Local administration type medicine for treating eye trauma complications and preparation method thereof

Abstract

The invention provides a local administration type medicine for treating ocular trauma complications and a preparation method thereof, it is proved in a traumatic mouse model that rapamycin can obviously promote absorption of vitreous haemoplasty; the symptoms of retina injury are improved, including reducing the range of retinal detachment, inhibiting proliferation of retinal cells, reducing deposition of extracellular matrix, inhibiting retinal fibrosis and reducing traction injury to the retina; therefore, the rapamycin shows a strong inhibition effect on the progress of eye trauma complications.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to the use of rapamycin in the preparation of a medicine for treating ocular trauma, an ophthalmic nanoemulsion preparation containing rapamycin, and a preparation method and use thereof. Background Art

[0002] Ocular trauma is a public health problem that seriously endangers eye health worldwide. In my country, an average of 5 to 12 million cases of ocular trauma occur each year, accounting for 16 to 35% of inpatients in ophthalmology departments, and is the leading cause of monocular blindness. Once an ocular trauma occurs, vision can immediately drop to a state of no light perception, and in severe cases, the eyeball may even need to be removed. Currently, the standard treatment for ocular trauma is to perform a primary suture surgery in the emergency department as soon as possible to restore the integrity of the eyeball structure, and to perform a secondary vitrectomy at an appropriate time to repair the damaged retina and save visual function to the greatest extent possible.

[0003] Vitreous hemorrhage is a common complication of decreased vision after ocular trauma. On the one hand, thick blood accumulation affects the transparency of the vitreous body, causing turbidity of the refractive medium, blocking light signals from reaching the retina, and seriously affecting visual function; on the other hand, if a large amount of blood accumulation is not absorbed for a long time, it may cause organization and cause fundus proliferation and fibrosis, which in turn leads to tractional retinal detachment, seriously affecting vision. In addition, inflammatory cells from the peripheral blood circulation release a large number of cytokines, and the wounds caused by ocular trauma are stimulated by inflammation and do not heal, which will further aggravate the damage to the retinal photoreceptor cells. Traumatic retinal detachment is often caused by ocular trauma accompanied by a large range of eyeball damage, full-thickness rupture of the retina or huge holes, and in the anatomical structure of the retina itself, the contact between the neuroepithelium and the pigment epithelium is not close, and there is a potential space (subretinal cavity) between the two, which makes it relatively easy for the two to separate during trauma. With the destruction of the retinal barrier, a large number of inflammatory cells gather in the subretinal cavity, hindering retinal reattachment. Long-term retinal detachment leads to the restriction of the normal metabolic circulation pathway of photoreceptor cells, causing apoptosis of photoreceptor cells and loss of visual function. Long-term organization of vitreous hemorrhage and wrinkles caused by retinal detachment will further produce a large number of proliferative cords, forming fundus hyperplasia and retinal fibrosis. Even if anatomical reduction is achieved through surgery, it is difficult to reverse and save the remaining visual function.

[0004] Rapamycin (also known as "sirolimus") is a secondary metabolite secreted by soil Streptomyces that was first discovered by scientists in the soil of Easter Island, Chile in 1975. Its chemical structure belongs to the "triene macrolide" compound. As a new type of highly effective immunosuppressant, it is clinically used for the treatment of organ transplant rejection and autoimmune diseases. Its molecular formula is C51H79NO13, and its structural formula is as follows:

[0005]

[0006] Studies have shown that rapamycin has inhibitory or therapeutic effects on a variety of diseases. For example, in a mouse pancreatic cancer model, rapamycin can inhibit cancer development and block tumor spread; in a mouse cerebral malaria model, the survival advantage of mice in the rapamycin treatment group is significant; during radiotherapy, rapamycin prevents radiation-induced tissue damage in mice by protecting normal stem cells that repair tissues. Summary of the invention

[0007] In view of this, the present invention aims to overcome the defects in the prior art and proposes the use of rapamycin in the preparation of drugs for treating ocular trauma, an ophthalmic nanoemulsion preparation containing rapamycin, and a preparation method and application thereof.

