Flurbiprofen axetil liposome and its preparation and application

Abstract

The invention belongs to the field of medical technology, relate to a kind of flurbiprofen axetil liposome and its preparation and as the application of anti-inflammatory preparation.The flurbiprofen axetil liposome of the present invention, by weight, 10 50 parts of flurbiprofen axetil, 300 600 parts of phospholipids, 5 60 parts of cholesterol, 2 20 parts of antioxidants, 500 1500 parts of hydration media, 500 1500 parts of absolute ethanol and 250 1000 parts of dichloromethane.The flurbiprofen axetil microneedle average length that the present invention combines microneedle with liposome technology to obtain is about 1000 μm, and microneedle spacing is about 350 μm, and shape is regular, and outward appearance is neat, and flurbiprofen axetil liposome can be directly pierced into skin and plays a role, further plays the role of storage and sustained-release effect in the skin of liposome, better plays the anti-inflammatory effect of flurbiprofen axetil. In addition, the hyaluronic acid contained in the microneedles is a widely used anti-inflammatory material with good biocompatibility, which further improves the biocompatibility of the microneedles.

Description

Technical Field

[0001] The invention belongs to the technical field of medicine and relates to flurbiprofen axetil liposomes and a preparation method thereof and an application thereof as an anti-inflammatory preparation. Background Art

[0002] Arthritis is a complex disease, classified into two main categories: osteoarthritis and rheumatoid arthritis. It is characterized by joint inflammation and irreversible damage. If left untreated, disease progression can lead to a complete loss of daily activities. Nearly 1% of the world's population is reportedly affected by arthritis. Currently, intra-articular injections are the fastest and most effective treatment for arthritis, minimizing adverse reactions. However, intra-articular injections are prone to pain, physical damage, local infection, poor patient compliance, and the generation of large amounts of medical sharps waste. Therefore, oral and transdermal administration have become the most widely used treatments for early-stage arthritis. However, arthritis is a localized, rather than systemic, disease, and oral administration also faces challenges such as low bioavailability, gastrointestinal damage, and even liver and kidney damage.

[0003] Transdermal drug delivery systems (TDDS) refer to formulations that use the skin as a drug delivery route, transporting drugs to local tissues or the systemic blood circulation for local or systemic effects. Compared to commonly used dosage forms, transdermal drug delivery systems can avoid the first-pass effect in the liver and the destruction of drugs by the gastrointestinal tract during oral administration. Transdermal drug delivery is safe and convenient, prolongs the duration of effective action, and can maintain constant blood drug concentrations. However, the barrier effect of the stratum corneum significantly affects transdermal drug absorption. With the development of nanocarrier technology, a variety of nanocarriers have been developed for delivering drug molecules and enhancing transdermal drug penetration, such as lipid nanoparticles, polymeric micelles, and vesicles. Vesicles include classic liposomes, transfersomes, ethosomes, ethosomes, lipid vesicles, and precursor vesicles. Among these carriers, liposomes offer numerous advantages. Composed of phospholipids, cholesterol, a hydrating medium, and antioxidants, lipophilic and hydrophilic regions can accommodate drug molecules with a wide range of solubility. Their excellent deformability allows for easy penetration into the skin, holding them up for broad application prospects in the transdermal field. However, transdermal drug delivery is limited by the stratum corneum, making it difficult to achieve therapeutic effects through the skin. Furthermore, chemical permeation enhancers can only increase the transdermal permeability of small-molecule drugs, making it difficult for large-molecule drugs and drug carriers to be delivered therapeutically through the skin. Microneedles, a novel transdermal dosage form, can penetrate the skin and thus cross the skin barrier.

[0004] Flurbiprofen axetil is a nonsteroidal anti-inflammatory drug with a strong anti-inflammatory and anti-inflammatory effect. Because its current dosage form only has an injection preparation, and is mostly used for clinical analgesia and anti-inflammatory. Multiple administrations are prone to produce toxic and side effects, and poor patient compliance, so it is made into topical preparations for the treatment of arthritis. The current commercially available preparation of flurbiprofen axetil is flurbiprofen axetil injection, which uses lipid microspheres as carriers to deliver the drug. The microspheres are a layer of oil-in-water membrane structure, and injection can cause many toxic and side effects and adverse reactions, and can produce a large amount of medical sharp waste, poor patient compliance, and has great defects. In order to solve the above problems, the present invention, based on liposome carrier technology, prepares flurbiprofen axetil liposome suspension and flurbiprofen axetil liposome microneedles, respectively, thereby providing a flurbiprofen axetil transdermal preparation with strong transdermal ability and good biocompatibility. Summary of the Invention

[0005] The purpose of the present invention is to provide a flurbiprofen axetil liposome and its preparation and application as an anti-inflammatory preparation.

