Intelligent shock wave response type nano-motor microneedle transdermal drug delivery system and preparation method thereof
Through the intelligent shock wave responsive nanomotor microneedle system, the shock wave drives the nanomotor to respond and move on the microneedle, solving the problem that drugs are difficult to penetrate deep tissue in the prior art, and achieving efficient drug delivery in bone tissue and Release effect.
Patent Information
- Application Number
- CN202411210978.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-08-30
- Publication Date
- 2024-12-06
AI Technical Summary
Existing transdermal drug delivery technology is difficult to effectively penetrate drugs into deep tissues, especially bone tissues, and the penetration efficiency and targeting of nanomotors are insufficient, resulting in poor penetration and release of drugs in deep disease areas.
Using intelligent shock wave responsive nanomotor microneedle system, hydroxyapatite is used as the skeleton of the nanomotor, and the nanomotor is induced to respond and move on the microneedle through shock wave, enhancing the penetration depth and targeting of the drug, and combining with microneedle technology to achieve Deep drug delivery.
The transdermal delivery efficiency of the drug is significantly improved. The nanomotor can penetrate deep tissue and move in a directional manner under the action of shock wave, achieving efficient release of drugs at deep targeted sites, especially in bone tissue to promote drug penetration and bone tissue repair.
Smart Images
Figure CN119074632A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of medical technology, and in particular relates to an intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system and a preparation method thereof. Background Art
[0002] Transdermal drug delivery refers to drug delivery through the skin surface, using chemical and physical means to release drugs through the skin at a certain rate into the body, and enter the systemic circulation to produce systemic or local therapeutic effects. Its advantages are: drug absorption is not affected by factors such as pH, food, and transit time in the digestive tract; avoid the first-pass effect of the liver; overcome adverse reactions caused by excessive blood drug concentration due to rapid absorption; sustainable control of drug delivery speed, flexible delivery, etc. The outermost layer of the epidermis in the skin, the stratum corneum, is currently the biggest obstacle to transdermal drug delivery. The stratum corneum is about 20um thick. The stratum corneum cells and the natural substances filling the stratum corneum cells, including lipids, free amino acids, pyrrolidone carboxylic acid and salts, combine together to play a natural barrier function to prevent drugs from entering the body through the skin.
[0003] In the early stage of transdermal drug delivery, patches were mainly used to allow lipophilic, low-molecular-weight drugs to enter the local area through the stratum corneum to exert their effects. With the in-depth study of skin structure, physical and chemical methods began to be used to destroy the stratum corneum, thereby removing the restrictions on the drug itself. As the third-generation transdermal drug delivery technology, microneedles load drugs on a micron-sized needle tip array and slowly release the drugs after being applied to the patient. Compared with other transdermal methods, microneedles can deliver drugs directly to the dermis of the skin, allowing the drugs to quickly enter the local area, or enter the capillary network under the dermis of the skin and enter the systemic blood circulation. In addition, microneedles cause less damage to the skin, and their convenience gives good patient compliance.
[0004] However, since the release of drugs from the microneedle tip is a passive, non-targeted process, the drugs still do not aggregate after being released from the needle tip, and cannot significantly increase the local drug concentration. Therefore, it is necessary to increase the drug penetration depth and promote the local aggregation of drugs. Nanomotor (NM) is a type of nano-scale micro-device composed of nanomaterials that can actively move and is powered by chemical energy, light energy, electrical energy, etc., and has good controllability and modifiability. [14,15]. Due to its wide diffusion range, efficient cargo transport and deep tissue penetration, nanomotors have good application prospects in biomedicine and other fields. Combining nanomotors with microneedles and using nanomotors as carriers to achieve active movement of drugs can meet the requirements of efficient deep delivery, improve drug distribution, and avoid damage to the skin during transdermal administration. Some scholars have proposed a series of nanomotors that can achieve self-driven movement through chemical fuels, but the toxic side effects of chemical fuels on living organisms limit the application of nanomotors. In addition, researchers have constructed nanomotors that can generate non-toxic gas drive autonomously for autonomous and active drug penetration. However, the irregular movement of gas-driven nanomotors reduces their targeting in solid tissues. Exogenous physical stimuli such as light and magnetic fields are another way to drive drug penetration, but these exogenous physical stimuli have low penetration efficiency in human tissues and require high energy. In addition, the penetration efficiency of these nanomotors in human tissues is low, the energy required is high, and there is a lack of relevant pathways and solutions to promote the penetration and release of nanomotors and drugs in deep disease areas, such as the penetration of bone tissue. There is no solution. Combining nanomotors with shock waves can solve these problems well.