[0008] To achieve the above object, the technical solution of the present invention is achieved as follows:

[0009] In a first aspect, the present invention provides use of rapamycin or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating complications of ocular trauma.

[0010] In a feasible embodiment, the complication of ocular trauma is vitreous hemorrhage, retinal detachment or retinal fibrosis.

[0011] Preferably, the treatment of complications of ocular trauma includes one or both of the following aspects:

[0012] a. Promote the absorption of vitreous hemorrhage;

[0013] b. Improve the symptoms of retinal damage.

[0014] Preferably, the improvement of the symptoms of retinal damage includes one or more of: reducing the scope of retinal detachment, inhibiting the proliferation of retinal cells, reducing the deposition of extracellular matrix, inhibiting retinal fibrosis, and reducing traction damage to the retina.

[0015] Preferably, the dosage form of the drug is an ophthalmic preparation;

[0016] Further preferably, the ophthalmic preparation is an ophthalmic nanoemulsion preparation;

[0017] More preferably, the ophthalmic preparation is eye drops, eye wash or intraocular injection, more preferably intravitreal injection.

[0018] In a second aspect, the present invention provides an ophthalmic nanoemulsion preparation, comprising: rapamycin or a pharmaceutically acceptable salt thereof, an oil phase, an emulsifier and a co-emulsifier; preferably, the ophthalmic nanoemulsion preparation further comprises an aqueous phase.

[0019] Preferably, the oil phase is selected from one or more of caprylic / capric triglyceride, tricaprylin, isopropyl myristate, and cinnamaldehyde, preferably tricaprylin.

[0020] Preferably, the emulsifier is selected from one or more of PEG-40 hydrogenated castor oil, tyloxapol, octylphenol polyether-40, poloxamer 188, and polysorbate 80, preferably PEG-40 hydrogenated castor oil.

[0021] Preferably, the co-emulsifier is selected from one or more of glycerol, ethanol, ethylene glycol, and propylene glycol, preferably ethanol.

[0022] Further preferably, the ophthalmic nanoemulsion preparation comprises, by mass percentage: the ophthalmic nanoemulsion preparation comprises, by mass percentage: 0.5-2 parts of rapamycin or a pharmaceutically acceptable salt thereof, 10-20 parts of an oil phase, 40-70 parts of an emulsifier, and 20-30 parts of an auxiliary emulsifier.

[0023] More preferably, the mass percentage of the emulsifier is 50-65 parts.

[0024] More preferably, the ophthalmic nanoemulsion preparation consists of the following components by mass: about 0.5 parts of rapamycin or a pharmaceutically acceptable salt thereof, about 15.1 parts of tricaprylin, about 60.3 parts of PEG-40 hydrogenated castor oil and about 24.1 parts of ethanol.

[0025] More preferably, the mass concentration of rapamycin in the ophthalmic nanoemulsion preparation is 5 mg / mL.

[0026] In a specific embodiment, the ophthalmic nanoemulsion preparation is eye drops, eye wash or intraocular injection, preferably intravitreal injection.

[0027] In a third aspect, the present invention provides a method for preparing the ophthalmic nanoemulsion preparation as described in the second aspect above, the preparation method comprising:

[0028] S1: mixing the emulsifier and the co-emulsifier to prepare a mixed emulsifier;

[0029] S2: Add rapamycin or a pharmaceutically acceptable salt thereof and the oil phase to the mixed emulsifier, mix well, and sonicate until completely dissolved.

[0030] Preferably, the preparation method further comprises: S3: adding deionized water to the mixed solution obtained in step S2, while stirring, until a clear, transparent liquid with uniform appearance is obtained.

[0031] Preferably, the ultrasonic conditions are:

[0032] Ultrasonic power 180-200W, frequency 40Khz;

[0033] Ultrasonic temperature: 4-25℃;

[0034] Ultrasonication time: 10-20 minutes.

