Eye drops for improving or preventing retinal circulation disorders and retinal neurovascular connection disorders

Abstract

An eye drop for improving or preventing retinal circulation disorder and retinal neurovascular connection disorder, comprising nanoparticles containing fibrate.

Description

Technical Field

[0001] The present invention relates to eye drops for improving or preventing retinal circulation disorders and retinal neurovascular connection disorders.

[0002] This application claims the benefit of Japanese Patent Application No. 2020-192736 filed on November 19, 2020, the entire contents of which are incorporated herein by reference. Background Art

[0003] It is known that fibrate has the effect of inhibiting retinal angiogenesis by oral administration. For example, Patent Document 1 reports that retinal angiogenesis is inhibited by forced oral administration of fenofibrate to mice. However, side effects (such as rhabdomyolysis) of fibrate combined with statins have been reported, and fibrate is not used in actual clinical practice to treat eye diseases.

[0004] As a method of administration of drugs for eye diseases, topical administration is preferred because of fewer systemic side effects compared to oral administration. For example, commonly used anti-VEGF agents for treating retinal diseases such as age-related macular degeneration and diabetic macular edema are mainly administered into the vitreous cavity. However, vitreous hemorrhage and lens damage have been reported, and serious eye complications such as endophthalmitis have also been reported. From the perspective of safety, it is preferred to administer the drug by eye drops. However, it is difficult to make the effective concentration of the drug reach the retinal tissue present at the back of the eyeball through the commonly used eye drops.

[0005] On the other hand, disorders in retinal circulation and retinal neural connections are caused earlier than the onset of obvious retinopathy and are considered to be one of the causes of retinal disease. However, eye drops effective for improving these disorders have not yet been put into practical use.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: JP2016-539098 Summary of the invention

[0009] Technical Problems to be Solved by the Invention

[0010] If retinal circulation disorders or retinal nerve connection disorders can be improved or prevented by eye drops, retinal diseases caused by these disorders can be safely and effectively prevented or treated.

[0011] Therefore, an object of the present invention is to provide an eye drop that can improve or prevent retinal circulation disorders and retinal neurovascular connection disorders.

[0012] Technical solutions for solving technical problems

[0013] The present invention includes the following embodiments.

[0014] [1] An eye drop for improving or preventing retinal circulation disorders and retinal neurovascular connection disorders, comprising nanoparticles containing fibrate.

[0015] [2] The eye drops according to [1], wherein the nanoparticles are wet-pulverized products of benzyl alcohol.

[0016] [3] The eye drops according to [1] or [2], further comprising a thickener.

[0017] [4] The eye drops according to [3], wherein the thickener is a cellulose-based thickener.

[0018] [5] The eye drops according to [4], wherein the cellulose thickener is methyl cellulose.

[0019] [6] The eye drop according to any one of [1] to [5], further comprising cyclodextrins.

[0020] [7] The eye drops according to [6], wherein the cyclodextrin is 2-hydroxypropyl-β-cyclodextrin.

[0021] Effects of the Invention

[0022] According to the present invention, an eye drop capable of improving or preventing retinal circulation disorders and retinal neurovascular connection disorders is provided. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The results of particle size distribution analysis of the beta nanoparticles prepared in the examples by laser diffraction / scattering method are shown.

[0024] Figure 2 This figure shows an atomic force microscope image of the beta nanoparticles prepared in the example.

[0025] Figure 3A The weight changes of diabetic model mice used for eye drop administration test over time. Mean ± SEM. Treatment group: administered with eye drops containing fibrate nanoparticles (n = 6). Control group: administered with Vehicle nanoparticle eye drops (n = 6). NS: no significant difference.

[0026] Figure 3B The figure shows the changes of real-time blood glucose values ​​of diabetic model mice used for eye drop administration test over time. Mean ± SEM. Treatment group: administered eye drops containing fibrate nanoparticles (n = 6). Control group: administered eye drops containing vehicle nanoparticles (n = 6). NS: no significant difference.

[0027] Figure 3C The figure shows the changes in intraocular pressure (left eye) of diabetic model mice used for eye drop administration test over time. Mean ± SEM. Treatment group: administered eye drops containing fibrate nanoparticles (n = 6). Control group: administered eye drops containing vehicle nanoparticles (n = 6). NS: no significant difference.

