Cyclovirobuxin D microneedle patch, preparation method and application thereof
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- BEIJING CAS MICRONEEDLE TECH LTD
- Filing Date
- 2025-10-10
- Publication Date
- 2026-07-21
Smart Images
Figure CN120960127B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of pharmaceutical preparations, specifically relating to a poorly soluble drug, Cyclovinaxyl D microneedle patch, its preparation method, and its application. Background Technology
[0002] Cardiovascular diseases (CVDs) are a leading cause of death worldwide, causing approximately 18.6 million deaths annually, accounting for 32.3% of all deaths globally. Currently, about 540 million people worldwide suffer from CVDs, with over 70% requiring lifelong multidrug therapy. However, medication adherence is generally low, especially among the elderly, where adherence rates are as low as 38%, leading to a significant 300% increase in readmission risk. This problem carries a heavy socioeconomic burden, with direct healthcare expenditures reaching US$1.2 trillion annually and a productivity loss equivalent to 370 million disability-adjusted life years (DALYs).
[0003] Cyclovitakine D (CVB-D), the main alkaloid component of Huangyangning tablets, is a significant botanical drug discovery in China following artemisinin and ephedrine. It possesses a dual mechanism of cardioprotection, regulating voltage-gated calcium channels and improving microcirculation. However, its low water solubility leads to low bioavailability during oral administration, primarily due to the first-pass effect and gastrointestinal degradation. Furthermore, the drug requires multiple daily doses (2–3 times daily), making adherence particularly challenging for elderly patients with cognitive decline and complex multidrug use. Over 60% of patients fail to maintain consistent dosing, severely impacting efficacy and increasing the risk of cardiovascular events. While transdermal delivery can improve patient adherence, traditional patches are limited by the stratum corneum's permeability barrier, hindering the achievement of sustained effective blood concentrations of CVB-D. Therefore, developing a non-invasive, patient-friendly drug delivery system capable of long-term, stable drug release has become an urgent clinical need.
[0004] Microneedle (MN) technology, as an emerging platform for transdermal drug delivery, can physically penetrate the stratum corneum to deliver drugs to the dermis, significantly improving drug penetration efficiency and bioavailability. However, the practical application of this technology heavily relies on the development of next-generation matrix materials that possess excellent mechanical properties, biocompatibility, and controllable degradation.
[0005] In recent years, polysaccharide components derived from traditional Chinese medicine have attracted widespread attention due to their pharmacological activity, functional properties, and potential as excipients. For example, Bletilla striata polysaccharide has been used in the preparation of soluble microneedles, exhibiting superior hemostatic and healing-promoting properties, as well as good needle-forming properties and skin permeability compared to traditional materials such as hyaluronic acid and polyvinyl alcohol. It also possesses adjuvant therapeutic functions such as anti-inflammatory, antioxidant, and antibacterial effects.
[0006] Laminarin (LAM), a natural marine polysaccharide, possesses biodegradability, biocompatibility, and multifunctional bioactivity, making it a highly promising biomedical material. This study utilizes LAM as the core matrix of a self-supporting dissolvable microneedle (LAM-DMN) for sustained transdermal delivery of cardiovascular blood vessel blockade (CVB-D). This not only addresses the compliance and pharmacokinetic challenges associated with oral administration but also enables a novel cardiovascular treatment modality based on drug-excipient synergy. Summary of the Invention
[0007] To address the aforementioned technical problems, this invention provides a method for preparing and applying a cyclovinylbuxusin D microneedle patch. The cyclovinylbuxusin D microneedle patch is used for the treatment of myocardial ischemia and is suitable for transdermal administration to achieve the synergistic cardioprotective effect of cyclovinylbuxusin D (CVB-D) and laminarin.
[0008] This invention is achieved through the following technical solution: A CVB-D microneedle patch, wherein the CVB-D microneedle patch comprises CVB-D solid dispersion and laminarin polysaccharide; the laminarin polysaccharide is used as the matrix material of the microneedles and plays a synergistic therapeutic role as a cardioprotective bioactive component, thereby enhancing the overall therapeutic effect.
[0009] Furthermore, the microneedle tip height of the cyclohexane star D microneedle patch is 200-500 μm (preferably 300-500 μm), and the tip distance is 150-600 μm (preferably 300-500 μm). In the cyclovinyl buxus microneedle patch, the mass ratio of the CVB-D solid dispersion to kelp polysaccharide is (1-3):1.
[0010] Furthermore, in the Cyclofibrillated Buxusin D Microneedle Patch, laminarin polysaccharide serves as an excipient and also as a bioactive ingredient with cardioprotective effects to enhance the overall therapeutic effect. The microneedle portion of the Cyclofibrillated Buxusin D Microneedle Patch completely dissolves within 30 minutes.