[0006] To achieve the above purpose, the present invention adopts the following technical solutions:

[0007] A flurbiprofen axetil liposome comprises, by weight, 10-50 parts of flurbiprofen axetil, 300-600 parts of phospholipids, 5-60 parts of cholesterol, 2-20 parts of antioxidant, 500-1500 parts of hydration medium, 500-1500 parts of anhydrous ethanol and 250-1000 parts of dichloromethane.

[0008] Preferably, the flurbiprofen axetil liposomes contain, by weight, 20-40 parts of flurbiprofen axetil, 300-400 parts of phospholipids, 10-20 parts of cholesterol, 5-10 parts of antioxidants, 500-1000 parts of hydration medium, 500-1000 parts of ethanol and 250-500 parts of dichloromethane.

[0009] The phospholipid is one or more of soybean lecithin, soybean lecithin, lecithin, egg yolk lecithin, phosphatidylcholine, dioleoyl lecithin, and dipalmitoyl phosphatidylethanolamine;

[0010] The antioxidant includes one or more of vitamin E, α-tocopherol, deoxycholic acid, and sodium deoxycholate; preferably vitamin E. When the antioxidant is vitamin E, the preparation is more stable and has less irritation;

[0011] The hydration medium includes one or more of PBS (potassium dihydrogen phosphate and sodium hydroxide), PBS (sodium dihydrogen phosphate and sodium hydroxide), PBS (sodium dihydrogen phosphate and disodium hydrogen phosphate), 0.9% normal saline, and deionized water. 0.9% normal saline is preferred because experiments have shown that when 0.9% normal saline is used as the hydration medium, the flurbiprofen axetil liposomes prepared are more stable, have a higher encapsulation efficiency, and are not subject to drug degradation.

[0012] A method for preparing flurbiprofen axetil liposomes, which adopts a passive drug loading method, wherein the formation of the liposomes and the loading of the drug are completed simultaneously, comprises the following steps:

[0013] (1) A liposome preparation is prepared by a thin film hydration-ultrasound method in passive drug loading. Flurbiprofen axetil, phospholipids, cholesterol and antioxidants are dissolved in anhydrous ethanol and dichloromethane according to the above proportions. A uniform thin film is obtained by rotary evaporation at 35-45°C (the rotary evaporation temperature is preferably 40°C, which is the most efficient). This solvent system is relatively safe and has better solubility for the film material. The solvent evaporates completely and has good safety. A hydration medium solution is added, and after rotary hydration and ultrasonication, a flurbiprofen axetil liposome mother solution is obtained.

[0014] (2) Extrusion-filtering the obtained mother liquor through a filter membrane to obtain flurbiprofen axetil liposomes.

[0015] In the step (1), a hydration medium solution is added, and the mixture is subjected to rotary hydration at room temperature for 30-60 minutes. After hydration, the mixture is ultrasonically treated in an ice bath for 10-15 minutes (the power is set to 300-750W).

[0016] An application of the flurbiprofen axetil liposome, and an application of the flurbiprofen axetil liposome in preparing an anti-inflammatory preparation.

[0017] The flurbiprofen axetil liposomes are used in the preparation of anti-inflammatory microneedle preparations. When the liposome microneedles are used as transdermal delivery carriers for flurbiprofen axetil, compared with aqueous solutions, the liposome microneedles of the present invention can increase the in vitro transdermal permeation release, and better improve the encapsulation efficiency and stability of the drug.

[0018] A flurbiprofen axetil liposome microneedle comprises a microneedle-shaped matrix and a mother liquid embedded in the matrix, wherein the mother liquid comprises, by weight, 0.1-30 parts of the flurbiprofen axetil liposome according to claim 1, 10-50 parts of sodium hyaluronate, 50-100 parts of hydroxypropyl-β-cyclodextrin, 100-300 parts of PVP K90, and 100-1000 parts of purified water.