[0035] In a fourth aspect, the present invention provides use of the ophthalmic nanoemulsion preparation as described in the second aspect above in the preparation of a medicament for treating complications of ocular trauma.

[0036] Preferably, the complication of ocular trauma is vitreous hemorrhage, retinal detachment or retinal fibrosis.

[0037] Preferably, the treatment of ocular trauma complications includes one or two of the following aspects:

[0038] a. Promote the absorption of vitreous hemorrhage;

[0039] b. Improve the symptoms of retinal damage.

[0040] Preferably, the improvement of the symptoms of retinal damage includes: reducing the scope of retinal detachment, inhibiting the proliferation of retinal cells, reducing the deposition of extracellular matrix, inhibiting retinal fibrosis or reducing one or more of the traction damage to the retina.

[0041] In a fifth aspect, the present invention provides a method for treating ocular trauma (particularly vitreous hemorrhage, retinal detachment and retinal fibrosis caused by ocular trauma), the method comprising: administering a preventive or therapeutically effective amount of rapamycin or a pharmaceutically acceptable salt thereof, or the ophthalmic nanoemulsion preparation as described in the second aspect above to a subject in need thereof.

[0042] Preferably, the subject is a mammal, preferably selected from humans, mice, rats, dogs, rabbits, pigs, monkeys, and more preferably humans.

[0043] Compared with the prior art, the present invention has the following advantages:

[0044] (1) The present invention provides the use of rapamycin or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating ocular trauma (particularly vitreous hemorrhage, retinal detachment and retinal fibrosis caused by ocular trauma). The present invention confirms in a traumatic mouse model that rapamycin can significantly promote the absorption of vitreous hemorrhage; improve the symptoms of retinal damage, including: reducing the scope of retinal detachment, inhibiting the proliferation of retinal cells, reducing the deposition of extracellular matrix, inhibiting retinal fibrosis, and reducing traction damage to the retina; thus, rapamycin exhibits a strong inhibitory effect on the progression of complications of ocular trauma.

[0045] (2) The present invention also provides a rapamycin ophthalmic nanoemulsion preparation and a preparation method and application thereof. The rapamycin ophthalmic nanoemulsion preparation has good intraocular safety and high drug loading, and can deliver a therapeutically effective amount of rapamycin to the eye, thereby achieving efficient treatment of ocular trauma (such as vitreous hemorrhage, retinal detachment and retinal fibrosis). The topical administration preparation can achieve a more effective therapeutic effect through topical administration; the inventors have confirmed through experiments that the rapamycin ophthalmic nanoemulsion preparation of the present invention has a good alleviating or therapeutic effect on mice with traumatic eye damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 The appearance comparison of the blank nanoemulsion, 5 mg / mL rapamycin nanoemulsion and 5 mg / mL rapamycin solution prepared in Example 1 of the present invention is shown;

[0047] Figure 2 A transmission electron micrograph (×40.0K) of the rapamycin nanoemulsion prepared in Example 1 of the present invention, as described in Example 2, is shown; the scale bar in the figure is 100 nm;

[0048] Figure 3 The figure shows the fundus of normal mice after intraocular injection of rapamycin nanoemulsion; Figure A is a color fundus photo and optical coherence tomography scan, and Figure B is a statistical chart measuring the thickness of the inner, outer and full layers of the retina;

[0049] Figure 4 The electroretinograms of normal mice injected with rapamycin nanoemulsions are shown, wherein Figure A is the electroretinograms of mice in the normal control group, the blank NE administration group, and the rapamycin-NE administration group, and Figure B is a statistical graph of the a and b wave amplitudes of low, medium, and high intensity flash stimulation;

[0050] Figure 5 The H&E staining results of the ocular tissue of normal mice after intraocular injection of rapamycin nanoemulsion are shown, and the scale bar in the figure is 20 μm;

