CN117731662A - Application and preparation method of a cholinergic receptor antagonist in preventing myopia - Google Patents

Abstract

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

Description

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

Technical field

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

Background technique

Myopia is one of the most common human eye diseases. In recent years, the incidence of myopia has increased rapidly and has become a global medical health problem. Myopia is a refractive disorder usually caused by a mismatch between the refractive power and axial length of the eye, causing images 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-staph infections.

Studies have found that cholinergic antagonists such as atropine and pirenzepone can effectively inhibit the progression of myopia, but studies have also proven that not all cholinergic antagonists have the ability to inhibit the progression of myopia.

The sclera is a connective tissue composed of fibroblasts embedded in an extracellular matrix (ECM) rich in type I collagen and proteoglycans, which determines the shape and axial length of the eye. Studies have shown that human scleral fibroblasts express five mAChRs receptor subtypes. Experiments with scleral thinning and weakening of the extracellular matrix suggest that scleral elongation in experimental myopia involves changes in scleral cells and molecules for tissue remodeling. The sclera is the final effector of the development of myopia. After the occurrence of myopia, the expression of scleral collagen decreases, the collagen fibers are arranged disorderly, the longitudinal fibers become thinner, and the transverse fibers separate or disappear, thus causing the thinning of the sclera.

Contents of the invention

In some embodiments, the present invention provides the use of a compound selected from phencyclononate hydrochloride or a salt thereof in the preparation of a medicament for preventing, delaying, inhibiting and/or treating myopia or myopia-related diseases.

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

In some embodiments, the phencyclononyl hydrochloride is selected from the group consisting of levorotatory phencyclonyl hydrochloride, dextrorotary phencyclononyl hydrochloride, or racemic phencyclononyl hydrochloride.

In some embodiments, the structural formula of the compound is as follows: Formula (I) L-phencyclononyl hydrochloride (or "(-) phencyclononyl hydrochloride"), Formula (II) D-phencyclononyl hydrochloride (or It is called "(+) phencyclononyl ester hydrochloride") or formula (III) racemic phencyclononyl ester hydrochloride as shown:

In some embodiments, the myopia is acquired myopia caused by an extension of the anterior-posterior axis of the eye.

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 of the eye.

In some embodiments, the present invention demonstrates that intravitreal injection of phencyclonate hydrochloride can modulate the synthesis of scleral extracellular matrix, thereby inhibiting scleral remodeling.

In some embodiments, the present invention confirms that phencyclonate hydrochloride may inhibit the remodeling of the sclera by inhibiting the synthesis of type I collagen, the main component of the scleral extracellular matrix, thereby inhibiting the development of myopia.

Atropine is a non-selective muscarinic receptor antagonist that binds to all five muscarinic receptor subtypes and prevents acetylcholine from binding to them. To date, atropine has been proven by existing technology to be a potential myopia treatment drug and has significantly inhibited the progression 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 that L-phencyclononate hydrochloride has an inhibitory effect on five muscarinic receptors. The subtype is relatively selective, has higher selectivity for M1, M3, and M4 receptors, may produce fewer side effects than atropine, and has very good application prospects.

In some embodiments, the medicament includes topical preparations, ophthalmic preparations, injections, oral preparations or freeze-dried powder injections.

In some embodiments, the ophthalmic preparations include eye drops, eye ointments, ophthalmic sprays, implants, ophthalmic gels, eye patches, ophthalmic microspheres, ophthalmic sustained-release formulations, or ophthalmic injection.

In some embodiments, the ophthalmic formulations include pharmaceutically acceptable excipients.

In some embodiments, the pharmaceutically acceptable excipients include pH adjusters, thickeners, osmotic pressure adjusters, or bacteriostatic agents.

In some embodiments, the pH adjusting agent is selected from the group consisting 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, One or more of sodium citrate; preferably, the thickening agent is selected from dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerin, glucosamine polymer, and cartilage sulfate Gluten, gum arabic, tragacanth, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropylmethylcellulose, One or more of polyvinyl alcohol, polyethylene glycol, dextran, polyvinylpyrrolidone, polypropylene alcohol, and sodium hyaluronate, but are not limited thereto.

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

In some embodiments, the bacteriostatic agent is selected from the group consisting of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosol, benzalkonium chloride One or more of ammonium or benzalkonium bromide.