[0028] Figure 4A The mean blood pressure of diabetic model mice used for eye drop administration test changes over time. Mean ± SEM. Treatment group: administered eye drops containing fibrate nanoparticles (n = 6). Control group: administered eye drops containing vehicle nanoparticles (n = 6). NS: no significant difference.

[0029] Figure 4B The figure shows the changes of ocular regurgitation pressure of diabetic model mice used for eye drop administration test over time. Mean ± SEM. Treatment group: administered eye drops containing fibrate nanoparticles (n = 6). Control group: administered eye drops containing vehicle nanoparticles (n = 6). NS: no significant difference.

[0030] Figure 5 The baseline optic nerve head blood flow of diabetic model mice used for eye drop administration test changes over time. Mean ± SEM. Treatment group: administered eye drops containing fibrate nanoparticles (n = 6). Control group: administered eye drops containing vehicle nanoparticles (n = 6). NS: no significant difference.

[0031] Figure 6 The figures show the changes in ocular blood flow after flicker stimulation in diabetic model mice at 8, 10, 14 and 12 weeks of age used for the eye drop administration test. Treatment group: administered with eye drops containing fibrate (n=6). Control group: administered with Vehicle eye drops (n=6).

[0032] Figure 7 The total blood flow changes after flicker stimulation at different ages of diabetic model mice used in the eye drop administration test are shown. Treatment group: administered with eye drops containing fibrate nanoparticles (n=6). Control group: administered with vehicle nanoparticle eye drops (n=6).

[0033] Figure 8 The figures show the changes in ocular blood flow after hyperoxia in diabetic model mice at 8, 10, 14 and 12 weeks of age for eye drop administration. Treatment group: administered with fibrate-containing eye drops (n=6). Control group: administered with Vehicle eye drops (n=6).

[0034] Fig. 9The total blood flow changes after high oxygen load stimulation in diabetic model mice of different ages used in the eye drop administration test are shown. Treatment group: administered with eye drops containing fibrate nanoparticles (n=6). Control group: administered with vehicle nanoparticle eye drops (n=6).

[0035] Fig.10 The results of evaluating the a-wave implicit time and b-wave implicit time in the electroretinogram of diabetic model mice used in the eye drop administration test are shown. Treatment group: administered with eye drops containing fibrate nanoparticles (n=6). Control group: administered with vehicle nanoparticle eye drops (n=6). DETAILED DESCRIPTION

[0036] [Eye drops]

[0037] In one embodiment, the present invention provides an eye drop for improving retinal circulation disorders and retinal neurovascular connection disorders, comprising nanoparticles containing fibrate.

[0038] "Retinal circulation" refers to the blood flow in the retina. "Retinal circulation disorder" refers to the obstruction of blood flow in the retina. "Retinal neurovascular connection" refers to the changes in blood flow in the retina according to neural activity. "Retinal neurovascular connection disorder" refers to the obstruction of the response of blood flow changes in the retina corresponding to neural activity.

[0039] When retinal blood vessels are blocked or narrowed due to thrombus or embolus, retinal circulation disorder occurs, hindering the supply of oxygen and nutrients to the optic nerve. In addition, since the increase in retinal blood flow corresponding to the increase in oxygen demand accompanying neural activity is hindered, the retinal tissue falls into relative hypoxia, resulting in retinal neurovascular connection disorder.

[0040] Whether retinal circulation disorders and retinal neurovascular connection disorders have occurred can be confirmed quantitatively and non-invasively by measuring the changes in retinal blood flow caused by flicker stimulation. In a normal retina where no disorder has occurred, flicker stimulation generally causes an increase in retinal blood flow. However, when retinal circulation disorders and retinal neurovascular connection disorders occur, the increase in retinal blood flow caused by flicker stimulation will be hindered. Therefore, in the case where flicker stimulation does not cause an increase in retinal blood flow, it can be evaluated that retinal circulation disorders and retinal neurovascular connection disorders have occurred. Furthermore, in the case where the state in which flicker stimulation does not increase retinal blood flow changes to a state in which an increase in retinal blood flow caused by flicker stimulation can be confirmed, it can be evaluated that the retinal circulation disorders and retinal neurovascular connection disorders have improved.

[0041] Retinal circulation disorders and retinal neurovascular connection disorders are one of the causes of various retinal vitreous diseases. Such retinal vitreous diseases include, but are not limited to, diabetic retinopathy, retinal vein occlusion, retinal artery occlusion, age-related macular degeneration, central serous choroidopathy, etc. The eye drops of this embodiment can improve retinal circulation disorders and retinal neurovascular connection disorders, and therefore can be used for the treatment or prevention of the above-mentioned retinal vitreous diseases.

