Application of a cholinergic receptor antagonist in preventing myopia and preparation method thereof

Abstract

The present invention belongs to the field of biomedicine and relates to the use of a cholinergic receptor antagonist in preventing or treating myopia. The present invention provides a compound for use in preparing a drug for preventing, delaying, inhibiting and / or treating myopia or a disease related to myopia, wherein the compound is selected from bencyclonate hydrochloride or a salt thereof. The present invention has found that bencyclonate hydrochloride can inhibit the remodeling of the sclera, thereby inhibiting the increase in axial length and controlling the development of myopia.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine and relates to the application of a cholinergic receptor antagonist in preventing or treating myopia. Background Art

[0002] Myopia is one of the most common human ophthalmic diseases. In recent years, the incidence of myopia has increased rapidly and has become a global medical health problem. Myopia is a refractive disorder that is usually caused by a mismatch between the refractive power and axial length of the eye, causing the image to fall in front of the retinal photoreceptors, resulting in blurred vision. High myopia can increase the risk of potentially blinding eye diseases such as glaucoma, cataracts, retinal detachment, and post-staphylococcal infection.

[0003] Studies have found that cholinergic antagonists such as atropine and pirenzepine can effectively inhibit the progression of myopia, but at the same time, studies have shown that not all cholinergic antagonists have the ability to inhibit the development of myopia.

[0004] The sclera is a connective tissue composed of fibroblasts embedded in an extracellular matrix (ECM) with a large amount of type I collagen and proteoglycans, which determines the shape and axial length of the eye. Studies have shown that five mAChRs receptor subtypes are expressed in human scleral fibroblasts. Experiments with scleral thinning and weakening of the extracellular matrix have shown that scleral elongation in experimental myopia involves tissue remodeling through changes in scleral cells and molecules. The sclera is the final effector of the development of myopia. After myopia occurs, the expression of scleral collagen is reduced, the collagen fibers are disordered, the longitudinal fibers become thinner, and the transverse fibers separate or disappear, causing scleral thinning. Summary of the invention

[0005] In some embodiments, the present invention provides a use of a compound in the preparation of a drug for preventing, delaying, inhibiting and / or treating myopia or a disease related to myopia, wherein the compound is selected from phencyclnonate hydrochloride or a salt thereof.

[0006] In some embodiments, the compound is selected from phencyclnonate hydrochloride.

[0007] In some embodiments, the phencyclonate hydrochloride is selected from levorotatory phencyclonate hydrochloride, dextrorotatory phencyclonate hydrochloride or racemic phencyclonate hydrochloride.

[0008] In some embodiments, the structural formula of the compound is as shown in formula (I) levorotatory benzylcyclononyl hydrochloride (or "(-) benzylcyclononyl hydrochloride"), formula (II) dextrorotatory benzylcyclononyl hydrochloride (or "(+) benzylcyclononyl hydrochloride") or formula (III) racemic benzylcyclononyl hydrochloride:

[0009]

[0010] In some embodiments, the myopia is myopia caused by an acquired elongation of the anterior-posterior axial length of the eye.

[0011] In some embodiments, the drug prevents, delays, inhibits and / or treats myopia or myopia-related diseases by inhibiting scleral remodeling or increasing scleral thickness or increasing the expression of collagen fibers, or inhibiting the elongation of the axial length.

[0012] In some embodiments, the present invention demonstrates that intravitreal injection of phencyclnonate hydrochloride can regulate the synthesis of scleral extracellular matrix, thereby inhibiting scleral remodeling.

[0013] In some embodiments, the present invention demonstrates that phencyclnonate hydrochloride may inhibit the development of myopia by inhibiting the synthesis of type I collagen, a major component of the scleral extracellular matrix, and inhibiting the remodeling of the sclera.

[0014] Atropine is a non-selective muscarinic receptor antagonist that can bind to all five muscarinic receptor subtypes and prevent acetylcholine from binding to them. So far, atropine has been proven by the prior art to be a potential myopia treatment drug and has significantly inhibited the development of myopia in multiple clinical trials. In some embodiments, the inventors of the present invention have found that phencyclonate hydrochloride, as a muscarinic receptor antagonist, also has the ability to inhibit the development of myopia, and L-phencyclonate hydrochloride has relative selectivity for the five muscarinic receptor subtypes, and has a higher selectivity for M1, M3, and M4 receptors, and may have smaller side effects than atropine, and has very good application prospects.