In some embodiments, the content of phencyclonyl hydrochloride in the ophthalmic formulation is 0.1 to 10% w/v.

In some embodiments, the content of phencyclonate hydrochloride in the ophthalmic formulation is 0.5-10% w/v.

In some embodiments, the content of phencyclonate hydrochloride in the ophthalmic formulation is 0.5 to 5% w/v.

In some embodiments, the myopia is selected from the group consisting of refractive myopia, axial myopia, congenital myopia, early-onset myopia, late-onset myopia, late-onset myopia, low myopia, moderate myopia, high myopia, Pseudomyopia, true myopia, juvenile myopia, adult myopia, elderly myopia, simple myopia, pathological myopia, primary myopia, or secondary myopia.

In some embodiments, the present invention provides the use of a compound selected from the group consisting of phencyclononate in the preparation of a medicament for inhibiting axial lengthening.

In some embodiments, the compound is selected from phencyclonyl hydrochloride.

In some embodiments, the phencyclononyl hydrochloride is selected from the group consisting of levorotatory phencyclonyl hydrochloride, dextrorotary phencyclononyl hydrochloride, or racemic phencyclononyl hydrochloride.

In some embodiments, the structural formula of the compound is as shown in formula (I) levorotatory phencyclononyl hydrochloride, formula (II) dextral phencyclononyl hydrochloride or formula (III) racemic phencyclononyl hydrochloride:

In some embodiments, the medicament includes topical preparations, ophthalmic preparations, injections, oral preparations or freeze-dried powder injections.

In some embodiments, the ophthalmic preparations include eye drops, eye ointments, ophthalmic sprays, implants, ophthalmic gels, eye patches, ophthalmic microspheres, ophthalmic sustained-release formulations, or ophthalmic injection.

In some embodiments, the ophthalmic formulations include pharmaceutically acceptable excipients.

In some embodiments, the pharmaceutically acceptable excipients include pH adjusters, thickeners, osmotic pressure adjusters, or bacteriostatic agents.

In some embodiments, the pH adjusting agent is selected from the group consisting 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, One or more of sodium citrate; preferably, the thickening agent is selected from dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerin, glucosamine polymer, and cartilage sulfate Gluten, gum arabic, tragacanth, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropylmethylcellulose, One or more of polyvinyl alcohol, polyethylene glycol, dextran, polyvinylpyrrolidone, polypropylene alcohol, and sodium hyaluronate.

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

In some embodiments, the bacteriostatic agent is selected from the group consisting of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosol, benzalkonium chloride One or more of ammonium or benzalkonium bromide.

In some embodiments, the content of phencyclonyl hydrochloride in the ophthalmic formulation is 0.1 to 10% w/v.

In some embodiments, the content of phencyclonate hydrochloride in the ophthalmic formulation is 0.5-10% w/v.

In some embodiments, the content of phencyclonate hydrochloride in the ophthalmic formulation is 0.5 to 5% w/v.

In some embodiments, the present invention provides a pharmaceutical composition for preventing and/or treating myopia, including the compound phencyclonate and one or more pharmaceutically acceptable substances.

In some embodiments, the compound is selected from phencyclonyl hydrochloride.

In some embodiments, the phencyclononyl hydrochloride is selected from the group consisting of levorotatory phencyclonyl hydrochloride, dextrorotary phencyclononyl hydrochloride, or racemic phencyclononyl hydrochloride.

In some embodiments, the structural formula of the compound is as shown in formula (I) levorotatory phencyclononyl hydrochloride, formula (II) dextral phencyclononyl hydrochloride or formula (III) racemic phencyclononyl hydrochloride:

In some embodiments, the pharmaceutically acceptable substance or substances include pharmaceutically acceptable excipients.

In some embodiments, the pharmaceutically acceptable excipients include pH adjusters, thickeners, osmotic pressure adjusters, or bacteriostatic agents.

In some embodiments, the pH adjusting agent is selected from the group consisting 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, One or more of sodium citrate; preferably, the thickening agent is selected from dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerin, glucosamine polymer, and cartilage sulfate Gluten, gum arabic, tragacanth, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropylmethylcellulose, One or more of polyvinyl alcohol, polyethylene glycol, dextran, polyvinylpyrrolidone, polypropylene alcohol, and sodium hyaluronate.

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

In some embodiments, the bacteriostatic agent is selected from the group consisting of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosol, benzalkonium chloride One or more of ammonium or benzalkonium bromide.

