WO2023226136A1

WO2023226136A1 - Microneedle system for diagnosis and treatment of brain diseases and preparation method therefor

Info

Publication number: WO2023226136A1
Application number: PCT/CN2022/101279
Authority: WO
Inventors: 盛宗海; 郑海荣; 胡德红; 高笃阳; 刘新
Original assignee: 深圳先进技术研究院
Priority date: 2022-05-26
Filing date: 2022-06-24
Publication date: 2023-11-30
Also published as: CN114948852A

Abstract

A microneedle system for diagnosis and treatment of brain diseases and a preparation method therefor. The microneedle system for diagnosis and treatment of brain diseases comprises 0.1-10 parts of an active pharmaceutical ingredient liposome and 1-10 parts of a dispersant, the active pharmaceutical ingredient liposome being prepared from the following raw materials comprising: 0.1-10 parts of an active pharmaceutical ingredient, 10-100 parts of phospholipid and 10-100 parts of cholesterol. The preparation method comprises: preparing same by means of thin film hydration, freeze thawing and a homogenization method. The microneedle system for diagnosis and treatment of brain diseases has the advantages of changing the mode of surgical implantation required by an existing microneedle for treatment of brain diseases, using transdermal drug delivery and thus requiring no surgery, and achieving a good treatment effect.

Description

Microneedle system for diagnosis and treatment of brain diseases and preparation method thereof

Technical field

The present application relates to the technical field of medical drug delivery, and more specifically, it relates to a microneedle system for diagnosis and treatment of brain diseases and a preparation method thereof.

Background technique

Brain diseases are a heterogeneous group of neurological and mental disorders that occur in the brain and can cause severe personal suffering and economic losses to patients. Among various types of brain diseases, brain tumors represented by glioblastoma multiforme and brain neurodegenerative diseases represented by Alzheimer's disease and Parkinson's disease have attracted particular attention. The number of patients worldwide is high, and the consequences of the disease are serious. However, the current drug diagnosis and treatment effects are not ideal. The reason is that the brain's natural defense system, the blood-brain barrier (BBB), while protecting the brain from harmful substances, also restricts the entry of most imaging probes and drug molecules, seriously affecting brain gliomas. diagnosis and treatment.

The lymphatic system is another liquid reticular circulatory system that is independent of the blood circulation system. It is spread throughout most tissues of the human body and helps remove metabolic waste from the interstitium to maintain body fluid homeostasis and play an immune response and immune surveillance role. Although meningeal lymphatic vessels have certain specificities in development time and morphology, similar to peripheral lymphatic vessels, they are highly expressed markers of mature lymphatic endothelial cells. The function of the cerebral lymphatic system is not only limited to the removal of metabolites, but its impact on cerebrospinal fluid circulation cannot be ignored. On January 15, 2020, the team of Professor Akiko Iwasaki of Yale University School of Medicine published an article in Nature magazine revealing that vascular endothelial growth factor C (VEGF-C) promotes the drainage of CD8 T cells from deep cervical lymph nodes into the brain, and promotes the initiation and migration of CD8 T cells. It reaches the tumor site, quickly clears glioblastoma, and exerts a long-lasting anti-tumor immune memory response. However, this anti-tumor effect of VEGF-C disappeared after the deep cervical lymph nodes were ligated, indicating that the elimination effect of VEGF-C on glioblastoma requires lymphatic drainage to the deep cervical lymph nodes. Therefore, the development of new safe and efficient brain drug delivery strategies through deep cervical lymph nodes has become an important research goal in the field of the central nervous system.

