Composite microneedle patch as well as preparation method and application thereof
By using hyaluronic acid-loaded paclitaxel in microneedle patches, the problem of drug delivery after CABG is solved, effective delivery and slow release of drugs are achieved, vascular stenosis is significantly reduced, and the administration process is simple and easy.
Patent Information
- Application Number
- CN202411287943.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-13
- Publication Date
- 2024-12-10
AI Technical Summary
The lack of suitable drug delivery methods for paclitaxel and/or paclitaxel derivatives after CABG surgery makes it difficult to effectively prevent vascular stenosis caused by endometrial hyperplasia.
Using microneedle patches with hyaluronic acid as the substrate and loaded with paclitaxel and/or paclitaxel derivatives, the biocompatibility of microneedles and drug release mechanisms can achieve localization and drug delivery and slow release of drugs in the skin to avoid Injury to the vein.
The effective delivery and slow release of drugs are achieved, which significantly reduces vascular stenosis of endometrial hyperplasia, and the administration process is simple and easy, without the need for specialized medical staff to operate.
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Figure CN119097591A_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the technical field of microneedle drug delivery, and in particular to a composite microneedle patch and a preparation method and application thereof. Background Art
[0002] After CABG (coronary artery bypass grafting) surgery, intimal hyperplasia (IH) of the vein graft is likely to occur, resulting in restenosis of the blood vessels.
[0003] Paclitaxel can bind to the microtubule protein of cells to promote aggregation and inhibit its dissociation, resulting in the inability of cells to divide normally, causing cell cycle arrest and apoptosis. It has a good effect on the treatment of vascular stenosis caused by intimal hyperplasia.
[0004] Experiments have shown that drug-eluting stents can prevent intimal hyperplasia after PCI (percutaneous coronary intervention), but there is a lack of suitable paclitaxel drug delivery methods for intimal hyperplasia after CABG. Summary of the invention
[0005] In view of this, the present application provides a composite microneedle patch and a preparation method and application thereof to solve the problem of lack of a suitable drug delivery method for paclitaxel and / or paclitaxel derivatives after CABG surgery.
[0006] In a first aspect, the present application provides a composite microneedle patch.
[0007] In an optional embodiment, the composite microneedle patch includes a needle seat and a needle body located on the needle seat, and the needle body includes a drug-loaded gel with hyaluronic acid as a substrate; the drug loaded in the drug-loaded gel includes paclitaxel and / or paclitaxel derivatives.
[0008] In order to improve the efficacy and accuracy of paclitaxel drug delivery in the vascular environment, this application uses a microneedle patch composed of microneedles with hyaluronic acid as the substrate and paclitaxel and / or paclitaxel derivatives as the delivery device. Paclitaxel can be dissolved in a large amount in the hyaluronic acid solution, which increases the drug loading; and hyaluronic acid will not affect the activity of paclitaxel. The paclitaxel loaded can pass through the epidermis of the skin to achieve intradermal localization and drug administration. The biocompatibility of the microneedle is good, and the patch is wrapped in the adventitia of the great saphenous vein without causing damage to the vein; the paclitaxel loaded in the microneedle is gradually released as the hyaluronic acid gel dissolves in the skin. The immunofluorescence results 48 hours after the operation show that the microneedle patch loaded with paclitaxel can smoothly penetrate the intima of the great saphenous vein and realize the distribution of drugs in the intima, and the drug absorption is good. At the same time, microneedle administration does not require special medical staff to operate, and patients can administer the drug by themselves, which is convenient to use.
[0009] In an optional embodiment, the height of the needle body is 700-800 μm;
[0010] And / or, the needle body is provided with at least one microhole for releasing the drug.
[0011] In a second aspect, the present application provides a method for preparing the composite microneedle patch, the method comprising: placing a drug-loaded gel solution in a microneedle mold, drying and demoulding, to obtain a composite microneedle patch.
[0012] In an optional embodiment, the method for preparing the drug-loaded gel solution comprises:
[0013] (1) dissolving paclitaxel and / or a paclitaxel derivative in a solvent to obtain a paclitaxel solution;
[0014] (2) Hyaluronic acid and water are mixed to form a gel solution;
[0015] (3) Mixing the paclitaxel solution obtained in step (1) and the gel solution obtained in step (2) to form a drug-loaded gel solution.
[0016] The preparation method of the drug-loaded gel in the present application is mild and will not affect the active ingredient of paclitaxel.
