Application of atropine in treatment of fundus lesion

Abstract

The invention discloses application of atropine in treating fundus lesion. It is found for the first time that atropine can relieve apoptosis of mouse retina ONL and RPE cells, it is indicated that atropine can relieve fundus lesion, especially diabetic retinopathy, a new thought is provided for treating fundus lesion, especially diabetic retinopathy, and the application has wide application prospects clinically.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to the application of atropine in the treatment of fundus lesions. Background Art

[0002] Fundus lesions include inflammations, tumors, various vascular diseases, various degenerative diseases, and ocular lesions caused by multi-system diseases of the retina, choroid, optic nerve, and vitreous. They not only have a wide variety but also cause great damage to visual function. Currently, common and severely vision-impairing fundus diseases include diabetic retinopathy, age-related macular degeneration, retinal vein occlusion, etc., and significant progress has been made in the clinical research of these diseases.

[0003] Diabetic retinopathy (DR) is one of the most common and severe microvascular complications of diabetes mellitus (DM) and has become the main cause of blindness in the working-age population worldwide. The retina is a complex system composed of the retinal pigment epithelium (RPE) and the retinal neurosensory layer, which from outside to inside are: retinal pigment epithelium (RPE), photoreceptor layer, outer limiting membrane (OLM), outer nuclear layer (ONL), outer plexiform layer (OPL), inner nuclear layer (INL), inner plexiform layer (IPL), ganglion cell layer (GCL), nerve fiber layer (NFL), and inner limiting membrane (ILM).

[0004] Traditional treatment methods include surgical treatments such as laser treatment and vitrectomy, and drug treatment. Surgical treatment has certain risks, while the cost of intravitreal injection of anti-VEGF drugs is high, and repeated injections may also cause retinal neurodegeneration, choroidal capillary atrophy, macular scar formation, and even seriously affect vision. Therefore, finding a safe and effective drug is of great significance for conquering this disease. Summary of the Invention

[0005] To make up for the deficiencies of the prior art, the present invention provides the application of atropine in the treatment of fundus lesions.

[0006] To achieve the above object, the present invention adopts the following technical solutions: The first aspect of the present invention provides the application of atropine in the preparation of a pharmaceutical composition for preventing / treating fundus lesions.

[0007] Further, the fundus lesion is diabetic retinopathy.

[0008] Further, the pharmaceutical composition further includes other drugs for treating fundus lesions.

[0009] Further, the other drugs for treating fundus lesions include anti-VEGF drugs, glucocorticoids, and antioxidants.

[0010] Further, the pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0011] Further, the pharmaceutically acceptable excipients include excipients, buffers, surfactants, and preservatives.

[0012] Further, the excipients include viscosity enhancers and antioxidants.

[0013] Further, the dosage form of the pharmaceutical composition includes solid preparations, semi-solid preparations, or liquid preparations.

[0014] Further, the solid preparations include tablets, troches, capsules, pills, or granules.

[0015] Further, the semi-solid preparations include creams, gels, ointments, or emulsions.

[0016] Further, the liquid preparations include solutions or suspensions.

[0017] Further, the administration method of the pharmaceutical composition includes systemic administration or local administration.

[0018] Further, the administration method is local administration.

[0019] Further, the local administration is ocular administration.

[0020] The second aspect of the present invention provides a pharmaceutical composition for preventing / treating fundus lesions, and the pharmaceutical composition includes atropine.

[0021] Further, the fundus lesion is diabetic retinopathy.

[0022] The third aspect of the present invention provides the use of atropine in the preparation of a drug for inhibiting the apoptosis of ONL cells and / or RPE cells.

[0023] The fourth aspect of the present invention provides a method for in vitro regulating the apoptosis of ONL cells and / or RPE cells, and the method includes administering atropine.

[0024] Further, the method is a method for non-therapeutic purposes.