[0019] The microneedle preparation method comprises the following steps: freeze-drying the flurbiprofen axetil liposomes, dissolving them with hyaluronic acid and hydroxypropyl-β-cyclodextrin in the above-mentioned proportion to obtain a flurbiprofen axetil liposome microneedle matrix mother solution, placing the mother solution in a microneedle mold, centrifuging the mold, removing the bottom matrix, scraping off the bottom matrix, adding PVP K90 as a base, and drying and demolding to obtain the flurbiprofen axetil liposome microneedle.

[0020] The centrifugal speed is 2500-3500 rpm / min, the centrifugal time is 5-15 min, and the drying time is 12-24 h. After drying, the flurbiprofen axetil liposome microneedles are obtained by automatic demoulding.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] 1. The present invention is directed to the problem of reduced encapsulation efficiency or drug leakage in liposomes, and utilizes the self-structural characteristics of the drug flurbiprofen axetil to combine liposomes with a passive drug loading method. The preparation of liposomes can be divided into two major categories, namely, passive drug loading method and active drug loading method, according to the different drug properties and the mechanism of being loaded. Traditionally, people have adopted more passive drug loading method. This method is to dissolve the drug in an aqueous phase or an organic phase, and then prepare drug-containing liposomes by a suitable method. This method is suitable for highly liposoluble or water-soluble drugs, and the resulting liposomes have a higher encapsulation efficiency and stability, while the active drug loading method is suitable for the liposome preparation of amphiphilic drugs.

[0023] 2. by screening prescription, the encapsulation efficiency and stability of the drug are preferably improved. And the preparation technology of the above-mentioned flurbiprofen axetil liposome is simple, the conditions are mild, and the flurbiprofen axetil liposome average particle diameter obtained is 100-200nm, narrow distribution, and good dispersion stability in aqueous medium. The flurbiprofen axetil liposome prepared by this prescription can improve the solubility of the drug, the retention amount of in vitro percutaneous penetration, higher encapsulation efficiency and stability.

[0024] 3. The flurbiprofen axetil microneedles produced by combining microneedle and liposome technology have an average length of approximately 1000 μm, a microneedle spacing of approximately 350 μm, and a regular shape and neat appearance. Flurbiprofen axetil liposomes can be directly inserted into the skin to exert their effect, further leveraging the liposomes' intradermal retention and sustained release properties, further enhancing the anti-inflammatory efficacy of flurbiprofen axetil. Furthermore, the hyaluronic acid contained in the microneedles is a widely used anti-inflammatory material with good biocompatibility, further enhancing the biocompatibility of the microneedles. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 A is the particle size diagram of flurbiprofen axetil liposomes prepared in Example 4, Figure 1 B is a scanning electron microscope (SEM) image;

[0026] Figure 2 The graph is a relationship between the in vitro release curves of flurbiprofen axetil liposomes and free drug and time prepared in Example 4;

[0027] Figure 3 A is an optical micrograph of the flurbiprofen axetil liposome microneedles prepared in Example 6, Figure 3 B is a scanning electron microscope image of the flurbiprofen axetil liposome microneedles prepared in Example 6

[0028] Figure 4 Graph showing the relationship between the in vitro cumulative transdermal penetration of the flurbiprofen axetil liposome microneedles prepared in Example 6 and time;

[0029] Figure 5 Photos showing the anti-inflammatory efficacy of the flurbiprofen axetil liposome microneedles and flurbiprofen axetil liposome solution prepared in Example 6 on a rat model;

[0030] Figure 6 AB are curves showing the changes in plantar thickness and ankle swelling of the flurbiprofen axetil liposome microneedles and flurbiprofen axetil liposome solution prepared in Example 6 in a rat model;

[0031] Figure 7 HE staining images of the skin of the flurbiprofen axetil liposome microneedle and flurbiprofen axetil liposome solution groups prepared in Example 6;

[0032] Figure 8 A is the blood compatibility data of the flurbiprofen axetil liposome microneedles prepared in Example 6, Figure 8 B is a group of pictures showing the blood compatibility of the flurbiprofen axetil liposome microneedles prepared in Example 6. DETAILED DESCRIPTION

[0033] The present invention is further described with reference to the accompanying drawings and examples, which are intended to illustrate the present invention but are not intended to limit the present invention in any way.