[0011] Furthermore, the application of the cycloviniferin D microneedle patch in the preparation of drugs for treating myocardial ischemia. This application achieves the synergistic effect of cycloviniferin D and laminarin through transdermal administration, and the therapeutic benefit provided by a single microneedle transdermal patch administration is greater than that of a twice-daily oral dose.
[0012] A method for preparing a cyclovin-based Buxus microneedle patch, the method comprising the following steps: CVB-D solid dispersion was prepared using cyclovinyl fulvidin D and a dispersant; The CVB-D solid dispersion was ultrasonically dissolved and then mixed with laminarin. After centrifugation to remove air bubbles, a dispersion-laminarin mixed solution was obtained. The dispersion-laminarin mixed solution was added into a microneedle mold, vacuum-suctioned, dried, and then the mold was peeled off to obtain the cyclovinyl fulva D microneedle patch.
[0013] Furthermore, the preparation method of the CVB-D solid dispersion is as follows: dissolve cyclovinyl flavonoid D and a dispersant in an organic solution, and prepare the CVB-D solid dispersion by ultrasonic dissolution and freeze drying.
[0014] Furthermore, the dispersant is povidone K30 or povidone K90; the organic solution is an ethanol solution with a volume concentration of 50% to 100%, including anhydrous ethanol and all intermediate concentrations between 50% and 100%.
[0015] The mass ratio of the cyclohexane D to the dispersant is 1:(2-10).
[0016] Furthermore, the mass ratio of the CVB-D solid dispersion to the kelp polysaccharide is (1-3):1.
[0017] Furthermore, the preparation method of the dispersion-laminarin mixed solution is as follows: CVB-D solid dispersion is ultrasonically dissolved in water. After complete dissolution, an aqueous solution of solid dispersion is obtained. Laminarin is added to the aqueous solution of solid dispersion, stirred evenly, and centrifuged to remove air bubbles to obtain the dispersion-laminarin mixed solution.
[0018] Furthermore, the vacuum suction conditions are: vacuum suction time of 10-30 min; the drying conditions are: drying temperature of 20-45℃ and drying time of 2-6 h.
[0019] Beneficial technical effects of the present invention: In this invention, laminarin is a bioactive material (excipient) with synergistic therapeutic effects. Through a "drug (CVB-D)-excipient (laminarin) fusion" strategy, laminarin functions as both an excipient and a bioactive cardioprotective agent, enhancing the overall therapeutic effect. Furthermore, this "drug-excipient fusion" mechanism allows laminarin to simultaneously exert its cardioprotective bioactivity and act as a self-releasing enhancer, enabling transdermal CVB-D administration (over 24 hours) without the need for electronic devices.
[0020] The method for preparing the Cyclofibrillaria dichotoma D microneedle patch provided by this invention uses a one-step microforming process, requires no additives, simplifies the process, and has good skin penetration efficiency and excellent mechanical strength.
[0021] The Cyclocarya spp. D microneedle patch provided by this invention, according to pharmacokinetic studies, achieves a systemic exposure equivalent to twice-daily oral administration with a single patch application. The synergistic effect of laminarin and CVB-D produces a superior anti-myocardial ischemia effect. The painless once-daily patch administration method solves the challenges of medication management (e.g., missed / duplicate doses), and is particularly beneficial for patients requiring long-term cardiovascular treatment.
[0022] The Cyclovinibari D microneedle patch provided by this invention offers a novel strategy for the active targeted therapy of CVB-D in clinical applications. Compared to gavage, the use of CVB-D-soluble microneedles avoids significant systemic toxicity, providing a new approach for painless and compliant treatment for cardiovascular patients. Furthermore, long-term administration via CVB-D microneedles offers advantages such as improved adherence, stable blood drug concentrations, reduced toxicity risks, and ease of use, providing a superior chronic disease management solution for cardiovascular patients. Moreover, the Cyclovinibari D microneedle patch provided by this invention allows for self-administration by patients, eliminating the need for any additional professional medical personnel, which greatly improves the convenience of treatment.
[0023] The Cyclofibrillated Buxus microneedle patch provided by this invention is an integrated needle, with the entire microneedle carrying the drug, resulting in a large drug loading capacity and a relatively simple manufacturing process. Attached Figure Description
[0024] Figure 1 Schematic diagram of the preparation of CVB-D@LAM DMNs microneedles and drug delivery platform.
[0025] Figure 2 Example 1: Preparation method of CVB-D solid dispersion.
[0026] Figure 3 Characterization of the CVB-D solid dispersion in Example 1, where a is the XRD result of different groups, b is the DSC result of different groups, and c is the scanning electron microscope image of different groups.
[0027] Figure 4 Morphological characterization of CVB-D@LAM DMNs: a) morphology of CVB-D@LAM DMNs magnified under a stereomicroscope; b) morphology of CVB-D@LAM DMNs magnified under a scanning electron microscope; c) overall view of the CVB-D@LAM DMNs with a needle height of 300µm; d) local morphology of the CVB-D@LAM DMNs with a needle height of 300µm.