In some embodiments, the content of phencyclonyl hydrochloride in the ophthalmic formulation is 0.1 to 10% w/v.

In some embodiments, the content of phencyclonate hydrochloride in the ophthalmic formulation is 0.5-10% w/v.

In some embodiments, the content of phencyclonate hydrochloride in the ophthalmic formulation is 0.5 to 5% w/v.

In some embodiments, the myopia is selected from the group consisting of refractive myopia, axial myopia, congenital myopia, early-onset myopia, late-onset myopia, late-onset myopia, low myopia, moderate myopia, high myopia, Pseudomyopia, true myopia, juvenile myopia, adult myopia, elderly myopia, simple myopia, pathological myopia, primary myopia, or secondary myopia.

In some embodiments, the present invention provides a pharmaceutical preparation for preventing and/or treating myopia, including phencyclonate and pharmaceutically acceptable excipients.

In some embodiments, the compound is selected from phencyclonyl hydrochloride.

In some embodiments, the phencyclononyl hydrochloride is selected from the group consisting of levorotatory phencyclonyl hydrochloride, dextrorotary phencyclononyl hydrochloride, or racemic phencyclononyl hydrochloride.

In some embodiments, the structural formula of the compound is as shown in formula (I) levorotatory phencyclononyl hydrochloride, formula (II) dextral phencyclononyl hydrochloride or formula (III) racemic phencyclononyl hydrochloride:

In some embodiments, the pharmaceutically acceptable excipients include pH adjusters, thickeners, osmotic pressure adjusters, or bacteriostatic agents.

In some embodiments, the pH adjusting agent is selected from the group consisting 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, One or more of sodium citrate; preferably, the thickening agent is selected from dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerin, glucosamine polymer, and cartilage sulfate Gluten, gum arabic, tragacanth, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropylmethylcellulose, One or more of polyvinyl alcohol, polyethylene glycol, dextran, polyvinylpyrrolidone, polypropylene alcohol, and sodium hyaluronate.

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

In some embodiments, the bacteriostatic agent is selected from the group consisting of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosol, benzalkonium chloride One or more of ammonium or benzalkonium bromide.

In some embodiments, the content of phencyclonate hydrochloride in the ophthalmic formulation is 0.1 to 10% w/v.

In some embodiments, the content of phencyclonate hydrochloride in the ophthalmic formulation is 0.5-10% w/v.

In some embodiments, the content of phencyclonate hydrochloride in the ophthalmic formulation is 0.5 to 5% w/v.

In some embodiments, the pharmaceutical preparations include topical preparations, ophthalmic preparations, injections, oral preparations or freeze-dried powder injections.

In some embodiments, the ophthalmic preparations include eye drops, eye ointments, ophthalmic sprays, implants, ophthalmic gels, eye patches, ophthalmic microspheres, ophthalmic sustained-release formulations, or ophthalmic injection.

In some embodiments, the eye drops further include water.

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

In some embodiments, the ophthalmic injection is selected from intravitreal injections.

In some embodiments, the present invention provides the use of the pharmaceutical composition or the pharmaceutical preparation in the preparation of products for relieving visual fatigue.

Description of drawings

Figure 1 shows a schematic diagram of the establishment of the guinea pig form deprivation myopia model; the left side is the blank control group of guinea pigs, and the right side is the form deprivation group guinea pigs.

Figure 2 shows the results of hematoxylin-eosin staining (HE) staining of the scleral tissue of guinea pig eyeballs after 14 days of experimental intervention, where n=3.

Figure 3 shows the results of Sirius red staining of the scleral tissue of guinea pig eyeballs after 14 days of experimental intervention, where n=3.

Figure 4A-Figure 4B show the expression changes of type Ⅰ collagen (Collagen-I) protein measured by Western Blotting, where Figure 4A shows the Western Blotting band, and Figure 4B shows the results of quantitative band analysis, where n=6. Note: FDM: form deprivation;

PCH: racemic phencyclononyl hydrochloride; S(+)-PCH: dextrorotatory phencyclononyl hydrochloride; R(-)-PCH: levorotatory phencyclononyl hydrochloride.

Detailed ways

The technical solutions of the present invention will be further described below through specific examples, which do not limit the scope of protection of the present invention. Some non-essential modifications and adjustments made by others based on the concept of the present invention still belong to the protection scope of the present invention.