[0051] Figure 6 The results of TUNEL staining to detect apoptosis and retinal toxicity of rapamycin nanoemulsion (RA-NE) injected intraocularly into normal mice; the positive control was the fluorescence of cells that were treated with the kit to cause DNA breakage and apoptosis, the negative control was the auto-apoptosis and background fluorescence of retinal cells under normal conditions (i.e., the normal control group), and the other two groups were mice in the blank NE administration group and the RA-NE administration group; the scale bar in the figure is 20 μm;

[0052] Figure 7The figure shows the therapeutic effect of rapamycin nanoemulsion (RA-NE) on vitreous hemorrhage mice; Figure A shows the fundus color photos of mice in each group, and Figure B shows the statistical situation of the absorption of vitreous hemorrhage in mice in each group;

[0053] Figure 8 The figure shows the therapeutic effect of rapamycin nanoemulsion (RA-NE) on retinal detachment mice; wherein Figure A shows the fundus color photos of each group of mice, Figure B shows the rating of retinal detachment of each group of mice (0-4 grades), and Figure C shows the quantitative statistics of the range (area) of retinal detachment of each group of mice;

[0054] Fig. 9 The therapeutic effect of rapamycin nanoemulsion (RA-NE) on Dispase-induced retinal fibrosis in mice is shown; wherein, Figures A and B show fundus color photographs and optical coherence tomography images of each group of mice (the area marked with yellow dotted lines is the fibrous membrane), Figure C shows the time curve of retinal fibrous membrane formation in each group of mice, Figure D shows the fundus hyperplasia grade, Figure E shows the changes in αSMA protein levels in the eyeball tissues of each group of mice and quantitative statistics, and Figure F shows the localization of Ki67 and αSMA in the frozen sections of the eyeballs of each group of mice (the scale is 100 μm, the white dotted line indicates the eyeball outline, and the area marked with yellow dotted lines is the fibroproliferative membrane) and quantitative statistics. DETAILED DESCRIPTION

[0055] Unless otherwise defined, the technical terms used in the following examples have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. The test reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the experimental methods, unless otherwise specified, are all conventional methods.

[0056] The present invention will be described in detail below with reference to the embodiments.

[0057] In the following examples, the experimental animals used and their feeding conditions are as follows:

[0058] Experimental animals: C57Bl / 6JNifdc, clean, male, 20-23 g, purchased from the Experimental Animal Center of Tianjin Medical University; kept in the clean animal room of the Experimental Animal Center of Tianjin Medical University, with a temperature of 21°C to 26°C and a relative humidity of 40% to 70%.

[0059] Example 1. Preparation and properties of rapamycin nanoemulsion

[0060] Formula: 17.5 mg of rapamycin drug powder (purchased from Solarbio, product number R8140), 0.5 g of tricaprylin, 2.0 g of PEG-40 hydrogenated castor oil, and 1.0 mL of ethanol.

[0061] Preparation method: 2.0 g of emulsifier PEG-40 hydrogenated castor oil and 1 mL of co-emulsifier ethanol are prepared into a mixed emulsifier, and then 17.5 mg of rapamycin drug powder and 0.5 g of oil phase tricaprylin are added to the mixed emulsifier, mixed evenly, and ultrasonicated until completely dissolved. The ultrasonic conditions are: ultrasonic power 180 W, frequency 40 Khz; ultrasonic temperature: 4°C; ultrasonic time: 15 minutes, until a clear and transparent liquid with a uniform appearance is obtained. The obtained preparation is a 5 mg / mL rapamycin nanoemulsion (abbreviated as RA-NE).

[0062] Meanwhile, a blank nanoemulsion (referred to as blank NE) was prepared according to the above formulation and procedure (except that rapamycin drug powder was not added, everything else was exactly the same as above); in addition, a 5 mg / mL rapamycin suspension aqueous solution (referred to as rapamycin solution) was prepared by directly adding rapamycin drug powder into deionized water.

[0063] The appearance pictures of the above three preparations are shown in Figure 1 ,like Figure 1 As shown, the blank nanoemulsion is a colorless, clear and transparent liquid, the 5 mg / mL rapamycin nanoemulsion is a colorless, clear and transparent liquid, and the 5 mg / mL rapamycin solution is a distinct white suspension, and rapamycin white powder is precipitated to form distinct layers. This result fully confirms that the rapamycin nanoemulsion preparation of the present invention is a homogeneous liquid preparation, in which rapamycin is fully dissolved.