Microneedles can overcome stratum corneum obstacles in a minimally invasive and painless way, and can effectively promote the transdermal penetration of drug molecules, especially in the transdermal delivery of macromolecular drugs. At the beginning of the 21st century, experiments gradually demonstrated that microneedles can significantly improve the transdermal delivery of insulin. With the increasing abundance of materials, more and more materials are being developed for the preparation of insulin microneedles. Among them, soluble insulin microneedles have received widespread attention because they can achieve drug delivery effects similar to those of injections. The team of researcher Tao Hu from the Shanghai Institute of Microsystems, Chinese Academy of Sciences, and the team of Professor Mao Ying from the Department of Neurosurgery, Huashan Hospital, Fudan University, developed a heterogeneous, heterogeneous and degradable microneedle patch based on silk protein for the treatment of brain glioma. This microneedle patch can carry three kinds of drugs at the same time. The release sequence and cycle of the drugs can match the different requirements of clinical medication standards. It has the functions of rapid intraoperative hemostasis, long-term postoperative chemotherapy to inhibit tumor cells, and timely start of targeted blood vessel inhibition as needed. key functions such as generation. During the surgical process of tumor resection, the patch is implanted in situ into the tumor cavity. After releasing the drug, it can be completely degraded and disappeared. There is no need for a second surgery to remove it, and the degradation products will not cause immune and inflammatory reactions. This method of intracranial local administration not only solves the problem of obstruction of drug molecules by the blood-brain barrier, but also reduces the toxic and side effects of conventional systemic high-dose administration. In animal experiments, compared with the blank group and the injection group, the treatment group using silk protein-loaded microneedle patches effectively suppressed tumor volume and significantly prolonged the survival time of mice. Professor Li Xingde of Johns Hopkins University in the United States reported an ultra-small (outer diameter 580 μm) therapeutic deep brain microneedle, combined with 800nm optical coherence tomography and laser ablation. The effectiveness of this method was demonstrated through in vivo ultra-high-resolution (axial 1.7 μm, lateral 5.7 μm), high-speed (20 frames per second) volumetric imaging and light attenuation coefficient measurement of mouse brain microstructure. Its translational potential was further demonstrated by in vivo tumor visualization in the deep brain of mice (imaging depth of 1.23mm) and efficient tissue resection (1448nm continuous wave laser at 350mw power).

However, all of the above methods require microneedles to be surgically implanted into the brain tissue. As we all know, there are certain risks in surgery regardless of size, not to mention that for brain surgery, the risks are even higher. Therefore, the inventor believes that the relevant technology has shortcomings in that the administration method for treating brain diseases is single and the risk is high.

Contents of the invention

In order to improve the shortcomings of the related technology of single microneedle administration method and high surgical risk for the diagnosis and treatment of brain diseases, this application provides a microneedle system for the diagnosis and treatment of brain diseases and a preparation method thereof.

In the first aspect, this application provides a microneedle system for diagnosis and treatment of brain diseases, using the following technical solutions:

A microneedle system for the diagnosis and treatment of brain diseases, including the following raw materials in parts by weight:

Active drug liposome 0.1-10 parts;

1-10 parts of dispersant;

Wherein, the active drug liposome includes the following raw materials:

0.1-10 parts of active drug;

Phospholipids 10-100 parts;

Cholesterol 10-100 servings.

By adopting the above technical solution, microneedles loaded with active drug liposomes can successfully deliver active drug liposomes to the brain through deep cervical lymph nodes to exert therapeutic effects. It can avoid the interference of the gastrointestinal environment on drug efficacy and the "first-pass effect" of the liver, maintain a constant optimal blood drug concentration or physiological effect, prolong the effective action time, reduce the number of medication times, and allow patients to self-administer drugs with better compliance.

Optionally, the microneedle system for diagnosis and treatment of brain diseases also includes NK cell membrane proteins.

By adopting the above technical solution, NK cells are natural killer cells (NK), which are important immune cells in the body. They are not only related to anti-tumor, anti-viral infection and immune regulation, but also participate in hypersensitivity reactions in some cases. and the occurrence of autoimmune diseases; using its membrane protein for bionics can effectively increase the rate at which active drug liposomes enter brain cells, thereby quickly exerting effects.

Optionally, the mass ratio of the active drug liposome to the NK cell membrane protein is 300:1.