[0017] In an optional embodiment, in step (1), paclitaxel or a paclitaxel derivative is dissolved in acetone to prepare a paclitaxel-acetone solution; and the paclitaxel-acetone solution is rapidly injected into rapidly stirred ultrapure water to form a paclitaxel solution.
[0018] The present invention quickly forms a milky white nanocrystalline suspension by rapidly dispersing paclitaxel and / or paclitaxel derivatives from non-polar acetone to a reverse polarity transition in a highly polar aqueous medium. The paclitaxel and / or paclitaxel derivatives can be evenly dispersed in the microneedles and continuously release the drug, solving the technical problem that poorly soluble drug molecules cannot be evenly dispersed in hyaluronic acid hydrogels. The drug simulation release experiment shows that paclitaxel can be released by more than 8% within 6 hours, achieving the effect of faster drug release in the early stage and sustained release in the later stage.
[0019] In an optional embodiment, the mass volume ratio of paclitaxel or paclitaxel derivative and acetone is (1.5 mg-2.5 mg): 1 mL;
[0020] And / or, the volume ratio of the paclitaxel-acetone solution to ultrapure water is (0.05-0.15):1;
[0021] And / or, the rotation speed of the rapid stirring is 800r / min-1000r / min.
[0022] In an optional embodiment, the mass volume ratio of the hyaluronic acid and water is (0.05g-0.15g): 10mL;
[0023] And / or, the volume ratio of the paclitaxel solution to the gel solution is (0.5-1.5):1.
[0024] In an optional embodiment, the drying is vacuum drying at 37°C-39°C;
[0025] And / or, the drying time is 8 hours to 10 hours.
[0026] In an optional embodiment, the microneedle mold is a dimethylsiloxane microneedle template.
[0027] In a third aspect, the present application provides a use of the composite microneedle patch or the composite microneedle patch prepared by the preparation method in preparing a drug for preventing or treating intimal hyperplasia;
[0028] Preferably, the composite microneedle patch is used in the preparation of a drug for preventing or treating intimal hyperplasia after coronary artery bypass grafting.
[0029] Compared with the prior art, the beneficial effects of this application are:
[0030] The composite microneedle patch of the present application includes a needle seat and a needle body located on the needle seat, wherein the needle body includes a drug-loaded gel with hyaluronic acid as a substrate; the drug loaded in the drug-loaded gel includes paclitaxel and / or paclitaxel derivatives. The microneedle patch is used as a delivery device to dissolve a large amount of paclitaxel in the hyaluronic acid solution, thereby increasing the drug loading amount; and hyaluronic acid will not affect the activity of paclitaxel.
[0031] The paclitaxel loaded in the composite microneedle patch can penetrate the epidermis of the skin to achieve targeted drug delivery within the skin. The microneedles have good biocompatibility, and the patch is wrapped in the adventitia of the great saphenous vein without causing damage to the vein.
[0032] The paclitaxel loaded in the microneedles was gradually released as the hyaluronic acid gel dissolved in the skin. The drug simulation release experiment showed that paclitaxel could be released by more than 8% within 6 hours, achieving the effect of rapid drug release in the early stage and sustained release in the later stage. Immunofluorescence results 48 hours after surgery showed that the microneedle patch loaded with paclitaxel could successfully penetrate the intima of the great saphenous vein and achieve drug distribution in the intima, with good drug absorption.
[0033] At the same time, microneedle drug delivery does not require specialized medical personnel to operate, and patients can administer the drug themselves, which is convenient to use. BRIEF DESCRIPTION OF THE DRAWINGS
[0034] In order to more clearly illustrate the specific implementation methods of the present application or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.
