KR20220085726A - Novel peptide of inhibiting bet protein and composition for preventing and treating ophthalmic diseases comprising the same - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of various eye diseases, such as glaucoma, uveitis, wet and dry macular degeneration, and age-related macular degeneration, containing the novel peptide and the composition as an active ingredient, and the novel peptide according to the present invention was confirmed to exhibit the effect of preventing and improving various eye diseases caused by retinal degeneration by inhibiting BET protein to improve inflammation caused by retinal damage caused by epigenetic changes , uveitis, wet and dry macular degeneration, age-related macular degeneration, etc. can be provided as a pharmaceutical composition for the prevention or treatment of various eye diseases.

Description

A novel peptide inhibiting BET protein and a composition for preventing and treating ophthalmic diseases comprising the same {NOVEL PEPTIDE OF INHIBITING BET PROTEIN AND COMPOSITION FOR PREVENTING AND TREATING OPHTHALMIC DISEASES COMPRISING THE SAME}

The present invention relates to a novel peptide exhibiting Bromodomain Extra-Terminal (BET) protein inhibitory ability and a composition for preventing, treating and improving ophthalmic diseases comprising the same as an active ingredient. Specifically, the present invention inhibits retinal degeneration by inhibiting inflammatory response through epigenetic changes through a novel peptide BET inhibitor, thereby preventing various ophthalmic diseases such as diabetic retinopathy, wet and dry macular degeneration, glaucoma, and uveitis. And it may provide a pharmaceutical composition for treatment.

The eye is composed of the outer membrane, the media, the inner membrane, and the refractive medium. The outer membrane is made up of the cornea, which is the anterior surface covering the black pupil, and the sclera following it. The middle membrane is composed of the iris, ciliary body, and choroid. , the inner membrane consists of the retina. The lens, vitreous, and aqueous humor are included in the refractive medium. Functional impairment or loss of the eye is one of the factors that greatly reduce the quality of life, and it is important to maintain and protect the health of the eye due to various factors that can adversely affect it, such as aging, disease, and vision. is getting As ophthalmic diseases, there are various diseases such as retinal diseases including retinal degenerative diseases and glaucoma, cataracts, corneal epithelial disorders or corneal epithelial wounds.

Currently, laser treatment, photocoagulation, cryocoagulation, and photodynamic therapy are known as treatments for these eye diseases. All of these treatments are surgery-based treatment methods, and treatment with drugs is still in the development stage. Treatment by surgery has the disadvantage that it cannot be applied to all patients, and the success rate is low and the cost is high, causing social and economic problems. Most of the patients who cannot be operated on, unfortunately, lead to blindness because there is currently no special treatment. As the lifespan of humans is extended, these eye diseases are continuously increasing, so there is an urgent need to develop an appropriate therapeutic agent for this.

Treatments for ophthalmic diseases currently under development are mainly steroids, matrix metalloproteinase (MMP) inhibitors, angiogenesis inhibitors, and antibodies to angiogenic growth factors.

The macula is a nervous tissue located in the center of the retina, and most of the photoreceptors are gathered here. Macular degeneration is an eye disease that most often progresses with age. It is one of the difficult-to-treat eye diseases that causes blurred vision and distorted vision in the early stages of the onset, but later causes blindness.

Age-related macular degeneration is recently known as the most common cause of blindness in the elderly, and about 30 million people worldwide suffer from this disease, and about 500,000 patients suffer from this disease every year. is losing It is one of the three major causes of blindness along with glaucoma and diabetic retinopathy in Korea, and its prevalence is gradually increasing with an increase in the elderly population. The onset period also tends to decrease from the 60s to the middle-aged people in their 40s and 50s.

Age-related macular degeneration is largely classified into two types: exudative (wet) macular degeneration and atrophic (dry) macular degeneration. It is accompanied by fundus findings such as rupture of the pigment epithelium, and it is known that 70-90% of blindness due to age-related macular degeneration is due to exudative lesions.