[0015] In some embodiments, the medicament includes an external preparation, an ophthalmic preparation, an injection, an oral preparation, or a lyophilized powder injection.

[0016] In some embodiments, the ophthalmic preparation includes eye drops, eye ointments, eye sprays, implants, eye gels, eye patches, eye microspheres, eye sustained-release preparations, or eye injections.

[0017] In some embodiments, the ophthalmic formulation includes a pharmaceutically acceptable excipient.

[0018] In some embodiments, the pharmaceutically acceptable excipients include a pH adjuster, a thickener, an osmotic pressure regulator, or a bacteriostatic agent.

[0019] In some embodiments, the pH adjuster is selected from one or more of disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, tartaric acid, acetic acid, sodium acetate, boric acid and its salts, borax, citric acid, and sodium citrate; preferably, the thickener is selected from one or more of dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerol, glucosamine polymer, chondroitin sulfate, gum arabic, tragacanth, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropyl methylcellulose, polyvinyl alcohol, polyethylene glycol, dextran, polyvinyl pyrrolidone, polypropylene alcohol, and sodium hyaluronate, but is not limited thereto.

[0020] In some embodiments, the osmotic pressure regulator is selected from one or more of mannitol, sorbitol, sodium chloride, potassium chloride, sodium sulfate, potassium sulfate, sodium nitrate, potassium nitrate, propylene glycol, glucose or glycerol.

[0021] In some embodiments, the antibacterial agent is selected from one or more of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosal, benzalkonium chloride, or benzalkonium bromide.

[0022] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.1-10% w / v.

[0023] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-10% w / v.

[0024] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-5% w / v.

[0025] In some embodiments, the myopia is selected from refractive myopia, axial myopia, congenital myopia, early-onset myopia, delayed myopia, late-onset myopia, low myopia, moderate myopia, high myopia, pseudomyopia, true myopia, myopia in minors, myopia in adults, myopia in the elderly, simple myopia, pathological myopia, primary myopia, or secondary myopia.

[0026] In some embodiments, the present invention provides a use of a compound in the preparation of a medicament for inhibiting axial elongation of the eye, wherein the compound is selected from phencyclnonate.

[0027] In some embodiments, the compound is selected from phencyclnonate hydrochloride.

[0028] In some embodiments, the phencyclonate hydrochloride is selected from levorotatory phencyclonate hydrochloride, dextrorotatory phencyclonate hydrochloride or racemic phencyclonate hydrochloride.

[0029] In some embodiments, the structural formula of the compound is as shown in Formula (I) levorotatory benzylcyclopentane hydrochloride, Formula (II) dextrorotatory benzylcyclopentane hydrochloride or Formula (III) racemic benzylcyclopentane hydrochloride:

[0030]

[0031] In some embodiments, the medicament includes an external preparation, an ophthalmic preparation, an injection, an oral preparation, or a lyophilized powder injection.

[0032] In some embodiments, the ophthalmic preparation includes eye drops, eye ointments, eye sprays, implants, eye gels, eye patches, eye microspheres, eye sustained-release preparations, or eye injections.

[0033] In some embodiments, the ophthalmic formulation includes a pharmaceutically acceptable excipient.

[0034] In some embodiments, the pharmaceutically acceptable excipients include a pH adjuster, a thickener, an osmotic pressure regulator, or a bacteriostatic agent.

[0035] In some embodiments, the pH adjuster is selected from one or more of disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, tartaric acid, acetic acid, sodium acetate, boric acid and its salts, borax, citric acid, and sodium citrate; preferably, the thickener is selected from one or more of dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerol, glucosamine polymer, chondroitin sulfate, gum arabic, tragacanth, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropyl methylcellulose, polyvinyl alcohol, polyethylene glycol, dextran, polyvinyl pyrrolidone, polypropylene alcohol, and sodium hyaluronate.

