materials: PNIPAAm, mass ratio of PLGA to PNIPAAm is 3:1.

[0099] Coating process: solution dropping speed 2ml / min, aqueous phase volume 50 times that of organic phase, stirring speed 500rpm.

[0100] Purification: rotary evaporation, temperature 45 °C.

[0101] Microneedle preparation:

[0102] Material: PLGA, microneedle length 500μm, diameter 100μm, density 1000 / cm 2 .

[0103] Preparation method: mold casting.

[0104] Summary: This example demonstrates a basic multifunctional nanocarrier transdermal drug delivery system, including nanocarrier preparation, targeted modification, and microneedle application.

[0105] Example 2: Enhanced targeting and transdermal absorption

[0106] Materials and Methods

[0107] Preparation of multifunctional nanocarriers:

[0108] Core material: PLGA, Shell material: gold nanoparticles.

[0109] Preparation method: double emulsion method. The mass ratio of PLGA to gold is 2:1, and the total concentration is 4% (w / v).

[0110] Encapsulated drug: doxorubicin, encapsulation rate 80%.

[0111] Matrix material mixing:

[0112] Matrix material: gel, concentration 12% (w / v), mixed with nanocarriers at a ratio of 1:8.

[0113] Surface modification:

[0114] Targeting ligand: RGD peptide, concentration 1.5 mg / ml.

[0115] Penetration enhancer: carboxymethyl chitosan, concentration 3% (w / v).

[0116] Smart Responsive Materials:

[0117] Materials: PNIPAAm, mass ratio of PLGA to PNIPAAm is 2:1.

[0118] Coating process: solution dropping speed 3ml / min, aqueous phase volume 70 times that of organic phase, stirring speed 700rpm.

[0119] Purification: Rotary evaporation, temperature 50°C.

[0120] Microneedle preparation:

[0121] Material: PVP, microneedle length 750μm, diameter 150μm, density 1500 / cm 2 .

[0122] Preparation method: 3D printing, photocuring stereolithography technology.

[0123] Summary: This example enhances targeting and transdermal absorption using gold nanoparticles as shell material and RGD peptide as targeting ligand.

[0124] Example 3: Intelligent responsive material improvement

[0125] Materials and Methods

[0126] Preparation of multifunctional nanocarriers:

[0127] Core material: PLA, Shell material: silica nanoparticles.

[0128] Preparation method: nanoprecipitation method. The mass ratio of PLA to silica is 1:1, and the total concentration is 6% (w / v).

[0129] Encapsulated drug: dexamethasone, encapsulation rate 85%.

[0130] Matrix material mixing:

[0131] Matrix material: cream, concentration 15% (w / v), mixed with nanocarriers at a ratio of 1:7.

[0132] Surface modification:

[0133] Targeting ligand: Antibody Her2, concentration 2 mg / ml.

[0134] Penetration enhancer: quaternized chitosan, concentration 4% (w / v).

[0135] Smart Responsive Materials:

[0136] Materials: PNIPAAm, the mass ratio of PLGA to PNIPAAm is 1:1.

[0137] Coating process: solution dropping speed 4ml / min, aqueous phase volume 80 times that of organic phase, stirring speed 800rpm.

[0138] Purification: rotary evaporation, temperature 40 °C.

[0139] Microneedle preparation:

[0140] Material: PLGA, microneedle length 1000μm, diameter 200μm, density 1200 / cm 2 .

[0141] Preparation method: mold casting.

[0142] Summary: This example achieves precise release of drugs under different environmental conditions by improving smart responsive materials.

[0143] Example 4: Optimization of drug release control

[0144] Materials and Methods

[0145] Preparation of multifunctional nanocarriers:

[0146] Core material: PLGA, Shell material: gold nanoparticles.

[0147] Preparation method: double emulsion method. The mass ratio of PLGA to gold is 3:2, and the total concentration is 7% (w / v).

