A medicament for treating ocular surface and corneal epithelial diseases

Abstract

The present application provides a medicine for treating ocular surface and corneal epithelial diseases, which has fibroblast growth factor 5 as an effective component. The medicine has the effects of promoting corneal epithelial repair and treating ocular surface diseases caused by fibroblast growth factor 5 (FGF5) gene mutation, and has no toxic side effects.

Description

TECHNICAL FIELD

[0001] The present application relates to a kind of drugs for treating ocular surface and corneal epithelial diseases, belong to the technical field of eye medicine. BACKGROUND

[0002] Fibroblast growth factor 5 (FGF5), initially identified based on a screening method for transforming oncogenes, is a member of the FGF family, is a protein with 268 amino acids, 29.1 kDa, FGF5 is closely related to the growth and development of epithelium, and is considered to be a regulator of hair growth cycle, leading to the appearance of angola phenotype in knockout mice. FGF5 is a myogenic trophic factor for motor neurons of the peripheral nervous system, and is also a regulator of brain neuron differentiation and survival. In addition, FGF5 affects astrocytes in vitro and shows neurotrophic activity in vivo. FGF5 affects the self-renewal of stem cells by binding to FGFR1, promoting the differentiation of more stable progenitor cells. Gallagher et al. confirmed the increased expression of FGF5 in the expanded corneal epithelium in amniotic membrane by qRT-PCR, suggesting that it may have an impact on the repair of corneal epithelial damage, but no drugs for treating ocular surface and corneal epithelial related diseases using FGF5 have been reported. SUMMARY

[0003] The present application provides a kind of drugs for treating ocular surface and corneal epithelial diseases, which can effectively solve the above problems.

[0004] The present application is realized as follows:

[0005] A kind of drugs for treating ocular surface and corneal epithelial diseases, the effective component is fibroblast growth factor 5.

[0006] In some embodiments, the concentration of the fibroblast growth factor 5 is 4 ng / mL to 4 mg / mL.

[0007] In some embodiments, the fibroblast growth factor 5 is a natural fibroblast growth factor 5 or a recombinant fibroblast growth factor 5.

[0008] In some embodiments, the drug is eye drops, eye ointment or eye gel preparation.

[0009] In some embodiments, the eye drops further comprise a dissolving agent, a pH adjusting agent, an osmotic pressure adjusting agent, a stabilizing agent and water.

[0010] In some embodiments, the eye ointment further comprises an eye ointment base, an emulsifying agent and / or an eye drug bacteriostatic agent.

[0011] In some embodiments, the eye gel preparation further comprises a gel base, a dissolving agent, an osmotic pressure adjusting agent, a pH adjusting agent, a preservative and water.

[0012] In some embodiments, the dissolving agent is propylene glycol and / or hydroxypropyl beta cyclodextrin.

[0013] In some embodiments, the eye drug bacteriostatic agent is hydroxyphenyl ethyl ester or benzalkonium chloride.

[0014] Use of fibroblast growth factor 5 in the preparation of a medicament for the treatment of ocular surface and corneal epithelial diseases.

[0015] The present application has the following advantages:

[0016] The medicament for treating ocular surface and corneal epithelial diseases of the present application uses fibroblast growth factor 5 (FGF5) as an effective component, and has the effects of promoting corneal epithelial repair and treating ocular surface diseases caused by fibroblast growth factor 5 (FGF5) gene mutation.

[0017] The medicament for treating ocular surface and corneal epithelial diseases of the present application has no toxic side effects on the cornea. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed in the embodiments will be briefly introduced below, and it should be understood that the following drawings only show some embodiments of the present application, and therefore should not be regarded as a limitation on the scope, and for those skilled in the art, other related drawings can also be obtained without creative labor on the basis of these drawings.

[0019] Figure 1 is an experimental effect diagram of corneal epithelial repair provided by example 1 of the present application.

[0020] Figure 2 is a repair rate diagram of corneal epithelial repair provided by example 1 of the present application.

[0021] Figure 3 is a cytotoxicity test diagram provided by example 2 of the present application. DETAILED DESCRIPTION

[0022] In order to make the purposes, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative effort should fall within the scope of the present application. Therefore, the following detailed description of the embodiments of the present application provided in the drawings is not intended to limit the scope of the claimed present application, but merely represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative effort should fall within the scope of the present application.