[0028] Figure 5 Example 1: Microneedle puncture verification: a) Morphology of CVB-D@LAM DMNs microneedles after implantation into pig skin; b) HE staining results after CVB-D@LAM DMNs microneedles puncture pig skin.
[0029] Figure 6 Morphological characterization of CVB-D microneedles in comparative examples: a) Morphology of CVB-D@LAM DMNs microneedles magnified under a stereomicroscope; b) Morphology of CVB-D@LAM DMNs microneedles magnified under a scanning electron microscope.
[0030] Figure 7 Comparative example: Verification of CVB-D microneedle puncture capability; a) Morphology of CVB-D-LAM-DMN microneedles after implantation into pig skin; b) HE staining results after CVB-D@LAM DMNs microneedles puncture pig skin.
[0031] Figure 8 Example 1 and the comparative example demonstrate the in vitro release kinetics of microneedles.
[0032] Figure 9 Example 1: Skin irritation and hemolysis of CVB-D@LAM DMNs microneedles; a is a picture of the actual product, b is a graph of the hemolysis rate; Figure 10 Example 1: Changes in the dissolution state of CVB-D@LAM DMNs microneedles after being inserted into isolated pig skin.
[0033] Figure 11 The amount of CVB-D residue on the skin surface and accumulation in the dermis delivered by CVB-D@LAM DMNs microneedles in Example 1.
[0034] Figure 12 Pharmacokinetic parameters of CVB-D@LAM DMNs microneedles and oral CVB-D in SD rats in Example 1: a is the 24-hour plasma concentration curve; b is the 72-hour plasma concentration curve; each point represents the mean ± SD (n = 6). Figure 13 Example 1: Verification of the effect of CVB-D@LAM DMNs microneedles on cardiac function in rats with myocardial ischemia: a) left ventricular ejection fraction, b) left ventricular fractional shortening, c) ventricular end-diastolic volume, d) left ventricular end-systolic volume, e) left ventricular end-diastolic diameter, f) myocardial oxygen consumption.
[0035] Figure 14 HE staining of rats with myocardial ischemia treated with CVB-D@LAM DMNs microneedles in Example 1; a is the sham-operated group, b is the model group, c is the CVB-D@LAM DMNs group, d is the gavage group; e is the LAM-DMN group.
[0036] Figure 15Masson staining of rats with myocardial ischemia treated with CVB-D@LAM DMNs microneedles in Example 1; a is the sham-operated group, b is the model group, c is the CVB-D@LAM DMNs group, d is the gavage group, and e is the blank-DMN group.
[0037] Figure 16 Example 1: Detection of biomarkers of myocardial injury in rats with myocardial ischemia treated with CVB-D@LAM DMNs microneedling: a is the CK-MB content in rat serum, b is the cTnT content in rat serum, c is the MDA content in rat serum, and d is the SOD content in rat serum. Detailed Implementation
[0038] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
[0039] Conversely, this invention encompasses any substitutions, modifications, equivalent methods, and solutions made within the spirit and scope of the invention as defined in the claims. Furthermore, to provide a better understanding of the invention, certain specific details are described in detail below. However, those skilled in the art will fully understand the invention even without these detailed descriptions.
[0040] To address the limitations of existing technologies such as poor water solubility and short half-life of CVB-D, which restrict its clinical application, this invention provides a cyclovinyl fulvic acid D microneedle patch for transdermal drug delivery. This aims to overcome the limitations of traditional oral formulations and interventional therapies, achieving painless and long-lasting treatment of myocardial ischemia and providing a new approach for painless and compliant treatment of clinical patients with myocardial ischemia. Furthermore, this invention elucidates the synergistic effect of bioactive laminarin and CVB-D through molecular biological analysis and pharmacokinetic studies in rats, providing new ideas for innovative research on modern drug delivery systems. By integrating the concepts of "drug-excipient integration" and "needle-drug integration," and elucidating the synergistic mechanism of polysaccharide-CVB-D through molecular biology and pharmacokinetic studies, this invention provides new strategies for the innovation and clinical translation of cardiovascular disease drug delivery systems.