[0025] Advantages and beneficial effects of the present invention: The present application for the first time discovers that atropine can reduce the apoptosis of ONL and RPE cells in the retina of mice, indicating that atropine can relieve fundus lesions, especially diabetic retinopathy. The present application provides a new idea for treating fundus lesions, especially diabetic retinopathy, and has broad application prospects in clinical practice. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a graph showing the correlation between low choroid - retina volume ratio and DR in DM mice. Among them, 1A is a schematic diagram of the animal modeling experiment process, 1B is the wide - angle color fundus image (CF), fundus autofluorescence image (AF) and OCTA 6×6 mm 2 scanning image (the yellow arrow indicates the temporal side, n = 7) of DM mice, 1C is a representative HE - stained image of the retina tissue of DM mice (n = 6), and 1D is a TUNEL - stained image for detecting apoptosis of the retina tissue of DM mice (n = 6). DETAILED DESCRIPTION OF THE EMBODIMENTS

[0027] The following provides definitions of some terms used in this specification. Unless otherwise specified, all technical and scientific terms used herein generally have the same meaning as commonly understood by those of ordinary skill in the art to which this invention pertains.

[0028] The present invention provides the use of atropine in the preparation of a pharmaceutical composition for preventing / treating fundus diseases.

[0029] In some embodiments, the atropine also includes pharmaceutically acceptable salts of atropine. Pharmaceutically acceptable salts of atropine refer to some salt forms of atropine used in pharmaceutical preparations, and these salts are considered safe, effective and suitable for use in preparations in pharmacy. They include, but are not limited to, one or more of atropine sulfate, atropine hydrobromide, atropine hydrochloride, and atropine phosphate.

[0030] In some embodiments, the fundus diseases include, but are not limited to, diabetic retinopathy (DR), age - related macular degeneration (AMD), retinal vein occlusion, retinitis pigmentosa (RP), and glaucoma - related retinopathy.

[0031] In a specific embodiment, the fundus disease is selected from diabetic retinopathy (DR).

[0032] The pharmaceutical composition further includes other drugs for treating fundus diseases.

[0033] In some embodiments, the other drugs for treating fundus diseases include anti - VEGF drugs, glucocorticoids, and antioxidants.

[0034] The anti - VEGF drugs include, but are not limited to, bevacizumab, ranibizumab, and conbercept; the glucocorticoids include, but are not limited to, triamcinolone acetonide and dexamethasone; the antioxidants include, but are not limited to, vitamin C and vitamin E.

[0035] The pharmaceutical composition further includes pharmaceutically acceptable excipients.

[0036] In some embodiments, pharmaceutically acceptable excipients are used to refer to a material that is compatible with a recipient, preferably a mammal, more preferably a human, and is suitable for delivering an active agent to a target site without terminating the activity of the agent. The toxicity or side effects (if any) associated with the pharmaceutically acceptable excipients are preferably commensurate with the reasonable risk / benefit ratio for the intended use of the active agent. Pharmaceutically acceptable excipients include, but are not limited to, excipients, buffers, surfactants, and / or preservatives.

[0037] Examples of excipients include one or more viscosity-imparting agents. Viscosity-imparting agents refer to one or more relatively non-toxic chemical compounds or agents that change the viscosity of a pharmaceutical ingredient and / or formulation. Representative examples of viscosity-imparting agents include petrolatum, liquid paraffin, light liquid paraffin, castor oil, mineral oil, cottonseed oil, soybean oil, sesame oil, corn oil, petroleum resin, polyethylene glycol, glycerol, polybutene, rosin, polyvinyl alcohol, polystyrene, polyacrylic acid, propylene glycol, piperonyl butoxide, hypromellose, talc, gelatin, hydrogenated rosin glyceride, aliphatic hydrocarbon resin, benzyl acetate, copal resin, silicic acid, silicone, dimethylpolysiloxane, magnesium aluminum silicate, xanthan gum, sodium chondroitin sulfate, cyclodextrin, carboxyvinyl polymer, sodium alginate, propylene glycol alginate, carrageenan, sodium carboxymethyl cellulose, glucono delta-lactone, squalene, stearyl alcohol, aluminum stearate, lanolin, cetyl alcohol, gelatin, sorbitol, dextran, dextrin, tragacanth, palmitic acid, hyaluronate, hydroxyethyl cellulose, hydroxyethyl methyl cellulose, hydroxypropyl cellulose, butylene glycol, polyoxyethylene polyoxypropylene glycol, polysorbate, sodium metaphosphate, methyl cellulose, methyl vinyl ether maleic anhydride copolymer, locust bean gum, or cellulose polymers, etc.