[0042] <Nanoparticles containing beta-tert-butyl>

[0043] The eye drops of the present embodiment contain nanoparticles containing fibrate (hereinafter also referred to as "fibrate nanoparticles"). The fibrate nanoparticles are solid nanoparticles, and fibrate is preferably nanoparticle-formed.

[0044] "Fibrate" is a general term for compounds that function as lipid-lowering drugs in amphiphilic carboxylic acid derivatives. As fibrates, for example, gemfibrozil, fenofibrate, bezafibrate, clofibrate, ciprofibrate, beclofibrate, binifibrate, ciprofibrate, clenofibrate, etofibrate, nicofibrate, pirofibrate, ronifibrate, simfibrate, teafibrate, pemafibrate, benbutic acid derivatives and pharmaceutically acceptable salts or esters of the above-mentioned fibric acid derivatives, but are not limited thereto. Among them, as fibrate, fenofibrate is preferred. One kind of fibrate can be used alone, or two or more kinds can be used in combination.

[0045] The content of benzyl alcohol in the eye drops of the present embodiment is not particularly limited, and can be, for example, 0.1 to 10% (w / v) relative to the total amount of the eye drops. The content of benzyl alcohol is preferably 0.5 to 5% (w / v) relative to the total amount of the eye drops, more preferably 1 to 4% (w / v), and even more preferably 1.5 to 3% (w / v).

[0046] "Nano particles" are particles having a particle diameter of 1 nm or more and less than 1000 nm. The beta nanoparticles contained in the eye drops of the present embodiment preferably have a particle diameter in the range of 10 to 300 nm, and more preferably have a particle diameter in the range of 30 to 150 nm, from the perspective of cell uptake efficiency. As beta nanoparticles, for example, it can be a material having a particle size distribution with an average diameter of 50 to 120 nm measured by a laser diffraction / scattering method or a dynamic light scattering method.

[0047] Bet nanoparticles can be obtained by wet grinding Bet. Wet grinding of Bet can be performed using a bead mill or a ball mill, etc. Before wet grinding, dry grinding can also be performed. By performing dry grinding before wet grinding, Bet can be effectively nano-particleized. Bet nanoparticles can be called wet grinding of Bet, wet bead mill grinding or wet ball mill grinding. The wet grinding of Bet can be a wet grinding of Bet dry grinding, a wet bead mill grinding of Bet dry grinding, or a wet ball mill grinding of Bet dry grinding.

[0048] <Water>

[0049] The eye drops of this embodiment contain water as a dispersion medium. The water may be water of a grade usable for eye drops, such as purified water, sterilized purified water, water for injection, and distilled water for injection.

[0050] <Optional Ingredients>

[0051] The eye drops of this embodiment may contain an optional component in addition to the beta nanoparticles. Examples of the optional component include a thickener, a preservative, a tonicity adjuster, cyclodextrins, a buffer, and a pH adjuster.

[0052] (Thickener)

[0053] The eye drops of this embodiment preferably contain a thickener. By containing a thickener in the eye drops, the formation of meringue or cream during the preparation of beta nanoparticles can be suppressed, and a beta nanoparticle dispersion in a good state can be obtained.

[0054] As the thickener, for example, it can be cellulose thickeners such as methyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, etc.; vinyl thickeners such as carboxyvinyl polymer, polyvinyl alcohol (completely or partially saponified) and polyvinyl pyrrolidone; sodium alginate, chondroitin sulfate, polyethylene glycol, etc., but it is not limited to these.

[0055] Among them, as the thickener, a cellulose-based thickener is preferred, and methylcellulose is more preferred, because the retention of beta nanoparticles is improved and the transferability into the eye is improved.

[0056] The thickener may be used alone or in combination of two or more.

[0057] The content of the thickener in the eye drops of the present embodiment is not particularly limited, and is, for example, preferably 0.01 to 3% (w / v), more preferably 0.05 to 2% (w / v), and even more preferably 0.1 to 1% (w / v) relative to the entire eye drops.

[0058] (preservative)

[0059] The eye drops of this embodiment may contain a preservative. By containing a preservative, the storage stability of the eye drops is improved.