The recently developed intravitreal anti-angiogenic growth factor antibody injection method, a treatment for exudative age-related macular degeneration, improved patients' vision and improved the prognosis of the disease. Due to the inconvenience of having to undergo injection and the procedure that requires direct intravitreal drug delivery, complications such as cataract, endophthalmitis, vitreous hemorrhage, and retinal detachment increase. There is an urgent need to develop therapeutic agents that take gender into account.

On the other hand, glaucoma is a disease that causes the loss of ganglion cells in the retina and is closely related to retinal diseases.

Retinal degenerative disease is known as a disease in which vision is damaged due to degeneration of photoreceptors as it progresses by various environmental factors such as genetic or oxidative stress, and in most cases, from the early stage of the disease, peripheral They complain of a decrease in visual field, but central visual acuity is relatively well preserved. In addition, glaucoma is a disease group consisting of various aspects showing various clinical and histopathological findings, and exhibits symptoms such as changes in the optic disc, damage to retinal ganglion cells, and visual field defects.

The retina is the innermost tissue of the eye belonging to the central nervous system. Retinal degeneration is a pathological phenomenon that is accompanied by retinal diseases such as age-related macular degeneration and retinitis pigmentosa and leads to blindness due to the death of photoreceptors. As with other neurodegenerative diseases, there is currently no way to suppress retinal degeneration, so retinal disease accompanied by retinal degeneration is an intractable disease. Recently, there is a report that the inflammatory response is very important in retinal degeneration, and that retinal degeneration can be suppressed by controlling such neuroinflammation. Many diseases, including neurodegenerative diseases, accompany epigenetic changes, and recent attempts to prevent and treat diseases by controlling them have been made. As an example, a substance called JQ, known as one of a wide range of BET protein inhibitors, showed the potential to treat degenerative retinal diseases through microglial cell activation in a mouse model of retinitis pigmentosa (Zhao et al, 2017, Photoreceptor protection via blockade of BET epigenetic readers in a murine model of inherited retinal degeneration. Journal of Neuroinflammation (2017) 14:14, 1-15).

Under this background, the present inventors confirmed that a novel peptide inhibitor exhibiting excellent inhibitory ability on BET protein, an epigenetic recognizer, has excellent retinal degeneration inhibitory efficacy to prevent and treat eye diseases caused by retinal degeneration, etc. invention was completed.

Zhao et al, 2017, Photoreceptor protection via blockade of BET epigenetic readers in a murine model of inherited retinal degeneration. Journal of Neuroinflammation (2017) 14:14, 1-15

The present invention provides a novel peptide having the ability to inhibit BET protein, and also uses the effect of the novel peptide to inhibit retinal degeneration such as reduction of inflammation through epigenetic changes, such as glaucoma, uveitis, wet and dry macular degeneration, age-related An object of the present invention is to provide a composition for preventing or treating ophthalmic diseases such as macular degeneration.

The present invention provides a novel peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2.

In addition, the present invention provides a pharmaceutical composition for preventing or treating ophthalmic diseases comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2.

The ophthalmic diseases of the present invention include endophthalmitis, keratitis, conjunctivitis, keratoconjunctivitis, uveitis, blepharitis, scleritis, iritis, glaucoma, retinopathy, retinitis pigmentosa, retinal detachment, retinal pigment epithelial detachment, retinal tear, diabetic retinopathy, Retinopathy of prematurity, polypoidal choroidal vasculopathy, ischemic proliferative retinopathy, cone dystrophy, proliferative vitreoretinopathy, retinal artery occlusion, retinal vein occlusion, Levers's optic neuropathy, corneal renal neovascularization, retinal choroidal neovascularization, wet and dry macular degeneration, and age-related macular degeneration.

In addition, the present invention provides a food composition for preventing or improving ophthalmic diseases comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2.

In addition, the present invention provides a feed composition for animals for preventing or improving ophthalmic diseases comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2.

The novel peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2 of the present invention alleviates the inflammatory response induced by retinal degeneration by epigenetic action that inhibits BET protein, thereby effectively inhibiting retinal degeneration. This effect of inhibiting retinal degeneration can be applied to retinal degeneration and other central nervous system diseases through neuroinflammation control. Therefore, the novel peptide of the present invention can be usefully used for preventing or treating various eye diseases, such as glaucoma, uveitis, wet and dry macular degeneration, and age-related macular degeneration.

