Ophthalmic external preparation and preparation method thereof

Abstract

The invention belongs to the technical field of medicine preparation, and provides an ophthalmic external preparation and a preparation method thereof. According to the ophthalmic external preparation, curcumin included by hydroxypropyl-beta-cyclodextrin loaded on exosome modified by polylysine is adopted as an effective component. The preparation can improve the drug concentration of cornea tissues and realize accurate delivery of ocular surfaces; meanwhile, the exosome and curcumin form a synergistic effect, so that the release of proinflammatory factors such as IL-1beta and TNF-alpha can be inhibited, the secretion of goblet cell mucoprotein and the proliferation of corneal epithelial cells can be promoted, and the tear film rupture time is prolonged; in addition, the preparation is excellent in biocompatibility, free of ocular surface stimulation reaction and high in safety.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine preparation, and relates to a drug containing exosomes. BACKGROUND

[0002] The information disclosed in this background section is intended to provide an overview of the general background of the application and is not necessarily a concession that the information was prior art to the present application.

[0003] Dry eye syndrome is a chronic ocular surface disease characterized by tear film homeostasis imbalance, ocular surface inflammation and epithelial damage. Its incidence continues to rise in screen-dependent and elderly populations, and has become one of the major global eye diseases. Existing treatment methods rely on artificial tears to relieve symptoms, but cannot fundamentally solve ocular surface inflammation and tissue damage. Traditional anti-inflammatory drugs such as glucocorticoids have side effects such as elevated intraocular pressure and cataracts, and have low bioavailability, making it difficult to penetrate the ocular surface barrier and exert long-term effects.

[0004] Mesenchymal stem cell exosomes, as natural bioactive vesicles, are rich in active ingredients such as cytokines and microRNAs, have low immunogenicity, anti-inflammatory repair and cell communication regulation functions, and exhibit unique advantages in tissue damage repair. Curcumin, as a natural active ingredient, can regulate multiple signaling pathways to exert antioxidant, anti-inflammatory and tissue repair effects, but has problems such as poor water solubility, low stability and insufficient ocular surface penetration, which limit its application in eye treatment.

[0005] In the prior art, although nanomicelles, cyclodextrin inclusion compounds and other carriers are used for curcumin delivery, there are defects such as insufficient targeting and easy elimination by the ocular surface. Simple mesenchymal stem cell exosome preparations lack targeted drug loading strategies, and the treatment effect is limited. Therefore, developing a preparation method that can achieve efficient loading of curcumin, targeted delivery to the ocular surface and synergistic repair effect has become a key technical requirement in the field of dry eye treatment. SUMMARY

[0006] In view of the problems of poor penetration, low curcumin loading rate and poor targeting of existing dry eye treatment drugs, the present application provides an ophthalmic external preparation with high loading rate and good targeting.

[0007] Another object of the present application is to provide a preparation method of the above-mentioned ophthalmic external preparation, which is simple in process and good in loading effect.

[0008] To achieve the above object, the present application adopts the following technical solutions.

[0009] A preparation method of curcumin-loaded exosomes, comprising the following steps: (1) drop the methanol solution of curcumin into the hydroxypropyl-β-cyclodextrin solution with pH of 1.5-2.5 under the protection of atmosphere and stirring to obtain a reaction solution; (2) centrifuge the reaction solution, dry the supernatant to obtain curcumin inclusion complex; (3) re-dissolve the curcumin inclusion complex, add the polylysine modified exosome suspension, stir and incubate to obtain curcumin loaded exosome suspension.

[0010] The preparation method of the polylysine modified exosome comprises the following steps: mix the exosome suspension and the polylysine solution, stir and react to obtain the polylysine modified exosome.

[0011] The ratio of the exosome and the polylysine is 5×10 10 -2×10 11 Particle number: 1 mg.

[0012] The average polymerization degree of the polylysine is preferably 10.

[0013] The mass ratio of the curcumin and the hydroxypropyl-β-cyclodextrin is 1:8-1:15. In order to adapt the release speed of curcumin, the hydroxypropyl-β-cyclodextrin is preferably a mixture of low-substitution and high-substitution hydroxypropyl-β-cyclodextrin, the low-substitution degree is 3.8-5.2, the high-substitution degree is 7.0-8.5; the mass ratio of the low-substitution and high-substitution hydroxypropyl-β-cyclodextrin is 1:6-1:12.