[0005] Shock waves are a special type of wave. Compared with ultrasound, shock waves have a more significant cavitation effect. They generate almost no heat during the penetration process and have little energy loss. The difference between the positive and negative pressures of shock waves is larger than that of ultrasound, so the pressure generated is greater and the penetration is farther, reaching bone tissue. After being stimulated by shock waves, some calcium salts are deposited and mineralized, and the increase in mineralized deposition is conducive to mediating the reconstruction of new bone, thereby completing the repair of bone tissue. At the same time, under the action of shock waves, shock wave-responsive drug-containing nanoparticles will absorb the energy of the shock waves, thereby promoting their radial movement, and as the number of shock waves increases, the drug particles will be destroyed, thereby releasing the drug.
[0006] There is little research on the technology of using shock waves and nanomotors to penetrate and release drugs into deep disease areas, so it is a pressing issue to be solved. Summary of the invention
[0007] The technical problem to be solved by the present invention is to provide an intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system and a preparation method thereof, which utilizes shock waves to increase the penetration of nanomotor into deep tissues and promote drug release in deep target tissues.
[0008] The technical solutions adopted are:
[0009] An intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system adopts shock wave induced response, comprising a microneedle patch, on which microneedles are arranged, wherein the microneedles comprise a microneedle matrix and a shock wave responsive nanomotor carrying a drug. The shock wave responsive nanomotor carrying a drug has hydroxyapatite as a skeleton.
[0010] The microneedle is conical in shape, 750 microns in height and 300 microns in bottom diameter.
[0011] Each microneedle array can be composed of 10*10 microneedles, and each microneedle array can be loaded with an average of about 1.3 mg of nanoparticles. The microneedle array can also be composed of 12*12, 16*16 microneedles, etc.
[0012] Preferably, a shock wave response is used, and the intensity of the shock wave is 2 to 4 bar. The intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system is used for transdermal drug delivery to deep tissues, especially to penetrate into bone tissue. Shock waves are applied to the part where the microneedle patch is applied. The intensity of the shock wave is 2 to 4 bar. The number of applications is 2000 times, once every other week, for a total of 4 times.
[0013] A method for preparing an intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system comprises the following steps:
[0014] (1) Preparation of nanomotor (ESW-NM), using micelles composed of PEG and Tween 80 as templates, stirring evenly, conjugating calcium chloride on the surface, adding disodium hydrogen phosphate solution, and continuing to stir; collecting the solution, removing the PEG and Tween80 templates from the remaining solution, and then centrifuging to obtain the nanomotor;
[0015] (2) incubating the nanomotor with the drug, stirring and centrifuging to obtain the nanomotor with the drug on its surface;
[0016] (3) Add the drug-loaded nanomotors into the grooves of the PDMS (polydimethylsiloxane) microneedle array mold, place it in a vacuum oven to evacuate, discard the excess suspension in the grooves, add hyaluronic acid solution to fill the grooves, and dry overnight to obtain a microneedle patch.
[0017] Preferably, in step (1), PEG is first dissolved in a Tris-HCl solution and then added dropwise to Tween 80 while stirring until the mixture is fully stirred.
[0018] Preferably, the mass ratio of PEG to Tris-HCl is 1:10-15 (g / ml), and the mass ratio of PEG to Tween 80 is 1:5-15; after being fully dissolved, ultrasonic treatment is required for 20-30 minutes in a room temperature water bath.
[0019] Preferably, the calcium chloride solution is added dropwise to the mixed solution of PEG, Tris-HCl and Tween 80 after ultrasonic treatment, stirred for 30 to 40 minutes, and then the disodium hydrogen phosphate solution is added dropwise, and stirring is continued for 1 hour; after centrifugation, the precipitate is washed with deionized water.
[0020] Preferably, in step (2), the co-incubation method is to place the nanomotor in a saturated aqueous solution of the drug for 1 to 2 hours. The drug includes common drugs such as bisphosphonates and sodium dichloride.
[0021] As a further preference, the stirring time is 40 to 80 minutes, the stirring rate is 100 to 400 r / min, and then the mixture is left to stand and wait.
[0022] In the step 3), the vacuum is drawn to -30 mmHg for 20 to 30 minutes; and the drying temperature is 40 to 60°C.
[0023] Compared with the prior art, the present invention has the following beneficial effects:
[0024] The intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system of the present invention utilizes shock waves to increase the penetration of drug-carrying nanomotor into deep tissues and release drugs in deep target tissues, which can significantly improve the transdermal delivery efficiency of drugs. Under the cavitation effect of shock waves, micro-nano bubbles are generated on the surface of the nanomotor. The energy generated after the bubbles burst pushes the nanomotor forward to make directional movements, thereby achieving deep tissue penetration of the nanomotor (ESW-NM) and directly releasing drugs at the targeted site.