1 to 5 show the ERG analysis results for intraperitoneal injection of the peptide (BBC0008) consisting of the amino acid sequence of SEQ ID NO: 2.
6 to 9 show the results of analyzing the retinal sections of mice administered with the peptide (BBC0008) consisting of the amino acid sequence of SEQ ID NO: 2 by histological staining and immunohistochemical analysis.
10 and 11 show ERG analysis results for intraocular injection of Eylea, intraocular injection and intraperitoneal injection of the peptide (BBC0002) consisting of the amino acid sequence of SEQ ID NO: 1.
12 and 13 show ERG analysis results for intraocular injection of Eylea, intraocular injection and intraperitoneal injection of the peptide (BBC0008) consisting of the amino acid sequence of SEQ ID NO: 2.

Hereinafter, embodiments and examples of the present invention will be described in detail with reference to the accompanying drawings so that those of ordinary skill in the art to which the present invention pertains can easily carry out. However, the present application may be embodied in various forms and is not limited to the embodiments and examples described herein.

Throughout this specification, when a part "includes" a component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

The present invention provides a novel peptide (BBC0002) consisting of the amino acid sequence of SEQ ID NO: 1. Specifically, SEQ ID NO: 1 has the sequence of CSIECW, C is cysteine, S is serine, I is isoleucine, E is glutamate, and W is tryptophan.

In addition, the present invention provides a novel peptide (BBC0008) consisting of the amino acid sequence of SEQ ID NO: 2. Specifically, SEQ ID NO: 2 has a sequence of CSWWCW, C is cysteine, S is serine, and W is tryptophan.

The novel peptide of the present invention can prevent or treat macular degeneration by inhibiting angiogenesis in the eye, and the macular degeneration may be wet and dry macular degeneration and age-related macular degeneration, but is not limited thereto. It can also act on various eye diseases such as glaucoma and uveitis.

In addition, the present invention provides a composition for the treatment, prevention or improvement of an eye disease comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2. The eye diseases include glaucoma, uveitis, wet and dry macular degeneration, age-related macular degeneration, and the like.

Specifically, the composition of the present invention may be a pharmaceutical composition for preventing or treating various eye diseases, such as glaucoma, uveitis, wet and dry macular degeneration, and age-related macular degeneration, including a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2 have.

The pharmaceutical composition is selected from the group consisting of suitable carriers, excipients, disintegrants, sweeteners, coating agents, swelling agents, lubricants, lubricants, flavoring agents, antioxidants, buffers, bacteriostats, diluents, dispersants, surfactants, binders and lubricants. One or more additives may be further included.

The preferred dosage of the peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2, or the composition comprising the peptide may vary depending on the subject's condition and weight, the type and degree of disease, drug form, administration route and period and may be appropriately selected by those skilled in the art.

In the present invention, the 'subject' may be a mammal including a human, but is not limited to these examples.

In addition, the composition of the present invention provides a food composition for preventing or improving various eye diseases, such as glaucoma, uveitis, wet and dry macular degeneration, and age-related macular degeneration, including a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2. .

The food composition may be a health functional food, dairy product, fermented product or food additive.

The food composition includes various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids , a protective colloidal thickener, a pH adjuster, a stabilizer, a preservative, glycerin, alcohol, a carbonation agent used in carbonated beverages, and the like.

The health functional food refers to a food manufactured and processed using raw materials or ingredients useful for the human body, and "functionality" is useful for health purposes such as regulating nutrients or physiological action for the structure and function of the human body. It means taking it for the purpose of obtaining an effect.

In addition, the composition of the present invention contains a peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2 for preventing or improving various eye diseases such as glaucoma, uveitis, wet and dry macular degeneration, and age-related macular degeneration. to provide.

The feed composition for animals can be ingested by all non-human animals, such as non-human primates, sheep, dogs, cattle, horses, and the like.