[0064] Example 2: Morphological Study of Rapamycin Nanoemulsion

[0065] Take an appropriate amount of the rapamycin nanoemulsion prepared in Example 1, dilute it 100 times with deionized water, and then add it dropwise to the carbon film-coated copper mesh. After incubation at room temperature for 10 minutes, the excess liquid is absorbed with filter paper, and then pH 6.8 phosphotungstic acid is added for staining for 5 minutes, oven dried, and observed under a transmission electron microscope after standing.

[0066] The results are as follows Figure 2 As shown by Figure 2 It can be seen that the rapamycin nanoemulsions are spherical under a transmission electron microscope, with an oil phase inside and a particle size ranging from 1 to 100 nm.

[0067] Example 3: Retinal safety assessment of intraocular injection of rapamycin nanoemulsion

[0068] Experimental animal grouping and treatment: 30 wild-type C57Bl / 6JNifdc mice, male, weighing 20-23 g, were randomly divided into 3 groups and treated as follows:

[0069] 1. Normal control group: no treatment was performed;

[0070] 2. Blank NE administration group: The blank NE prepared in Example 1 was diluted 100 times with deionized water and then injected into the vitreous body, 1 μl / mouse, and observed for 7 days after administration;

[0071] 3. RA-NE administration group: The RA-NE prepared in Example 1 was diluted 100 times with deionized water (0.05 mg / mL) and injected into the vitreous body, 1 μL / mouse, and observed for 7 days after administration.

[0072] After the above treatments, the mice in each group were photographed with fundus color and detected by optical coherence tomography (OCT) using a small animal retinal imager. The results are shown in Figure 3 , Figure 3 In the figure, Figure A shows the fundus conditions of mice in each group, which shows that there is no significant difference in the fundus conditions of the three groups of mice, all of which are as follows: the fundus red light reflex is normal, the blood vessels are roughly normal, and there is no obvious bleeding, exudation, and degeneration. The black circle in the figure is the OCT scanning line, and the corresponding scanning image of this layer shows that the retinal structure is clear and there is no obvious disorder; Figure B shows the statistical results of OCT measurement of the inner, outer, and full-layer retinal thickness of mice in each group, which shows that there is no significant difference between the groups, all of which are as follows: there is no obvious edema, thickening, or atrophy and thinning of the retina. The above results show that there is no significant difference in the fundus and retina conditions between the RA-NE administration group and the normal control group and the blank NE administration group.

[0073] In addition, the Diagnosys small animal visual electrophysiology platform was used to perform electroretinogram testing. Figure 4 Figure A shows the waveform of the dark-adapted flash-stimulated electroretinogram of each group of mice, and Figure B shows the statistical graph of the a and b wave amplitude measurements. The results showed that there was no significant difference among the groups, which indicates that the retina of the mice in the rapamycin nanoemulsion (RA-NE) administration group had no obvious functional changes compared with the normal control mice.

[0074] The ocular histopathological results of mice in each group are shown in Figure 5 , Figure 5 Shown are the results of H&E staining of eyeball sections of mice in each group. The results showed that the retinal structure of mice in the rapamycin nanoemulsion (RA-NE) administration group was clear, and the thickness of the inner nuclear layer and the outer nuclear layer did not change significantly compared with the normal control mice.

[0075] In addition, the retinal toxicity of rapamycin nanoemulsion was detected by TUNEL staining of cell apoptosis. Figure 6 , Figure 6Shown in the figure are immunofluorescence images of mouse eye sections of the TUNEL staining positive control (provided by the TUNEL kit, after DNase treatment), negative control (normal control group), blank NE administration group and RA-NE administration group. The results show that there are no TUNEL-positive cells in the retinal layers of the mice in the rapamycin nanoemulsion (RA-NE) administration group, that is, no obvious cell apoptosis and retinal toxicity were observed.