[0014] The centrifugal speed is 20000 rpm.

[0015] The ratio of the polylysine modified exosome and the curcumin inclusion complex is 5×10 10 -2×10 11 Particle number: 1 mg.

[0016] The application further provides a curcumin loaded exosome obtained by the above preparation method.

[0017] The above curcumin loaded exosome can be used for preparing an ophthalmic external preparation.

[0018] A medicine comprising the above curcumin loaded exosome. The medicine is preferably an ophthalmic external medicine.

[0019] The medicine further comprises a medically or pharmaceutically acceptable carrier or excipient.

[0020] Preferably, the medicine is eye drops, eye drop gel or atomized liquid.

[0021] The medicine comprises 2×10 9 -6×10 9The above-mentioned curcumin-loaded exosomes were loaded with particles per mL.

[0022] The mechanism / synergistic effect of this invention is as follows: Compared to long-chain polylysine, K10-Exo modified with short-chain polylysine exhibits higher biocompatibility. Furthermore, the modified Exo is less prone to aggregation, resulting in a low leakage rate after curcumin loading, thus ensuring formulation homogeneity. The corneal epithelium is a lipophilic barrier; long-chain lysine, with its strong positive charge, is easily retained by the corneal mucus layer, reducing bioavailability. In contrast, the weakly positively charged modification of K10-Exo allows for gentle binding and efficient penetration, enabling more curcumin to reach the corneal stroma and lacrimal gland tissue.

[0023] During the inclusion of curcumin, using hydroxypropyl-β-cyclodextrin solution in an acidic range can prevent the oxidation, isomerization, and aggregation of curcumin under neutral / alkaline conditions, and curcumin has higher solubility within this pH range. At acidic pH, hydroxypropyl-β-cyclodextrin exhibits a weakly negative zeta potential, allowing for efficient binding with positively charged polylysine-modified exosomes via electrostatic interactions. The combined effect of these factors effectively improves the inclusion rate.

[0024] The present invention has the following advantages: This invention provides curcumin-loaded exosomes and their preparation as eye drops. The curcumin-loaded exosomes are first positively charged through polylysine surface modification, forming a targeted adsorption with the negative potential of the ocular surface, thereby increasing the drug concentration in the corneal tissue and achieving precise delivery to the ocular surface. Simultaneously, the anti-inflammatory and repairing activity of the exosomes synergistically works with the antioxidant and anti-inflammatory functions of curcumin, not only inhibiting the release of pro-inflammatory factors such as IL-1β and TNF-α, but also promoting goblet cell mucin secretion and corneal epithelial cell proliferation, prolonging tear film breakup time, and exhibiting significantly better repair effects than single-component formulations. Furthermore, the formulation demonstrates excellent biocompatibility, no ocular surface irritation, and high safety. Attached Figure Description

[0025] Figure 1 The results are the identification results of the modified exosomes; among which, (A) particle size distribution; (B) CD9⁺CD81⁺ positive detection; (C) purity detection; Figure 2 The stability (A), solubility (B), and time-release rate curves (C, D) of curcumin in the curcumin inclusion complex HP-β-CD-Cur are shown. Figure 3 The effect of Exo-HP-β-CD-Cur on the proliferation of human corneal epithelial cells; Figure 4 The effect of Exo-HP-β-CD-Cur on the secretion of inflammatory factors in human corneal epithelial cells; Figure 5is the tear volume measurement of benzalkonium chloride mice before and after different eye drop interventions; Figure 6 is the tear film break-up time measurement of benzalkonium chloride mice before and after different eye drop interventions; Figure 7 is the fluorescein staining score of the corneal tissue of benzalkonium chloride mice after different eye drops are dropped. DETAILED DESCRIPTION

[0026] The application will be further described below in conjunction with the examples and drawings, but the application is not limited by the following examples.

[0027] Example 1 Preparation of polylysine modified exosomes Cultivate umbilical cord-derived mesenchymal stem cells, and when the cell confluence reaches 80%-90%, collect the culture supernatant to obtain exosomes by density gradient centrifugation.