The present invention will be described in more detail through the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[Example 1]

preparation of the sample

About 120 peptides were prepared by requesting synthesis companies such as Chengdu Youngshe Chemical Co., Ltd (China).

[Example 2]

Evaluation of BET (Bromodomain-containing protein 4) protein binding inhibition ability

In order to evaluate the ability to inhibit the interaction between the bromodomain and the tetraacetylated histone H4 peptide, the following experiment was performed.

The inhibitory ability test for BRD4 (BD1+BD2) is as follows.

Compounds were diluted 1:5 serial dilutions in assay buffer from a 10 mM stock in DMSO (100 μM starting concentration) in white OptiPlate-384 (PerkinElmer). A mixture of 100 nM GST-BRD4 (BD1 + BD2) and 100 nM biotinylated acetyl-histone H4 (Lys5, 8, 12, 16) peptides was added in assay buffer (50 mM HEPES pH 7.4; 25 mM NaCl; 0,05%). Tween 20; 0.1% bovine serum albumin (BSA); 10 mM dithiothreitol (DTT)). 6 μl of the mixture was added to the dilution followed by 6 μl of premixed AlphaLISA Glutathione Acceptor Beads from PerkinElmer (each in assay buffer at a concentration of 10 μg/ml) and AlphaScreen strep. Tavidin donor beads (AlphaScreen Streptavidin Donor Beads) were added, and samples were incubated with shaking at 300 rpm for 30 minutes at room temperature in the dark. Thereafter, signals were measured with a PerkinElmer Envision HTS Multilabel Reader using PerkinElmer's Alpha Screen protocol. Each plate contained a negative control in which biotinylated acetyl-histone H4 peptide and GST-BRD4 (BD1 + BD2) were drained and replaced with assay buffer. When the software GraphPad Prism was used for calculation, a negative control value was entered as a low reference value. A positive control (probe molecule I-BET762 with protein/peptide mixture) was also pipetted. Determination of IC50 values was performed using GraphPad Prism 3.03 software (or an updated version thereof).

As a result of confirming the BRD4 binding inhibitory efficacy for a total of about 120 peptides synthesized in Example 1, the peptide consisting of the amino acid sequence of SEQ ID NO: 1 of the present invention represents 56%, and the peptide consisting of the amino acid sequence of SEQ ID NO: 2 is 47 %, it was confirmed that it was the most excellent material among all the peptide materials used in the test.

[Example 3]

Analysis of the treatment efficacy of macular degeneration in a mouse model of retinal degeneration.

For the production of the retinal degeneration model, 7-week-old male albino BALB/c mice were used. Three rats were randomly assigned to each test group, and the contrast was maintained at 12-hour intervals. After each mouse was dark-adapted for 24 h, the pupils were dilated with 0.5% tropicamide and 0.5% phenylephrine hydrochloride eye drops (Santen, Osaka, Japan) 30 min before blue LED exposure. Unanesthetized mice were exposed to a blue LED (460 ± 10 nm) of 2,000 lux for 2 hours in a cage with reflective interiors. The light intensity was measured using an LED photometer (model TM-201L, TENMARS Electronics, Taipei, Taiwan). After exposure to blue LEDs, they were placed in the dark for 24 hours and then the 12-hour light-dark cycle was resumed for 3 days.

Each test group was divided into an untreated control group (retinal degeneration untreated control group), a group administered with PBS to retinal degeneration mice, and a peptide (BBC0008) administration group (5 mg/kg) consisting of the amino acid sequence of SEQ ID NO: 2, 3 for each test group were randomly assigned.

Each sample was administered 1 hour after LED exposure, and the PBS-treated group and the peptide (BBC0008) administration group consisting of the amino acid sequence of SEQ ID NO: 2 were first injected into the abdominal cavity immediately before LED exposure, and the second injection was performed 3 days later. It was administered intraperitoneally.

Electroretinography (ERG)

Electroretinal angiography (ERG) recordings were performed by Kim et al. (Kim, G. H., Kim, H. I., Paik, S. S., Jung, S. W., Kang, S., and Kim, I. B. (2016). Functional and morphological evaluation of blue light-emitting diode. -induced retinal degeneration in mice, Graefes Arch. Clin. Exp. Ophthalmol. 254, 705-716) was followed.