Optionally, the active drug liposomes and NK cell membrane proteins are prepared through a homogenization method to produce imitated active drug liposomes. The specific operations of the homogenization method are:

1) Perform 3-10 homogenization operations at an air pressure of 20 psi;

2) Adjust the air pressure to 40psi and continue the homogenization operation for 3-10 times to obtain the bionic active drug liposome.

By adopting the above technical solution, active drug liposomes and NK cell membrane proteins are mixed with high-pressure homogenization method, so that the final particle size of the biomimetic active drug liposome is suitable, uniform, and the surface is smooth and free of crystals.

Optionally, the active drug includes a water-soluble active drug and a fat-soluble active drug; when the active drug is a fat-soluble active drug, the preparation of the liposome of the fat-soluble active drug includes the following steps:

S1 Pretreatment of fat-soluble active drugs and preparation of fat-soluble active drug carriers,

Preparation of fat-soluble active drug carrier: Take 10-100 parts of phospholipids and 10-100 parts of cholesterol, dissolve them in chloroform to obtain a mixed solution I for later use;

The pretreatment method for fat-soluble drugs is as follows: take 0.1-10 parts of fat-soluble active drugs and dissolve them in anhydrous methanol or ether to obtain a mixed solution II for later use;

S2 rotary evaporation mixture,

Mix Mixed Liquid I and Mixed Liquid II, and then evaporate to dryness to obtain a mixture;

Preparation of S3 active drug liposomes,

1) Rinse the mixture with PBS buffer, then freeze and thaw at -65°C in liquid nitrogen, and cycle 4-8 times to obtain an active drug liposome solution;

2) Perform dialysis or perform extrusion filtration first and then dialysis to prepare active drug liposomes, and store them at 4°C in the dark;

When the active drug is a water-soluble active drug, the preparation of the water-soluble active drug liposome includes the following steps:

Preparation of S1 water-soluble active pharmaceutical carrier: Take 10-100 parts of phospholipids and 10-100 parts of cholesterol, dissolve them in chloroform to obtain a mixed solution I for later use;

S2 rotary evaporation mixture: rotary evaporate mixture I to dryness to obtain a mixture;

Preparation of S3 active drug liposomes,

1) Rinse the mixture with PBS buffer and add 0.1-10 parts of water-soluble active drug to fully dissolve it, then freeze and thaw at -65°C in liquid nitrogen, and cycle 4-8 times to obtain an active drug liposome solution;

2) Perform dialysis or perform extrusion filtration first and then dialysis to prepare active drug liposomes, and store them at 4°C in the dark.

By adopting the above technical solution, first use chloroform to dissolve phospholipids and cholesterol, use anhydrous methanol and ether to dissolve the active drugs, and then mix, so that the active drugs can be fully mixed with the phospholipids and cholesterol, so that the active drugs can be evenly distributed in the liposomes Secondly, PBS is used to rinse the mixture to remove the mixture from the container wall. The last step of S3 is dialysis to remove free active drugs; among them, fat-soluble drugs such as curcumin, nimodipine, and flunarizine , tetramethylpyrazine, papaverine, vinpocetine, etc., need to be dissolved in anhydrous methanol and then fully dispersed in the liposome. For example, water-soluble drugs such as betahistine, sodium valproate, phenytoin, etc., then Just use PBS buffer for dissolution.