[0076] The above results show that the rapamycin nanoemulsion preparation of the present invention has high safety for the retina.

[0077] Example 4. Evaluation of the therapeutic effect of intraocular injection of rapamycin nanoemulsion on vitreous hemorrhage

[0078] Experimental animal grouping and treatment: 24 wild-type C57Bl / 6JNifdc mice, male, weighing 20-23 g, were randomly divided into 8 groups and subjected to the following treatments:

[0079] 1. Blank NE administration group: The blank NE prepared in Example 1 was injected into the vitreous cavity at 1 μl / mouse, and then 0.25, 0.5, 0.75 and 1 μl of autologous blood were injected into the vitreous cavity of the mice, respectively, to induce vitreous hemorrhage model. One day after modeling, fundus color photography was performed using a small animal retinal imager, and flake or lumpy blood accumulation was observed in front of the retina and in the vitreous cavity, indicating that the modeling was successful; 7 days after modeling, fundus color photography was performed using a small animal retinal imager to evaluate the absorption of blood accumulation.

[0080] 2. RA-NE administration group: The rapamycin nanoemulsion (RA-NE) prepared in Example 1 was injected into the vitreous cavity at 1 μl / mouse, and then 0.25, 0.5, 0.75 and 1 μl of autologous blood were injected into the vitreous cavity of the mice, respectively, to induce vitreous hemorrhage model. After 1 and 7 days of modeling, fundus color photography was performed using a small animal retinal imager to evaluate the absorption of vitreous hemorrhage.

[0081] The results are as follows Figure 7 As shown. Figure A shows the fundus conditions of each group of mice 1 day and 7 days after modeling, which shows that: the mice given blank NE still have blood accumulation in the eyes, and the absorption is not obvious; while the vitreous hemorrhage of mice treated with rapamycin nanoemulsion (RA-NE) is significantly reduced, and a large amount of absorption is achieved. Figure B shows the absorption rate of blood accumulation in each group of mice. The statistical method is blood accumulation absorption rate (%) = (blood volume on the first day - blood volume on the seventh day) / blood volume on the first day * 100%, which shows that compared with the fundus images of mice given blank NE, the fundus of mice treated with RA-NE shows obvious absorption of vitreous hemorrhage.

[0082] Example 5. Evaluation of the therapeutic effect of intraocular injection of rapamycin nanoemulsion on retinal detachment

[0083] Experimental animal grouping and treatment: 20 wild-type C57Bl / 6JNifdc mice, male, weighing 20-23 g, were randomly divided into 2 groups and subjected to the following treatments:

[0084] 1. Blank NE administration group: The blank NE prepared in Example 1 was injected into the vitreous, 1 μl / mouse, and then 2 μl of balanced salt solution was injected subretinaally to induce retinal detachment model. One week after modeling, fundus color photography was performed using a small animal retinal imager. If bulge or detachment of at least one quadrant of the retina was observed, it represented that the modeling was successful;

[0085] 2. RA-NE administration group: The RA-NE prepared in Example 1 was injected into the vitreous body at 1 μl / mouse, and then 2 μl of balanced salt solution was injected subretinaally to induce a retinal detachment model. One week after modeling, fundus color photography was performed using a small animal retinal imager to evaluate the retinal position or detachment.

[0086] The results are as follows Figure 8 As shown, Figure A shows the fundus conditions of each group of mice after modeling, which shows that the retina of mice given blank NE is grayish, with large-scale bulges and tortuous blood vessels; while the retina of mice treated with rapamycin nanoemulsion (RA-NE) is normal in color, the range of detachment is significantly reduced, and the blood vessels are relatively straight. Figure B shows the grading of the degree of retinal detachment of each group of mice (i.e., retinal detachment quadrant), which shows that compared with the fundus images of mice given blank NE, the retinal detachment grade of mice treated with RA-NE is significantly reduced. Figure C shows the quantitative analysis of the range of retinal detachment of each group of mice (i.e., retinal detachment area), which shows that compared with the fundus images of mice given blank NE, the range of retinal detachment of mice treated with RA-NE is significantly reduced.