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

Furthermore, the use of "or" means "and/or" unless stated otherwise. Similarly, "includes" and "includes" are interchangeable and are not intended to be limiting. It will be further understood that where the term "comprises" is used to describe various embodiments, those skilled in the art will understand that in some specific circumstances the language "consisting essentially of" or "consisting of" may alternatively be used. Composition" to describe the embodiments.

Unless otherwise defined, 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.

It is to be understood that this disclosure is not limited to the specific methods, 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 disclosure.

As used herein, the term "pharmaceutically acceptable" refers to those materials (so far as reasonable medical judgment is concerned) that are suitable for contact with tissue of a patient without causing additional toxicity, irritation, allergic reactions, or other problems or complications. (i.e. have a reasonable risk/benefit ratio) compounds, materials, compositions and/or dosage forms.

The term "prevention" has its art-recognized meaning when used in connection with a condition, disease, syndrome or any other medical condition and includes the administration of a drug to a patient who is administered the drug relative to a patient who does not receive the drug. , the frequency of their medical conditions will decrease, or their onset and symptoms will be delayed.

As used herein, the term "treatment" 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.

The term "prevention or treatment" has its art-recognized meaning and includes administering to a recipient one or more agents of the present invention. Treatment is prophylactic (i.e., prevents the recipient from developing the undesirable condition) if it is given before clinical manifestation of the undesirable condition (e.g. disease or other undesirable condition in the recipient animal), whereas if it is given after clinical manifestation of the undesirable condition , then the treatment is therapeutic (i.e. used to reduce, improve or stabilize an existing adverse condition or its side effects).

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

Main reagents :

Racemic phencyclonyl hydrochloride, left-handed phencyclonyl hydrochloride (A) and dextrorotary phencyclonyl hydrochloride (B) [synthesized by our unit, see Chinese patent 97125424.9 and Chinese patent application 200410029597.9]; sodium chloride injection (Shijiazhuang Fourth Medicine Co., Ltd.); Orbuvacaine 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.).

Among them, phencyclonyl ester hydrochloride (2′-cyclopentyl-2′-phenyl-2′-glycolic acid-9α-[3-methyl-3-azabicyclo(3.3.1)nonyl] ester hydrochloride salt) and its isomers, the structures of left-handed phencyclononyl ester hydrochloride (Ⅰ), dextrorotatory phencyclononyl ester hydrochloride (Ⅱ) and racemic phencyclononyl ester hydrochloride (Ⅲ) are shown in the following chemical structural formulas.

Chemical formula C ₂₂ H ₃₂ ClNO ₃ ; molecular weight is 393.9.

Method steps :

1. Reagent preparation

(1) Preparation of atropine intravitreal injection: Dissolve 1g of atropine powder in physiological saline, dissolve completely, adjust the volume to 100 mL, and prepare a 1% w/v atropine solution.

(2) Preparation of Levophenyl Hydrochloride Intravitreal Injection: Dissolve 1g of Levophenyl Hydrochloride powder in physiological saline, dissolve completely, adjust the volume to 100 mL, and prepare 1% w/v Levophenyl Hydrochloride. Nonyl ester solution.

(3) Preparation of dextrocycline hydrochloride intravitreal injection: 1g of dextrocyclonate hydrochloride powder is dissolved in physiological saline, completely dissolved, diluted to 100 mL, and prepared into 1% w/v dextrocycline. Phencyclonyl hydrochloride solution.

(4) Preparation of racemic phencyclonyl hydrochloride intravitreal injection: 1g of racemic phencyclonyl hydrochloride powder was dissolved in physiological saline, completely dissolved, diluted to 100 mL, and prepared into 1% w/v racemic phencyclonyl hydrochloride. Phencyclonyl hydrochloride solution.

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

(1) Take tricolor guinea pigs (young guinea pigs of 2 to 3 weeks old) weighing about 120g, and randomly divide them into 7 groups, with 9 animals in each group: blank control group, single-eye blindfold shape deprivation group, single-eye blindfold shape deprivation + normal saline group, monocular eyepatch form deprivation + atropine group, monocular eyepatch form deprivation + L-phencyclonyl hydrochloride group, monocular eyepatch form deprivation + dextro-phencyclonyl hydrochloride group, and monocular eyepatch shape deprivation + racemic benzene hydrochloride group. Cyclononyl ester group. Except for the blank control, the other six groups of guinea pigs used translucent goggles to cover their right eyes and expose their left eyes. Check the eye patch coverage once a day to ensure that the experimental eye is completely covered and does not cause pressure on the eyeball. The blank control group received no treatment.