That is, mice were kept in a completely dark room for 16 h prior to ERG recording. All animals were intraperitoneally anesthetized with zolazepam (20 mg/kg) and xylazine (7.5 mg/kg). The cornea was coated with hydroxypropyl methylcellulose gel and covered with a gold ring contact electrode. A ground electrode and a reference electrode were placed subcutaneously on the tail and ears, respectively. Stimulation was a short white flash delivered through a Ganzfeld stimulator (UTAS-3000; LKC Technologies, Gaithersburg, MD, USA). The signal was amplified and filtered through a digital bandpass filter ranging from 5 to 300 Hz to generate a wave and b wave. Scotopic ERG, rod-mediated responses were obtained at increasing light intensities of 0.025 and 3.96 cd/s · m ² . Photopick, cone-mediated responses were obtained after 5 min light adaptation to background luminosity. Recordings were taken at a luminous intensity of 6.28 cd/s m ² . Each recording was the average of three responses obtained within the 15 s interstimulation interval. The amplitude of the A-wave was measured from the baseline to the maximum a-wave peak, and the b-wave was measured from the maximum a-wave peak to the maximum b-wave peak.

The results of ERG analysis are shown in FIGS. 1 to 5 . 1 to 5, the peptide (BBC0008) administration group (5 mg/kg) consisting of the amino acid sequence of SEQ ID NO: 2 is significantly superior to both a-wave and b-wave compared to other administration groups wavelength (P<0.05).

tissue staining method

The tissue staining method was performed as follows. After 3 days of blue LED exposure, the eyecup was removed and fixed in 4% paraformaldehyde for 2 h. After fixation, for frozen tissue specimens (TissueTek OCT compound; Sakura Finetek, Alphen aan den Rijn, The Netherlands), eye cups were washed with 0.1 M phosphate buffer (PB; pH 7.4), transferred to 30% sucrose, and infiltrated overnight.

Retinal sections (thickness 7 μm) were cut in the sagittal plane and stained with hematoxylin and eosin (H & E). Quantitative analysis was performed on the stained sections. Outer nuclear layer (ONL) thickness was measured at 240 μm spacing (top to bottom) in vertical strips of the retina.

The analysis results are shown in FIGS. 6 to 9 .

As shown in FIGS. 6 to 9 , in the retinal degeneration animal model, it was confirmed that the number of microglia increasing retinal inflammation increases while retinal degeneration occurs due to retinal damage. That is, looking at the outer nuclear layer (ONL) with photoreceptors, the photoreceptors are degraded by blue LED stimulation. However, it was confirmed that the ONL of the peptide (BBC0008) administration group consisting of the amino acid sequence of SEQ ID NO: 2 exhibited a significantly thicker outer nuclear layer (ONL) than the PBS administration group. Therefore, it can be seen that the peptide (BBC0008) treatment group consisting of the amino acid sequence of SEQ ID NO: 2 of the present invention significantly improves retinal function degeneration.

In addition, Muller glial cells express GFAP in response to retinal damage or stress. GFAP expression is initiated towards the ONL in the ganglion cell layer (GCL). It was confirmed that the peptide (BBC0008) administration group consisting of the amino acid sequence of SEQ ID NO: 2 had significantly less GFAP expression than the PBS administration group. Therefore, it can be seen that the peptide (BBC0008) treatment group consisting of the amino acid sequence of SEQ ID NO: 2 of the present invention significantly improves retinal damage.

In addition, microglia are mainly observed at the boundary of the outer nuclear plexus (OPL), inner nuclear layer (INL), and inner plexiform layer (IPL) in normal cells. However, the outer nuclear layer (ONL) is very thin in the degenerative cells in which the retinal degeneration has occurred, and microglia are widely distributed in the degenerating outer nuclear layer (OPL) and the subretinal space. Thus, microglia are activated and migrated in response to retinal damage and inflammation. It was confirmed that the group administered with the peptide (BBC0008) consisting of the amino acid sequence of SEQ ID NO: 2 had significantly fewer IBA-1 labeled microglia in the outer nuclear layer (OPL) than the group administered with PBS.