[0087] Example 6. Evaluation of the therapeutic effect of intraocular injection of rapamycin nanoemulsion on retinal fibrosis

[0088] Experimental animal grouping and treatment: 30 wild-type C57Bl / 6JNifdc mice, male, weighing 20-23 g, were randomly divided into 3 groups and treated as follows:

[0089] 1. Normal control group: no treatment was performed, and fundus color photography and optical coherence tomography (OCT) were performed directly using a small animal retinal imager;

[0090] 2. Blank NE administration group: The blank NE prepared in Example 1 was injected into the vitreous cavity, 1 μl / mouse, and then 1 μl of Dispase enzyme was injected into the vitreous cavity of the mice to induce a retinal fibrosis model. After 7 days of modeling, fundus color photography and optical coherence tomography (OCT) were performed using a small animal retinal imager. If retinal wrinkles or gray-white fibrous tissue were observed, it represented that the modeling was successful;

[0091] 3. RA-NE administration group: The RA-NE prepared in Example 1 was injected into the vitreous cavity at 1 μl / mouse, and then 1 μl of Dispase enzyme was injected into the vitreous cavity of the mice to induce a fibrosis model. After 7 days of modeling, fundus color photography and optical coherence tomography (OCT) were performed using a small animal retinal imager.

[0092] The results are as follows Fig. 9 As shown in the figure, Figure A and Figure B show the fundus conditions of each group of mice, which show that: the model mice given blank NE have a large range of retinal proliferation cords in the eyes, and OCT shows high-intensity subretinal reflection signals indicating the formation of fibrous membranes, while the retinal wrinkles of the mice treated with RA-NE are significantly reduced, and OCT shows that the formation of subretinal fibrosis is significantly delayed; Figure C shows the time curve of subretinal fibrous membrane formation in each group of mice, which shows that: compared with the eyeballs of mice given blank NE, the eyeballs of mice treated with RA-NE show a milder fibrotic phenotype; Figure D shows the fundus proliferation grade of each group of mice (used to evaluate the severity of retinal fibrosis), which shows that: compared with the eyeballs of mice given blank NE, the eyeballs of mice treated with RA-NE show a lower proliferation grade; Figure E shows the changes in the αSMA protein level in the eyeball tissues of each group of mice (αSMA is a marker of fibroblasts or myofibroblasts, representing fibrosis). The progression level of αSMA protein level increases with the progression of the disease, that is, the degree of fibrosis is positively correlated with the αSMA protein level), which shows that: compared with the eyeballs of mice given blank NE, the eyeballs of mice treated with RA-NE show lower αSMA protein levels; Figure F shows the localization and relative expression of αSMA and Ki67 in the frozen sections of the eyeballs of mice in each group (αSMA protein is generally located at the location of retinal fibrosis formation, which is used to indicate the severity of the lesion, that is, the degree of retinal fibrosis is positively correlated with the αSMA fluorescence density; Ki67 nuclear protein is generally located at the location of fundus proliferation, which is used to indicate the fundus proliferation coefficient, that is, the degree of fundus proliferation is positively correlated with the number of Ki67 positive cells per unit), which shows that: compared with the eyeballs of mice given blank NE, the fundus proliferation coefficient of mice treated with RA-NE is significantly reduced, the degree of retinal fibrosis is significantly reduced, and the tractional damage to the retina is lighter.

[0093] The above results all indicate that intraocular injection of rapamycin nanoemulsion can significantly reduce various complications caused by ocular trauma, inhibit fundus hyperplasia and retinal fibrosis, thereby alleviating damage to the retina and protecting visual function.

[0094] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. Use of rapamycin or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating complications of ocular trauma.

2. The use according to claim 1, characterized in that: The ocular trauma complication is vitreous hemorrhage, retinal detachment or retinal fibrosis.