[0060] Preservatives include, but are not limited to, inverse soaps such as benzalkonium chloride, benzethonium chloride, and chlorhexidine gluconate; parabens such as methyl paraben, ethyl paraben, propyl paraben, and butyl paraben; and alcohols such as chlorobutanol, phenethyl alcohol, and benzyl alcohol.

[0061] Among them, as the preservative, in view of being widely used as eye drops and having a high preservative effect, a cationic surfactant such as a reversible soap is preferred, and benzalkonium chloride is more preferred.

[0062] The preservatives may be used alone or in combination of two or more.

[0063] The content of the preservative in the eye drops of the present embodiment is not particularly limited, and is, for example, preferably 0.0005 to 0.5% (w / v), more preferably 0.001 to 0.1% (w / v), and even more preferably 0.001 to 0.01% (w / v), relative to the entire eye drops.

[0064] (Tonomicity adjuster)

[0065] The eye drops of this embodiment may contain a tension adjuster. By containing a tension adjuster, irritation to the eyes can be reduced.

[0066] Examples of the tonicity adjusting agent include, but are not limited to, sugars such as glucose; polyols such as propylene glycol, glycerol, mannitol, sorbitol, and xylitol; and inorganic salts such as sodium chloride and potassium chloride.

[0067] Among them, as the tension adjusting agent, polyhydric alcohols are preferred, and mannitol is more preferred, from the viewpoint of being able to protect the cornea from stimulation by the protective agent or the like.

[0068] The tension adjuster may be used alone or in combination of two or more.

[0069] The content of the tension adjusting agent in the eye drops of the present embodiment is not particularly limited, and is, for example, preferably 0.01 to 3% (w / v), more preferably 0.05 to 2% (w / v), and even more preferably 0.1 to 1% (w / v) relative to the entire eye drops.

[0070] (Cyclodextrins)

[0071] The eye drops of this embodiment preferably contain cyclodextrins. Cyclodextrins can suppress the aggregation of beta-nano particles and make the beta-nano particles uniformly dispersed.

[0072] "Cyclodextrins" refers to cyclodextrins or cyclodextrin derivatives. Examples of cyclodextrins include α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, and derivatives thereof. Examples of cyclodextrin derivatives include alkyl derivatives, hydroxyalkyl derivatives, sulfoalkyl ether derivatives, and sugar bond derivatives.

[0073] Among them, as cyclodextrins, hydroxyalkyl derivatives are preferred, and 2-hydroxypropyl-β-cyclodextrin is more preferred, from the viewpoint of excellent dispersibility of beta nanoparticles.

[0074] The cyclodextrins may be used alone or in combination of two or more.

[0075] The content of cyclodextrins in the eye drops of the present embodiment is not particularly limited, and is, for example, preferably 0.1 to 15% (w / v), more preferably 1 to 10% (w / v), and even more preferably 3 to 8% (w / v) relative to the entire eye drops.

[0076] (pH adjuster)

[0077] The eye drops of this embodiment may contain a pH adjuster. Examples of the pH adjuster include dilute hydrochloric acid and sodium hydroxide.

[0078] (Buffer)

[0079] The eye drops of this embodiment may also contain a buffer, such as sodium citrate hydrate, sodium acetate hydrate, sodium bicarbonate, tromethamine, boric acid, borax, sodium hydrogen phosphate hydrate, sodium dihydrogen phosphate, and the like.

[0080] The eye drops of this embodiment may be eye drops containing beta nanoparticles, water and a thickener; eye drops containing beta nanoparticles, water, a thickener and cyclodextrins; eye drops containing beta nanoparticles, water, a thickener, cyclodextrins and a preservative; and eye drops containing beta nanoparticles, water, a thickener, cyclodextrins, a preservative and a tension regulator, etc.

[0081] More specifically, it can be eye drops containing beta nanoparticles, water and cellulose thickeners; eye drops containing beta nanoparticles, water, cellulose thickeners and cyclodextrins; eye drops containing beta nanoparticles, water, cellulose thickeners, cyclodextrins and benzalkonium chloride; and eye drops containing beta nanoparticles, water, cellulose thickeners and cyclodextrins, benzalkonium chloride and mannitol, etc.