Immunohistochemical analysis

Immunohistochemical analysis was performed as follows. After washing with 0.01 M PBS, retinal sections were pre-incubated in 10% normal donkey serum for 1 h at room temperature. The sections were then divided into a rabbit polyclonal anti-ionizable calcium binding adapter molecule 1 (Iba-1) antibody (1:1000; Wako Pure Chemical Industries, Osaka, Japan), a rabbit polyclonal anti-glial fibrotic acid protein (GFAP) antibody ( 1:1000; Chemicon, Temecula, CA, United States) was diluted in PBS at 4 °C for 1 day. Sections were then washed with PBS and Cy3-conjugated (dilution, 1:3,000; Jackson ImmunoResearch, West Grove, PA, USA) or Alexa 488-conjugated donkey anti-rabbit IgG (1:3000; Molecular Probes, Eugene, OR, United States). States) and incubated for 2 hours at room temperature. After rinsing several times with PBS, the cell nuclei were counterstained with DAPI for 10 minutes as a fluorescent specimen, and then mounted with anti-fading mounting media (Vector Laboratories, Burlingame, CA, US). Images were obtained using a Zeiss LSM 510 meta confocal microscope (Carl Zeiss Co., Ltd.). Quantitative image analysis was performed using Zen 2.3 software (Blue Edition, Carl Zeiss). A region of interest 300 μm away from the optic disc of each retinal section was selected and the intensity of Iba-1 and GFAP immunoreactivity was automatically measured.

As shown in Figures 6 to 9, the peptide (BBC0008) administration group consisting of the amino acid sequence of SEQ ID NO: 2 was confirmed that Iba-1 (red) and GFAP (green) expression in the outer nuclear layer was significantly reduced than the PBS administration group.

[Example 4]

Analysis of the treatment efficacy of macular degeneration according to the route of administration in a retinal degeneration mouse model, and comparative analysis with a commercial preparation (Eylia)

Analyzing the difference in macular degeneration treatment efficacy according to the administration route of intraperitoneal injection (IP) and intravitreal injection (IVT), Eyelea (aflibercept) and drugs In order to compare the efficacy, mouse retinal degeneration was induced in the same manner as in Example 3.

Each test group was an untreated control group (retinal degeneration untreated control group), a group administered with Normal Saline (NS) to retinal degeneration mice, a group administered with Eylea IVT (administration dose 25 mg/kg), and a peptide consisting of the amino acid sequence of SEQ ID NO: 1 ( BBC0002) and the peptide (BBC0008) consisting of the amino acid sequence of SEQ ID NO: 2 were divided into an IVT administration group (administration dose 0.5 mg/kg) and an IP administration group (10 mg/kg), and three animals were randomly assigned to each test group.

Each sample was administered 1 hour after the LED exposure, the IVT group was administered once by intravitreal injection, and the PBS treated group and the IP group were injected twice in the abdominal cavity immediately before exposure to the LED and 2 times after 3 days. It was administered intraperitoneally. After administration, the ERG was analyzed in the same manner as in Example 3, and the results are shown in FIGS. 10 to 13 .

As shown in FIGS. 10 and 11, the peptide (BBC0002) IP administration group (10 mg/kg) consisting of the amino acid sequence of SEQ ID NO: 1 had a-wave, b-wave compared with other administration groups including the Eylea IVT administration group All of them showed significantly superior wavelengths.

In addition, the peptide (BBC0002) IVT administration group (0.5 mg/kg) consisting of the amino acid sequence of SEQ ID NO: 1 was 20 times higher than that of the IP administration group (10 mg/kg) and the Eylia IVT administration group (administration dose 25 mg/kg) and It showed better wavelength at 50 times lower concentration.

12 and 13, the peptide (BBC0008) IP administration group (10 mg/kg) consisting of the amino acid sequence of SEQ ID NO: 2 had a-wave, b-wave compared with other administration groups including the Eylia IVT administration group. All of them showed significantly superior wavelengths.