3. The use according to claim 1, characterized in that: The treatment of complications of eye trauma includes one or more of the following aspects: a. promoting the absorption of vitreous hemorrhage; b. improving symptoms of retinal damage; preferably, the improvement of symptoms of retinal damage includes: reducing the scope of retinal detachment, inhibiting the proliferation of retinal cells, reducing the deposition of extracellular matrix, inhibiting retinal fibrosis, and reducing one or more of the traction damage to the retina.

4. The use according to any one of claims 1 to 3, characterized in that: The dosage form of the drug is an ophthalmic preparation; preferably, the ophthalmic preparation is an ophthalmic nanoemulsion preparation; more preferably, the ophthalmic preparation is eye drops, eye wash or intraocular injection, and further preferably, the ophthalmic preparation is an intravitreal injection.

5. An ophthalmic nanoemulsion preparation, characterized in that: The ophthalmic nanoemulsion preparation comprises: rapamycin or a pharmaceutically acceptable salt thereof, an oil phase, an emulsifier and a co-emulsifier; preferably, the ophthalmic nanoemulsion preparation further comprises an aqueous phase.

6. The ophthalmic nanoemulsion preparation according to claim 5, characterized in that: The oil phase is selected from one or more of caprylic / capric triglyceride, tricaprylin, isopropyl myristate, and cinnamaldehyde, preferably tricaprylin; Preferably, the emulsifier is selected from one or more of PEG-40 hydrogenated castor oil, tyloxapol, octylphenol polyether-40, poloxamer 188, and polysorbate 80, preferably PEG-40 hydrogenated castor oil; Preferably, the co-emulsifier is selected from one or more of glycerol, ethanol, ethylene glycol, and propylene glycol, preferably ethanol.

7. The ophthalmic nanoemulsion preparation according to claim 5, characterized in that: The ophthalmic nanoemulsion preparation comprises, by weight: 0.5-2 parts of rapamycin or a pharmaceutically acceptable salt thereof, 10-20 parts of an oil phase, 40-70 parts of an emulsifier, and 20-30 parts of an auxiliary emulsifier; Preferably, the mass percentage of the emulsifier is 50-65 parts; More preferably, the ophthalmic nanoemulsion preparation is composed of the following components by mass: about 0.5 parts of rapamycin or a pharmaceutically acceptable salt thereof, about 15.1 parts of tricaprylin, about 60.3 parts of PEG-40 hydrogenated castor oil, and about 24.1 parts of ethanol; More preferably, the mass concentration of rapamycin in the ophthalmic nanoemulsion preparation is 5 mg / mL.

8. The ophthalmic nanoemulsion preparation according to claim 5, characterized in that: The ophthalmic nanoemulsion preparation is eye drops, eye wash or intraocular injection, preferably intravitreal injection.

9. The method for preparing the ophthalmic nanoemulsion according to any one of claims 5 to 8, characterized in that: The preparation method comprises the following steps: S1: mixing the emulsifier and the co-emulsifier to prepare a mixed emulsifier; S2: adding rapamycin or a pharmaceutically acceptable salt thereof and an oil phase to the mixed emulsifier, mixing evenly, and sonicating until completely dissolved; Preferably, the ultrasonic conditions are: ultrasonic power 180-200W, frequency 40Khz; ultrasonic temperature: 4-25°C; ultrasonic time: 10-20 minutes.

10. Use of the ophthalmic nanoemulsion preparation according to any one of claims 5 to 8 in the preparation of a medicament for treating complications of ocular trauma; Preferably, the ocular trauma complication is vitreous hemorrhage, retinal detachment or retinal fibrosis; Preferably, the treatment of ocular trauma complications includes one or two of the following aspects: a. promoting the absorption of vitreous hemorrhage; b. improving symptoms of retinal damage; preferably, the improvement of symptoms of retinal damage includes: One or more of the following: reducing the scope of retinal detachment, inhibiting the proliferation of retinal cells, reducing the deposition of extracellular matrix, inhibiting retinal fibrosis or reducing traction damage to the retina.

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