[0041] Example 1: A method for preparing a cyclovin-based Buxus microneedle patch (e.g.) Figure 1 As shown), the preparation method includes the following steps: (1) The solid dispersion of CVB-D was prepared by dissolving cyclovinyl sulfadiazine D and a dispersant in an ethanol solution, followed by ultrasonic dissolution and freeze-drying. Figure 2 (as shown) Specifically, in this step, the dispersant used is povidone K30 (PVP K30). Weigh 1.0 g of CVB-D and 3.5 g of PVP K30, add 60 mL of 50% ethanol solution, sonicate (40 kHz) for 15 minutes, freeze at -80℃ for 6 hours, and vacuum dry for 24 hours to obtain an amorphous powder, which is the CVB-D solid dispersion. Characterization of the CVB-D solid dispersion powder by XRD, DSC, and scanning electron microscopy confirmed complete drug dispersion, as shown in the following results. Figure 3 As shown, XRD ( Figure 3 As shown in (a), its amorphous state was confirmed by the absence of crystallization in CVB-D or PVP K30 diffraction peaks. DSC thermogram ( Figure 3 As shown in Figure b), the glass transition temperature (Tg) is 32.5°C, and no CVB-D crystallinity was detected, which is confirmed by the absence of a melting endothermic peak. SEM imaging ( Figure 3 Figure c shows the uniform dispersion of CVB-D particles in the PVP matrix, with a smooth surface and no residual drug crystals, thus confirming effective physical stabilization. These multimodal analyses collectively validate the formation of a physicochemically stable amorphous system through intermolecular interactions between CVB-D and PVP K30, demonstrating enhanced drug dispersion efficiency and physical stability, and providing structural evidence for potential pharmacokinetic advantages in drug delivery applications.
[0042] (2) Preparation of Cyclofibrillated Buxus D Microneedle Patches: Accurately weigh 0.3 g of CVB-D solid dispersion into a 5 ml centrifuge tube. After sonication to completely dissolve the CVB-D solid dispersion in 4.45 ml of deionized water, add 0.25 g of laminarin, stir well, and centrifuge to remove air bubbles to obtain a dispersion-laminarin mixed solution. Use a pipette to add 80 μL of solution to each well of the microneedle mold, and vacuum aspirate for 30 min. After vacuum aspiration, dry at room temperature, peel off the mold, and obtain the cyclofibrillated Buxus D microneedle patches. The morphology of the cyclofibrillated Buxus D microneedle patches is as follows: Figure 4 As shown in a and b, the prepared microneedles have complete tips and good morphology.
[0043] The puncture performance of the microneedles was then tested. Using excised pig skin, the Cyclofibrillated Buxus microneedle patch was attached to a hydrogel backing. A needle inserter was then used to assist insertion. After 20 seconds, the patch was removed, observed, and photographed. The puncture results are as follows: Figure 5 As shown in Figure a, a clear array of micropores is visible on the pig skin, indicating that the prepared microneedles have good puncture performance. Furthermore, the punctured pig skin was fixed with tissue fixative and stained with hematoxylin and eosin (HE) to observe the microneedle puncture performance. The puncture results are as follows: Figure 5 As shown in Figure b, the microneedles exhibit good puncture performance.
[0044] The drug loading and uniformity of the microneedles were then tested, and the specific CVB-D content in the microneedles was analyzed using high-performance liquid chromatography (HPLC). The main analytical steps included patch dissolution: one CVB-D microneedle patch was randomly placed in a 15 mL centrifuge tube, and 7.5 mL of acetate buffer solution (pH = 5.0) was added to each tube. The tube was then shaken to aid dissolution for 1 hour, followed by the addition of 7.5 mL of methanol and another 1 hour of shaking. After complete dissolution, the patch was centrifuged, filtered, and 20 μL of the supernatant was injected into the HPLC system for analysis. The 1 cm... 2 The average CVB-D content of the cyclovinyl fulvidracoside D microneedle patch was 1560 ± 7.02 μg / patch, with an RSD of 3.07%.
[0045] Example 2: Preparation of cyclovinylbuxus D microneedle patches with a needle height of 300 µm: The preparation method of CVB-D solid dispersion is the same as in Example 1. 0.3 g of CVB-D solid dispersion was accurately weighed into a 5 mL centrifuge tube. After sonication to completely dissolve the CVB-D solid dispersion in 4.45 mL of deionized water, 0.25 g of laminarin was added, stirred evenly, and centrifuged to remove air bubbles, obtaining a dispersion-laminarin mixed solution. 80 μL of the solution was added to each well of a microneedle mold (300 µm needle height) using a pipette, and vacuum aspiration was performed for 30 min. After vacuum aspiration, the solution was dried at room temperature, and the mold was peeled off to obtain cyclovinylbuxus D microneedle patches. The morphology of the cyclovinylbuxus D microneedle patches is as follows: Figure 4 As shown in Figures c and d, the prepared microneedles have complete tips and good morphology.
[0046] The drug loading and uniformity of the microneedles were then tested, and the specific CVB-D content in the microneedles was analyzed using high-performance liquid chromatography (HPLC). The main analytical steps included patch dissolution: one CVB-D microneedle patch was randomly placed in a 15 mL centrifuge tube, and 7.5 mL of acetate buffer solution (pH = 5.0) was added to each tube. The tube was then shaken to aid dissolution for 1 hour, followed by the addition of 7.5 mL of methanol and another 1 hour of shaking. After complete dissolution, the patch was centrifuged, filtered, and 20 μL of the supernatant was injected into the HPLC system for analysis. The 1 cm... 2 The average CVB-D content of the cyclovinyl fulvidracoside D microneedle patch was 1570.4 ± 8.52 μg / patch, with an RSD of 5.63%.