Preferably, the active drug includes a water-soluble active drug and a fat-soluble active drug; when the active drug is a fat-soluble active drug, the preparation of the liposome of the fat-soluble active drug includes the following steps:

Preparation of fat-soluble active drug carrier: Take 50 parts of phospholipids and 25 parts of cholesterol, dissolve them in chloroform to obtain a mixture I for later use;

The pretreatment method for fat-soluble drugs is as follows: take 0.25 parts of fat-soluble active drugs and dissolve them in anhydrous methanol or ether to obtain a mixed solution II for later use;

S2 rotary evaporation mixture,

Preparation of S3 active drug liposomes,

1) Rinse the mixture with PBS buffer, then freeze and thaw at -65°C in liquid nitrogen, and cycle 6 times to obtain an active drug liposome solution;

Preparation of S1 water-soluble active drug carrier: Take 50 parts of phospholipids and 25 parts of cholesterol, dissolve them in chloroform to obtain a mixture I for later use;

Preparation of S3 active drug liposomes,

1) Rinse the mixture with PBS buffer and add 0.25 parts of water-soluble active drugs to fully dissolve the water-soluble active drugs, then freeze and thaw at -65°C in liquid nitrogen, and cycle 5 times to obtain an active drug liposome solution;

Optionally, the phospholipid is soy lecithin.

Optionally, the extrusion operation in step 2) in S3 is specifically: repeatedly extruding 20 times using a liposome extruder equipped with filter membranes of different pore sizes, and the pore sizes of the filter membranes are successively reduced.

Optionally, the filter membrane is a polycarbonate membrane, and the pore diameters of the polycarbonate membrane are 200 nm, 100 nm and 50 nm.

Specifically, the extrusion operation in step 2) in S3 is to sequentially use a liposome extruder equipped with polycarbonate membranes with pore diameters of 200nm, 100nm and 50nm for extrusion filtration. The number of extrusion filtrations is 20 times respectively.

By adopting the above technical solution and repeated extrusion and filtration, the particles in the liposomes are crushed and passed through 200nm, 100nm and 50nm filter membranes in sequence to make the particle size of the liposomes uniform, and finally obtain active drugs with a particle size of 50nm. Liposomes.

Optionally, the dispersing agent is one or a combination of sodium hyaluronate, hyaluronic acid, dextran or polyethylene glycol (PEG).

Preferably, the dispersing agent is sodium hyaluronate and dextran.

Optionally, the active drugs are curcumin, nimodipine, flunarizine, betahistine, ligustrazine, butylphthalide, papaverine, ginkgo leaf extract, vinpocetine, quintiaphen fumarate Citalopram, olanzapine, citalopram, alprazolam, oxazepam (norazepam), lorazepam (Lola), triazolam (Hyloshen), Madopar, Taserda , Senfurol, Xining, Antam, Selegiline, Rasagiline, Tolcapone, Ropinirole, Antam, Ambroxol, Codan, Amantadine, Carbamazepine, Valpro Sodium phenytoin, gabapentin, lamotrigine, oxcarbazepine, phenobarbital, topiramate, vigabatrin, levetiracetam, clonazepam, diazepam, midazole maleate One of the following: ethosuximide, primidone, zonisamide, pregabalin, or retigabine.

In a second aspect, this application provides a method for preparing a microneedle system for diagnosis and treatment of brain diseases, adopting the following technical solution: a method for preparing a microneedle system for diagnosis and treatment of brain diseases, including Following preparation steps:

1) Take the active drug liposomes or imitation active drug liposomes prepared above and freeze-dry them to obtain white powder;

2) Disperse 0.1-10 parts of active drug liposomes or imitation active drug liposomes and 1-10 parts of dispersant in 1) in water, and stir until fully dissolved to obtain a matrix liquid;

3) Inject the matrix liquid obtained in 2) into the microneedle mold, and perform centrifugal inversion to make the matrix liquid evenly distributed in the mold and fill the microneedle tip in the mold;

4) Then take 1-10 parts of the dispersant and disperse it in water to obtain a dispersion; then add the dispersion to the microneedle tip in step 3), centrifuge and dry to obtain active drug liposome microneedles or imitation active drug lipids. plastid microneedle;

5) Attach a pressure-sensitive adhesive lining to the back of the microneedle base and release the mold to obtain an active drug liposome microneedle patch or a bionic active drug liposome microneedle patch.