[0035] Figure 1 This is the TEM characterization result of the milky white nanocrystalline suspension containing paclitaxel in Example 1 of the present application;
[0036] Figure 2 is a graph showing the particle size distribution of nanoparticles in the milky white nanocrystalline suspension of Example 1 of the present application;
[0037] Figure 3 This is the microscopic morphology of the nanoparticles in Example 1 of the present application;
[0038] Figure 4 is a technical morphology diagram of the microneedle array of Example 1 of the present application;
[0039] Figure 5 This is an appearance diagram of the microneedle patch of Example 1 of the present application;
[0040] Figure 6 This is the drug-loaded microneedle patch under a 60x magnifying glass of Example 1 of the present application;
[0041] Figure 7 The drug-loaded microneedle patch under a scanning electron microscope of Example 1 of the present application;
[0042] Figure 8 is a comparison of infrared spectra of two microneedle patches in Experimental Example 1 of the present application;
[0043] Fig. 9 In Experimental Example 2 of the present application, the composite microneedle patch is wrapped around the adventitia of the great saphenous vein, A is before wrapping, and B is after wrapping;
[0044] Fig.10 The great saphenous vein after the drug-loaded microneedle of Experimental Example 2 of the present application is dissolved;
[0045] Fig.11 is the immunofluorescence test result of Experimental Example 2 of the present application;
[0046] Fig.12 The drug release of the composite microneedle patch of Example 1 of the present application under in vitro simulated conditions;
[0047] Fig.13 The drug release of the composite microneedle patch of Example 2 of the present application under in vitro simulated conditions;
[0048] Fig.14 This is the drug release of the composite microneedle patch of Example 3 of the present application under in vitro simulated conditions. DETAILED DESCRIPTION
[0049] The following examples are provided for a better understanding of the present application, but are not limited to the best implementation mode described, and do not limit the content and protection scope of the present application. Any product identical or similar to the present application obtained by anyone under the inspiration of the present application or by combining the features of the present application with other prior arts shall fall within the protection scope of the present application.
[0050] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:
[0051] In this application, the selection scope of "and / or" includes any one of two or more related listed items, and also includes any and all combinations of related listed items, and the said any and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. For example, "A and / or B" includes three parallel solutions of A, B and A+B.
[0052] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.
[0053] In the present application, the percentage content, unless otherwise specified, refers to mass percentage for solid-liquid mixing and solid-solid mixing, and refers to volume percentage for liquid-liquid mixing.
[0054] In this application, when it comes to percentage concentration, unless otherwise specified, it refers to the final concentration. The final concentration refers to the proportion of the added component in the system after the addition of the component.
[0055] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.
[0056] The present application is further described in detail below in conjunction with specific embodiments. These embodiments should not be construed as limiting the scope of protection claimed in the present application.
[0057] Paclitaxel in the following experiments was purchased from MedChemexpress (MCE China) with a purity of 99.98%.
[0058] If no specific experimental steps or conditions are specified in the following experiments, the conventional experimental steps or conditions described in the literature in this field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be purchased commercially.
[0059] Example 1
[0060] This embodiment provides a composite microneedle patch including a needle seat and a needle body located on the needle seat, wherein the needle body includes a drug-loaded gel with hyaluronic acid as a substrate; the drug loaded in the drug-loaded gel includes paclitaxel and / or paclitaxel derivatives; the height of the needle body is 750 μm; and a plurality of micropores are provided on the needle body.
[0061] The preparation method of the composite microneedle patch includes:
[0062] (1) Paclitaxel was dissolved in acetone to prepare a 2 mg / mL paclitaxel-acetone solution; 0.2 mL of the paclitaxel-acetone solution was then rapidly injected into 2 mL of ultrapure water that was rapidly stirred (at a speed of 800 r / min) to form a paclitaxel solution (a milky white nanocrystalline suspension containing paclitaxel), which was then placed at 4° C. for later use; a transmission electron microscope (TEM) and a laser particle size analyzer were used to characterize the microstructure. The TEM characterization results of the milky white nanocrystalline suspension containing paclitaxel are shown in FIG. Figure 1 As can be seen from the figure, nanoparticles have been formed, and their solution morphology is as follows Figure 3 It can be seen that the nanoparticles are evenly distributed in the milky white nanocrystal suspension, and the particle size distribution is as follows Figure 2 ,The hydrated particle size of the preparation was 99 nm, and the particle size dispersion index was 0.08, indicating that the monodispersity of the drug was good.
[0063] (2) Weigh 0.1 g of hyaluronic acid and mix with 10 mL of water to form a gel solution;
[0064] (3) Mix 10 ml of the paclitaxel solution obtained in step (1) and 10 ml of the gel solution obtained in step (2) to form a drug-loaded gel solution.
[0065] (4) Spread the drug-loaded gel solution obtained in step (3) on a dimethylsiloxane microneedle template (poly PDMS mold), place the mold in a drying oven, and vacuum dry it at 38°C for 9 hours. After it is completely dried, demold it to obtain a composite microneedle patch. Microneedle array technology morphology Figure 4 The microneedle patch array after demoulding is a square piece with a side length of 1.5 cm. Its base is flat, the microneedles are uniform in length, the shape is good, and there is no obvious breakage. Figure 5 , the drug-loaded microneedle patch under a 60x magnifying glass Figure 6, the drug-loaded microneedle patch under a scanning electron microscope Figure 7 The single microneedle is in the shape of a quadrangular pyramid, with an average height of 750 μm and an average bottom width of 300 μm, which is consistent with the size of the PDMS template. This indicates that the microneedle patch has good physical and mechanical properties.