[0028] Adjust the exosome concentration to 1×10 11 Partices / mL, polylysine with an average degree of polymerization of 10 is prepared as a mother liquor, and then mixed with the exosome suspension to make the exosome concentration in the mixed solution 5×10 10 Partices / mL, and the final concentration of polylysine is 0.1 mg / mL. After 1 h of gentle stirring at 37°C and 80 rpm, sterilization is performed by filtering through a 0.22 μm filter membrane to obtain the modified exosomes, which are named K10-Exo. The nanoflow result is shown in Figure 1 Exosome particle size is 60-120 nm, with a median particle size of 76.2 nm and an average particle size of 82.2 nm. The CD9⁺CD81⁺ positive rate is 79.7%, and the purity is 86.4%.

[0029] Example 2 Preparation of curcumin-loaded exosomes 1. Preparation of curcumin inclusion complex (1) Preparation of hydroxypropyl-β-cyclodextrin (HP-β-CD) acid solution: 0.1 g of HP-β-CD with a degree of substitution of 4.5 and 0.9 g of HP-β-CD with a degree of substitution of 7.8 are mixed and dissolved in 100 mL of phosphate buffer (pH 2.0 phosphate buffer solution adjusted by 0.1 mol / L hydrochloric acid) to prepare a 5.0 wt% HP-β-CD acid solution; (2) Gradientwise adding: 0.1 g of curcumin is dissolved in 10 mL of anhydrous methanol, and the solution is added to the HP-β-CD acid solution in a gradient decreasing manner within 30 min under nitrogen protection and dark stirring for 4 h; (3) Purification and freeze-drying: the reaction solution is centrifuged at 20,000 rpm and 4°C for 20 min, and the supernatant is vacuum freeze-dried to obtain yellow powder curcumin inclusion complex (HP-β-CD-Cur).

[0030] The same method as above, in step (1), 5.0wt% HP-β-CD solution was prepared using buffer solution with pH 6.0 and 7.4 respectively, and different HP-β-CD-Cur was obtained; In step (1), two kinds of HP-β-CD were weighed according to the mass ratio of single low degree of substitution 4.5, single high degree of substitution 7.8, low degree of substitution 4.5 and high degree of substitution 7.8 HP-β-CD 1:9, 3:7, 5:5 respectively, and different 5.0wt% HP-β-CD solution was prepared in turn to obtain different HP-β-CD-Cur.

[0031] 2. Stability of curcumin in HP-β-CD-Cur In order to compare the stability of HP-β-CD-Cur and free Cur, HP-β-CD-Cur and free Cur were dissolved in 2mL PBS (pH 7.4) at the same concentration of 1.5μg / mL, and incubated in a 37°C water bath in the dark. The fluorescence intensity values of HP-β-CD-Cur and free Cur were detected at 528nm at time points of 0min, 30min, 60min, 90min, 120min and 150min respectively. The results are shown in Figure 2 As shown in Figure A, free curcumin retains about 70% in aqueous solution after 150 minutes, and Cur in HP-β-CD-Cur is relatively stable, retaining 98% after 150 minutes of incubation at 37°C, which proves that the inclusion structure effectively inhibits the oxidation and hydrolysis of curcumin. The residual rate (RR) was calculated according to the following formula: RR%=F t / F0×100% F t F0 is the fluorescence intensity value of Exo-Cur or free Cur at the beginning.

[0032] 3. Solubility of curcumin In order to determine the increase of solubility of curcumin encapsulated in HP-β-CD, excess HP-β-CD-Cur and free Cur were added to 2mL PBS (pH 7.4) respectively, and incubated in a 37°C water bath in the dark for 30min with multiple shaking, then centrifuged at 15000rpm for 15min, the supernatant was absorbed, filtered with 0.45μm microporous filter, and the fluorescence intensity of the two curcumin solutions was detected by fluorescence spectrophotometry, and the concentration of the two was determined according to the standard curve of curcumin solution, which was the maximum solubility of HP-β-CD-Cur and free Cur.

[0033] The results are shown in Figure 2B showed that the solubility of curcumin in deionized water was (5.67 ± 0.347) μg / mL, while the solubility of curcumin after inclusion with HP-β-CD was (61.63 ± 2.947) μg / mL, which was about 11 times higher than that of curcumin in deionized water, indicating that HP-β-CD significantly improved the water solubility of curcumin by forming host-guest inclusion complexes.

