Novel peptide and use thereof

Abstract

The present invention relates to: a novel peptide which has a prophylactic, ameliorative or therapeutic effect on amyloidosis and / or macular degeneration, is safe to a living body, and has few side effects including abnormal reactions; a pharmaceutical composition comprising the same; and a health functional food.

Description

TECHNICAL FIELD

[0001] The present application relates to a novel peptide and use thereof. BACKGROUND

[0002] Amyloidosis is a general term for a group of diseases caused by accumulation of abnormal proteins called amyloid fibrils in tissues.

[0003] Alzheimer's disease (AD) is the most common type of amyloidosis in humans. Alzheimer's disease is also the most common degenerative brain disease associated with dementia, which can be accompanied by memory loss, language ability, spatial perception ability, personality disorder, and personality changes.

[0004] Alzheimer's disease is characterized by pathological infiltration of beta amyloid plaques composed of abnormal amyloid proteins, apoptotic nerve cells, and inflammatory cells outside the nerve cells, and gliosis. Various inflammatory processes and cytokines also play an important role in the pathology of Alzheimer's disease, and inflammatory cytokines can be an indicator of secondary or immunological reactions in tissue damage in Alzheimer's disease.

[0005] Due to aging, the prevalence of Alzheimer's disease is on the rise, and the resulting social and economic costs are also increasing.

[0006] However, no treatment method capable of curing Alzheimer's disease has been developed, and only AchE (acetylcholinesterase) inhibitors or N-methyl-D-aspartate (NMDA) antagonists, which are considered to be effective in slowing down the speed of memory loss, are used as therapeutic drugs.

[0007] Age-related macular degeneration (AMD) is an ophthalmic disease in which the function of the center of the retina for imaging, i.e., the macula, is impaired, resulting in gradual vision loss from the center of the visual field and eventually blindness. Age-related macular degeneration can be divided into dry AMD and wet AMD.

[0008] Dry AMD occurs as a result of apoptosis and degeneration of retinal cells due to continuous exposure to stress conditions such as oxidative stress. Dry AMD can develop into wet AMD, and thus treatment at the dry AMD stage is the most important.

[0009] In the case of wet macular degeneration, neovascularization caused by vascular endothelial growth factor (VEGF) is considered to be a direct cause, and drugs such as Eylea ® (Eylea ® ) targeting VEGF are currently used as therapeutic agents.

[0010] Unlike wet macular degeneration, which has a variety of commercial therapeutic agents, dry macular degeneration has only one drug approved by the Food and Drug Administration (FDA) so far. Therefore, there is an urgent need to develop a therapeutic agent for dry macular degeneration that can regulate the early progression stage of age-related macular degeneration.

[0011] On the other hand, GV1001 consisting of 16 amino acids selected from the reverse transcriptase of human telomerase (hTERT) has been reported to have not only anticancer, anti-inflammatory, and antioxidant effects, but also to be effective for the treatment of Alzheimer's disease. In addition, GV1001 has completed toxicity tests in various clinical trials targeting various diseases, and the safety in terms of side effects has been confirmed.

[0012] However, although GV1001 has excellent efficacy, it is easily cleaved within the body and has a short half-life, and thus there is a problem that frequent administration is inevitable when used as a drug.

[0013] Therefore, the present inventors have intensively researched to improve the stability of GV1001 peptide, and as a result, have invented a novel peptide having not only high stability but also more excellent therapeutic efficacy in amyloidosis and macular degeneration. SUMMARY

[0014] TECHNICAL PROBLEM

[0015] The present application aims to provide a novel peptide having preventive or therapeutic efficacy in amyloidosis and macular degeneration.

[0016] The present application aims to provide a pharmaceutical composition for preventing or treating Alzheimer's disease.

[0017] The present application aims to provide a health functional food for preventing or improving Alzheimer's disease.

[0018] The present application aims to provide a pharmaceutical composition for preventing or treating Cerebral amyloid angiopathy (CAA).