[0148] Encapsulated drug: Vitamin C, encapsulation rate 90%.

[0149] Matrix material mixing:

[0150] Matrix material: gel, concentration 20% (w / v), mixed with nanocarriers at a ratio of 1:6.

[0151] Surface modification:

[0152] Targeting ligand: glycoprotein, concentration 2.5 mg / ml.

[0153] Penetration enhancer: methylated chitosan, concentration 3.5% (w / v).

[0154] Smart Responsive Materials:

[0155] Materials: PNIPAAm, the mass ratio of PLGA to PNIPAAm is 1:2.

[0156] Coating process: solution dropping speed 5ml / min, aqueous phase volume 60 times that of organic phase, stirring speed 900rpm.

[0157] Purification: rotary evaporation, temperature 45 °C.

[0158] Microneedle preparation:

[0159] Material: PVP, microneedle length 1250μm, diameter 250μm, density 1800 / cm 2 .

[0160] Preparation method: 3D printing, photocuring stereolithography technology.

[0161] Summary: This example optimizes drug release control and further improves the accuracy of drug release under specific circumstances by adjusting the proportion of smart materials and nanocarrier design.

[0162] Example 5: Multi-drug simultaneous delivery system

[0163] Materials and Methods

[0164] Preparation of multifunctional nanocarriers:

[0165] Core material: PLA, Shell material: silica nanoparticles.

[0166] Preparation method: nanoprecipitation method. The mass ratio of PLA to silica is 1:2, and the total concentration is 8% (w / v).

[0167] Encapsulated drugs: Penicillin and triptorelin, with encapsulation rates of 95%.

[0168] Matrix material mixing:

[0169] Matrix material: cream, concentration 18% (w / v), mixed with nanocarriers at a ratio of 1:5.

[0170] Surface modification:

[0171] Targeting ligand: polypeptide chain, concentration 3 mg / ml.

[0172] Penetration enhancer: carboxymethyl chitosan, concentration 5% (w / v).

[0173] Smart Responsive Materials:

[0174] Materials: PNIPAAm, mass ratio of PLA to PNIPAAm is 1:3.

[0175] Coating process: solution dropping speed 6ml / min, aqueous phase volume 90 times that of organic phase, stirring speed 1000rpm.

[0176] Purification: Rotary evaporation, temperature 50°C.

[0177] Microneedle preparation:

[0178] Material: PLGA, microneedle length 1500μm, diameter 300μm, density 2000 / cm 2 .

[0179] Preparation method: mold casting.

[0180] Summary: This embodiment achieves the simultaneous delivery and controlled release of multiple drugs, ensuring that different drugs act accurately in the same environment and improving the overall therapeutic effect.

[0181] Summary:

[0182] These five examples respectively demonstrate the gradual deepening process from the basic multifunctional nanocarrier transdermal drug delivery system to enhancing targeting, improving intelligent responsive materials, optimizing drug release control, and achieving multi-drug simultaneous delivery. Each example gradually improves the transdermal absorption efficiency, targeting, and release control accuracy of the drug by adjusting the materials, preparation methods, and parameters, solving the challenges of traditional transdermal drug delivery systems in terms of drug stability, lack of targeting, and release control.

[0183] Experimental design: verify the effectiveness and superiority of the transdermal drug delivery technology of the present invention

[0184] Purpose:

[0185] Verify the effectiveness of the multifunctional nanocarrier, smart response material and microneedle array in the present invention in improving drug transdermal absorption efficiency, targeting and release control. Compare with the prior art and evaluate the drug delivery performance of the present invention under different conditions.