[0038] Other representative examples of excipients include one or more antioxidants (such as thiosulfates, sodium thiosulfate, sodium formaldehyde sulfoxylate, sodium formaldehyde sulfoxylate dihydrate, etc.) and tonicity adjusters (such as sodium chloride, etc.).

[0039] Examples of buffers include, but are not limited to, acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, hydrochloric acid - potassium chloride, glycine, aconitic acid, citric acid - phosphoric acid, succinic acid, phthalic acid, maleic acid, cacodylic acid, tris (tris (hydroxymethyl) aminomethane), barbituric acid, borax, 2 - amino - 2 - methyl - 1,3 - propanediol (Ammediol), sodium carbonate - sodium bicarbonate, HEPES (4 - (2 - hydroxyethyl) - 1 - piperazineethanesulfonic acid), ACES (N - (2 - acetamido) - 2 - aminoethanesulfonic acid), ADA (N - (2 - acetamido)iminodiacetic acid), BES (N,N - bis(2 - hydroxyethyl) - 2 - aminoethanesulfonic acid), Bicine (N,N - bis(2 - hydroxyethyl)glycine), Bis - tris (bis(2 - hydroxyethyl)iminotris(hydroxymethyl)methane), CAPS (N - cyclohexyl - 3 - aminopropanesulfonic acid), CAPSO (N - cyclohexyl - 2 - hydroxy - 3 - aminopropanesulfonic acid), CHES (N - cyclohexyl - 2 - aminoethanesulfonic acid), DIPSO (3 - [N,N - bis(2 - hydroxyethyl)amino] - 2 - hydroxypropanesulfonic acid), EPPS (3 - [4 - (2 - hydroxyethyl) - 1 - piperazinyl]propanesulfonic acid), HEPES - Na (2 - [4 - (2 - hydroxyethyl) - 1 - piperazinyl]ethanesulfonate), HEPPSO (2 - hydroxy - 3 - [4 - (2 - hydroxyethyl) - 1 - piperazinyl]propanesulfonic acid monohydrate), MES (2 - morpholinoethanesulfonic acid monohydrate), MOPS (3 - morpholinopropanesulfonic acid), MOPSO (2 - hydroxy - 3 - morpholinopropanesulfonic acid), PIPES (piperazine - 1,4 - bis(2 - ethanesulfonic acid)), POPSO (piperazine - 1,4 - bis(2 - hydroxy - 3 - propanesulfonic acid) dihydrate), TAPSO (2 - hydroxy - N - tris(hydroxymethyl)methyl - 3 - aminopropanesulfonic acid), TES (N - tris(hydroxymethyl)methyl - 2 - aminoethanesulfonic acid), Tricine (N - [tris(hydroxymethyl)methyl]glycine), hydrochloric acid; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, and sodium lactate; and buffers such as citrate / glucose, sodium bicarbonate, and ammonium chloride, and citrate, phosphate, borate, bicarbonate, sodium salts, or potassium salts, including combinations thereof.

[0040] Examples of surfactants include, but are not limited to, sorbitan fatty acid esters of oleic acid (such as polysorbate 80 or Tween 20 and 80), polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, cremophor, sodium alkylbenzene sulfonate, glycerol, lecithin, sucrose esters, polyoxyethylene alkyl ethers, polyoxyethylene stearates, polyoxyl 40 stearate, ethylene glycol monostearate, polyethylene glycol monostearate, polymers of oxyethylated octylphenol (tyloxapol), propylene glycol, benzyl alcohol, macrogol, cyclodextrin, dibutylhydroxytoluene, sorbitol, trometamol, propylene glycol, mannitol, and polyoxyethylene polyoxypropylene glycols (such as polyoxyethylene (160) polyoxypropylene (30) glycol, or polyoxyethylene (200) polyoxypropylene (70) glycol), or combinations thereof. In some specific embodiments, the ophthalmic composition of the present invention comprises, consists essentially of, or further consists of: polysorbate 80, polyoxyethylene hydrogenated castor oil, lecithin, or combinations thereof.