Preferably, a method for preparing a microneedle system for diagnosis and treatment of brain diseases includes the following preparation steps:

2) Disperse 2 parts of the active drug liposomes or imitation active drug liposomes and 4 parts of the dispersant in 1) in water, and stir until fully dissolved to obtain a matrix liquid;

4) Disperse 1 part of dextran and 2 parts of sodium hyaluronate in water to obtain a dispersion; then add the dispersion to the microneedle tip in step 3), centrifuge and dry to obtain active drug liposomes Microneedles or liposome microneedles imitating active drugs;

Optionally, in step 4), the drying temperature is 4°C and the drying time is 1-100h; preferably, the drying time is 24h.

To sum up, this application has the following beneficial effects: the microneedle system used in the diagnosis and treatment of brain diseases does not require surgery, is simple in process, highly safe, and does not require long-term application. Implantable sustained-release microneedles. Transdermal drug delivery (attached to the head or neck) allows the drug to pass through the deep cervical lymph nodes and meningeal lymphatic vessels, bypassing the blood-brain barrier and reaching the brain tissue to exert therapeutic effects.

Description of the drawings

Figure 1 is a TEM image of curcumin liposomes of a microneedle system disclosed in the present application for diagnosis and treatment of brain diseases;

Figure 2 is a particle size diagram of curcumin liposomes of a microneedle system disclosed in the present application for diagnosis and treatment of brain diseases;

Figure 3 is a distribution diagram of curcumin in mouse brain slices in Example 1 of the present application;

Figure 4 is a comparison table of the efficiency of curcumin entering the brain at different time points in Application Examples 1-3 of the present application.

Detailed ways

The purpose of the present invention is to overcome the above-mentioned shortcomings of the prior art and provide an in vitro brain or neck microneedle patch for treating brain diseases. This method loads active drugs for treating brain diseases into microneedles, and explores the transdermal delivery effect of microneedles loaded with active drugs (attached to the head or neck). The active drug in the embodiments of this application takes curcumin as an example. In other embodiments, the active drug can also be nimodipine, flunarizine, betahistine, ligustrazine, butylphthalide, papaverine, Ginkgo biloba extract, vinpocetine, quetiapine fumarate, olanzapine, citalopram, alprazolam, oxazepam (norazepam), lorazepam (lorola), triazole Lun (Helena), Madopar, Taserta, Senfuluo, Xining, Antam, Selegiline, Rasagiline, Tolcapone, Ropinirole, Antam, Ambroxol , Codan, amantadine, carbamazepine, sodium valproate, phenytoin, gabapentin, lamotrigine, oxcarbazepine, phenobarbital, topiramate, vigabatrin, levetiracetam, chloride Nitrazepam, diazepam, midazolam maleate, ethosuximide, primidone, zonisamide, pregabalin, retigabine and other drugs for the treatment of brain diseases.

The results showed that microneedles loaded with curcumin liposomes could successfully deliver curcumin liposomes to the brain through deep cervical lymph nodes, exerting therapeutic effects. The present invention is of great significance in promoting the application transformation of microneedle in clinical treatment of brain diseases, and provides new methods and new technologies for the treatment of brain diseases.

The present application will be further described in detail below in conjunction with Figures 1 to 4 and examples. It should be noted that if specific conditions are not specified in the following examples, the conditions should be carried out according to conventional conditions or conditions recommended by the manufacturer. Unless otherwise specified, the raw materials used in the following examples can be obtained from ordinary commercial sources.