[0066] Example 2
[0067] This embodiment provides a method for preparing a composite microneedle patch, comprising:
[0068] (1) dissolving paclitaxel in acetone to prepare a 1.5 mg / mL paclitaxel-acetone solution; then rapidly injecting the paclitaxel-acetone solution (0.1 mL) into 2 mL of ultrapure water that is rapidly stirred (speed of 800 r / min) to form a paclitaxel solution (a milky white nanocrystal suspension containing paclitaxel), and placing it at 4° C. for use;
[0069] (2) Weigh 0.05 g of hyaluronic acid and mix with 10 mL of water to form a gel solution;
[0070] (3) Mix 15 ml of the paclitaxel solution obtained in step (1) and 10 ml of the gel solution obtained in step (2) to form a drug-loaded gel solution.
[0071] (4) Spread the drug-loaded gel solution obtained in step (3) on a dimethylsiloxane microneedle template (poly-PDMS mold), place the mold in a drying oven, and vacuum dry it at 37° C. for 10 hours. After it is completely dried, demold the mold to obtain a composite microneedle patch.
[0072] Example 3
[0073] This embodiment provides a method for preparing a composite microneedle patch, comprising:
[0074] (1) dissolving paclitaxel in acetone to prepare a 2.5 mg / mL paclitaxel-acetone solution; then rapidly injecting the paclitaxel-acetone solution (0.3 mL) into 2 mL of ultrapure water that is rapidly stirred (speed of 800 r / min) to form a paclitaxel solution (a milky white nanocrystal suspension containing paclitaxel), and placing it at 4° C. for use;
[0075] (2) Weigh 0.15 g of hyaluronic acid and mix with 10 mL of water to form a gel solution;
[0076] (3) Mix 5 ml of the paclitaxel solution obtained in step (1) and 10 ml of the gel solution obtained in step (2) to form a drug-loaded gel solution.
[0077] (4) Spread the drug-loaded gel solution obtained in step (3) on a dimethylsiloxane microneedle template (poly-PDMS mold), place the mold in a drying oven, and vacuum dry it at 39° C. for 8 hours. After it is completely dried, demold the mold to obtain a composite microneedle patch.
[0078] Comparative Example 1
[0079] This comparative example provides a method for preparing a composite microneedle patch, which is as follows:
[0080] (1) Weigh 0.05 g to 0.15 g of hyaluronic acid and mix with 10 mL of water to form a gel solution;
[0081] (2) Spreading the gel solution on a dimethylsiloxane microneedle template (poly-PDMS mold), placing the mold in a drying oven, and vacuum drying at 37° C. to 39° C. for 8 to 10 hours. After being completely dried, demolding is performed to obtain a hyaluronic acid microneedle patch without paclitaxel loading.
[0082] Test Example 1
[0083] The composite microneedle patch loaded with paclitaxel (drug-loaded microneedles) prepared in Example 1 was compared with the hyaluronic acid microneedle patch not loaded with paclitaxel (blank microneedles) prepared in Comparative Example 1. The infrared spectra of the two microneedle patches are shown in FIG. Figure 8 , drug-loaded microneedle patch at 1728, 2834, 2915cm -1 It showed obvious differentiated infrared absorption peaks belonging to the drug, and the results showed that paclitaxel was successfully doped into the hyaluronic acid microneedle patch (HA-MN).
[0084] Test Example 2
[0085] The paclitaxel-loaded composite microneedle patch prepared in Example 1 was placed on Fig. 9 The wrapping method is to completely and evenly wrap the adventitia of the great saphenous vein. Fig. 9 In the figure, A is before wrapping, and B is after wrapping. After 5 minutes, the test was carried out, and the results are as follows Fig.10 The picture shows that the microneedle patch was completely dissolved and the veins expanded well, indicating that the microneedle patch did not damage the veins. Immunofluorescence testing was performed 48 hours after surgery. The results are as follows Fig.11 As shown, the composite microneedle patch carrying paclitaxel can successfully penetrate the intima of the great saphenous vein and achieve drug distribution in the intima.