[0025] The intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system of the present invention is more easily broken under the action of shock waves to release relevant drugs, including but not limited to zoledronic acid, calcium ions, etc., thereby playing relevant roles, such as inhibiting osteoclasts, promoting osteoblasts, and promoting bone tissue repair, etc., providing a new idea for transdermal drug delivery to deep tissues.
[0026] The preparation method of the invention is simple, the preparation conditions are easy to achieve, and it can be promoted on a large scale. BRIEF DESCRIPTION OF THE DRAWINGS
[0027] Figure 1 This is a TEM photo of the nanomotor particles prepared in Example 1 of the present invention.
[0028] Figure 2 This is a photograph of the microneedle under a fluorescence microscope after the nanomotors carrying drugs labeled with different fluorescent substances of the present invention are loaded into the microneedle.
[0029] Figure 3 Cell viability graphs of bone marrow macrophages (a) and bone marrow mesenchymal stem cells (b) after incubation with different concentrations of nanomotors.
[0030] Figure 4 The figure shows the movement of the drug-carrying nanomotor of the present invention under the action of shock waves. DETAILED DESCRIPTION
[0031] The drawings are only used for illustrative purposes; for those skilled in the art, some well-known structures and their descriptions in the drawings may be omitted, and therefore, they should not be understood as limitations on the present invention.
[0032] The technical solutions and effects of the present invention are described in detail below with reference to the accompanying drawings.
[0033] Example 1
[0034] A method for preparing an intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system, using hydroxyapatite as a skeleton to prepare a shock wave responsive nanomotor carrying a drug, comprising the following steps:
[0035] (1) Preparation of hydroxyapatite nanomotors by template method to form spherical particles.
[0036] a. Prepare Tris-HCl solution: weigh 0.6057 g Tris-Base, dissolve it in 10 mL of secondary water, and adjust the pH to 8.0 with concentrated hydrochloric acid.
[0037] Prepare CaCl2 solution: weigh 0.19425 g CaCl2 and dissolve it in 10 mL secondary water.
[0038] b. Weigh 0.15g PEG, dissolve it in 3mL Tris-HCl, add it dropwise into 2g Tween 80, stir while adding, stir thoroughly and ultrasonicate in a water bath at 25℃ for 20min.
[0039] c. Take 3.5mL of CaCl2 solution and add it dropwise to the system in step b. After stirring for 30min, add disodium hydrogen phosphate solution dropwise and continue stirring for 60min.
[0040] d. Collect the reaction system, centrifuge repeatedly at 4°C, centrifugal force 10000g, and the precipitate is the prepared nanomotor. Figure 1 As shown, the prepared nanomotor is spherical in shape and has a size of 96.34±8.54nm under an electron microscope.
[0041] (2) incubating the nanomotor with the drug for 1.5 h, stirring for at least 30 min, and stirring at a rate of 100 to 400 r / min; then allowing the nanomotor to stand, and then centrifuging to remove excess drug to obtain a nanomotor with the drug loaded on its surface;
[0042] (4) Add the drug-loaded nanomotors into the grooves of the PDMS microneedle array mold, place it in a vacuum oven and evacuate to -30 mmHg for 30 min, discard the excess suspension in the grooves, add hyaluronic acid solution to fill the grooves, and dry at 40°C overnight to obtain a microneedle patch.
[0043] The prepared microneedle patch has a conical microneedle tip with a height of 750 microns and a bottom diameter of 300 microns. Each array consists of 10*10 microneedles.
[0044] like Figure 2 As shown, the surface-loaded drug nanomotors labeled with calcein fluorescently were loaded into the microneedles. Figure 2 It can be seen that the nanomotors with drugs loaded on the surface are dispersed in the prepared soluble microneedle matrix.
[0045] The microneedle patch prepared in Example 1 of the present invention was tested.
[0046] Test Example 1, cytotoxicity of nanomotors.
[0047] After incubating bone marrow macrophages (BMM) and bone marrow mesenchymal cells (BMSC) with nanomotors of different concentrations, the cell viability was detected by CCK-8 method. A blank was also used for comparison.
[0048] A sterile 100 μM nanomotor suspension was prepared and added dropwise into the culture medium of bone marrow macrophages (BMM) and bone marrow mesenchymal cells (BMSC), respectively, to make the nanomotor concentrations 1, 2, 5, and 10 μM. After incubation for 24 h, the cell viability was detected using an ELISA instrument according to the instructions of the commercial product of the CCK-8 reagent.