[0047] Example 3: A method for preparing a cyclohexane-based Buxus microneedle patch, the method comprising the following steps: (1) The solid dispersion of CVB-D was prepared by dissolving cyclovinyl flavonoid D and a dispersant in an ethanol solution, followed by ultrasonic dissolution and freeze-drying. Specifically, in this embodiment, the dispersant used is povidone K90, the mass ratio of cyclovinyl fulvic acid D to povidone K90 is 1:3, and an ethanol solution with a volume fraction of 40-70% is used. (2) Weigh the CVB-D solid dispersion into a centrifuge tube, sonicate the CVB-D solid dispersion to completely dissolve in deionized water, add laminarin, stir evenly, centrifuge to remove air bubbles, and obtain a dispersion-laminarin mixed solution. Use a pipette to add liquid into the microneedle mold and vacuum suction. After vacuum suction, dry at room temperature, peel off the mold, and obtain the cyclovinyl fulva D microneedle patch. Specifically, in this embodiment, the mass ratio of CVB-D solid dispersion to laminarin is 3:2.
[0048] Comparative Example: This embodiment provides a CVB-D microneedle formulation, the preparation method of which is as follows: Raw material ratio: CVB-D 5wt%, laminarin 5wt%, trehalose 2wt%, deionized water 88wt%; Preparation of CVB-D integrated needles: Accurately weigh 0.1g CVB-D, 0.25g laminarin, and 0.1g trehalose into a mortar, grind thoroughly for 10min, transfer the powder to a 10ml centrifuge tube, add ultrapure water to dissolve, stir thoroughly, centrifuge to remove air bubbles, add 80 μL of liquid to each well using a pipette, and vacuum aspirate for 30min. After completion, dry at room temperature. The microneedle morphology is as follows. Figure 6 As shown, the prepared microneedles have complete tips and good morphology.
[0049] The puncture performance of the microneedles was then tested. Using excised pig skin, the microneedles were attached to a hydrogel backing, and then an inserter was used to assist insertion. After 20 seconds, the skin was removed, observed, and photographed. The puncture results are as follows: Figure 6 As shown, a clear array of micropores is visible on the pig skin, indicating that the prepared microneedles have good puncture performance. Furthermore, the punctured pig skin was fixed with tissue fixative and stained with hematoxylin and eosin (HE) to observe the microneedle puncture performance. The puncture results are as follows: Figure 7 As shown, the microneedles have good puncture performance.
[0050] The drug loading and uniformity of the microneedles in this comparative example were then tested, and the specific CVB-D content in the microneedles was analyzed using high-performance liquid chromatography (HPLC). The main analytical steps included patch dissolution: one CVB-D microneedle patch was placed in a 15 ml centrifuge tube, and 7.5 mL of acetate buffer solution (pH = 5.0) was added to each tube. The tube was then shaken to aid dissolution for 1 hour, followed by the addition of 7.5 mL of methanol and another 1 hour of shaking. After complete dissolution, the patch was centrifuged, filtered, and 20 μL of the supernatant was analyzed by HPLC. The 1 cm... 2The average CVB-D content in the microneedle patches was 895 ± 0.47 μg / patch, with an RSD of 2.32%. This indicates that solid dispersion technology increases the solubility of CVB-D, resulting in higher drug loading. The solubility of CVB-D by laminarin alone is not as high as that achieved with solid dispersion.
[0051] The release kinetics of CVB-D@LAM DMNs were tested using the methods provided in Example 1 and the comparative example to prepare CVB-D-LAM-DMN microneedles. The release experiment was conducted according to the guidelines of (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0931, Method VII). The specific experimental steps are as follows: First, prepare the dissolution medium for the release experiment, requiring 2L of 0.1M acetate-acetate buffer (pH=5.0) as the microneedle release medium. Then, attach three CVB-D microneedles to the lower edge of the inner wall of the reciprocating cylinder. The temperature was then set to 32±0.5℃; the reciprocating cylinder speed to 25 dpm; the reciprocating stroke to 10 cm; the flushing volume to 10 mL; the total receiving liquid volume to 220 mL; the sampling time points were: 5 min; 10 min; 30 min; 1 h; 2 h; 3 h; 4 h; 6 h; the sampling volume to 2 mL; and the replenishment volume to 2 mL. The solutions collected at each time point were filtered through a 0.22 μm aqueous filter into 1.5 mL liquid chromatography vials, and then sent for liquid chromatography analysis. The test results are as follows: Figure 8 As shown (the microneedle preparation method corresponding to the CVB-D solid dispersion curve is the same as in Example 1, and the microneedle preparation method corresponding to the CVB-D crystal curve is the same as in the comparative example), the release curves show that the CVB-D on both microneedle patches is completely released within 24 h. However, the CVB-D microneedles prepared by the method in Example 1 have a significantly faster release rate, indicating that the CVB-D solid dispersion increases the dissolution of CVB-D.