[0034] 4. Drug release rate of HP-β-CD-Cur (1) The cumulative release amount of curcumin was determined by fluorescence spectrophotometry, and the conditions were as follows: excitation wavelength 425 nm, emission wavelength 530 nm, and slit width 5 nm. Different HP-β-CD-Cur samples were placed in dialysis bags and dialyzed in buffer solutions with pH values of 2.0, 6.7, and 7.4. The release was carried out at 37°C with constant temperature oscillation at 100 r / min. Samples were taken at regular intervals, and the cumulative release amount was calculated according to the standard curve by fluorescence spectrophotometry.

[0035] The results are shown in Figure 2 C showed that the release of curcumin from HP-β-CD was significantly affected by different pH values, and the release efficiency increased significantly with increasing pH. The cumulative release amount in pH 2.0 medium was only 52.3%, and the stability of the inclusion system was excellent, which could effectively avoid drug precipitation during preparation. In pH 6.0 and 7.4 media, the cumulative release amounts at 24 h were 88.4% and 95.2%, respectively, which proved that the inclusion of curcumin at pH 2.0 could increase the inclusion amount, and the release was efficient and stable after adjusting to 6.0-8.0, which met the needs of ocular drug delivery.

[0036] (2) The dialysis bag method (molecular weight cutoff 8000 Da) was used. HP-β-CD-Cur solutions with mixed substitution degree, single substitution degree, low and high substitution degree, and different mass ratios (low substitution degree: high substitution degree = 1:9, 3:7, 5:5) were placed in dialysis bags, and 50 mL of pH 7.4 phosphate buffer (simulating physiological environment) was immersed at 37°C with constant temperature oscillation at 100 rpm. Samples were taken at 0.5, 1, 2, 4, 8, 12, and 24 h, and 5 mL of fresh buffer was added at the same time. The concentration of curcumin was determined by fluorescence spectrophotometry, and the cumulative release rate was calculated.

[0037] The results are shown in Figure 2D shows: different substitution degree and ratio of HP-β-CD have significant effect on the release behavior of curcumin, and the release efficiency increases obviously with the increase of the proportion of high-substitution 7.8 HP-β-CD in the system; the single high-substitution 7.8 group releases the fastest, and the 24h cumulative release rate reaches 96.9%, with obvious burst release and sufficient drug release in the early stage; the single low-substitution 4.5 group releases the slowest, and the 24h cumulative release rate is only 81.2%, with strong inclusion stability and outstanding controlled release effect; the mixed ratio group has a release rate between the two single substitution groups, and gradually slows down with the increase of the proportion of low-substitution, among which the mass ratio 1:9 group has both high-efficiency release and gentle controlled release characteristics, with a 24h cumulative release rate of 94.5%, which not only retains the advantages of high-substitution high-efficiency dissolution, but also slows down the burst release through the regulation of a small amount of low-substitution, realizes the long-acting and stable release of curcumin, and meets the drug delivery needs in physiological environment.

[0038] 5. Preparation of curcumin-loaded exosomes The curcumin inclusion compound was reconstituted with sterile normal saline, and the exosome suspension prepared in Example 1 was added to make the final concentration of exosomes 60 μg / mL and the final concentration of curcumin 10 μg / mL. The mixture was incubated at 4°C and 100 rpm for 2 hours to obtain a curcumin-loaded exosome suspension.

[0039] The absorbance at 419 nm was detected by ultraviolet spectrophotometry, and the supernatant concentration and encapsulation efficiency were calculated according to the curcumin standard curve. The encapsulation efficiency calculation formula is as follows: Encapsulation efficiency (%) = [(total curcumin mass - free curcumin mass) / total curcumin mass] × 100% After testing and calculation, the encapsulation efficiency of curcumin in the prepared modified exosomes was 91.5%.

[0040] Example 3 Preparation of eye drops The curcumin-loaded exosome suspension prepared in Example 2 was added with sodium hyaluronate and hydroxypropyl methyl cellulose to a concentration of 0.1 wt%, and the pH was adjusted to 7.2 with phosphate buffer after complete mixing. The mixture was filtered through a 0.22 μm filter membrane to obtain eye drops Exo-HP-β-CD-Cur. The viscosity of the eye drops was 20 mPa·s, and the curcumin loading rate was 86.3%.