The schematic diagram of establishing the guinea pig form deprivation myopia model is shown in Figure 1. It can be seen from Figure 1 that the guinea pig form deprivation myopia model was successfully constructed.

Guinea pigs were anesthetized with intraperitoneal injection of sodium pentobarbital (1 mL/kg) and local ocular anesthesia with oxybuvacaine eye drops (0.01 mL). The guinea pigs were fixed and routine disinfection was performed. Different drugs were injected intravitreally into the right eyes of the guinea pigs in the experimental group under a stereomicroscope (5 minutes after topical anesthesia with oxybuvacaine, and immediately after sodium pentobarbital anesthesia).

Single eye patch shape deprivation + normal saline group: Use a microsyringe to inject 10 μL of normal saline into the vitreous cavity.

Monocular eyepatch form deprivation + atropine group: Use a microsyringe to inject 10 μL of 1% w/v atropine solution into the vitreous cavity.

Monocular eyepatch shape deprivation + L-phencyclonate hydrochloride group: Use a microsyringe to inject 10 μL of 1% w/v L-phencyclonate hydrochloride solution into the vitreous cavity.

Monocular eyepatch form deprivation + dextrocycline hydrochloride group: Use a microsyringe to inject 10 μL of 1% w/v dextrocyclononate hydrochloride solution into the vitreous cavity.

Monocular eyepatch form deprivation + racemic phencyclonate hydrochloride group: Use a microsyringe to inject 10 μL of 1% w/v racemic phencyclononate hydrochloride solution into the vitreous cavity.

Example 1 Effect of phencyclonate on diopter and axial length

Diopter measurement: Use a strip retinoscope (YZ24 type, Suzhou Liuliu Company, China) to measure the diopter of the left and right eyes of the guinea pig.

Measurement of axial length: STRONG 6000A ophthalmic A-ultrasound measuring instrument is used. The A-ultrasound frequency is 11MHz. Use oxybuvacaine for corneal surface anesthesia before measurement. Measurement starts after 5 minutes. During measurement, the probe is aligned with the center of the cornea and perpendicular to the corneal plane. . The ophthalmic A-ultrasound measuring instrument measures the axial length of the eye, taking the average of 10 measurements each time, and is accurate to 0.01mm.

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 diopter of the right eye of the guinea pigs with one eye covered was (-3.05±0.24) D, which was negative, resulting in myopia; compared with the guinea pigs with one eye covered, The diopter reduction in the right eye of guinea pigs after administration of atropine and phencyclonyl hydrochloride and its optical isomers was mild, and the degree of myopia was mild. And after 14 days of intervention, the axial length of the right eye of guinea pigs with one eye covered was significantly longer than that of the guinea pigs in the control group; the axial length of the right eye of guinea pigs administered atropine, phencyclonyl hydrochloride and its optical isomers was (9.83±0.21) mm respectively. , (9.92±0.32)mm, (9.91±0.26)mm, (9.98±0.41)mm. Compared with the axial length of the single-eye covered guinea pig [(10.09±0.12)mm], the axial length of the right eye is shorter.

To sum up, after administration of atropine and phencyclonate hydrochloride and its optical isomers, the diopter of the right eye of guinea pigs decreased slowly, and the axial growth of the eye was slow. Among them, dexphencyclonate had the best effect in delaying the development of myopia, and it was the same as atropine. The effect is quite good.

Table 1

Note: Table 1 shows the measurement data of diopter and eyeball biology (axial length) after 0d and 14d of experimental intervention, where n=8.

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

(1) After 14 days of experimental intervention (after administration) according to the above "method steps", the guinea pigs were euthanized, the right eyeball was removed, placed in eyeball tissue fixative, embedded in paraffin, and serially sectioned along the sagittal axis. , the slice thickness is 6μm;

(2) Preheat the baking machine to 98°C, put the paraffin slices into the baking machine, wait for 20 minutes, wait until the paraffin on the surface of the slide melts, use xylene to dewax twice, 10 minutes each;

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

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

(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;

(6) 70%, 80%, and 90% ethanol are dehydrated once each, 100% ethanol is dehydrated twice, and xylene is transparent;

(7) Neutral resin sealing;

(8) Measure the thickness of the posterior pole sclera under an optical microscope, and take the thickness of the central area within 3 mm of the optic nerve on the eyeball wall under the microscope as the measurement area.