[0052] To investigate the safety of the CVB-D-LAM-DMN microneedle patch, the irritation effects on the skin of New Zealand albino rabbits were evaluated. (The difference between the blank DMN and the CVB-D-LAM-DMN microneedle patch is that the blank DMN is without the drug, containing only laminarin matrix material.) Figure 9 As shown in Figure a, no obvious erythema, edema, or other irritation reactions were observed in either the blank DMN group or the cyclophosphamide D microneedle patch group. However, it should be noted that in both DMN groups, slight redness appeared locally on the skin due to the puncture effect on the stratum corneum, which is a typical reaction to a 500 μm high DMN. Furthermore, a small amount of hair regrowth was observed at the DMN application site 24 hours later, indicating the skin compatibility and biosafety of the DMN patch, making it suitable for treating diseases requiring long-term and frequent administration.
[0053] To further verify the safety of the Cyclofibrillated Buxus Microneedle Patch, a red blood cell hemolysis experiment was conducted using fresh rabbit blood. Figure 9 As shown in b, CVB-D microneedles do not cause hemolysis of erythrocytes during use, and the drug-loaded formulations all exhibit good erythrocyte compatibility.
[0054] In vitro solubility test of cyclovinylbuxusin D microneedle patch: The cyclovinylbuxusin D microneedle patch prepared in Example 1 was attached to a hydrogel backing. Then, a needle inserter was used to puncture the microneedles into isolated pig skin. The microneedles were removed at 0 min, 1 min, 3 min, 5 min, 10 min, and 30 min, respectively, and the solubility was observed and photographed under a stereomicroscope. The experimental results are as follows: Figure 10 As shown, the results indicate that the cyclovinyl flavonoid D microneedle patch has good solubility, and the microneedle body is completely dissolved within 30 minutes.
[0055] Intradermal solubility test of the cyclovinyl fulva D microneedle patch. The preparation method of the cyclovinyl fulva D microneedle patch is the same as in Example 1, as follows: Figure 11 As shown, the skin surface residual ratio and dermal accumulation of CVB-D delivered via the laminarin microneedle system exhibited an inverse dynamic correlation. During the 0–10 h period after administration, the skin surface residual ratio (y = 96.87–3.556x, R² = 0.9759) showed first-order kinetic decay, decreasing from 100% to 59.4%; the dermal accumulation (y = 35.99 + 52.85x, R² = 0.9835) showed zero-order release characteristics, with a constant administration rate of 59.4 μg / h. During this phase, drug permeability was linearly negatively correlated with residual amount (R² = 0.982), indicating that matrix dissolution played a dominant role in drug release. After 12 h, the system reached steady-state equilibrium, with the residual ratio fluctuating within ±2.1%, and the cumulative skin permeation amount reaching 757.02 ± 79.99 μg. This indicates a significantly improved transdermal efficiency (TE = 50.47%) compared to traditional transdermal formulations. Biphasic monitoring data collectively demonstrate that this microneedle system maintained an effective dosing window of 12 hours while achieving continuous and stable transdermal administration. These findings provide crucial experimental evidence for optimizing dosing time in subsequent efficacy assessments, suggesting continuous dosing for ≥12 hours to maximize therapeutic efficacy.
[0056] Pharmacokinetic validation of cycloviniferin D microneedle patch: The preparation method of cycloviniferin D microneedle patch is the same as in Example 1, as follows... Figure 12As shown, the pharmacokinetic characteristics of CVB-D in SD rats after administration of CVB-D@LAM DMNs via transdermal microneedle patch (CVB-D@LAM DMNs) differed significantly in key kinetic parameters after single oral administration and two oral administrations, highlighting the superior therapeutic potential of transdermal delivery. The Cmax of CVB-D@LAM DMNs was 135.74 ng / mL, comparable to a single oral dose, but the Tmax was delayed by 10 hours, indicating a slower but more controllable release rate. Furthermore, it showed a significantly prolonged half-life (28.84 h) and mean residence time (MRT, 25.06 h), indicating a prolonged systemic exposure time. AUC 0-∞ The concentration reached 5696.11 ng·h / ml, approximately three times that of the oral single-dose group, exhibiting high systemic bioavailability with a relative bioavailability (RB) of 307.11%. In contrast, the oral single-dose group had the shortest Tmax (3.83 h) and the highest Cmax (135.17 ng / mL), but a shorter half-life of only 14.30 h and a lower AUC (1854.77 ng·h / mL), resulting in rapid drug clearance and a short treatment duration. While fractionated oral administration improved T1 / 2 (24.80 h) and MRT (29.80 h), it resulted in a lower Cmax, a significantly delayed Tmax (18 h), and an AUC still lower than that of CVB-D@LAM DMNs. This indicates that fractionated administration reduced fluctuations compared to single-dose administration, but drug utilization was still not optimal. The microneedle group continuously released CVB-D for over 24 hours without electronic devices, maintaining a relatively stable and elevated plasma concentration over 10 to 48 hours, demonstrating characteristics of sustained release. In both oral administration groups, plasma drug levels dropped sharply within 24 hours. The microneedle patch formulation significantly improved systemic exposure and bioavailability of CVB-D, prolonged drug action, and avoided first-pass metabolism. Compared with oral administration, microneedle delivery offers advantages such as sustained release, effective absorption, and improved patient compliance, making it a more ideal method of drug delivery.