Preparation example of curcumin liposomes

Preparation Example 1

S1 curcumin dissolution and preparation of curcumin carrier,

Preparation of curcumin carrier: Dissolve 50 mg of phospholipid and 25 mg of cholesterol in 2 ml of chloroform to obtain a mixture I for later use;

Curcumin pretreatment: Take 0.25 mg curcumin and dissolve it in 2 ml of anhydrous methanol to obtain a mixed solution II for later use;

S2 rotary evaporation mixture,

Mix Mixed Liquid I and Mixed Liquid II in a round-bottom flask, and then place it on a rotary evaporator (model IKA-RV10) for rotary evaporation. The rotation speed of the rotary evaporator is 60 rpm, the temperature is 27°C, and the round bottom is evaporated to dryness. A film of mixture forms on the walls of the flask;

Preparation of S3 curcumin liposomes,

1) Use 5 ml of PBS to rinse the film mixture on the wall of the flask to make it completely fall off, then freeze and thaw at -65°C in liquid nitrogen, and cycle 5 times to obtain a liposome solution;

2) Use a liposome extruder equipped with a polycarbonate membrane (model Avanti liposome extruder 610023) to repeatedly squeeze 20 times and filter in sequence; the pore diameters of the polycarbonate membrane are 200nm, 100nm and 50nm; the pore size of the filter membrane used during squeeze filtration is gradually reduced in order to obtain liposomes with uniform particle size.

3) Finally, dialyze to prepare curcumin liposomes and store them at 4°C in the dark.

Preparation Example 2-3 is different from Preparation Example 1 in that the amounts of raw materials used are different, as shown in the table below.

Preparation example of bionic curcumin liposomes

Preparation Example 4

Put 30 mg of the curcumin liposome prepared in Preparation Example 1 into a syringe, and then add 0.1 mg of NK cell membrane protein into the syringe.

Turn on the air pump of the high-pressure homogenizer, set the pressure to 20 psi, open the homogenization valve to fill the pressure; load the syringe containing the sample into the homogenizer, repeat the homogenization operation 5 times, then set the pressure to 40 psi and continue homogenization The operation was repeated 5 times, and finally bionic curcumin liposomes were obtained.

Example

Example 1

1) Take the curcumin liposome prepared in Preparation Example 1 and freeze-dry it to obtain white powder;

2) Disperse 2g of curcumin liposomes or bionic curcumin liposomes and 4g of sodium hyaluronate in 1) in 4ml of water, and stir until fully dissolved to obtain a matrix solution;

3) Inject the matrix liquid obtained in 2) into the microneedle mold, and perform centrifugation and inversion 6 times to make the matrix liquid evenly distributed in the mold and fill the microneedle tip in the mold;

4) Then take 1g of dextran and 2g of sodium hyaluronate and disperse it in 7ml of water to obtain a dispersion; then add the dispersion to the microneedle tip in step 3), centrifuge and dry for 24 hours to obtain curcumin lipids body microneedle;

5) Attach a pressure-sensitive adhesive lining to the back of the microneedle base and demould to obtain a curcumin liposome microneedle patch.

Example 2-3

The difference between Example 2-3 and Example 1 is that the amounts of raw materials used are different, as shown in the table below.

Example 4

The difference between this example and Example 1 is that the bionic curcumin liposome prepared in Preparation Example 4 is used. The obtained bionic curcumin liposomes were scanned under an electron microscope to obtain the TEM image of the biomimetic curcumin liposomes as shown in Figure 1. From the TEN image, it can be seen that the particle size of the biomimetic curcumin liposomes is approximately 50 nm. , the dispersion is better. In addition, the particle size distribution of bionic curcumin liposomes was measured using a particle size analyzer. As shown in Figure 2, the hydrated particle size of bionic curcumin liposomes is approximately 50 nm, and the particles are relatively uniform.

Application examples

Application Example 1: First, take the experimental mouse and remove the hair on the mouse's neck and head; then stick the curcumin liposome microneedle patch prepared in Example 1 on the mouse's neck (curcumin liposome group); finally, the small animal in vivo imaging system (IVIS) was used to track the distribution of curcumin in the mouse brain, as shown in Figures 3 and 4.

Application Example 2: The difference from Application Example 1 is that the bionic curcumin liposome microneedle patch prepared in Example 4 was applied to the neck of mice (biomimetic curcumin liposome group). In vivo imaging system (IVIS) tracked the distribution of curcumin in the mouse brain, as shown in Figure 4.