[0082] More specifically, it can be eye drops containing beta nanoparticles, water and methylcellulose; eye drops containing beta nanoparticles, water, methylcellulose and 2-hydroxypropyl-β-cyclodextrin; eye drops containing beta nanoparticles, water, methylcellulose and 2-hydroxypropyl-β-cyclodextrin and benzalkonium chloride; and eye drops containing beta nanoparticles, water, methylcellulose, 2-hydroxypropyl-β-cyclodextrin, benzalkonium chloride and mannitol, etc.

[0083] <Manufacturing method>

[0084] The eye drops of the present embodiment can be prepared by appropriately adding any component such as a thickener to Bet and mixing, preferably dispersing it in an aqueous dispersion medium containing cyclodextrins, and performing wet grinding. Before performing wet grinding, dry grinding can also be performed. "Dry grinding" refers to grinding in a gas. "Wet grinding" refers to grinding in a liquid.

[0085] Hereinafter, a specific example of the method for producing the eye drops according to the present embodiment will be described, but the present invention is not limited thereto.

[0086] A thickener is appropriately added to Bet for mixing, mixing and fine grinding. At this time, as a component mixed with Bet, components other than cyclodextrins and water can be cited. These components can be added after dry grinding and before wet grinding, and preferably the thickener and tension regulator are mixed with Bet before fine grinding. Mixing and fine grinding can also be first carried out with an agate mortar, etc., and then dry grinding is carried out using a bead mill. Mixing and grinding are preferably carried out under low temperature conditions (e.g., 4°C). In the dry grinding performed using a bead mill, zirconia beads with a diameter of about 2 mm can be used.

[0087] The finely pulverized material obtained by dry pulverization is dispersed in an aqueous solution containing cyclodextrins. At this time, any component such as a preservative may be appropriately added. Next, wet pulverization is performed using a bead mill or the like. In the wet pulverization using a bead mill, zirconium oxide beads with a diameter of about 0.1 mm may be used. The wet pulverization is preferably performed under low temperature conditions (e.g., 4° C.). In the case where bubbles are generated during wet pulverization, or the material becomes a creamy state, it can be restored to a suspension state by performing centrifugal separation while adding a small amount of dispersion medium.

[0088] After wet grinding, the beads are removed and the mixture is sterilized by filtration using a 0.2 μm filter. Thus, the eye drops of this embodiment can be obtained. When the eye drops are used, the concentration of the beta nanoparticles can be adjusted by appropriately using a diluent containing components other than the beta nanoparticles.

[0089] <How to use>

[0090] The eye drops of this embodiment can be used to improve retinal circulation disorders and retinal neurovascular connection disorders. For example, the eye drops of this embodiment can be administered to patients who suffer from or are likely to suffer from retinal vitreous diseases such as diabetic retinopathy, retinal vein occlusive disease, retinal artery occlusive disease, age-related macular degeneration, and central serous choroideremia due to retinal circulation disorders and retinal neurovascular connection disorders. The eye drops of this embodiment can be used to improve retinal circulation disorders and retinal neurovascular connection disorders, and it is expected that these retinal vitreous diseases will be cured, alleviated, or prevented from developing.

[0091] The eye drops of this embodiment can be used as eye drops for treating or preventing retinal vitreous diseases. The retinal vitreous diseases that are the treatment targets of the eye drops of this embodiment can be mild, moderate or severe. In the case where the retinal vitreous disease is mild, the cure of the retinal vitreous disease can be expected by administering the eye drops of this embodiment. In the case where the retinal vitreous disease is moderate or severe, the eye drops of this embodiment can also be used in combination with existing medical treatments (anti-vascular endothelial growth factor VEGF inhibitors or steroid drugs) or surgical treatments (retinal photocoagulation, vitrectomy, etc.).

[0092] The eye drops of this embodiment can be used to prevent retinal circulation disorders and retinal neurovascular connection disorders. The occurrence of retinal circulation disorders and retinal neurovascular connection disorders can be prevented by preventively administering the eye drops of this embodiment to subjects at high risk of developing retinal circulation disorders and retinal neurovascular connection disorders.

[0093] The administration interval of the eye drops of the present embodiment can be appropriately determined according to the patient's symptoms, body weight, age, sex, etc. The administration interval can be set to, for example, 2 to 3 times a day.

[0094] [Other embodiments]

[0095] In one embodiment, the present invention provides a method for improving or preventing retinal circulation disorders and retinal neurovascular connection disorders, the method comprising administering the eye drops of the above embodiment to a subject.