[0036] In some embodiments, the osmotic pressure regulator is selected from one or more of mannitol, sorbitol, sodium chloride, potassium chloride, sodium sulfate, potassium sulfate, sodium nitrate, potassium nitrate, propylene glycol, glucose or glycerol.

[0037] In some embodiments, the antibacterial agent is selected from one or more of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosal, benzalkonium chloride, or benzalkonium bromide.

[0038] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.1-10% w / v.

[0039] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-10% w / v.

[0040] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-5% w / v.

[0041] In some embodiments, the present invention provides a pharmaceutical composition for preventing and / or treating myopia, comprising the compound phencyclnonate and one or more pharmaceutically acceptable substances.

[0042] In some embodiments, the compound is selected from phencyclnonate hydrochloride.

[0043] In some embodiments, the phencyclonate hydrochloride is selected from levorotatory phencyclonate hydrochloride, dextrorotatory phencyclonate hydrochloride or racemic phencyclonate hydrochloride.

[0044] In some embodiments, the structural formula of the compound is as shown in Formula (I) levorotatory benzylcyclopentane hydrochloride, Formula (II) dextrorotatory benzylcyclopentane hydrochloride or Formula (III) racemic benzylcyclopentane hydrochloride:

[0045]

[0046] In some embodiments, the pharmaceutically acceptable substance or substances include a pharmaceutically acceptable excipient.

[0047] In some embodiments, the pharmaceutically acceptable excipients include a pH adjuster, a thickener, an osmotic pressure regulator, or a bacteriostatic agent.

[0048] In some embodiments, the pH adjuster is selected from one or more of disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, tartaric acid, acetic acid, sodium acetate, boric acid and its salts, borax, citric acid, and sodium citrate; preferably, the thickener is selected from one or more of dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerol, glucosamine polymer, chondroitin sulfate, gum arabic, tragacanth, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropyl methylcellulose, polyvinyl alcohol, polyethylene glycol, dextran, polyvinyl pyrrolidone, polypropylene alcohol, and sodium hyaluronate.

[0049] In some embodiments, the osmotic pressure regulator is selected from one or more of mannitol, sorbitol, sodium chloride, potassium chloride, sodium sulfate, potassium sulfate, sodium nitrate, potassium nitrate, propylene glycol, glucose or glycerol.

[0050] In some embodiments, the antibacterial agent is selected from one or more of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosal, benzalkonium chloride, or benzalkonium bromide.

[0051] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.1-10% w / v.

[0052] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-10% w / v.

[0053] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-5% w / v.

[0054] In some embodiments, the myopia is selected from refractive myopia, axial myopia, congenital myopia, early-onset myopia, delayed myopia, late-onset myopia, low myopia, moderate myopia, high myopia, pseudomyopia, true myopia, myopia in minors, myopia in adults, myopia in the elderly, simple myopia, pathological myopia, primary myopia, or secondary myopia.

[0055] In some embodiments, the present invention provides a pharmaceutical preparation for preventing and / or treating myopia, comprising phencyclnonate and a pharmaceutically acceptable excipient.

[0056] In some embodiments, the compound is selected from phencyclnonate hydrochloride.

[0057] In some embodiments, the phencyclonate hydrochloride is selected from levorotatory phencyclonate hydrochloride, dextrorotatory phencyclonate hydrochloride or racemic phencyclonate hydrochloride.

[0058] In some embodiments, the structural formula of the compound is as shown in Formula (I) levorotatory benzylcyclopentane hydrochloride, Formula (II) dextrorotatory benzylcyclopentane hydrochloride or Formula (III) racemic benzylcyclopentane hydrochloride:

[0059]

[0060] In some embodiments, the pharmaceutically acceptable excipients include a pH adjuster, a thickener, an osmotic pressure regulator, or a bacteriostatic agent.

[0061] In some embodiments, the pH adjuster is selected from one or more of disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, tartaric acid, acetic acid, sodium acetate, boric acid and its salts, borax, citric acid, and sodium citrate; preferably, the thickener is selected from one or more of dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerol, glucosamine polymer, chondroitin sulfate, gum arabic, tragacanth, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropyl methylcellulose, polyvinyl alcohol, polyethylene glycol, dextran, polyvinyl pyrrolidone, polypropylene alcohol, and sodium hyaluronate.