[0023] In the description of the present application, the terms "first", "second" are only used for descriptive purposes, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined with "first", "second" can explicitly or implicitly include one or more of the features. In the description of the present application, the meaning of "a plurality of" is two or more, unless otherwise explicitly and specifically limited.

[0024] The embodiment of the present application provides a medicine for treating ocular surface and corneal epithelial diseases, and an effective component of the medicine is fibroblast growth factor 5. The fibroblast growth factor 5 has the functions of promoting corneal epithelial proliferation repair and treating ocular surface diseases caused by fibroblast growth factor 5 (FGF5) gene mutation.

[0025] In some embodiments, the concentration of the fibroblast growth factor 5 is 4 ng / mL to 4 mg / mL.

[0026] In some embodiments, the fibroblast growth factor 5 is a natural fibroblast growth factor 5 or a recombinant fibroblast growth factor 5.

[0027] In some embodiments, the medicine is eye drop, eye ointment or eye gel preparation.

[0028] In some embodiments, the eye drop further comprises a dissolving agent, a pH adjusting agent, an osmotic pressure adjusting agent, a stabilizing agent and water.

[0029] In some embodiments, the eye ointment further comprises an eye ointment base, an emulsifying agent and / or an eye drug bacteriostatic agent.

[0030] In some embodiments, the eye gel preparation further comprises a gel base, a dissolving agent, an osmotic pressure adjusting agent, a pH adjusting agent, a preservative and water.

[0031] In some embodiments, the solubilizing agent is propylene glycol and / or hydroxypropyl beta cyclodextrin.

[0032] In some embodiments, the bacteriostatic agent for ophthalmic drugs is hydroxypropyl benzoate or benzalkonium chloride.

[0033] In some embodiments, the pH adjusting agent is at least one of sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, sodium hydroxide, potassium hydroxide, hydrochloric acid, phosphoric acid, boric acid, borax, acetic acid, sodium acetate, citric acid, sodium citrate, tartaric acid, sodium tartrate, sodium carbonate, potassium carbonate, sodium bicarbonate and potassium bicarbonate.

[0034] In some embodiments, the osmotic pressure adjusting agent is at least one of sodium chloride, potassium chloride, boric acid, borax, sodium sulfate, potassium sulfate, sodium nitrate, potassium nitrate, sodium acetate, mannitol, glycerol, propylene glycol and glucose.

[0035] In some embodiments, the petrolatum base, liquid paraffin and anhydrous lanolin are included in the ophthalmic ointment base.

[0036] In some embodiments, the emulsifying agent is carbomer, propylene glycol or polysorbate 80.

[0037] In some embodiments, the gel base is at least one of carbomer, hydroxypropyl cellulose, sodium hyaluronate and xanthan gum.

[0038] In some embodiments, the preservative is at least one of benzalkonium chloride, benzalkonium bromide, chlorobutanol and nipagin.

[0039] The embodiments of the present application also provide a use of fibroblast growth factor 5 in the preparation of a medicament for treating ocular surface and corneal epithelial diseases.

[0040] Embodiment 1

[0041] The preparation method of eye drops comprises the following steps:

[0042] 1. The natural FGF5 protein is dissolved with a solubilizing agent, stirred and mixed, the pH range is adjusted to 7.0-8.0 with a pH adjusting agent, the osmotic pressure is adjusted to 295-309 mOsm / l with an osmotic pressure adjusting agent, and the volume is adjusted to 1000 ml with water for injection to obtain a liquid medicine;

[0043] 2. The liquid medicine obtained in step 1 is filtered 1-5 times through a microporous filter membrane in a hundred-level environment;

[0044] 3. The liquid medicine is filled into single-dose packaging containers in a hundred-level environment, sealed, and the finished product is obtained.

[0045] The dissolving agent is propylene glycol; the pH regulator is sodium dihydrogen phosphate; and the osmotic pressure regulator is sodium chloride. The final concentration of FGF5 protein is 4 ng to 4 mg / mL.

[0046] Example 2

[0047] The preparation method of the eye ointment comprises the following steps:

[0048] 1. Sterilize the mixture of anhydrous lanolin and white petrolatum by heating and melting, filtering through sterile gauze, and drying at 150 degrees for 1 hour;

[0049] 2. Sterilize the liquid paraffin by drying at 150 degrees for 1 hour;

[0050] 3. Weigh an appropriate amount of natural FGF5 protein, add an appropriate amount of emulsifier, add a small amount of liquid paraffin, grind into a fine paste, add a small amount of sterilized remaining eye ointment base, grind evenly, add the remaining base in several portions, and add an eye drug preservative to make the total amount, and then sterilize and package to obtain the FGF5 protein eye ointment.