[0096] In one embodiment, the present invention provides use of nanoparticles containing fibrate in the preparation of eye drops for improving or preventing retinal circulation disorders and retinal neurovascular connection disorders.

[0097] In one embodiment, the present invention provides the eye drops of the above embodiment for improving or preventing retinal circulation disorders and retinal neurovascular connection disorders.

[0098] In one embodiment, the present invention provides use of the eye drops of the above embodiment for improving or preventing retinal circulation disorders and retinal neurovascular connection disorders.

[0099] In one embodiment, the present invention provides a method for treating or preventing a retinal vitreous disease, comprising administering the eye drops of the above embodiment to a subject.

[0100] In one embodiment, the present invention provides use of nanoparticles comprising fibrate in the preparation of eye drops for treating or preventing retinal vitreous diseases.

[0101] In one embodiment, the present invention provides the eye drops of the above embodiment for treating or preventing retinal vitreous diseases.

[0102] In one embodiment, the present invention provides use of the eye drops of the above embodiment for treating or preventing retinal vitreous diseases.

[0103] Example

[0104] Hereinafter, the present invention will be described by way of examples, but the present invention is not limited to the following examples.

[0105] [Preparation of eye drops]

[0106] <Preparation of eye drops containing beta nanoparticles>

[0107] The eye drops containing fibrate were prepared by the following steps.

[0108] 1) 450 mg of fenofibrate (Takeda Teva), 112.5 mg of methylcellulose and 112.5 mg of D-mannitol were mixed and ground in an agate mortar. (Low temperature (4°C), 1 hour).

[0109] 2) Recover the finely pulverized material in 1) and perform dry pulverization (low temperature (4°C)) using a bead cell disruptor (Micro Smash MS-100R, Tommy Seiko Co., Ltd.) (twice with 2 mm zirconia beads, 3000 rpm each time, 30 seconds) or a multi-specimen cell disruptor (Sheikmaster Neo, Biomedical Science Co., Ltd.) (500 rpm, 10 minutes with zirconia beads).

[0110] 3) Weigh 450 mg of the dry pulverized product obtained in 2) and adjust the volume to 15 mL with a 5% (w / v) 2-hydroxypropyl-β-cyclodextrin aqueous solution, and then add 75 μL of a 1% (w / v) benzalkonium chloride (BAC) aqueous solution.

[0111] 4) The mixture obtained in 3) was subjected to 30 wet pulverizations (30 times with 0.1 mm zirconium oxide beads, 15,000 rpm, 30 seconds each time, at low temperature (4°C)) using a bead cell disruptor (Micro Smash MS-100R, manufactured by TOMY SEIKO Co., Ltd.). If bubbles were generated during the wet pulverization or the mixture became a meringue, centrifugal separation (800×g, 60 seconds, 4°C) was performed appropriately to restore the suspension state.

[0112] 5) The wet-pulverized product obtained in 4) was wet-pulverized using a multi-specimen cell disruptor (Sheikmaster Neo, Biomedical Science Co., Ltd.) (with 0.1 mm zirconia beads, 1500 rpm, 1 hour, low temperature conditions (4° C.)).

[0113] 6) The zirconium oxide beads were removed from the ultra-crushed product obtained in 5), and the product was sterilized by filtration using a 0.2 μm filter, and used as an eye drop containing fibrate. If necessary, the concentration was appropriately adjusted using the following vehicle (dispersion medium) before use.

[0114] Vehicle components

[0115] Benzalkonium chloride 0.005% (w / v)

[0116] Mannitol 0.5% (w / v)

[0117] Methylcellulose 0.5% (w / v)

[0118] 2-Hydroxypropyl-β-cyclodextrin 5% (w / v)

[0119] The final concentration of each drug in the eye drops prepared as described above is as follows.

[0120] Fenofibrate 2% (w / v)

[0121] Benzalkonium chloride 0.005% (w / v)

[0122] Mannitol 0.5% (w / v)

[0123] Methylcellulose 0.5% (w / v)

[0124] 2-Hydroxypropyl-β-cyclodextrin 5% (w / v)

[0125] (Particle size distribution analysis by laser diffraction / scattering method)

[0126] The particle size distribution of the fibrate-containing eye drops prepared in the above manner was analyzed using a laser diffraction particle size distribution measuring apparatus (SALD-7100, Shimadzu Corporation). Figure 1As shown in Table 1. The BET nanoparticles have a particle size of 34 nm to 244 nm. Figure 2 An atomic force microscope image showing the BET nanoparticles.