[0062] In some embodiments, the osmotic pressure regulator is selected from one or more of mannitol, sorbitol, sodium chloride, potassium chloride, sodium sulfate, potassium sulfate, sodium nitrate, potassium nitrate, propylene glycol, glucose or glycerol.

[0063] In some embodiments, the antibacterial agent is selected from one or more of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosal, benzalkonium chloride, or benzalkonium bromide.

[0064] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.1-10% w / v.

[0065] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-10% w / v.

[0066] In some embodiments, the content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-5% w / v.

[0067] In some embodiments, the pharmaceutical preparation includes an external preparation, an ophthalmic preparation, an injection, an oral preparation, or a lyophilized powder injection.

[0068] In some embodiments, the ophthalmic preparation includes eye drops, eye ointments, eye sprays, implants, eye gels, eye patches, eye microspheres, eye sustained-release preparations, or eye injections.

[0069] In some embodiments, the eye drops further include water.

[0070] In some embodiments, the eye drops include 1% w / v phencyclnonate, 0.1% w / v hypromellose, an appropriate amount of dihydrogen phosphate, and an appropriate amount of sterile water for injection.

[0071] In some embodiments, the ophthalmic injection is selected from intravitreal injection.

[0072] In some embodiments, the present invention provides use of the pharmaceutical composition or the pharmaceutical preparation in preparing a product for relieving visual fatigue. BRIEF DESCRIPTION OF THE DRAWINGS

[0073] Figure 1 Schematic diagram of establishing the form deprivation myopia model in guinea pigs; the left side shows the guinea pigs in the blank control group, and the right side shows the guinea pigs in the form deprivation group.

[0074] Figure 2 Figure 2 shows the results of hematoxylin-eosin staining (HE) of guinea pig eye sclera tissue after 14 days of experimental intervention, where n=3.

[0075] Figure 3 Figure 2 shows the results of Sirius red staining of guinea pig eye sclera tissue after 14 days of experimental intervention, n=3.

[0076] Figure 4A-4B The expression of collagen type I was determined by Western Blotting. Figure 4A Shows Western Blotting bands, Figure 4B The results of quantitative analysis of the bands are shown, where n = 6. Note: FDM: form deprivation;

[0077] PCH: racemic phencyclonate hydrochloride; S(+)-PCH: dextrorotatory phencyclonate hydrochloride; R(-)-PCH: levorotatory phencyclonate hydrochloride. DETAILED DESCRIPTION

[0078] The technical solution of the present invention is further described below by specific embodiments, which do not limit the protection scope of the present invention. Some non-essential modifications and adjustments made by others based on the concept of the present invention still fall within the protection scope of the present invention.

[0079] As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0080] In addition, the use of "or" means "and / or" unless otherwise stated. Similarly, "comprising" and "including" are interchangeable and are not intended to be limiting. It should be further understood that where the term "comprising" is used to describe various embodiments, those skilled in the art will understand that in some specific cases, the language "consisting essentially of" or "consisting of" may be alternatively used to describe the embodiments.

[0081] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although many methods and reagents are similar or equivalent to those described herein, exemplary methods and materials are disclosed herein.

[0082] It is to be understood that the present disclosure is not limited to the particular methodology, protocols, reagents, etc. described herein and as such may vary. The terminology used herein is for the purpose of describing particular embodiments or aspects only and is not intended to limit the scope of the present disclosure.

[0083] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms that are suitable (within the scope of reasonable medical judgment) for use in contact with the tissues of patients without causing excessive toxicity, irritation, allergic response or other problems or complications (i.e., with a reasonable risk / benefit ratio).

[0084] The term "prevent" has an art-recognized meaning and, when used in connection with a condition, disease, syndrome or any other medical condition, its meaning is well known in the art and includes the administration of a drug, which reduces the frequency of occurrence of the medical condition in a patient who does not receive the drug, or delays the onset or symptoms of the condition.

[0085] As used herein, the term "treating" includes reversing, alleviating or inhibiting the symptoms, clinical features and underlying pathology of a disease in a manner that improves or stabilizes the patient's condition.