[0019] The present application aims to provide a health functional food for preventing or improving cerebral amyloid angiopathy.

[0020] The present application aims to provide a pharmaceutical composition for preventing or treating age-related macular degeneration.

[0021] The present application aims to provide a health functional food for preventing or improving age-related macular degeneration.

[0022] Technical Solution

[0023] 1. A peptide of the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof:

[0024] [Chemical Formula 1]

[0025]

[0026] 2. A pharmaceutical composition for preventing or treating amyloidosis, comprising the peptide of the above 1 or a pharmaceutically acceptable salt thereof.

[0027] 3. The pharmaceutical composition for preventing or treating amyloidosis according to the above 2, wherein the amyloidosis is Alzheimer's disease or cerebral amyloid angiopathy.

[0028] 4. The pharmaceutical composition for preventing or treating amyloidosis according to the above 2, wherein the prevention or treatment of amyloidosis is achieved by reducing deposition of amyloid beta (Aβ).

[0029] 5. A pharmaceutical composition for preventing or treating macular degeneration, comprising the peptide of the above 1 or a pharmaceutically acceptable salt thereof.

[0030] 6. The pharmaceutical composition for preventing or treating macular degeneration according to the above 5, wherein the macular degeneration is age-related macular degeneration.

[0031] 7. The pharmaceutical composition for preventing or treating macular degeneration according to the above 5, wherein the prevention or treatment of macular degeneration is achieved by reducing degeneration of retinal pigment epithelial cells (RPE cells).

[0032] 8. A health functional food for preventing or improving amyloidosis, comprising a peptide of the following Chemical Formula 1 or a food-acceptable salt thereof:

[0033] [Chemical Formula 1]

[0034]

[0035] 9. The health functional food for preventing or improving amyloidosis according to claim 8, wherein the amyloidosis is Alzheimer's disease or cerebral amyloid angiopathy.

[0036] 10. The health functional food for preventing or improving amyloidosis according to claim 8, wherein the prevention or improvement of amyloidosis is achieved by reducing β-amyloid protein deposition.

[0037] 11. A health functional food for preventing or improving macular degeneration comprising a peptide of the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof:

[0038] [Chemical Formula 1]

[0039]

[0040] 12. The health functional food for preventing or improving macular degeneration according to claim 11, wherein the macular degeneration is age-related macular degeneration.

[0041] 13. The health functional food for preventing or improving macular degeneration according to claim 11, wherein the prevention or improvement of macular degeneration is achieved by reducing retinal pigment epithelial cell degeneration.

[0042] Effects of the Invention

[0043] The peptide of Chemical Formula 1 and the pharmaceutically acceptable salt thereof according to the present invention have an excellent effect of reducing β-amyloid plaque deposition.

[0044] The peptide of Chemical Formula 1 and the pharmaceutically acceptable salt thereof according to the present invention have an effect of reducing β-amyloid protein level.

[0045] The peptide of Chemical Formula 1 and the pharmaceutically acceptable salt thereof according to the present invention have an effect of reducing inflammatory cytokine production.

[0046] The peptide of Chemical Formula 1 and the pharmaceutically acceptable salt thereof according to the present invention have an effect of reducing neuroglial proliferation.

[0047] The peptide of Chemical Formula 1 and the pharmaceutically acceptable salt thereof according to the present invention have an effect of reducing reactive oxygen species (ROS).

[0048] The peptide of Chemical Formula 1 and the pharmaceutically acceptable salt thereof according to the present invention have an effect of reducing retinal pigment epithelial cell degeneration.

[0049] The pharmaceutical composition and the health functional food comprising the peptide of Chemical Formula 1 or the salt thereof according to the present invention can exhibit an effect of preventing, improving and / or treating Alzheimer's disease, cerebral amyloid angiopathy and age-related macular degeneration.

[0050] The peptide, the pharmaceutical composition, and the health functional food of the present application can exhibit a pharmaceutical effect in a living body for a long time.