[0057] To investigate the therapeutic effect of Cyclofibrul Ginkgo biloba D microneedle patch on myocardial ischemia, a Sprague-Dawley rat model of myocardial ischemia was established for pharmacological evaluation. The preparation method of Cyclofibrul Ginkgo biloba D microneedle patch was the same as in Example 1, and the experimental scheme is as follows: Animal model: Myocardial ischemia model was established by ligation of the left anterior descending coronary artery in SD rats; Grouped drug administration: Sham surgery group: open chest suturing only; Model group (MI): ischemic treatment; Microneedle group (CVB-D-LAMDMN): daily application of CVB-D microneedles; Oral group: CVB-D solution administered by gavage; Blank control group (Blank MNs): drug-free laminarin microneedles; Detection indicators: Cardiac function: echocardiography (LVEF, SV, FS, CO, LVSP, LVEDV); Myocardial injury: serum CK-MB, cTnT, MDA and SOD; Pathology: HE staining (inflammatory infiltration), masseter staining.
[0058] The results are as follows Figure 13 As shown, after four weeks of continuous administration of CVB-D@LAM microneedle patches, the left ventricular end-diastolic diameter (LVIDd) and left ventricular end-systolic diameter (LVIDd) in the microneedle-treated rats were significantly improved compared with the model group. The efficacy of the CVB-D microneedle-treated group and the gavage-treated group was comparable, and the kelp polysaccharide microneedle-treated group also showed a synergistic therapeutic effect. The CVB-D@LAM microneedle group showed significant improvement in cardiac function, including increased left ventricular ejection fraction (EF), decreased left ventricular systolic pressure (LVSP), left ventricular fractional shortening (FS%), stroke volume (SV), cardiac output (CO), and left ventricular end-diastolic volume (LVEDV). The blank microneedle-treated group also showed improvement in the above indicators, demonstrating a synergistic therapeutic effect on myocardial ischemia. The CVB-D@LAM microneedling group showed better results compared to the gavage control group. These findings indicate that CVB-D@LAM microneedling serves as a platform for myocardial repair through synergistic drug delivery and mechanical tissue remodeling, with the aim of providing long-term drug delivery for the treatment of chronic diseases such as myocardial ischemia. Histopathological evaluation showed ( Figure 14 and Figure 15 CVB-D@LAM microneedles promoted myocardial structural repair and reduced inflammation (hematoxylin-eosin [HE] staining), while the model group showed severe myocardial fibrosis and inflammatory infiltration. Furthermore, serum myocardial injury biomarkers (CK-MB, cTnT, MDA, and SOD) were lower in the microneedle administration group compared to the model group. Figure 16 Compared with the gavage control group, the results were better. The blank microneedle group also showed improvement in the above indicators, and CVB-D played a synergistic therapeutic role. These findings indicate that CVB-D@LAM microneedles are a platform for myocardial repair through synergistic drug delivery and mechanical tissue remodeling, with the aim of providing a platform for long-term drug delivery for the treatment of chronic diseases such as myocardial infarction.
[0059] Based on experimental data, laminarin and CVB-D provide multi-layered synergistic treatment for myocardial ischemia within a laminarin-based dissolving microneedle (DMN) platform through a "pharmacoexisting synergistic effect." The main mechanism of this synergistic effect is as follows: laminarin not only acts as a sustained-release carrier, ensuring continuous delivery and stable plasma CVB-D levels while avoiding first-pass metabolism in the liver associated with oral administration, but also possesses significant intrinsic biological activities, such as antioxidant and anti-inflammatory properties. It directly enhances endogenous antioxidant defense capabilities by activating the Nrf2 pathway, thereby increasing SOD activity and reducing MDA levels. CVB-D, as the key therapeutic molecule, significantly improves cardiac function, repairs myocardial tissue, and reduces fibrosis. Their synergistic effects are evident in two key aspects: 1) The properties of the laminarin delivery system optimize the pharmacokinetics of CVB-D, resulting in more stable treatment and better LDH, cTnT, MDA, and SOD outcomes in the CVB-D@LAM DMNs group compared to the oral group; 2) The natural activities of laminarin (such as antioxidant and anti-inflammatory effects) and the mechanisms of CVB-D (such as anti-apoptosis and pro-repair effects) work synergistically across multiple disease pathways, providing additional protection. For example, the LAM-DMN group showed that laminin alone reduced oxidative damage (↑SOD / ↓MDA) and inflammation, while the CVB-D@LAM DMNs group achieved a greater reduction in chronic fibrosis through a combined effect and promoted myocardial regeneration of cTnT. Compared to monotherapy, this comprehensive approach provides superior cardioprotection, ultimately restoring cardiac function (↑EF / FS), reducing acute injury, and preventing long-term remodeling.