[0127] [Table 1]

[0128]

[0129] [Preparation of Vehicle eye drops]

[0130] Prepare the Vehicle described in <Preparation of eye drops containing BET> and use it as the Vehicle eye drops.

[0131] [Eye drop administration test]

[0132] As shown in Table 2, a female diabetic model mouse (C57BL / 6J db / db) at 6 weeks of age was used for the eye drop administration test. The eye drops were administered twice a day, morning and evening.

[0133] [Ocular blood flow measurement test]

[0134] The ocular blood flow measurement was performed under isoflurane inhalation anesthesia (induction 3.5 - 4.0% / min, maintenance 1.5 - 2.5% / min). After maintaining the body temperature of the mouse at 36 - 37 °C with a warming pad, the experiment was conducted. The measurement of each evaluation item started at 8 weeks of age. The measurements were performed every other week.

[0135] [Table 2]

[0136] Treatment Group Control group Eye drops Eye drops containing beta nanoparticles Vehicle eye drops Number of medications 6 6 Frequency of eye drops 2 times in the morning and evening + 1 hour before measurement 2 times in the morning and evening + 1 hour before measurement Eye drops application site Left Eye Left Eye

[0137] [Measurement of systemic parameters]

[0138] As systemic parameters, the body weight, real-time blood glucose value, intraocular pressure (left eye), mean blood pressure, and eye reflux pressure were measured every other week starting from 8 weeks of age. The between-group analysis between the treatment group and the control group was performed by two-way repeated measured ANOVA (Two-way Repeated Measured ANOVA Sidak’s multiple test). The analysis between different weeks of age was performed by non-parametric test for multiple independent samples (Jonckheere-Terpstra test).

[0139] The results are as Figure 3A to Figure 4B shown (mean ± SEM).

[0140] The body weight showed a tendency to increase with age in both the treatment group and the control group. There was no significant difference in body weight gain between the treatment group and the control group ( Figure 3A ).

[0141] Real-time blood glucose levels were maintained at high values ​​in both the treatment and control groups. In all groups, there were no significant differences in real-time blood glucose levels between the ages of the children ( Figure 3B ).

[0142] intraocular pressure( Figure 3C ), mean blood pressure( Figure 4A ) and ocular reflux pressure ( Figure 4B ) There was no significant difference between the treatment group and the control group. There was no significant difference in age among all groups.

[0143] <Determination of Baseline Optic Nerve Head Blood Flow>

[0144] Baseline optic nerve head (ONH) blood flow was measured every other week starting from 8 weeks of age using a laser speckle blood flow measurement device (LSFG-Micro, Softcare (available)). The measurement was performed in the left eye. Inter-group analysis between the treatment group and the control group was performed by Two-way Repeated Measured ANOVA. Inter-age analysis was performed by the Johckheere-Terpstra test.

[0145] The results are as follows Figure 5 The mean ± SEM is shown. There was no significant difference in resting blood flow before stimulation between the treatment group and the control group. There was no significant difference in age between all groups.

[0146] <Measurement of changes in ocular blood flow caused by flicker stimulation>

[0147] Changes in ocular blood flow induced by flicker stimulation were measured every other week starting from 8 weeks of age. Flicker stimulation was performed at a frequency of 12 Hz for 3 minutes, and ocular blood flow was measured every 20 seconds using LSFG-Micro. Measurements were performed in the left eye. Intergroup analysis between the treatment group and the control group was performed using Two-way Repeated Measured ANOVA Sidak's multiple test.

[0148] The results are as follows Figure 6-7 Shown are (mean ± SEM). Figure 6 The graphs represent the blood flow changes after flicker stimulation at 8 weeks of age, 10 weeks of age, 14 weeks of age, and 12 weeks of age, respectively. Figure 7 This is a graph summarizing the changes in total blood flow at each age. Figure 6 The “Base” on the horizontal axis of represents the baseline optic nerve head blood flow before loading.

[0149] In the control group, the obstruction of the blood flow increase response after flicker stimulation was confirmed. On the other hand, in the treatment group, the blood flow increase response after flicker stimulation was maintained without obstruction. After 10 weeks of age, there was a significant difference in the change of blood flow after flicker stimulation between the treatment group and the control group. From this result, it can be confirmed that the obstruction of retinal neurovascular connection can be avoided by administering eye drops containing beta nanoparticles.