[0086] The term "prevention or treatment" has a meaning recognized in the art and includes administering one or more of the present invention to a recipient. If administered before the adverse condition has clinical manifestations (e.g., a disease or other adverse condition in the recipient animal), such treatment is preventive (i.e., to avoid the development of adverse conditions in the recipient), whereas, if administered after the adverse condition has clinical manifestations, such treatment is therapeutic (i.e., to reduce, improve or stabilize existing adverse conditions or their side effects).

[0087] Patent application 200410029597.9 is incorporated herein by reference in its entirety.

[0088] Main reagents :

[0089] Racemic benzylcyclohexane hydrochloride, levorotatory benzylcyclohexane hydrochloride (A) and dextrorotatory benzylcyclohexane hydrochloride (B) [synthesized by our company, see Chinese Patent 97125424.9 and Chinese Patent Application 200410029597.9]; sodium chloride injection (Shijiazhuang No. 4 Pharmaceutical Co., Ltd.); oxybuprocaine hydrochloride (Shanghai McLean Biochemical Technology Co., Ltd.); tropicamide (Shanghai McLean Biochemical Technology Co., Ltd.); chloramphenicol (Shanghai McLean Biochemical Technology Co., Ltd.); atropine sulfate (Shanghai McLean Biochemical Technology Co., Ltd.).

[0090] Among them, the structures of benzylcyclononyl hydrochloride (2′-cyclopentyl-2′-phenyl-2′-hydroxyacetic acid-9α-[3-methyl-3-azabicyclo(3.3.1)nonyl] ester hydrochloride) and its isomers, levorotatory benzylcyclononyl hydrochloride (I), dextrorotatory benzylcyclononyl hydrochloride (II) and racemic benzylcyclononyl hydrochloride (III) are shown in the following chemical structural formula.

[0091] Chemical formula C22 H 32 ClNO3; molecular weight is 393.9.

[0092] Method steps :

[0093] 1. Reagent preparation

[0094] (1) Preparation of atropine intravitreal injection: Dissolve 1 g of atropine powder in normal saline until it is completely dissolved, and dilute to 100 mL to prepare a 1% w / v atropine solution.

[0095] (2) Preparation of L-phencyclnonate hydrochloride intravitreal injection: Dissolve 1 g of L-phencyclnonate hydrochloride powder in physiological saline until completely dissolved, and dilute to 100 mL to prepare a 1% w / v L-phencyclnonate hydrochloride solution.

[0096] (3) Preparation of dextrorotatory phencyclonate hydrochloride intravitreal injection: Dissolve 1 g of dextrorotatory phencyclonate hydrochloride powder in physiological saline until completely dissolved, and dilute to 100 mL to prepare a 1% w / v dextrorotatory phencyclonate hydrochloride solution.

[0097] (4) Preparation of racemic phencyclonate hydrochloride intravitreal injection: Dissolve 1 g of racemic phencyclonate hydrochloride powder in physiological saline until it is completely dissolved, and dilute to 100 mL to prepare a 1% w / v racemic phencyclonate hydrochloride solution.

[0098] 2. Establishment and grouping of guinea pig form deprivation myopia model

[0099] (1) Tricolor guinea pigs weighing about 120 g (young guinea pigs of 2 to 3 weeks old) were randomly divided into 7 groups, each with 9 animals, including a blank control group, a monocular eye mask form deprivation group, a monocular eye mask form deprivation + saline group, a monocular eye mask form deprivation + atropine group, a monocular eye mask form deprivation + L-phencyclonate hydrochloride group, a monocular eye mask form deprivation + D-phencyclonate hydrochloride group, and a monocular eye mask form deprivation + racemic phencyclonate hydrochloride group. Except for the blank control group, the other six groups of guinea pigs were covered with a translucent eye mask on the right eye, leaving the left eye exposed. The eye mask covering was checked once a day to ensure that the experimental eye was completely covered and no pressure was caused on the eyeball. No treatment was given to the blank control group.

[0100] The schematic diagram of the guinea pig form deprivation myopia model is as follows: Figure 1 As shown, from Figure 1 It can be seen that the guinea pig form deprivation myopia model was successfully constructed.