[0086] Test Example 3
[0087] The paclitaxel in the vascular homogenate of the great saphenous vein wrapped by the composite microneedle patch loaded with paclitaxel prepared in Examples 1-3 was continuously measured by LC-MS / MS method for 24 hours. The dissolution and release curve of paclitaxel in the composite microneedle patch of Example 1 over time is shown in FIG. Fig.12 The drug release of the composite microneedle patch of Example 2 under in vitro simulated conditions is shown in Fig.13 The drug release of the composite microneedle patch of Example 3 under in vitro simulated conditions is as follows Fig.14 .
[0088] As can be seen from the figure, the drug release is faster in the early stage and faster within 6 hours ( Fig.12 The drug release rate was 8% within 6 hours, slowed down in 6-12 hours, and slowed down in 12-24 hours, and the concentration had entered a plateau phase. The results showed that the drug could be continuously and slowly released from the microneedle patch and exert its efficacy. With the dissolution of the needle tip, the drug was completely released and the contained drug would slowly diffuse into the vascular endothelium.
[0089] In summary, the composite microneedle patch of the present application can penetrate the epidermis of the skin to achieve intradermal localized drug delivery, the biocompatibility of the microneedles is good, and the patch is wrapped in the adventitia of the great saphenous vein without causing damage to the vein. The paclitaxel loaded in the microneedles is gradually released as the hyaluronic acid gel dissolves in the skin. The drug simulation release experiment shows that paclitaxel can be released by more than 8% within 6 hours, achieving the effect of faster drug release in the early stage and sustained release in the later stage. The immunofluorescence results 48 hours after the operation showed that the microneedle patch loaded with paclitaxel can smoothly penetrate the intima of the great saphenous vein and achieve drug distribution in the intima, complete drug release, and good drug absorption.
[0090] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection created by this application.
Claims
1. A composite microneedle patch, comprising a needle seat and a needle body located on the needle seat, characterized in that: The needle body comprises a drug-loaded gel with hyaluronic acid as a base material; the drug loaded in the drug-loaded gel comprises paclitaxel and / or paclitaxel derivatives.
2. The composite microneedle patch according to claim 1, characterized in that: The height of the needle body is 700-800 μm; And / or, the needle body is provided with at least one microhole.
3. A method for preparing the composite microneedle patch according to claim 1 or 2, characterized in that: The preparation method comprises: placing a drug-loaded gel solution in a microneedle mold, drying and demoulding, and obtaining a composite microneedle patch.
4. The preparation method according to claim 3, characterized in that: The preparation method of the drug-loaded gel solution comprises: (1) dissolving paclitaxel and / or a paclitaxel derivative in a solvent to obtain a paclitaxel solution; (2) Hyaluronic acid and water are mixed to form a gel solution; (3) Mixing the paclitaxel solution obtained in step (1) and the gel solution obtained in step (2) to form a drug-loaded gel solution.
5. The preparation method according to claim 4, characterized in that: In step (1), paclitaxel or a paclitaxel derivative is dissolved in acetone to prepare a paclitaxel-acetone solution; and the paclitaxel-acetone solution is rapidly injected into rapidly stirred ultrapure water to form a paclitaxel solution.
6. The preparation method according to claim 5, characterized in that: The mass volume ratio of paclitaxel or paclitaxel derivative and acetone is (1.5 mg-2.5 mg):1 mL; And / or, the volume ratio of the paclitaxel-acetone solution to ultrapure water is (0.05-0.15):1; And / or, the rotation speed of the rapid stirring is 800r / min-1000r / min.
7. The preparation method according to claim 4, characterized in that: The mass volume ratio of the hyaluronic acid and water is (0.05g-0.15g): 10mL; And / or, the volume ratio of the paclitaxel solution to the gel solution is (0.5-1.5):
1.
8. The preparation method according to claim 3, characterized in that: The drying is vacuum drying at 37°C-39°C; And / or, the drying time is 8 hours to 10 hours.
9. The preparation method according to claim 5, characterized in that: The microneedle mold is a dimethylsiloxane microneedle template.
10. Use of the composite microneedle patch according to claim 1 or 2 or the composite microneedle patch prepared by the preparation method according to any one of claims 3 to 9 in the preparation of a drug for preventing or treating intimal hyperplasia; Preferably, the composite microneedle patch is used in the preparation of a drug for preventing or treating intimal hyperplasia after coronary artery bypass grafting.