[0186] Experimental design:

[0187] Experimental group: five embodiments of the present invention are used, namely:

[0188] Experimental Group A: Example 1

[0189] Experimental Group B: Example 2

[0190] Experimental Group C: Example 3

[0191] Experimental Group D: Example 4

[0192] Experimental Group E: Example 5

[0193] Control group:

[0194] Control group 1: traditional transdermal patch (no nanocarrier, no microneedles)

[0195] Control group 2: Ordinary nanocarrier (monolayer, without targeting ligand and smart material)

[0196] Control group 3: multilayer nanocarrier without smart response material

[0197] Experimental procedures

[0198] Preparation of pharmaceutical preparations:

[0199] Prepare the drug preparations for the experimental and control groups to ensure consistent dosages.

[0200] Animal model selection and treatment:

[0201] Healthy adult female mice were selected, with 5 mice in each group. All mice were adaptively raised for one week, fasted for 12 hours before the experiment, and had free access to water.

[0202] Drug Delivery and Sampling:

[0203] Each group of drug preparations were applied to the shaved back skin of mice. The experimental group used the corresponding microneedle device combined with the matrix material, and the control group directly applied the drug preparation.

[0204] After drug administration, blood samples were collected at 0, 1, 2, 4, 6, 8, 12, and 24 hours, with approximately 0.2 ml of blood collected each time.

[0205] Drug concentration detection:

[0206] High performance liquid chromatography (HPLC) was used to determine the drug concentration in plasma at each time point.

[0207] Skin permeability assessment:

[0208] The drug residue in the skin was evaluated by skin punch method.

[0209] Drug release control evaluation

[0210] The drug release rate was evaluated under different temperature conditions (25°C and 40°C) and pH conditions (pH 5.5 and pH 7.4).

[0211] Table 1:

[0212]

[0213]

[0214] Experimental summary:

[0215] The experimental results show that the embodiments of the present invention significantly improve the transdermal absorption efficiency, maximum blood drug concentration (Cmax) and bioavailability (AUC0-24) of the drug, which are significantly improved compared with the control group. Experimental group E (multi-drug synchronous delivery system) showed the highest drug absorption and controlled release effect, verifying that the combination of multifunctional nanocarriers, multilayer structures, smart response materials and biodegradable microneedles is significantly better than traditional transdermal drug delivery systems. At the same time, the experiment also verified the dynamic control ability of smart responsive materials under temperature and pH changes, ensuring the precise release of drugs under specific pathological conditions.

[0216] The overall results show that the present invention has obvious innovation and superiority in transdermal drug delivery technology, and can effectively solve the problems of insufficient drug absorption, poor targeting and inaccurate release control in the prior art, providing a reliable solution for efficient and safe drug delivery.

[0217] Experimental design: verifying the transdermal absorption efficiency of the present invention under different skin conditions

[0218] Purpose:

[0219] The effects of the multifunctional nanocarriers and intelligent responsive materials of the present invention on drug transdermal absorption efficiency under different skin conditions (such as healthy skin and damaged skin) were further verified and compared with the prior art to demonstrate the superiority and wide applicability of the present invention under various skin conditions.

[0220] Experimental design:

[0221] Experimental group: using the best embodiment E of the present invention, divided into:

[0222] Experimental group E1: healthy skin

[0223] Experimental group E2: Mildly damaged skin (simulated mild abrasion)

[0224] Experimental group E3: moderately damaged skin (simulating mild burns)

[0225] Control group:

[0226] Control group 1: Use of traditional transdermal patch on healthy skin

[0227] Control group 2: Use of conventional transdermal patch on mildly damaged skin

[0228] Control group 3: Use of conventional transdermal patch on moderately damaged skin

[0229] Experimental steps:

[0230] Preparation of pharmaceutical preparations:

[0231] Prepare drug preparations for the experimental and control groups to ensure consistent drug concentrations.

[0232] Animal model selection and treatment:

[0233] Healthy adult female mice were selected, with 10 mice in each group.

[0234] Preparation of damaged skin model:

[0235] Mildly damaged skin: Lightly sand the mouse back skin with fine sandpaper.

[0236] Moderately damaged skin: Use a temperature-controlled hot plate in contact with the skin to simulate a mild burn.