[0034] The present invention utilizes a passive drug loading method to prepare flurbiprofen axetil liposomes, and simultaneously combines with a specific system to thereby improve the encapsulation efficiency and stability of the drug. The liposomes of the present invention have a double-layer membrane structure and have better stability. In vitro release and transdermal delivery studies have shown that the preparation has pH responsiveness. The prepared liposomes are further combined with microneedles to reduce skin irritation, improve biocompatibility, and better exert the storage and sustained-release effects of the liposomes in the skin, further improving the anti-inflammatory effect.

[0035] Example 1:

[0036] Weigh 300 mg of phospholipids, 30 mg of flurbiprofen axetil, 10 mg of cholesterol and 5 mg of vitamin E into an eggplant-shaped bottle, add 10 mL of anhydrous ethanol and 5 mL of dichloromethane to dissolve them, and evaporate under reduced pressure at 40°C to obtain a uniform film; then add 10 mL of 0.9% normal saline and rotate to hydrate for 1 hour, and ultrasonicate the probe for 10 minutes under ice bath conditions (power 450 W, working 3 seconds, rest 2 seconds). The intermittent method can avoid the impact of local high temperature on the preparation. The obtained flurbiprofen axetil-loaded liposomes are then squeezed and filtered through a 0.22 μm filter membrane to obtain flurbiprofen axetil liposomes, which are stored at 4°C. The particle size of the flurbiprofen axetil liposomes was determined to be 145 nm, and the encapsulation efficiency was 75%. Particle size Figure 1 As shown in A, the morphology of flurbiprofen axetil liposomes is shown in Figure 1 As shown in B. The particle size and PDI of the preparation were stable after being stored at 4°C for 30 days.

[0037] Example 2:

[0038] 400 mg of phospholipids, 30 mg of flurbiprofen axetil, 10 mg of cholesterol, and 5 mg of vitamin E were weighed into an eggplant-shaped bottle, 10 mL of absolute ethanol and 5 mL of dichloromethane were added to dissolve them, and rotary evaporation under reduced pressure at 40° C. was performed to obtain a uniform film; 10 mL of 0.9% normal saline was then added for rotary hydration for 1 hour, and probe ultrasound was applied under ice bath conditions for 10 minutes (power 450 W, working for 3 seconds, rest for 2 seconds) to obtain flurbiprofen axetil-loaded liposomes, which were then extruded and filtered through a 0.22 μm filter membrane to obtain flurbiprofen axetil liposomes, which were then stored at 4° C. The particle size of the flurbiprofen axetil liposomes was measured to be 165 nm, and the encapsulation efficiency was 75%. Excessive membrane material did not increase the encapsulation efficiency and could also cause waste.

[0039] Example 3:

[0040] (1) 300 mg of phospholipids, 30 mg of flurbiprofen axetil, 10 mg of cholesterol, and 5 mg of vitamin E were weighed into an eggplant-shaped flask, 10 mL of anhydrous ethanol and 5 mL of dichloromethane were added to dissolve them, and rotary evaporation under reduced pressure at 40°C was performed to obtain a uniform film. 10 mL of 0.9% normal saline was then added to rotate and hydrate for 1 h. The probe was ultrasonicated for 10 min (power 450 W, working for 3 s, rest for 2 s) under ice bath conditions to obtain flurbiprofen axetil-loaded liposomes. The liposomes were then extruded and filtered through a 0.22 μm filter membrane to obtain flurbiprofen axetil liposomes, which were then stored at 4°C. The particle size of the flurbiprofen axetil liposomes was determined to be 145 nm, and the encapsulation efficiency was 75%.

[0041] (2) Weigh 35 mg of sodium hyaluronate, 70 mg of hydroxypropyl-β-cyclodextrin, and 30 mg of flurbiprofen axetil freeze-dried liposomes (prepared in a freeze drying oven) into a 5 mL beaker, add 1 mL of ultrapure water to dissolve them, and then mix the resulting solution evenly.