[0041] Examples of preservatives include, but are not limited to, imidazolidinyl urea, methylparaben, propylparaben, phenoxyethanol, disodium EDTA, benzalkonium chloride, thimerosal, chlorobutanol, sorbic acid, and combinations.

[0042] In some embodiments, the dosage form of the pharmaceutical composition includes solid preparations, semi-solid preparations or liquid preparations. The solid preparations include tablets, troches, capsules, pills or granules; the semi-solid preparations include creams, gels, ointments or emulsions; the liquid preparations include solutions or suspensions.

[0043] In some embodiments, the administration route includes systemic administration or local administration.

[0044] In a preferred embodiment, the administration route is local administration. Further, the local administration is ophthalmic administration, and the ophthalmic administration methods include, but are not limited to, ocular surface administration (eye drops), injection administration (subconjunctival injection, intravitreal injection, periocular injection).

[0045] In a preferred embodiment, the dosage forms for ophthalmic administration include, but are not limited to, eye drops, eye ointments, ophthalmic sprays, implantable tablets, ophthalmic gels, eye patches, ophthalmic microspheres, ophthalmic sustained-release preparations, periocular injections, and intraocular injections.

[0046] In some embodiments, the pharmaceutical composition further comprises instructions for administering the ophthalmic preparation to the eyes of a subject in need thereof. In some embodiments, the pharmaceutical composition is provided or packaged in multiple dosage forms. In other embodiments, the pharmaceutical composition contains a preservative that prevents microbial contamination during use (i.e., repeated use). Among them, the administration instructions provide dosing instructions. In various embodiments, the instructions may be to administer the ophthalmic preparation once, twice, or three times a day. In embodiments where the pharmaceutical composition is a liquid preparation, the administration may be to place one, two, three, or more drops in one or both eyes (e.g., if one eye is affected by an ocular condition, both eyes may be treated, or if both eyes are affected by the condition), once, twice, three times, or more times a day.

[0047] In some embodiments, the appropriate dosing of the pharmaceutical composition can be prescribed in a variety of ways depending on factors such as the formulation method, the mode of administration, the age, weight, sex, morbidity, diet, administration time, administration route, excretion rate, and sensitivity of the patient. Skilled physicians can usually easily determine the prescription and the dosing that is effective for the desired treatment or prevention. The therapeutically effective amount and specific treatment regimen for a particular subject (such as a mammal: human) can be affected by many factors, including the pharmacodynamic activity of the drug used, the age, weight, general condition, sex, diet, administration time, disease susceptibility, disease progression, and the judgment of the treating physician. In addition, those skilled in the art are aware that the mode of administration, dosage form, and dosing of a drug are affected by many factors such as the age, weight, sex, morbidity, diet, administration time, excretion rate, and sensitivity of the patient. Therefore, the mode of administration, dosage form, and dosing of the ophthalmic preparation or pharmaceutical composition described in this application are not limited to those described in the embodiments of this application.

[0048] The present invention will be further illustrated below with specific examples. It should be understood that the specific embodiments described herein are presented by way of example and are not intended to limit the present invention. Without departing from the scope of the present invention, the main features of the present invention can be used in various embodiments.

[0049] Examples

[0050] 1. Materials and Methods 1.1 Experimental Animals Three-week-old male C57BL / 6J mice were purchased from Guangzhou Regene Biotech Co., Ltd. Atropine eye drops at a concentration of 2% were given to one eye twice a day, while the contralateral eye received saline eye drops as a control. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ) at 55 mg / kg, dissolved in sodium citrate buffer (pH 4.5), for 5 consecutive days. Blood glucose was measured 1 week after induction, and blood glucose ≥ 16.7 mmol / L confirmed diabetes.