[0237] Drug Delivery and Sampling:

[0238] Each group of drug preparations was applied to the back skin of mice. The experimental group used a combination of nanocarriers and microneedles, and the control group used traditional patches.

[0239] Blood samples were taken at 0, 1, 2, 4, 6, 8, 12, and 24 hours, with approximately 0.2 ml of blood being taken each time.

[0240] Drug concentration detection:

[0241] The drug concentration in plasma was determined by HPLC.

[0242] Skin penetration and distribution assessment:

[0243] Skin sampling was performed to measure the drug concentration distribution in different skin layers.

[0244] Skin Inflammatory Response and Repair Assessment:

[0245] The inflammatory response and healing of the damaged skin were evaluated using histopathological methods.

[0246] Table 2:

[0247]

[0248]

[0249] Experimental summary:

[0250] The experimental results show that the present invention significantly improves the transdermal absorption efficiency of drugs under different skin conditions, and the maximum blood drug concentration (Cmax) and area under the curve (AUC0-24) are significantly higher than those of the control group, especially on damaged skin, showing stronger penetration ability and stable drug release. In addition, the multifunctional nanocarrier and intelligent response material of the present invention effectively reduce the inflammatory response of damaged skin and accelerate the repair process of the skin, showing good biocompatibility and safety. These results further verify the applicability and superiority of the present invention under various skin conditions, and provide an effective solution for drug delivery under different pathological conditions.

[0251] Although 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 the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. Methods of enhancing transdermal drug delivery permeability using biomaterials. It is characterized in that The following steps are involved: S1. preparing a multifunctional nanocarrier comprising a polymer core and a functionalized shell; S2, mixing the multifunctional nanocarrier with the matrix material; S3, modifying the surface of the nanocarrier and adding a targeting ligand or a penetration enhancer; S4. Application of intelligent responsive materials, which are temperature-sensitive materials or pH-sensitive materials, to control drug release when temperature or pH changes; S5, adding an enhancer to the matrix material, wherein the enhancer is selected from hyaluronic acid, chitosan or other natural polysaccharides; S6. Prepare a microneedle array, wherein the microneedles are made of a biodegradable material selected from PLGA or PVP, and combine the biodegradable material with a carrier matrix.

2. The method for enhancing the permeability of transdermal drug delivery using biomaterials according to claim 1, It is characterized in that In the step S1, the core is selected from one of PLGA and PLA, the shell layer is selected from one of gold nanoparticles and silicon dioxide nanoparticles, and in the step S2, the matrix material is selected from one of cream, gel or patch.

3. The method for enhancing the permeability of transdermal drug delivery using biomaterials according to claim 1, It is characterized in that The preparation method of the core material in step S1 can be any of nanoprecipitation method and double emulsion method. The encapsulation technology used for the multifunctional nanocarrier in step S1 is to form a multilayer structure on the surface of the nanocarrier, each layer encapsulating different drugs or protective agents.

4. The method for enhancing the permeability of transdermal drug delivery using biomaterials according to claim 1, It is characterized in that In the step S3, the targeting ligand is selected from an antibody or a peptide chain, and the penetration enhancer is selected from a chitosan derivative. The chitosan derivative is optionally selected from methylated chitosan, carboxymethyl chitosan or quaternized chitosan.

5. The method for enhancing the permeability of transdermal drug delivery using biomaterials according to claim 1, It is characterized in that The S3 step specifically includes the following steps: S3.