In addition, the peptide (BBC0008) IVT administration group (0.5 mg/kg) consisting of the amino acid sequence of SEQ ID NO: 2 was 20 times higher than that of the IP administration group (10 mg/kg) and the Eylia IVT administration group (administration dose 25 mg/kg) and It showed better wavelength at 50 times lower concentration.

Therefore, it can be seen that the novel peptide of the present invention having excellent BET inhibitory ability exhibits excellent efficacy in the various biomarkers in the retinal degeneration-induced mouse model, thereby exhibiting an excellent retinal degeneration inhibitory effect, and the novel peptide is glaucoma, uveitis , wet and dry macular degeneration, age-related macular degeneration, etc. can be used as pharmaceuticals for the prevention and treatment of various eye diseases.

<110> BENOBIO Co., Ltd. <120> NOVEL PEPTIDE OF INHIBITING BET PROTEIN AND COMPOSITION FOR PREVENTING AND TREATING OPHTHALMIC DISEASES COMPRISING THE SAME <130> BNB20P-0001-KR-PRI <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> BBC0002 <400> 1 Cys Ser Ile Glu Cys Trp 1 5 <210> 2 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> BBC0008 <400> 2 Cys Ser Trp Trp Cys Trp 1 5

Claims (7)

A peptide consisting of the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2.
A pharmaceutical composition for preventing or treating ophthalmic diseases comprising the peptide of claim 1.
The method of claim 2, wherein the ophthalmic disease is endophthalmitis, keratitis, conjunctivitis, keratoconjunctivitis, uveitis, blepharitis, scleritis, iritis, glaucoma, retinopathy, retinitis pigmentosa, retinal detachment, retinal pigment epithelial detachment, retinal tear, diabetes mellitus. Retinopathy, retinopathy of prematurity, polypoidal choroidal vasculopathy, ischemic proliferative retinopathy, cone dystrophy, proliferative vitreoretinopathy, retinal artery occlusion, retinal vein occlusion, Levers's optic nerve atrophy , Corneal neovascularization, retinal choroidal neovascularization, wet and dry macular degeneration, or age-related macular degeneration, characterized in that, ophthalmic disease prevention or treatment pharmaceutical composition.
A food composition for preventing or improving ophthalmic diseases comprising the peptide of claim 1.
5. The method of claim 4, wherein the ophthalmic disease is endophthalmitis, keratitis, conjunctivitis, keratoconjunctivitis, uveitis, blepharitis, scleritis, iritis, glaucoma, retinopathy, retinitis pigmentosa, retinal detachment, retinal pigment epithelial detachment, retinal tear, diabetes mellitus. Retinopathy, retinopathy of prematurity, polypoidal choroidal vasculopathy, ischemic proliferative retinopathy, cone dystrophy, proliferative vitreoretinopathy, retinal artery occlusion, retinal vein occlusion, Levers's optic nerve atrophy , Corneal neovascularization, retinal choroidal neovascularization, wet and dry macular degeneration, or age-related macular degeneration, characterized in that the food composition for the prevention or treatment of ophthalmic diseases.
An animal feed composition for preventing or improving ophthalmic diseases comprising the peptide of claim 1.
The method of claim 6, wherein the ophthalmic disease is endophthalmitis, keratitis, conjunctivitis, keratoconjunctivitis, uveitis, blepharitis, scleritis, iritis, glaucoma, retinopathy, retinitis pigmentosa, retinal detachment, retinal pigment epithelial detachment, retinal tear, diabetes mellitus. Retinopathy, retinopathy of prematurity, polypoidal choroidal vasculopathy, ischemic proliferative retinopathy, cone dystrophy, proliferative vitreoretinopathy, retinal artery occlusion, retinal vein occlusion, Levers's optic nerve atrophy , Corneal neovascularization, retinal choroidal neovascularization, wet and dry macular degeneration, or age-related macular degeneration, characterized in that the ophthalmic disease prevention or improvement animal feed composition.

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