[0051] The mass ratio of anhydrous lanolin, white petrolatum and liquid paraffin is 8:1:1 to form the eye ointment base, the content of the drug preservative is 0.05wt%, the content of the emulsifier is 0.2wt%, and the final concentration of FGF5 protein is 4 ng to 4 mg / mL.

[0052] The emulsifier is carbomer, and the eye drug preservative is hydroxyethyl benzoate.

[0053] Example 3

[0054] The preparation method of the eye gel preparation comprises the following steps:

[0055] 1. First, dissolve the natural FGF5 protein with a small amount of dissolving agent;

[0056] 2. Adjust the osmotic pressure to 295 to 309 mOsm / l with an osmotic pressure regulator, adjust the pH to 6.8 to 8.2 with a pH regulator, add 0.05wt% of a preservative, mix and heat to dissolve, and swell 1.5wt% to 3.0wt% of a gel base to form a transparent cream, and then mix with the above-mentioned materials to obtain the eye gel preparation. The final concentration of FGF5 protein in the eye gel preparation is 4 ng to 4 mg / mL.

[0057] The gel base is carbomer, the dissolving agent is hydroxypropyl β-cyclodextrin, the osmotic pressure regulator is sodium chloride, the pH regulator is sodium hydroxide aqueous solution, and the preservative is benzalkonium chloride.

[0058] Test Example 1

[0059] Take 8 C57 mice of the same age, remove the central corneal epithelium, and prepare a corneal epithelial injury mouse model. Randomly divide into FGF5 administration group and control group. In the FGF5 administration group, drop the eye drops prepared in Example 1 (the concentration of FGF5 is 400 ng / ml), administer once every 4 hours, continuously administer for 48 hours, and at 0h, 12h, 24h, 36h, 48h; the control group is given an equal dose of PBS. The cornea is colored with fluorescein sodium, the area of injury and repair is measured, and the corneal epithelial repair rate is calculated. The experimental results are shown in Figure 1 and Figure 2 .

[0060] From Figure 1 and Figure 2 it can be seen that the eye drops prepared in Example 1 have the effect of promoting the repair of corneal epithelium.

[0061] Test Example 2

[0062] Cytotoxicity test

[0063] According to the instructions, the cell viability was determined using the Cell Counting Kit-8. The corneal epithelial cell line (TKE2) was seeded in a 96-well plate at 1×10 cells / well overnight, and then treated with different concentrations of rhFGF5 (0, 2.5, 5, 10, 20, 40, 80, 160 ng / ml). After 24 hours, the medium containing 10% CCK8 was replaced and incubated at 37°C in the dark for 1-4 hours. The absorbance at 450 nm was measured using a microplate reader. The experimental results are shown in Figure 3 .

[0064] From Figure 3 the results can be seen that FGF5 has a cell proliferation-promoting effect and is not cytotoxic.

[0065] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. For those skilled in the art, the present application can have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. The use of fibroblast growth factor 5 in the preparation of a medicament for treating corneal epithelial damage, characterized in that, The concentration of the fibroblast growth factor 5 is 4 ng / mL to 4 mg / mL; the fibroblast growth factor 5 is natural fibroblast growth factor 5 or reconstituted fibroblast growth factor 5.

2. The application according to claim 1, characterized in that, The drug is an eye drop, eye ointment, or ophthalmic gel preparation.

3. The application according to claim 2, characterized in that, The eye drops also include a solvent, a pH adjuster, an osmotic pressure adjuster, a stabilizer, and water.

4. The application according to claim 2, characterized in that, The ointment also includes an ointment base, an emulsifier, and / or an ophthalmic antibacterial agent.

5. The application according to claim 2, characterized in that, The ophthalmic gel formulation also includes a gel matrix, a solvent, an osmotic pressure regulator, a pH regulator, a preservative, and water.

6. An application according to claim 5, characterized in that, The solvent is propylene glycol and / or hydroxypropyl β-cyclodextrin.

7. The application according to claim 4, characterized in that, The antibacterial agent in the ophthalmic medication is ethylparaben or benzalkonium chloride.

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