1. Targeted ligand modification Ligand selection and connection method: First, according to the specific antigens or receptors of the target tissue or cells, appropriate targeting ligands are selected. The methods for connecting these ligands to the surface of nanocarriers include chemical bonding and physical adsorption; Optimization and post-processing: To achieve the best effect, the concentration of the targeting ligand needs to be controlled within the range of 0.1-2 mg / ml, and the reaction time needs to be controlled within the range of 1-3 h, the temperature needs to be 4-25 °C, and the pH value needs to be controlled within the range of 7-8. After the reaction is completed, the unreacted ligand should be removed by dialysis, centrifugation or chromatography; Testing and verification: Enzyme-linked immunosorbent assay and surface plasmon resonance technology are used to determine whether the ligand binding efficiency is between 70-90%. Cell experiments are also needed to verify the targeting ability of the nanocarrier; S3.2, Penetration Enhancer Modification: Enhancer selection and modification method: According to the requirements of drug transdermal absorption, select appropriate penetration enhancers. Modification methods include coating, blending or covalent bonding; Concentration control and post-processing: During the implementation process, the concentration of the enhancer needs to be controlled within the range of 0.1-5% w / v, and the solution is prepared according to the target concentration. After the modification is completed, the unbound enhancer is removed by dialysis or ultrafiltration.

6. The method for enhancing the permeability of transdermal drug delivery using biomaterials according to claim 1, It is characterized in that In the step S4, the temperature-sensitive material is optionally one of N-isopropylacrylamide, polyurethane or lactide-glycolide, and the pH-sensitive material is optionally one of polyacrylic acid, sodium polyacrylate or polyethyleneimine.

7. The method for enhancing the permeability of transdermal drug delivery using biomaterials according to claim 1, It is characterized in that The specific steps of applying the smart response material in step S4 are as follows: S4.

1. Intelligent material selection: one of the temperature-sensitive material and pH-sensitive material can be selected; S4.2, Material integration: In the process of preparing nanocarriers, PNIPAAm is blended with polymer materials to form nanoparticles with core-shell structure. PNIPAAm can be used as an outer layer to coat the core drug or carrier. The mass ratio of PNIPAAm to PLGA is between 1:4 and 1:

1. S4.3, preparation process: PNIPAAm and PLGA are dissolved in an organic solvent to form a uniform mixed solution, and the solution is added dropwise to the aqueous phase using a nanoprecipitation method, the dropping speed is controlled at 0.5-5 ml / min, the volume of the aqueous phase is 10-100 times that of the organic phase, the stirring speed is controlled at 300-1000 rpm, and nanoparticles are formed by solvent evaporation, and PNIPAAm is evenly distributed in the outer layer of the nanoparticles; S4.

4. Solvent removal and purification: Remove the organic solvent by rotary evaporation at 40-50°C, or use ultrafiltration with a membrane with a molecular weight cutoff point of 10-100 kDa to remove free low molecular weight components. Nanoparticle purification is performed by dialysis with a dialysis bag with a molecular weight cutoff point of 10-100 kDa and a dialysis time of 4-24 h.

8. The method for enhancing the permeability of transdermal drug delivery using biomaterials according to claim 1, It is characterized in that The step S6 specifically includes the following steps: S6.

1. Microneedle material selection and manufacturing: Pour the PLGA or PVP polymer solution into the prefabricated microneedle mold, dry it at room temperature or under vacuum conditions, further solidify the molded microneedles, build the microneedle structure layer by layer using high-precision 3D printing technology, and make the PLGA or PVP material into a liquid or powder form suitable for 3D printing; S6.

2. Microneedle parameter design Length range: 150-1500μm; Diameter range: 50-500μm; Density range: 200-2000 pieces / cm 2 ; S6.3, Microneedle and matrix combination: Drug loading: During the process of manufacturing microneedles, drugs are embedded directly into the polymer; Binding to a carrier matrix: Binding the prepared microneedle array to a carrier matrix such as a cream, gel or patch; Binding method: The microneedle array is fixed to the substrate by physical attachment or chemical bonding.

9. The method for enhancing the permeability of transdermal drug delivery using biomaterials according to claim 8, It is characterized in that The high-precision 3D printing technology in step S6.1 may be photo-curing stereolithography or fused deposition modeling technology, and the matrix material in step S6.2 may be water-based, oil-based or mixed matrix.

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