[0150] <Measurement of changes in ocular blood flow caused by high oxygen load stimulation>

[0151] Changes in ocular blood flow caused by hyperoxia were measured every other week starting from 8 weeks of age. Flicker stimulation was performed at a frequency of 12 Hz for 3 minutes, and measurements were taken every 20 seconds using LSFG-Micro. Measurements were made in the left eye. Intergroup analysis between the treatment group and the control group was performed using Two-way Repeated Measured ANOVA Sidak's multiple test.

[0152] The results are as follows Figure 8-Figure 9 Shown are (mean ± SEM). Figure 8 The graphs represent the changes in blood flow after high oxygen load stimulation at 8 weeks, 10 weeks, 14 weeks and 12 weeks of age, respectively. Fig. 9 This is a graph summarizing the changes in total blood flow at each age. Figure 8 The “Base” on the horizontal axis of represents the baseline optic nerve head blood flow before loading.

[0153] In the control group, the blood flow reduction reaction after high oxygen load stimulation was confirmed to be impaired. On the other hand, in the treatment group, the blood flow reduction reaction after high oxygen load stimulation was not impaired and maintained. From this result, it can be confirmed that the obstruction of glial cells and vascular endothelial cells can be avoided by administering eye drops containing beta nanoparticles.

[0154] <Electroretinogram (ERG)>

[0155] Electroretinogram was performed every other week starting from 8 weeks of age. The left eye was used for examination. Intergroup analysis between the treatment group and the control group was performed by Two-way Repeated Measured ANOVA Sidak's multiple test.

[0156] The results are as follows Fig.10 As shown (mean ± SEM). In the treatment group, the extension of the b-wave latency observed in the control group was improved. After 12 weeks of age, the b-wave latency in the treatment group was significantly shortened compared to the control group. From this result, it can be confirmed that retinal dysfunction can be suppressed by administering eye drops containing beta nanoparticles.

[0157] Possibility of Industrial Application

[0158] According to the present invention, an eye drop capable of improving or preventing retinal circulation disorders and retinal neurovascular connection disorders is provided.

[0159] Although the preferred embodiments of the present invention have been described and illustrated above, it should be understood that these are examples of the present invention and should not be considered as limiting examples. Additions, omissions, substitutions and other variations may be made without departing from the spirit or scope of the present invention. Therefore, the present invention should not be considered to be limited by the above description, but only by the scope of the attached technical solutions.

Claims

1. An eye drop comprising: nanoparticles containing fenofibrate, benzalkonium chloride, mannitol, methylcellulose, and 2-hydroxypropyl-β-cyclodextrin, The nanoparticles are wet-pulverized fenofibrate. The nanoparticles have a particle diameter ranging from 10 to 300 nm.

2. The eye drops according to claim 1, wherein In the eye drops, the concentration of fenofibrate is 2% (w / v).

3. The eye drops according to claim 1 or 2, wherein In the eye drops, the concentration of benzalkonium chloride is 0.005% (w / v).

4. The eye drops according to claim 1 or 2, wherein In the eye drops, the concentration of mannitol is 0.5% (w / v).

5. The eye drops according to claim 1 or 2, wherein In the eye drops, the concentration of methylcellulose is 0.5% (w / v).

6. The eye drops according to claim 1 or 2, wherein In the eye drops, the concentration of 2-hydroxypropyl-β-cyclodextrin is 5% (w / v).

7. Use of nanoparticles containing fenofibrate in the preparation of eye drops for retinal vitreous diseases, characterized in that: The eye drops further contain: benzalkonium chloride, mannitol, methylcellulose, and 2-hydroxypropyl-β-cyclodextrin, The nanoparticles are wet-pulverized fenofibrate. The nanoparticles have a particle diameter ranging from 10 to 300 nm.

8. The use according to claim 7, wherein the cause of the retinal vitreous disease is retinal circulation disorder and retinal neurovascular connection disorder.

9. The use according to claim 7 or 8, wherein in the eye drops, the concentration of fenofibrate is 2% (w / v), the concentration of benzalkonium chloride is 0.005% (w / v), the concentration of mannitol is 0.5% (w / v), the concentration of methylcellulose is 0.5% (w / v), and the concentration of 2-hydroxypropyl-β-cyclodextrin is 5% (w / v).

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