[0042] (3) Pipette an appropriate amount of the solution prepared in step (2) into the PDMS microneedle mold so that the liquid evenly covers the mold. Then place the mold in a 50 mL centrifuge tube (a flat surface is required in the centrifuge tube) and centrifuge at 3000 rpm / min for 10 min. Remove the mold and scrape off excess liquid.

[0043] (4) Take 0.3g PVP K90, add 1.0mL ultrapure water to dissolve it and mix it evenly, fill it into the PDMS microneedle mold as a base, wait for the microneedle to dry, and remove it after automatic demoulding to obtain the flurbiprofen axetil liposome microneedle. The obtained flurbiprofen axetil liposome microneedle was observed by optical microscopy, and the obtained micrographs and scanning electron micrographs are shown in FIG. Figure 3 As shown, the flurbiprofen axetil liposome microneedles prepared by the above method have an average length of about 1000 μm, a regular quadrangular pyramidal shape, and a neat appearance. The mechanical properties of the obtained flurbiprofen axetil liposome microneedles were measured using a universal testing machine. When the travel distance was 1.6 mm, the mechanical strength of each needle was 4.5 N.

[0044] Example 4

[0045] (1) 300 mg of phospholipid, 30 mg of flurbiprofen axetil, and 10 mg of cholesterol were weighed into an eggplant-shaped flask, 10 mL of anhydrous ethanol and 5 mL of dichloromethane were added to dissolve them, and rotary evaporation under reduced pressure at 40°C was performed to obtain a uniform film. 10 mL of 0.9% normal saline was then added to rotate and hydrate for 1 h. The probe was ultrasonicated for 10 min (power 500 W, working for 3 s, rest for 2 s) in an ice bath to obtain flurbiprofen axetil-loaded liposomes. The liposomes were then extruded and filtered through a 0.22 μm filter membrane to obtain flurbiprofen axetil liposomes, which were stored at 4°C. The particle size of the flurbiprofen axetil liposomes was determined to be 140 nm, and the encapsulation efficiency was 75%.

[0046] (2) Weigh 50 mg of sodium hyaluronate, 50 mg of hydroxypropyl-β-cyclodextrin, and 30 mg of flurbiprofen axetil freeze-dried liposomes into a 5 mL beaker, add 1 mL of ultrapure water to dissolve them, and then mix the resulting solution evenly.

[0047] (3) Pipette an appropriate amount of the solution prepared in step (2) into the PDMS microneedle mold so that the liquid evenly covers the mold. Then place the mold in a 50 mL centrifuge tube (a flat surface is required in the centrifuge tube) and centrifuge at 4000 rpm / min for 5 min. Remove the mold and scrape off excess liquid.

[0048] (4) 0.3 g of PVP K90 was dissolved and mixed evenly with 1.0 mL of ultrapure water. The PDMS microneedle mold was filled with the PVP K90 as a substrate. The microneedles were dried and automatically demolded to obtain the flurbiprofen axetil liposome microneedles. The mechanical properties of the obtained flurbiprofen axetil liposome microneedles were measured using a universal testing machine. When the moving distance was 1.6 mm, the mechanical strength of each needle was 3.0 N.

[0049] Comparative Example 1:

[0050] 200 mg of phospholipids, 20 mg of flurbiprofen axetil, 20 mg of cholesterol, and 5 mg of vitamin E were weighed into an eggplant-shaped flask and dissolved in 10 mL of anhydrous ethanol and 5 mL of dichloromethane. The mixture was then rotary evaporated at 40°C to obtain a uniform film. 10 mL of 0.9% saline was then added for 1 hour of rotary hydration. Ultrasound was applied to the flask at 300W for 20 minutes in an ice bath. The resulting flurbiprofen axetil-loaded liposomes were then filtered through a 0.22 μm filter membrane to obtain the flurbiprofen axetil liposomes, which were then stored at 4°C. The flurbiprofen axetil liposomes were measured to have a particle size of 105 nm and an encapsulation efficiency of 50%. The particle size and PDI of the formulation remained stable after 30 days of storage at 4°C. Low film material usage and prolonged ultrasonication resulted in a low liposome encapsulation efficiency, indicating that high encapsulation efficiency can only be achieved when the film material usage and process parameters are within a specific range.