[0101] The guinea pigs were anesthetized by intraperitoneal injection of sodium pentobarbital (1 mL / kg) and local ocular anesthesia with oxybuprocaine eye drops (0.01 mL). The guinea pigs were fixed and routinely disinfected, and different drugs were injected into the vitreous cavity of the right eyes of the guinea pigs in the experimental group under a stereomicroscope (5 minutes after oxybuprocaine topical anesthesia and immediately after sodium pentobarbital anesthesia).

[0102] Monocular form deprivation + saline group: 10 μL of saline was injected into the vitreous cavity using a microsyringe.

[0103] Monocular form deprivation + atropine group: 10 μL of 1% w / v atropine solution was injected into the vitreous cavity using a microsyringe.

[0104] Monocular form deprivation + L-phencyclnonate hydrochloride group: 10 μL of 1% w / v L-phencyclnonate hydrochloride solution was injected into the vitreous cavity using a microsyringe.

[0105] Monocular form deprivation + dextrorotatory phencyclonate hydrochloride group: 10 μL of 1% w / v dextrorotatory phencyclonate hydrochloride solution was injected into the vitreous cavity using a microsyringe.

[0106] Monocular form deprivation + racemic phencyclonate hydrochloride group: 10 μL of 1% w / v racemic phencyclonate hydrochloride solution was injected into the vitreous cavity using a microsyringe.

[0107] Example 1 Effect of Phencynonate on Refraction and Axial Length

[0108] Refractive power measurement: A strip retinoscope (model YZ24, Liuliu Company, Suzhou, China) was used to measure the refractive power of the left and right eyes of guinea pigs.

[0109] Axial length measurement: STRONG 6000A ophthalmic A-ultrasound measuring instrument was used, with an A-ultrasound frequency of 11MHz. Oxybuprocaine was used for corneal surface anesthesia before measurement, and measurement began 5 minutes after the effect. During measurement, the probe was aimed at the center of the cornea and perpendicular to the corneal plane. The ophthalmic A-ultrasound measuring instrument measured the axial length, and the average value was taken for 10 measurements each time, with an accuracy of 0.01mm.

[0110] The refractive power and axial length of each group of experimental animals were measured using a strip retinoscope and an ophthalmic A-ultrasound measuring instrument. The results are shown in Table 1. After 14 days of experimental intervention, compared with untreated guinea pigs, the refractive power of the right eye of the monocularly covered guinea pigs was (-3.05±0.24)D, which was a negative value, forming myopia; compared with the guinea pigs after monocularly covered, the refractive power of the right eye of the guinea pigs administered with atropine and phencyclonate hydrochloride and its optical isomers decreased less, and the degree of myopia was weaker. After 14 days of intervention, the axial length of the right eye of the monocularly covered guinea pigs was significantly longer than that of the control group. The axial lengths of the right eyes of the guinea pigs treated with atropine, phencyclnonate hydrochloride and its optical isomers were (9.83±0.21) mm, (9.92±0.32) mm, (9.91±0.26) mm, and (9.98±0.41) mm, respectively, which were shorter than the axial length of the monocularly covered guinea pigs [(10.09±0.12) mm].

[0111] In summary, after administration of atropine, phencyclnonate hydrochloride and its optical isomers, the refractive power of the guinea pig's right eye decreased slowly, and the axial length grew slowly. Among them, dextrorotatory phencyclnonate had the best effect in delaying the progression of myopia, which was equivalent to that of atropine.

[0112] Table 1

[0113]

[0114] Note: Table 1 shows the measurement data of refraction and ocular biology (axial length) at 0 and 14 days after the experimental intervention, where n=8.

[0115] Example 2 HE staining to detect the effect of phencyclonate hydrochloride on scleral thickness

[0116] (1) 14 days after the experimental intervention (after drug administration) according to the above-mentioned “Methods and Procedures”, the guinea pigs were euthanized, the right eyeballs were removed and placed in eyeball tissue fixative, embedded in paraffin, and serially sectioned along the sagittal axis with a section thickness of 6 μm;

[0117] (2) Preheat the slide oven to 98°C, place the paraffin sections in the oven, and wait for 20 min until the paraffin on the slide surface melts. Dewax the slides twice with xylene, 10 min each time.