[0041] The eye drops were stored at 4°C, and the curcumin content was detected by ultraviolet spectrophotometry on the day of preparation, the first week, the second week, the first month, the second month, and the third month, respectively, to calculate the curcumin retention rate.

[0042] Table 1 Change of curcumin content in Exo-HP-β-CD-Cur with time As shown in Table 1, the curcumin retention rate was 93.6% after 3 months, and the stability was good.

[0043] 1. The effect of eye drops on HCE-T proliferation 1 × 10 4 / cm 2 Human corneal epithelial cells HCE-T (100 μL / well) were divided into four groups and treated with 10 μL of DMEM complete culture medium as control (PBS), DMEM complete culture medium + 10 μL K10-Exo, DMEM complete culture medium + 10 μL HP-β-CD-Cur, and DMEM complete culture medium + 10 μL Exo-HP-β-CD-Cur, respectively, for 72 h for cell proliferation detection, with 6 replicates in each group. When the culture plates were incubated in the incubator for 72 h, 10 μL of CCK-8 solution was added to each well, and the culture plates were incubated in the incubator for 2 h. The absorbance at 450 nm was determined by an enzyme label instrument. The cell proliferation rate of each treatment group relative to the control group was calculated.

[0044] The results are shown in Table 1. Figure 3 As shown in Table 1, the proliferation rate of the eye drop Exo-HP-β-CD-Cur group was increased by 50%, and the proliferation rate was significantly higher than that of the control group, indicating that the eye drop of the application can significantly promote the in vitro proliferation of human corneal cells.

[0045] 2. The effect of eye drops on HCE-T inflammatory factor secretion Logarithmic growth phase HCE-T was taken, and 3 × 10 5 / cm 2 was inoculated in the culture plate and cultured to a confluence of 70%-80%. An inflammatory injury model was constructed by treating with 500 mOsm / L hypertonic solution or 1 ng / mL IL-1β combined with 20% ethanol for 30 s, and the inflammatory factor secretion was induced by continuing to culture for 24 h. The cells were divided into a blank control group, a model control group, a K10-Exo group, a HP-β-CD-Cur group, and an Exo-HP-β-CD-Cur group. Except for the blank control group, the other groups were added with corresponding drug intervention and incubated for 24 h. Then, the cell culture supernatant of each group was collected, centrifuged at 1000 rpm for 10 min, and the supernatant was stored at -80°C for standby. According to the instructions of the kit, the standard and sample were sequentially incubated, washed, and then added with enzyme conjugate for incubation. After washing again, the substrate was colored, and the absorbance value at 450 nm was determined by an enzyme label instrument after the reaction was terminated. The IL-1β and TNF-α concentrations were calculated according to the standard curve, and the inhibition rate of each eye drop preparation on HCE-T cell inflammatory factor secretion was calculated.

[0046] The results are shown in Table 2. Figure 4 As shown in Table 2, the eye drop Exo-HP-β-CD-Cur group inhibited the secretion of IL-1β and TNF-α, and the inhibition rates were 73.8% and 75.3%, respectively. The above results show that the eye drops prepared by surface modification of polylysine (K 10 ) with an average degree of polymerization of 10 and using a mixture of hydroxypropyl-β-cyclodextrin with mixed substitution degrees (4.5 and 7.8) as an inclusion carrier can greatly improve the inclusion rate of curcumin, significantly improve the stability and solubility of curcumin, and significantly promote the in vitro proliferation of human corneal cells and inhibit the secretion of inflammatory factors.

[0047] Example 1 Treatment of eye drops on dry eye mice Female C57BL / 6 mice were randomly divided into 3 groups: control group, model group, K10-Exo exosome (Example 1) treatment group, HP-β-CD-Cur (Example 2) treatment group, Exo-HP-β-CD-Cur (Example 3) treatment group. The control group was normal mice, and the other groups of mice were given 0.2% benzalkonium chloride solution to induce dry eye model, 5 μL of benzalkonium chloride was given to each eye, twice a day, for 5 consecutive days. On D5, the tear secretion was detected by phenol red cotton thread method. If the tear secretion of the model group is significantly lower than that of the control group (P<0.05), the modeling is successful. After the model is successfully constructed, the treatment group mice start to receive 5 μL of eye drops twice a day, for a total of 4 days of drug treatment. After treatment, the mice were injected intraperitoneally with immediate-acting anesthetic tribromoethanol at a specification of 250 μL per mouse, and then the following items were detected: 1. Tear secretion Phenol red cotton thread was placed in the lower eyelid outer 1 / 3 conjunctival sac for 30 s, and the length of the red part of the phenol red cotton thread soaked with tears was measured in millimeters. The results are shown in Figure 5 Exo-HP-β-CD-Cur eye drops greatly increased the tear secretion of dry eye model mice.