Application Example 3: Using the curcumin liposome obtained in Preparation Example 1, injecting it into the tail vein of mice (tail vein group), and then using a small animal in vivo imaging system (IVIS) to track the movement of curcumin in the mouse brain The distribution of parts is shown in Figure 4.

For other applications, the curcumin liposomes or bionic curcumin liposomes prepared by the preparation method of the present application can also be made into a gel and applied to the head or neck, or introduced into the head or neck using methods such as ultrasound and subcutaneous injection. neck.

Combining the embodiments and application examples, Figures 1 to 4, it can be seen that the microneedle patch prepared using the preparation method of the microneedle system provided in this application shows that the curcumin liposomes or bionic curcumin are loaded Liposome microneedles can successfully deliver curcumin liposomes or bionic yellowstone liposomes to the brain through deep cervical lymph nodes to exert therapeutic effects. The present invention is of great significance in promoting the clinical application and transformation of microneedles of curcumin liposomes, and provides new methods and technologies for the treatment of brain diseases. Compared with traditional treatment methods for brain diseases, it does not require surgery and reduces treatment risks. It is an implantable sustained-release microneedle with a simple process, high safety, and no need for long-term application. It can avoid the interference of the gastrointestinal environment on drug efficacy and the "first-pass effect" of the liver, maintain a constant optimal blood drug concentration or physiological effect, prolong the effective action time, and reduce the frequency of medication. The patient can self-administer the drug, and the compliance is better. The principle It uses transdermal administration (attached to the head or neck), allowing the drug to pass through the deep cervical lymph nodes and meningeal lymphatic vessels, bypassing the blood-brain barrier and reaching the brain tissue to exert its therapeutic effect.

This specific embodiment is only an explanation of the present application, and it is not a limitation of the present application. After reading this specification, those skilled in the art can make modifications to this embodiment without creative contribution as needed, but as long as the rights of this application are All requirements are protected by patent law.

Claims

A microneedle system for the diagnosis and treatment of brain diseases, which is characterized in that it is made of the following raw materials by weight:

Active drug liposome 0.1-10 parts;

1-20 parts of dispersant;

Wherein, the active drug liposome is made of the following raw materials in parts by weight:

0.1-10 servings of active drug

Phospholipids 10-100 parts;

Cholesterol 10-100 servings.
A microneedle system for diagnosis and treatment of brain diseases according to claim 1, characterized in that the microneedle system for diagnosis and treatment of brain diseases further includes NK cell membrane proteins.
A microneedle system for diagnosis and treatment of brain diseases according to claim 2, characterized in that the mass ratio of the active drug liposome to the NK cell membrane protein is 300:1.
A microneedle system for diagnosis and treatment of brain diseases according to claims 2-3, characterized in that the active drug liposomes and NK cell membrane proteins are prepared by a homogenization method to imitate active drug liposomes , the specific operations of the homogenization method are: 1) Under the air pressure of 20 psi, perform 3-10 homogenization operations; 2) Adjust the air pressure to 40 psi, continue 3-10 homogenization operations to obtain the imitation active drug lipid body.
The microneedle system for disease diagnosis and treatment according to any one of claims 1 to 3, wherein the active drug includes a water-soluble active drug and a fat-soluble active drug; when the active drug is a fat-soluble active drug , the preparation of the liposomes of lipophilic active drugs includes the following steps:

S1 Pretreatment of fat-soluble active drugs and preparation of fat-soluble active drug carriers,

Preparation of fat-soluble active drug carrier: Take 10-100 parts of phospholipids and 10-100 parts of cholesterol, dissolve them in chloroform to obtain a mixed solution I for later use;

The pretreatment method for fat-soluble drugs is as follows: take 0.1-10 parts of fat-soluble active drugs and dissolve them in anhydrous methanol or ether to obtain a mixed solution II for later use;

S2 rotary evaporation mixture,

Mix Mixed Liquid I and Mixed Liquid II, and then evaporate to dryness to obtain a mixture;