The results are shown in Figure 2. HE staining results showed that the thickness of the sclera of the guinea pigs in the form deprivation group was significantly reduced, and the fiber arrangement was disordered and sparse. After being given atropine, phencyclonyl hydrochloride and its optical isomers, compared with the form deprivation group, , significantly increase the scleral thickness, among which dexphencyclonate has the best effect on improving scleral thickness, which is equivalent to atropine. Therefore, administration of atropine and each optical isomer of phencyclononate hydrochloride inhibited the development of myopia in guinea pigs.

Example 3 Sirius Red Staining to Detect the Effect of Phencyclonate on Collagen Fibers

(1) The preliminary dewaxing operation is the same as the HE dyeing step of Example 2 (steps (1)-(3) of Example 2);

(2) Place the sections into Sirius Red staining solution, stain for 8 minutes, and then dehydrate in two absolute ethanols for 1 minute;

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

(4) Neutral resin sealing;

(5) Microscopic examination to observe the condition of type I collagen fibers.

The results are shown in Figure 3. Sirius red staining results showed that the content of type I collagen fibers in the form deprivation group of guinea pigs was reduced. Compared with the form deprivation group, the administration of atropine, phencyclonyl hydrochloride and its optical isomers showed that The content of type I collagen fibers increased after treatment. Among them, the collagen fibers increased the most after administration of dex-phencyclonyl hydrochloride, which was equivalent to atropine.

Example 4 Western blot detection of the effect of phencyclonyl hydrochloride on the expression of type Ⅰ collagen (Collagen-I) protein

(1) After 14 days of experimental intervention according to the above "method steps", the guinea pigs were euthanized, the eyeballs were quickly removed, the connective tissue wrapped in the ball wall was removed, and the equatorial part of the eyeball was cut open circularly to remove the anterior segment and vitreous body. , take the scleral tissue within 4 to 5 mm of the posterior pole;

(2) Use a high-speed low-temperature grinder to grind the tissue to form a tissue homogenate, and use RIPA lysis buffer to extract the total protein of each group of tissue;

(3) Configure 10% polyacrylamide gel;

(4) Sample loading: Insert the gel plate into the electrophoresis tank, add electrophoresis buffer to cover the gel plate, and then attach the sample to each hole of the gel plate;

(5) Electrophoresis: Cover the electrophoresis tank, insert the electrode, and set the electrophoresis conditions: the stacking gel constant voltage is 60V; the separation gel constant voltage is 150V. Observe the marker to determine the electrophoresis time;

(6) Membrane transfer: Cut a 2cm × 8cm 0.45 pore size PVDF membrane, process it by wet transfer method, remove the stacking gel, clamp the separation gel with filter paper and PVDF membrane and place it in the transfer electrophoresis tank, set the transfer conditions to 200mA constant current ,90min;

(7) Blocking: Completely immerse the membrane in the quick blocking solution and incubate on a horizontal shaker for 20 minutes;

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

(9) Secondary antibody incubation: dilute the secondary antibody and incubate at room temperature for 1 hour, wash 5 minutes × 3 times;

(10) Development: Use a gel imager to develop and observe statistical protein expression.

The results of Western blot are shown in Figure 4A. The Western blot measurement results were analyzed with one-way analysis of variance using Image J software. The results are shown in Figure 4B. As can be seen from Figure 4A and Figure 4B, the protein expression of type I collagen increased significantly after administration of atropine and phencyclonate hydrochloride and its optical isomers compared with the form deprivation group. Among them, dextrophencyclonate and Atropine has the same effect on increasing the expression of type Ⅰ collagen, but is more significant.

In summary, phencyclonate hydrochloride can inhibit the remodeling of the scleral extracellular matrix by up-regulating the expression of type I collagen in the sclera, thereby delaying the extension of the axial length of the eye and achieving the purpose of inhibiting the development of myopia.