[0051] Comparative Example 2:

[0052] 200 mg of phospholipids, 20 mg of flurbiprofen axetil, 10 mg of cholesterol, and 5 mg of vitamin E were weighed into an eggplant-shaped flask, dissolved in 10 mL of anhydrous ethanol and 5 mL of dichloromethane, and rotary evaporated at 40°C to obtain a uniform film; 10 mL of 0.9% normal saline was then added for rotary hydration for 1 hour, and probe ultrasonication was performed for 10 minutes (300W) under ice bath conditions. The resulting flurbiprofen axetil-loaded liposomes were then extruded and filtered through a 0.22 μm filter membrane to obtain flurbiprofen axetil liposomes, which were then stored at 4°C. The particle size of the flurbiprofen axetil liposomes was determined to be 165 nm, and the encapsulation efficiency was 55%. The particle size and PDI of the preparation were stable after being stored at 4°C for 30 days. By reducing the ultrasonication time, the encapsulation efficiency increased to a certain extent, but because the amount of membrane material components was not added at a specific amount, the encapsulation efficiency remained at a low level.

[0053] Comparative Example 3:

[0054] 200 mg of phospholipids, 20 mg of flurbiprofen axetil, 10 mg of cholesterol, and 5 mg of vitamin E were weighed into an eggplant-shaped flask, dissolved in 10 mL of anhydrous ethanol and 5 mL of dichloromethane, and rotary evaporated at 40°C to obtain a uniform film. 10 mL of 0.9% saline was then added for 1 hour of rotary hydration. Ultrasonic probe was ultrasonicated for 10 minutes (450W) in an ice bath. The resulting flurbiprofen axetil-loaded liposomes were then extruded and filtered through a 0.22 μm filter membrane to obtain flurbiprofen axetil liposomes, which were then stored at 4°C. The particle size of the flurbiprofen axetil liposomes was determined to be 135 nm, with an encapsulation efficiency of 55%. The particle size and PDI of the preparation were stable after 30 days at 4°C. Increasing the ultrasonic power slightly reduced the particle size, which is more beneficial for the stability of the preparation. However, since the amount of membrane material was not added at the specified dosage, the encapsulation efficiency remained low.

[0055] Test Example 1:

[0056] The pre-treated ex vivo mouse skin was placed on a Franz diffusion cell (stratum corneum facing up, dermis facing down). The experiment was divided into two groups, namely the flurbiprofen axetil liposomes prepared in Example 1 and the flurbiprofen axetil liposome microneedle group prepared in Example 3. The receiving medium was a 1% Tween 80-PBS solution with pH = 5.0 and 7.0, the rotation speed was 100 rpm / min, and the receiving liquid temperature was (37±0.5)°C. 1 mL of the receiving liquid was collected at 2, 4, 6, 8, 10, 12, 24, and 48 hours. The obtained liquid was centrifuged at a speed of 13000 rpm / min for 5 minutes, and the supernatant was measured by HPLC. The relationship between the cumulative drug permeation amount and time of each group was calculated, and a broken line graph was prepared as shown below. Figure 4 shown.

[0057] Liposomes are different from the physical penetration-enhancing method of microneedles. They mainly achieve chemical penetration-enhancing effects by virtue of the good biocompatibility between the material and the skin tissue. Human skin tissue is mainly composed of a lipophilic matrix composed of cholesterol, cholesterol esters, free fatty acids, phospholipids, etc. The similarity of lipid structure enables liposomes to better fuse with the epidermal lipid barrier, and improve its permeability by interacting with keratinocytes or intercellular lipids, making it easier for the encapsulated drugs to enter the deep skin, but this method is less efficient. The liposome-loaded microneedle matrix with good biocompatibility can dissolve rapidly after penetrating the skin. The liposomes encapsulated by the microneedles are distributed to various areas of the skin by free diffusion and continuously release drugs, thereby positioning the drugs in the epidermis and continuously penetrating to the site of action. Microneedles are directly inserted into the epidermis in a minimally invasive and painless manner, forming hundreds of microchannels in the skin to improve the efficiency of transdermal drug delivery. The dual penetration promotion of "liposome-microneedle" can greatly increase the transdermal penetration of drugs. According to Figure 2The liposome solution itself is pH-responsive, and the liposome-loaded microneedles also have this effect. However, compared with the liposome solution group, the amount of flurbiprofen axetil permeated through the skin in the microneedle treatment group was significantly increased.