[0118] (3) Place the paraffin sections in 100%-60% ethanol, debenzene for 1 min, and rinse with distilled water 3-4 times;

[0119] (4) Soak in hematoxylin for 5 minutes and rinse with running water for 5 minutes;

[0120] (5) 1% hydrochloric acid alcohol and 1% ammonia water for 1-2 seconds each, soak in eosin for 5 minutes, and then rinse with distilled water 3-4 times;

[0121] (6) Dehydrate with 70%, 80%, and 90% ethanol once each, dehydrate with 100% ethanol twice, and make transparent with xylene;

[0122] (7) Neutral resin sealing;

[0123] (8) The thickness of the posterior sclera was measured under an optical microscope. The thickness of the central area within 3 mm of the optic nerve of the eyeball wall was taken as the measurement area.

[0124] The results are as follows Figure 2 The results of HE staining showed that the scleral thickness of guinea pigs in the form deprivation group was significantly reduced, and the fibers were arranged in disorder and sparse. After the administration of atropine, phencyclonate hydrochloride and its optical isomers, the scleral thickness was significantly increased compared with the form deprivation group. Among them, dextrorotatory phencyclonate had the best effect on improving scleral thickness, which was equivalent to atropine. Therefore, the administration of atropine and each optical isomer of phencyclonate hydrochloride inhibited the development of myopia in guinea pigs.

[0125] Example 3 Sirius red staining to detect the effect of phencyclonate on collagen fibers

[0126] (1) The preliminary dewaxing operation is the same as the HE staining step of Example 2 (steps (1) to (3) of Example 2);

[0127] (2) Place the sections in picrosirius red staining solution for 8 min, and then dehydrate in two anhydrous ethanol solutions for 1 min each;

[0128] (3) Place the slices in xylene and make them transparent for 5 minutes;

[0129] (4) Neutral resin sealing;

[0130] (5) Observe the type I collagen fibers under a microscope.

[0131] The results are as follows Figure 3 The results of picrosirius red staining showed that the content of type I collagen fibers in guinea pigs in the form deprivation group was reduced. After administration of atropine, phencyclnonate hydrochloride and its optical isomers, the content of type I collagen fibers increased compared with the form deprivation group. Among them, the increase in collagen fibers after administration of dextrorotatory phencyclnonate hydrochloride was the greatest, which was equivalent to that of atropine.

[0132] Example 4 Western blot detection of the effect of phencyclonate hydrochloride on the expression of type I collagen (Collagen-I) protein

[0133] (1) After 14 days of experimental intervention according to the above-mentioned “Methods and Procedures”, the guinea pigs were euthanized, the eyeballs were quickly removed, the connective tissue wrapped in the eyeball wall was removed, the eyeballs were cut open at the equator behind the eyeballs, the anterior segment and vitreous body were removed, and the scleral tissue within 4 to 5 mm of the posterior pole was obtained;

[0134] (2) Grind the tissues with a high-speed cryogenic grinder to form tissue homogenates, and use RIPA lysis buffer to extract the total protein of each group of tissues;

[0135] (3) preparing 10% polyacrylamide gel;

[0136] (4) Sample loading: Insert the gel plate into the electrophoresis tank, add electrophoresis buffer to cover the gel plate, and then load the sample onto the wall of each well of the gel plate;

[0137] (5) Electrophoresis: Cover the electrophoresis tank, insert the electrodes, and set the electrophoresis conditions: constant voltage 60 V for the stacking gel and 150 V for the separation gel. Observe the marker to determine the electrophoresis time.

[0138] (6) Transfer: Cut a PVDF membrane with a size of 2 cm × 8 cm and a pore size of 0.45, and perform wet transfer. Remove the concentrated gel, and place the separation gel, filter paper, and PVDF membrane in a transfer electrophoresis tank. Set the transfer conditions to a constant current of 200 mA for 90 min.