Preparation of S3 active drug liposomes,

1) Rinse the mixture with PBS buffer, then freeze and thaw at -65°C in liquid nitrogen, and cycle 4-8 times to obtain an active drug liposome solution;

2) Perform dialysis or perform extrusion filtration and then dialysis to prepare active drug liposomes, and store them in the dark at 4°C; when the active drug is a water-soluble active drug, the water-soluble active drug liposomes The preparation includes the following steps:

Preparation of S1 water-soluble active pharmaceutical carrier: Take 10-100 parts of phospholipids and 10-100 parts of cholesterol, dissolve them in chloroform to obtain a mixed solution I for later use;

S2 rotary evaporation mixture: rotary evaporate mixture I to dryness to obtain a mixture;

Preparation of S3 active drug liposomes,

1) Rinse the mixture with PBS buffer and add 0.1-10 parts of water-soluble active drugs to fully dissolve the water-soluble active drugs, then freeze and thaw at -65°C in liquid nitrogen, and cycle 4-8 times to obtain active drug liposomes solution;

2) Perform dialysis or perform extrusion filtration first and then dialysis to prepare active drug liposomes, and store them at 4°C in the dark.
A microneedle system for the diagnosis and treatment of brain diseases according to claim 5, characterized in that the S3 also includes squeeze filtration, and the squeeze filtration operation is specifically: using devices with different pore sizes in sequence. The liposome extruder of the filter membrane is repeatedly squeezed 20 times, and the pore size of the filter membrane is gradually reduced.
A microneedle system for diagnosis and treatment of brain diseases according to claim 5, characterized in that the filter membrane is a polycarbonate membrane, and the pore diameters of the polycarbonate membrane are 200nm, 100nm and 50nm. .
A microneedle system for diagnosis and treatment of brain diseases according to claim 1, characterized in that the dispersant is one of sodium hyaluronate, hyaluronic acid, dextran or PEG, or Various combinations.
A microneedle system for diagnosis and treatment of brain diseases according to claim 1, characterized in that the active drug is a drug for treating brain diseases, specifically curcumin, nimodipine, flunarin Azine, betahistine, ligustrazine, butylphthalide, papaverine, ginkgo leaf extract, vinpocetine, quetiapine fumarate, olanzapine, citalopram, alprazolam, oxazepam (norazepam), lorazepam (Lola), triazolam (Hyloshen), Madopar, Taserta, Senfrox, Xinine, Antam, Selegiline, Rasagiline Lan, tolcapone, ropinirole, ambroxol, ambroxol, codan, amantadine, carbamazepine, sodium valproate, phenytoin, gabapentin, lamotrigine, oxcarbazepine, phenol Bital, topiramate, vigabatrin, levetiracetam, clonazepam, diazepam, midazolam maleate, ethosuximide, primidone, zonisamide, pregabalin, One of the retigabine.
The preparation method of a microneedle system for the diagnosis and treatment of brain diseases according to claims 1-9, characterized in that it includes the following preparation steps:

1) Take the active drug liposomes or imitation active drug liposomes prepared above and freeze-dry them to obtain white powder;

2) Disperse 0.1-10 parts of active drug liposomes or imitation active drug liposomes and 1-10 parts of dispersant in 1) in water, and stir until fully dissolved to obtain a matrix liquid;

3) Inject the matrix liquid obtained in 2) into the microneedle mold, and perform centrifugal inversion to make the matrix liquid evenly distributed in the mold and fill the microneedle tip in the mold;

4) Then take 1-10 parts of the dispersant and disperse it in water to obtain a dispersion; then add the dispersion to the microneedle tip in step 3), centrifuge and dry to obtain active drug liposome microneedles or imitation active drug liposomes plastid microneedle;

5) Attach a pressure-sensitive adhesive lining to the back of the microneedle base and release the mold to obtain an active drug liposome microneedle patch or a bionic active drug liposome microneedle patch.