[0051] 1.2 OCTA Image Acquisition and Analysis All optical coherence tomography angiography (OCTA) images were taken using a BM-400K (BMizar, TowardPi Medical Technology Co., Ltd., Beijing, China) at the same time period of the day (8:00 - 17:00). For C57BL / 6J mice, after anesthesia by intraperitoneal injection of 2% tribromoethanol at a dose of 300 mg / kg, OCTA imaging was performed with the optic nerve head (ONH) as the center using a 6×6 mm 2 scanning mode.

[0052] 1.3 Fundus Photography and Autofluorescence Images After induction of anesthesia in mice, wide-angle color fundus imaging and autofluorescence images centered on the ONH were obtained using an Optos 200 Tx (Optos PLC, Dunfermline, United Kingdom).

[0053] 1.4 H&E Staining and TUNEL Assay The mice were euthanized, and the eyeballs were removed and fixed with FAS eye fixative (Servicebio) for 24 hours. Tissue sections of 4 μm were prepared. Subsequently, hematoxylin and eosin (H&E) staining was performed using an H&E staining kit (Solarbio) according to the manufacturer's instructions.

[0054] 1.5 Statistical Analysis Paired t-tests were used to analyze the animal experiment data. The P-values were two-sided, and P < 0.05 was considered statistically significant.

[0055] 2. Experimental Results While maintaining atropine or saline eye drops, diabetes was induced by intraperitoneal injection of STZ ( Figure 1 A). Color fundus photography showed no significant difference between the two eyes. However, fundus autofluorescence imaging showed that there were fewer punctate hyperfluorescent lesions in the experimental eyes compared with the control group, indicating that atropine reduced the number of abnormal RPE cells in DM mice. In addition, OCTA showed that the choroid in the eyes treated with atropine was thicker than that in the control group ( Figure 1B). HE staining showed that the outer nuclear layer (ONL) of DM mice was arranged loosely and disorderly, and this situation was alleviated after atropine treatment ( Figure 1 C).

[0056] Increased apoptosis of retinal cells is a key feature of early DR. Here, this application demonstrated that atropine could reduce apoptosis of the outer retina ONL and RPE in DM mice ( Figure 1 D). Generally speaking, these data indicate that atropine can inhibit the progression of DR in DM mice.

[0057] The description of the above embodiments is only for understanding the method and its core idea of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. Use of atropine in the preparation of a pharmaceutical composition for preventing / treating fundus lesions; Preferably, the fundus lesion is diabetic retinopathy.

2. The use according to claim 1, characterized in that: The pharmaceutical composition also includes other drugs for treating fundus lesions; Preferably, the other drugs for treating fundus lesions include anti-VEGF drugs, glucocorticoids, and antioxidants.

3. The use according to claim 1, characterized in that: The pharmaceutical composition also includes pharmaceutically acceptable excipients.

4. The use according to claim 3, characterized in that: The pharmaceutically acceptable excipients include excipients, buffers, surfactants, and preservatives; Preferably, the excipients include a viscosity imparting agent and an antioxidant.

5. The use according to claim 1, characterized in that: The dosage form of the pharmaceutical composition includes solid preparation, semisolid preparation or liquid preparation.

6. The use according to claim 5, characterized in that: The solid preparations include tablets, lozenges, capsules, pills or granules; Preferably, the semisolid preparation comprises a cream, a gel, an ointment or an emulsion; Preferably, the liquid preparation comprises a solution or a suspension.

7. The use according to claim 1, characterized in that: The administration of the pharmaceutical composition includes systemic administration or local administration; Preferably, the administration method is topical administration; Preferably, the topical administration is ocular administration.

8. A pharmaceutical composition for preventing / treating fundus lesions, characterized in that: The pharmaceutical composition includes atropine; Preferably, the fundus lesion is diabetic retinopathy.

9. Use of atropine in the preparation of drugs for inhibiting apoptosis of ONL cells and / or RPE cells.

10. A method for regulating apoptosis of ONL cells and / or RPE cells in vitro, characterized in that: The method comprises administering atropine.

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