[0139] (7) Blocking: The membrane was completely immersed in the rapid blocking solution and incubated on a horizontal shaker for 20 min;

[0140] (8) Antibody incubation: incubate with diluted primary antibody at 4°C overnight, wash for 5 min × 3 times;

[0141] (9) Secondary antibody incubation: dilute secondary antibody and incubate at room temperature for 1 h, wash for 5 min × 3 times;

[0142] (10) Development: Use a gel imager to develop and observe the statistical status of protein expression.

[0143] Western blot results are shown in Figure 4A The Western blot results were analyzed using Image J software with one-way ANOVA. Figure 4B As shown. Figure 4A , Figure 4B It can be seen that the protein expression of type I collagen was significantly increased after administration of atropine, phencyclnonate hydrochloride and its optical isomers compared with the form deprivation group, among which dextrorotatory phencyclnonate and atropine had similar effects on increasing the expression of type I collagen, and the effects were more significant.

[0144] In summary, phencyclnonate hydrochloride can inhibit the remodeling of the scleral extracellular matrix by upregulating the expression of type I collagen in the sclera, thereby delaying the extension of the axial length and achieving the purpose of inhibiting the progression of myopia.

Claims

1. Use of a compound or a salt thereof in the preparation of a drug for preventing, delaying, inhibiting and / or treating myopia, characterized in that: The compound is L-type phencyclonate hydrochloride, D-type phencyclonate hydrochloride or racemic phencyclonate hydrochloride.

2. The use according to claim 1, characterized in that The structural formula of the compound is shown as formula (I) levorotatory benzylcyclohexane hydrochloride, formula (II) dextrorotatory benzylcyclohexane hydrochloride or formula (III) racemic benzylcyclohexane hydrochloride: or 。 3. The use according to claim 1, characterized in that The myopia is myopia caused by the elongation of the anterior-posterior axial length before the eyes that develops after birth.

4. The use according to claim 1, characterized in that The drug prevents, delays, inhibits and / or treats myopia by inhibiting scleral remodeling, increasing scleral thickness, increasing the expression of collagen fibers, or inhibiting the elongation of the eye axis.

5. The use according to claim 4, characterized in that The collagen fibers are selected from type I collagen fibers.

6. The use according to claim 1, characterized in that The medicine is an ophthalmic preparation.

7. The use according to claim 6, characterized in that The ophthalmic preparations include eye drops, eye ointments, eye sprays, implants, eye gels, eye patches, eye microspheres, eye sustained-release preparations or eye injections.

8. The use according to claim 6, characterized in that The ophthalmic preparation includes pharmaceutically acceptable excipients.

9. The use according to claim 8, characterized in that The pharmaceutically acceptable excipients include pH regulators, thickeners, osmotic pressure regulators or antibacterial agents.

10. The use according to claim 9, characterized in that The pH regulator is selected from one or more of disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, tartaric acid, acetic acid, sodium acetate, boric acid and its salts, borax, citric acid or sodium citrate.

11. The use according to claim 9, characterized in that The thickener is selected from one or more of dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerol, glucosamine polymer, chondroitin sulfate, gum arabic, tragacanth gum, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropyl methylcellulose, polyvinyl alcohol, polyethylene glycol, dextran, polyvinyl pyrrolidone, polypropylene alcohol or sodium hyaluronate.

12. The use according to claim 9, characterized in that The osmotic pressure regulator is selected from one or more of mannitol, sorbitol, sodium chloride, potassium chloride, sodium sulfate, potassium sulfate, sodium nitrate, potassium nitrate, propylene glycol, glucose or glycerol.

13. The use according to claim 9, characterized in that The antibacterial agent is selected from one or more of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thiothion, benzalkonium chloride or benzalkonium bromide.

14. The use according to claim 6, characterized in that The content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.1-10% w / v.

15. The use according to claim 6, characterized in that The content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-10% w / v.

16. The use according to claim 6, characterized in that The content of phencyclnonate hydrochloride in the ophthalmic preparation is 0.5-5% w / v.

17. The use according to claim 1, characterized in that The myopia is selected from refractive myopia, axial myopia, congenital myopia, early myopia, delayed myopia, late myopia, low myopia, moderate myopia, high myopia, pseudomyopia, true myopia, myopia in minors, myopia in adults, myopia in the elderly, simple myopia, pathological myopia, primary myopia or secondary myopia.

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