[0048] 2. Tear film break-up time 3 μL of 2% liquid fluorescein sodium was dropped into the conjunctival sac of the mouse, and when the first black spot appeared on the corneal surface under cobalt blue light using a slit lamp image system, the tear film break-up time was recorded in seconds by pausing the stopwatch. Each group of 6 right eyes was measured 3 times, and the average value was analyzed. The results are shown in Figure 6 Exo-HP-β-CD-Cur eye drops greatly prolonged the tear film break-up time of dry eye model mice.

[0049] 3. Corneal fluorescein sodium staining 1 μL of 0.5% liquid fluorescein sodium was dropped into the conjunctival sac of the mouse, and the eyes were closed manually for 3-4 times. After 90 seconds, the corneal epithelial damage was observed under the small animal slit lamp microscope with a cobalt blue filter, and scored according to the following scoring criteria: 0: no green staining; 0.5: micro green point staining; 1: Diffuse punctate green staining; 2: The area of ​​green staining on the cornea is less than one-third; 3: The area of ​​the cornea stained green exceeds one-third; 4: The area of ​​green staining on the cornea exceeds two-thirds.

[0050] The results are as follows Figure 7 As shown, after 4 days of treatment with Exo-HP-β-CD-Cur eye drops, the corneal opacity and score of mice decreased. The Exo-HP-β-CD-Cur group showed a significant difference compared with the model group, suggesting that the eye drops significantly promoted the recovery of the corneal epithelium.

[0051] The above description is merely an embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principle of this application should be included within the scope of the claims of this application.

Claims

1. A method for preparing curcumin-loaded exosomes, characterized in that, Includes the following steps: (1) Add the methanol solution of curcumin dropwise to the hydroxypropyl-β-cyclodextrin solution with a pH of 1.5-2.5 and stir under a protective atmosphere to obtain the reaction solution; (2) After centrifuging the reaction solution, the supernatant was dried to obtain curcumin inclusion complex; (3) The curcumin inclusion complex was reconstituted, and polylysine-modified exosome suspension was added. The mixture was stirred and incubated to obtain curcumin-loaded exosome suspension.

2. The preparation method according to claim 1, characterized in that, The preparation of polylysine-modified exosomes includes the following steps: mixing exosome suspension and polylysine solution and stirring to react, thereby obtaining polylysine-modified exosomes. The ratio of exosomes to polylysine is 5 × 10⁻⁶. 10 -2×10 11 Number of particles: 1mg; The average degree of polymerization of polylysine is 10.

3. The preparation method according to claim 1, characterized in that, The mass ratio of curcumin to hydroxypropyl-β-cyclodextrin is 1:8 to 1:15; The ratio of polylysine-modified exosomes to curcumin inclusion complex was 5 × 10⁻⁶. 10 -2×10 11 Number of particles: 1mg.

4. The preparation method according to claim 3, characterized in that, The hydroxypropyl-β-cyclodextrin is a mixture of low-substituted and high-substituted hydroxypropyl-β-cyclodextrins, with the low-substituted degree being 3.8-5.2 and the high-substituted degree being 7.0-8.

5.

5. The preparation method according to claim 4, characterized in that, The mass ratio of low-substituted and high-substituted hydroxypropyl-β-cyclodextrin is 1:6-1:

12.

6. A curcumin-loaded exosome obtained by the preparation method according to any one of claims 1-5.

7. The application of curcumin-loaded exosomes as described in claim 6 in the preparation of ophthalmic topical medications and the resulting drugs.

8. The medicament according to claim 7, characterized in that, The drug also includes medically or pharmaceutically acceptable carriers or excipients.

9. The drug according to claim 7, characterized in that, The medication is an eye drop, eye gel, or nebulizer.

10. The medicament according to claim 9, characterized in that, The drug contains 2×10 9 -6×10 9 The above-mentioned curcumin-loaded exosomes were loaded with particles per mL.

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