[0058] Test Example 2:

[0059] The flurbiprofen axetil liposome microneedle group prepared in Example 3 was treated for 5 minutes, and the flurbiprofen axetil liposome prepared in Example 1 was used as a subcutaneous injection group to verify the anti-inflammatory properties of the drug-loaded microneedles by examining and verifying the results.

[0060] After 7 days of adaptive feeding, male SD rats were randomly divided into 4 groups: (1) blank control group (Control); (2) model group (AIA); (3) flurbiprofen axetil liposome microneedle group (FA@Lipo-DMNs); (4) flurbiprofen axetil liposome subcutaneous injection group (FA@Lipo-SC); After 7 days of adaptive feeding, the adjuvant-induced arthritis (AIA) rat model was established. AIA group rats and drug group rats were injected with 0.15mL complete Freund's adjuvant (CFA) into the plantar surface of the right hind paw by subcutaneous injection. After 2 days, obvious changes were seen. The right hind paw of the rats showed redness and thickening of the sole. After 7 days, it was seen that the inflammation had spread to the ankles and joints of the rats, and the rats had difficulty moving. The rats' diseased joints were measured to verify the theoretical feasibility of the model. To ensure the parallel consistency of the experiment, the blank control group rats were injected with 0.15mL normal saline. The specific operation was exactly the same as the modeling method. After the model was successfully established, drug administration began on the eighth day at a dose of 1.35 mg·kg -1 , once every three days, for a total of 6 times.

[0061] The appearance of the paws Figure 5 As shown, the foot swelling curve changes as Figure 6 As shown in the figure, compared with the control group rats, the ankle joints and plantars of the rats in the model group and the treatment group showed obvious redness, swelling and inflammation; after 6 doses of administration, it was observed that the redness and swelling of the ankle joints and plantars of the rats in the FA@Lipo-DMNs group and the FA@Lipo-SC group were significantly reduced compared with those in the model group, while there was almost no significant difference between the FA@Lipo-DMNs group and the FA@Lipo-SC group, indicating that both treatment groups had significant effects on AIA rats, but the patients in the FA@Lipo-DMNs group had good compliance, which also proved the anti-inflammatory superiority of the flurbiprofen axetil liposome preparation.

[0062] Histopathological studies were performed on rats in each group to evaluate the level of inflammation. Figure 7As shown, the control group showed no redness, swelling, or inflammation, while the AIA group showed significant redness, swelling, and inflammation. Treatment with FA@Lipo-SC and FA@Lipo-DMNs reduced inflammation, demonstrating the drug's potent anti-inflammatory effects. There were no significant differences between the two groups, with the FA@Lipo-DMNs group demonstrating better patient compliance. Further H&E staining and pathological examination revealed no inflammatory infiltration in the control group, while the AIA group showed significant inflammatory cell infiltration, indicating progression of arthritis. Treatment with FA@Lipo-SC and FA@Lipo-DMNs significantly reduced inflammatory cell infiltration. Therefore, FA@Lipo-DMNs can inhibit joint inflammation in rats, providing an ideal drug delivery strategy for reducing side effects caused by intra-articular injections.

[0063] Test Example 3:

[0064] The flurbiprofen axetil liposome microneedles prepared in Example 3 were used to investigate the safety of FA@Lipo-DMNs at different doses using a hemolysis test. As shown in Fig. 8, normal saline was used as a negative control, deionized water was used as a positive control, and FA@Lipo-DMNs were used as a test sample to investigate the blood compatibility of FA@Lipo-DMNs at different doses. When the dose of FA was 100, 200, and 300 μg, no obvious hemolysis (<5%) was produced. This may be because the microneedle matrix gradually formed a gel and co-precipitated with blood cells to the bottom, indicating that the microneedle matrix can maintain the structural integrity of blood cells, showing low hemolytic toxicity and good material safety. The results show that FA@Lipo-DMNs have good biocompatibility at normal or higher doses and can be safely used for further research.