Claims (10)

1. The use of a compound in the preparation of a medicament for preventing, delaying, inhibiting and/or treating myopia or myopia-related diseases, characterized in that the compound is selected from phencyclononate or a salt thereof. 2. Use as claimed in claim 1, characterized in that the compound is selected from phencyclononyl hydrochloride; The phencyclononyl hydrochloride is selected from the group consisting of left-handed phencyclononyl hydrochloride, dextrorotary phencyclononyl hydrochloride or racemic phencyclononyl hydrochloride. 3. Use as claimed in claim 1, characterized in that the structural formula of the compound is such as formula (I) levorotatory phencyclononyl ester hydrochloride, formula (II) dextrorotatory phencyclononyl ester hydrochloride or formula (III) racemic Phencyclonyl hydrochloride is shown as: 4. The use according to claim 1, characterized in that the myopia is myopia caused by an acquired extension of the anterior and posterior axis of the eye; Preferably, 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 of the eye; Preferably, the collagen fibers are selected from type I collagen fibers; Preferably, the medicine includes external preparations, ophthalmic preparations, injections, oral preparations or freeze-dried powder injections; Preferably, the ophthalmic preparation includes eye drops, eye ointments, ophthalmic sprays, implants, ophthalmic gels, eye patches, ophthalmic microspheres, ophthalmic sustained-release agents, or ophthalmic injections; Preferably, the ophthalmic preparation includes pharmaceutically acceptable excipients; Preferably, the pharmaceutically acceptable excipients include pH adjusters, thickeners, osmotic pressure adjusters, or bacteriostatic agents; Preferably, the pH adjuster is selected from the group consisting 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. One or more of them; preferably, the thickening agent is selected from the group consisting of dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerin, glucosamine polymer, chondroitin sulfate, arabic Gum, tragacanth gum, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropylmethylcellulose, polyvinyl alcohol , one or more of polyethylene glycol, dextran, polyvinylpyrrolidone, polypropylene alcohol, and sodium hyaluronate; Preferably, 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. Preferably, the bacteriostatic agent is selected from the group consisting of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosol, benzalkonium chloride, or One or more benzalkonium bromide; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.1 to 10% w/v; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.5-10% w/v; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.5 to 5% w/v. 5. The use according to claim 1, wherein the myopia is selected from the group consisting of refractive myopia, axial myopia, congenital myopia, early-onset myopia, late-onset myopia, late-onset myopia, low-grade myopia, and late-onset myopia. Myopia, moderate myopia, high myopia, pseudomyopia, true myopia, juvenile myopia, adult myopia, elderly myopia, simple myopia, pathological myopia, primary myopia, or secondary myopia. 6. The use of a compound in the preparation of a drug for inhibiting axial lengthening, the compound being selected from phencyclononate or a salt thereof; Preferably, the compound is selected from phencyclonyl hydrochloride; Preferably, the phencyclononyl hydrochloride is selected from the group consisting of levorotatory phencyclononyl hydrochloride, dextral phencyclononyl hydrochloride or racemic phencyclononyl hydrochloride; Preferably, the structural formula of the compound is as shown in formula (I) left-handed phencyclononyl hydrochloride, formula (II) dextrorotatory phencyclononyl hydrochloride or formula (III) racemic phencyclononyl hydrochloride: Preferably, the medicine includes external preparations, ophthalmic preparations, injections, oral preparations or freeze-dried powder injections; Preferably, the ophthalmic preparation includes eye drops, eye ointments, ophthalmic sprays, implants, ophthalmic gels, eye patches, ophthalmic microspheres, ophthalmic sustained-release agents, or ophthalmic injections; Preferably, the ophthalmic preparation includes pharmaceutically acceptable excipients; Preferably, the pharmaceutically acceptable excipients include pH adjusters, thickeners, osmotic pressure adjusters, or bacteriostatic agents; Preferably, the pH adjuster is selected from the group consisting 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. One or more of them; preferably, the thickening agent is selected from the group consisting of dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerin, glucosamine polymer, chondroitin sulfate, arabic Gum, tragacanth gum, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropylmethylcellulose, polyvinyl alcohol , one or more of polyethylene glycol, dextran, polyvinylpyrrolidone, polypropylene alcohol, and sodium hyaluronate; Preferably, 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. Preferably, the bacteriostatic agent is selected from the group consisting of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosol, benzalkonium chloride, or One or more benzalkonium