[0060] The synergistic mechanism of the CVB-D microneedle patch provided by this invention is as follows: CVB-D regulates myocardial ion channels, laminarin scavenges reactive oxygen species, and jointly inhibits cardiomyocyte apoptosis; optimized percutaneous penetration: microneedle penetration depth -400 μm, the microneedle patch has sustained-release properties, and plasma concentration is stably maintained for >24 hours (C max / C min The ratio was 1.8, which was significantly different from the rapid decline (within 10 hours) in the oral group.
[0061] The advantages of the cyclosporum tobira D microneedle patch provided by this invention are as follows: Minimally invasive: avoids surgical trauma, resulting in high patient compliance; Sustained-release properties: Continuously releases CVB-D for over 24 hours without the need for electronic devices; Multi-target synergy: Simultaneously improves cardiac function, anti-oxidation, anti-inflammation and anti-fibrosis (reduced collagen deposition); Safety: CVB-D@LAM DMNs showed good erythrocyte compatibility, and skin irritation tests showed no adverse reactions such as erythema or edema.
[0062] The above description is merely a specific embodiment of the present invention, and the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope defined in the claims.
Claims
1. A type of cyclic-fiber Buxus microneedle patch, characterized in that, The cyclovinylbuxus microneedle patch comprises CVB-D solid dispersion and laminarin polysaccharide; the laminarin polysaccharide is used as the sole matrix material of the microneedles and as a bioactive component with cardioprotective properties. In the cyclovinyl bupleurum D microneedle patch, the mass ratio of the CVB-D solid dispersion to laminarin is (1-3):1; The specific preparation method of the CVB-D solid dispersion is as follows: cyclovinyl flavonoid D and a dispersant are dissolved in an organic solution, and the CVB-D solid dispersion is obtained by ultrasonic dissolution and freeze-drying; the dispersant is povidone K30 or povidone K90; the organic solution is an ethanol solution with a volume concentration of 50% to 100%; the mass ratio of cyclovinyl flavonoid D to the dispersant is 1:(2-10).
2. The cyclic-fiber boxwood star D microneedle patch according to claim 1, characterized in that, The microneedle tip height of the cyclohexane star D microneedle patch is 200-500 μm, and the tip distance is 150-600 μm.
3. The use of the cyclovinyl flavonoid D microneedle patch as described in claim 1 or 2 in the preparation of drugs for treating myocardial ischemia.
4. A method for preparing a cyclohexane-based Buxus microneedle patch, characterized in that, The preparation method includes the following steps: CVB-D solid dispersion was prepared using cyclovinyl fulvidin D and a dispersant; The CVB-D solid dispersion was ultrasonically dissolved and then mixed with laminarin. After centrifugation to remove air bubbles, a dispersion-laminarin mixed solution was obtained. The dispersion-laminarin mixed solution was added into a microneedle mold, vacuum-suctioned, dried, and then the mold was peeled off to obtain a cyclovinyl fulva D microneedle patch. The specific method for preparing the CVB-D solid dispersion is as follows: dissolving cyclovinyl flavonoid D and a dispersant in an organic solution, followed by ultrasonic dissolution and freeze-drying to obtain the CVB-D solid dispersion. The dispersant is povidone K30 or povidone K90; the organic solution is an ethanol solution with a volume concentration of 50% to 100%. The mass ratio of the cyclohexane D to the dispersant is 1:(2-10). The mass ratio of the CVB-D solid dispersion to the kelp polysaccharide is (1-3):
1.
5. The method for preparing the cyclohexane-based Buxus microneedle patch according to claim 4, characterized in that, The preparation method of the dispersion-laminarin mixed solution is as follows: CVB-D solid dispersion is ultrasonically dissolved in water. After complete dissolution, an aqueous solution of solid dispersion is obtained. Laminarin is added to the aqueous solution of solid dispersion, stirred evenly, and centrifuged to remove air bubbles to obtain the dispersion-laminarin mixed solution.
6. The method for preparing the cyclohexane-based Buxus microneedle patch according to claim 4, characterized in that, The vacuum suction conditions are: vacuum suction time of 10-30 min; the drying conditions are: drying temperature of 25-45℃ and drying time of 2-6 h.