[0049] The experimental results are shown in Figure 3 .like Figure 3 As shown, (a) shows the cell viability of bone marrow macrophages after incubation with different concentrations of nanomotors, and (b) shows the cell viability of bone marrow mesenchymal stem cells after incubation with different concentrations of nanomotors. There is no obvious difference in cytotoxicity of nanomotors at different concentrations.
[0050] Test Example 2: testing the penetration effect of the nanomotor.
[0051] First, a 5% gelatin gel was prepared. Before the gel was cooled, the nanomotor dispersion system was mixed with the gel and stored at 4° C. until the gel was formed.
[0052] The color developer was injected into the gel as a reference, and ESW was applied different times from the same direction. The relative displacement of the nanomotor was observed and recorded under a dark-field microscope.
[0053] The experimental results are shown in Figure 4 As shown, under the microscope, it can be found that with the application of ESW, the nanomotor gradually shows relative displacement.
[0054] The meanings of the English abbreviations used in this invention are:
[0055] HA-hyaluromic acid
[0056] ESW (Extracorporeal shock wave)-Extracorporeal shock wave.
[0057] ESW-NM shock wave responsive nanomotor particles
[0058] PEG-polyethylene glycol;
[0059] Tris-HCl-tris(hydroxymethyl)aminomethane hydrochloride;
[0060] Tris-Base
[0061] PDMS-polydimethylsiloxane;
[0062] Calcein-Calcium cyanine.
[0063] Of course, the above description is not a limitation of the present invention, and the present invention is not limited to the above examples. Changes, modifications, additions or substitutions made by technicians in this technical field within the essential scope of the present invention should also fall within the protection scope of the present invention.
Claims
1. An intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system, which adopts shock wave induced response, comprises a microneedle patch, on which microneedles are arranged, It is characterized in that The microneedle comprises a microneedle matrix and a shock wave responsive nanomotor carrying drugs.
2. The intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system according to claim 1, It is characterized in that The microneedle is conical in shape, 750 microns in height and 300 microns in bottom diameter.
3. The intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system according to claim 1, It is characterized in that Shock wave response is adopted, and the intensity of the shock wave is 2 to 4 bar.
4. A method for preparing an intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system as described in claims 1-3, It is characterized in that The steps include: (1) Preparation of nanomotors: using micelles composed of PEG and Tween 80 as templates, stirring evenly, conjugating calcium chloride on the surface, adding disodium hydrogen phosphate solution, and continuing stirring; The solution is then collected, and the PEG and Tween 80 templates are removed from the remaining solution, followed by centrifugation to obtain the nanomotor; (2) co-incubating the nanomotor with the drug, stirring and centrifuging to obtain the nanomotor with the drug on its surface; (3) Add the drug-loaded nanomotors into the grooves of the PDMS microneedle array mold, place it in a vacuum oven to evacuate, discard the excess suspension in the grooves, add hyaluronic acid solution to fill the grooves, and dry overnight to obtain a microneedle patch.
5. A method for preparing an intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system according to claim 4, It is characterized in that In the step (1), PEG is first dissolved in a Tris-HCl solution and then added dropwise to Tween 80 while stirring until the mixture is fully stirred.
6. A method for preparing an intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system according to claim 5, It is characterized in that The mass ratio of PEG to Tris-HCl is 1:10-15 (g / ml), and the mass ratio of PEG to Tween 80 is 1:5-15. After being fully dissolved, ultrasonic treatment is required for 20-30 minutes in a room temperature water bath.
7. A method for preparing an intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system according to claim 6, It is characterized in that Add the calcium chloride solution dropwise to the mixed solution of PEG, Tris-HCl and Tween 80 after ultrasonic treatment, stir for 30-40 minutes, then add disodium hydrogen phosphate solution dropwise and continue stirring for 1 hour; centrifuge and wash the precipitate with deionized water.
8. The method for preparing the intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system according to claim 4, It is characterized in that In the step (2), the co-incubation method is to place the nanomotor into a saturated aqueous solution of the drug, and the co-incubation time is 1 to 2 hours.
9. The method for preparing the intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system according to claim 8, It is characterized in that The stirring time is 40 to 80 minutes, the stirring rate is 100 to 400 r / min, and then stand and wait.
10. The method for preparing the intelligent shock wave responsive nanomotor microneedle transdermal drug delivery system according to claim 4, It is characterized in that In the step (4), the vacuum is drawn to -30 mmHg for 20 to 30 minutes; and the drying temperature is 40 to 50°C.