bromide; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.1 to 10% w/v; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.5-10% w/v; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.5 to 5% w/v. 7. A pharmaceutical composition for preventing and/or treating myopia, characterized by comprising the compound phencyclonate or a salt thereof and one or more pharmaceutically acceptable substances; Preferably, the compound is selected from phencyclonyl hydrochloride; Preferably, the phencyclononyl hydrochloride is selected from the group consisting of levorotatory phencyclononyl hydrochloride, dextral phencyclononyl hydrochloride or racemic phencyclononyl hydrochloride; Preferably, the structural formula of the compound is as shown in formula (I) left-handed phencyclononyl hydrochloride, formula (II) dextrorotatory phencyclononyl hydrochloride or formula (III) racemic phencyclononyl hydrochloride: Preferably, the pharmaceutically acceptable one or more substances include pharmaceutically acceptable excipients; Preferably, the pharmaceutically acceptable excipients include pH adjusters, thickeners, osmotic pressure adjusters, or bacteriostatic agents; Preferably, the pH adjuster is selected from the group consisting 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. One or more of them; preferably, the thickening agent is selected from the group consisting of dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerin, glucosamine polymer, chondroitin sulfate, arabic Gum, tragacanth gum, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropylmethylcellulose, polyvinyl alcohol , one or more of polyethylene glycol, dextran, polyvinylpyrrolidone, polypropylene alcohol, and sodium hyaluronate; Preferably, 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. Preferably, the bacteriostatic agent is selected from the group consisting of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosol, benzalkonium chloride, or One or more benzalkonium bromide; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.1 to 10% w/v; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.5-10% w/v; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.5 to 5% w/v; Preferably, the myopia is selected from the group consisting of refractive myopia, axial myopia, congenital myopia, early-onset myopia, late-onset myopia, late-onset myopia, low myopia, moderate myopia, high myopia, and pseudomyopia. , true myopia, juvenile myopia, adult myopia, elderly myopia, simple myopia, pathological myopia, primary myopia, or secondary myopia. 8. A pharmaceutical preparation for preventing and/or treating myopia, characterized by comprising phencyclonate hydrochloride and pharmaceutically acceptable excipients; Preferably, the phencyclononyl hydrochloride is selected from the group consisting of levorotatory phencyclononyl hydrochloride, dextral phencyclononyl hydrochloride or racemic phencyclononyl hydrochloride; Preferably, the structural formula of the compound is as shown in formula (I) left-handed phencyclononyl hydrochloride, formula (II) dextral phencyclononyl hydrochloride or formula (III) racemic phencyclononyl hydrochloride: Preferably, the pharmaceutically acceptable excipients include pH adjusters, thickeners, osmotic pressure adjusters, or bacteriostatic agents; Preferably, the pH adjuster is selected from the group consisting 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. One or more of them; preferably, the thickening agent is selected from the group consisting of dextrin and its derivatives, sodium alginate and its derivatives, sodium hyaluronate, glycerin, glucosamine polymer, chondroitin sulfate, arabic Gum, tragacanth gum, guar gum, xanthan gum, chitosan, carbomer, lectin, methylcellulose, carboxymethylcellulose, carboxypropylmethylcellulose, polyvinyl alcohol , one or more of polyethylene glycol, dextran, polyvinylpyrrolidone, polypropylene alcohol, and sodium hyaluronate; Preferably, the isotonic 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. ; Preferably, the bacteriostatic agent is selected from the group consisting of boric acid, ethylparaben, methylparaben, propylparaben, phenylmercuric acetate, sorbic acid, chlorobutanol, thimerosol, benzalkonium chloride, or One or more benzalkonium bromide; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.1 to 10% w/v; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.5-10% w/v; Preferably, the content of phencyclonyl hydrochloride in the ophthalmic preparation is 0.5 to 5% w/v. 9. The pharmaceutical preparation according to claim 8, characterized in that it includes external preparations, ophthalmic preparations, injections, oral preparations or freeze-dried powder for injection; Preferably, the ophthalmic preparation includes eye drops, eye ointments, ophthalmic sprays, implants, ophthalmic gels, eye patches, ophthalmic microspheres, ophthalmic sustained-release agents, or ophthalmic injections; Preferably, the eye drops also include water; Preferably, the ophthalmic injection is selected from intravitreal injection. 10. Application of the pharmaceutical composition according to claim 7 or the pharmaceutical preparation according to any one of claims 8 to 9 in the preparation of a product for relieving visual fatigue.