Claims

1. A flurbiprofen axetil liposome, characterized in that: By weight, 10-50 parts of flurbiprofen axetil, 300-600 parts of phospholipids, 5-60 parts of cholesterol, 2-20 parts of antioxidants, and 500-1500 parts of hydration medium; The antioxidant includes one or more of vitamin E, α-tocopherol, deoxycholic acid, and sodium deoxycholate; The passive drug loading method is used for preparation, and the formation of liposomes and the loading of drugs are completed simultaneously, including the following steps: (1) A liposome preparation is prepared by a thin film hydration-ultrasound method in passive drug loading, wherein flurbiprofen axetil, phospholipids, cholesterol and antioxidant are dissolved in anhydrous ethanol and dichloromethane according to the above proportions, and rotary evaporation is performed under reduced pressure at 35-45° C. to obtain a uniform thin film; a hydration medium solution is added, and after rotary hydration and ultrasonication, a flurbiprofen axetil liposome mother solution is obtained; (2) extruding and filtering the obtained mother liquor through a filter membrane to obtain flurbiprofen axetil liposomes; In the step (1), a hydration medium solution is added, and the mixture is subjected to rotary hydration at room temperature for 30-60 minutes, and then ultrasonicated in an ice bath for 10-15 minutes; The hydration medium includes one or more of potassium dihydrogen phosphate and sodium hydroxide, sodium dihydrogen phosphate and sodium hydroxide, sodium dihydrogen phosphate and disodium hydrogen phosphate, 0.9% normal saline, and deionized water.

2. The flurbiprofen axetil liposome according to claim 1, wherein: By weight, the invention comprises 20-40 parts of flurbiprofen axetil, 300-400 parts of phospholipids, 10-20 parts of cholesterol, 5-10 parts of antioxidant and 500-1000 parts of hydration medium.

3. A method for preparing the flurbiprofen axetil liposome according to claim 1, characterized in that: The passive drug loading method is used for preparation, and the formation of liposomes and the loading of drugs are completed simultaneously, including the following steps: (1) A liposome preparation is prepared by a thin film hydration-ultrasound method in passive drug loading, wherein flurbiprofen axetil, phospholipids, cholesterol and antioxidant are dissolved in anhydrous ethanol and dichloromethane according to the above proportions, and rotary evaporation is performed under reduced pressure at 35-45° C. to obtain a uniform thin film; a hydration medium solution is added, and after rotary hydration and ultrasonication, a flurbiprofen axetil liposome mother solution is obtained; (2) Extrusion-filtering the obtained mother liquor through a filter membrane to obtain flurbiprofen axetil liposomes.

4. The preparation method according to claim 3, characterized in that: In the step (1), a hydration medium solution is added, and the solution is subjected to rotary hydration at room temperature for 30-60 minutes. After hydration, the solution is ultrasonicated in an ice bath for 10-15 minutes.

5. A use of the flurbiprofen axetil liposome according to claim 1, characterized in that: Application of the flurbiprofen axetil liposome in preparing an anti-inflammatory preparation.

6. The use of the flurbiprofen axetil liposome according to claim 5, wherein: Application of the flurbiprofen axetil liposomes in the preparation of anti-inflammatory microneedle preparations.

7. A flurbiprofen axetil liposome microneedle comprising a microneedle-shaped matrix and a mother solution embedded in the matrix, characterized in that: The mother solution comprises, by weight, 0.1-30 parts of the flurbiprofen axetil liposome according to claim 1, 10-50 parts of sodium hyaluronate, 50-100 parts of hydroxypropyl-β-cyclodextrin, 100-300 parts of PVP K90, and 100-1000 parts of purified water.

8. A method for preparing the flurbiprofen axetil liposome microneedle according to claim 7, characterized in that: The flurbiprofen axetil liposomes according to claim 1 are freeze-dried and mixed with sodium hyaluronate and hydroxypropyl-β-cyclodextrin in proportion to obtain a flurbiprofen axetil liposome microneedle matrix mother solution, which is placed in a microneedle mold. The mold is removed after centrifugation, the bottom matrix is ​​scraped off, and PVP K90 is added as a base. The flurbiprofen axetil liposome microneedles are obtained after drying and demolding.

9. The method for preparing the flurbiprofen axetil liposome microneedle according to claim 8, characterized in that: The centrifugal speed is 2500-3500 rpm / min, the centrifugal time is 5-15 minutes, the drying time is 12-24 hours, and the flurbiprofen axetil liposome microneedles are obtained after automatic demoulding after drying.

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