[0051] The peptide of the present application is more stable than GV1001 and has a more excellent therapeutic effect on amyloidosis and macular degeneration.

[0052] Compared to GV1001, the peptide of the present application can exhibit a therapeutic effect on amyloidosis or macular degeneration even with a smaller dose or a smaller number of administrations. BRIEF DESCRIPTION OF DRAWINGS

[0053] Figure 1 Experimental results showing confirmation of stability of GV2001 in human plasma are shown.

[0054] Figure 2 Experimental results showing confirmation of pharmacokinetic profiles (PK profiles) of GV2001 or GV1001 after intravenous administration to rats are shown.

[0055] Figure 3 Experimental results showing confirmation of pharmacokinetic profiles of GV2001 or GV1001 after subcutaneous administration to rats are shown.

[0056] FIG. 4 shows experimental results of an evaluation of improvement in spatial learning ability of an Alzheimer's disease mouse model.

[0057] FIGS. 5 and Figure 6 Experimental results showing evaluation of the efficacy of GV2001 in reducing β-amyloid protein by immunoblotting and enzyme-linked immunosorbent assay (ELISA) are shown.

[0058] FIG. 7 shows experimental results of evaluation of the efficacy of GV2001 in reducing inflammatory cytokines.

[0059] FIGS. 8 to 11 show experimental results of evaluation of the efficacy of GV2001 in reducing β-amyloid plaques and amyloid protein by histological analysis.

[0060] FIGS. 12 and 13 show experimental results of evaluation of the efficacy of GV2001 in reducing glial proliferation.

[0061] Figure 14 Experimental results showing evaluation of the efficacy of GV2001 in reducing reactive oxygen species are shown.

[0062] Figure 15 Experimental results showing evaluation of the efficacy of GV2001 in reducing retinal pigment epithelial cell degeneration are shown. DETAILED DESCRIPTION

[0063] The present application provides a peptide of the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof:

[0064] [Chemical Formula 1]

[0065]

[0066] In the present application, the peptide of Chemical Formula 1 includes functional equivalents thereof. The "functional equivalents" mean a peptide that exhibits substantially identical physiological activity to the peptide of Chemical Formula 1.

[0067] The present application provides a pharmaceutical composition for preventing or treating amyloidosis, comprising a peptide of Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an effective ingredient.

[0068] In the present application, the "pharmaceutically acceptable" means a non-toxic property exhibited to cells or individuals exposed to the composition.

[0069] In the present application, the "amyloidosis" is a general term for diseases caused by deposition of β-amyloid protein, including Alzheimer's disease, cerebral amyloid angiopathy, and Down's syndrome, etc.

[0070] In one embodiment, the amyloidosis can be Alzheimer's disease or cerebral amyloid angiopathy.

[0071] In the present application, the "preventing amyloidosis" means all actions of inhibiting, delaying, or reducing amyloidosis.

[0072] In the present application, the "improving amyloidosis" and "treating amyloidosis" mean all actions of improving or turning to a favorable direction of symptoms of individuals suspected of having or having amyloidosis.

[0073] In one embodiment, the prevention, improvement, and / or treatment of amyloidosis can be achieved by reducing β-amyloid protein.

[0074] In one embodiment, the prevention, improvement, and / or treatment of amyloidosis can be achieved by reducing β-amyloid protein plaques.

[0075] In one embodiment, the prevention, improvement, and / or treatment of amyloidosis can be achieved by reducing inflammatory cytokines.

[0076] In one embodiment, the prevention, improvement, and / or treatment of amyloidosis can be achieved by reducing neuroglial proliferation.

[0077] In one embodiment, the prevention, improvement, and / or treatment of amyloidosis can be improving spatial learning ability of individuals suspected of having or having amyloidosis.

[0078] The present application provides a pharmaceutical composition for preventing or treating macular degeneration, comprising a peptide of Chemical Formula 1 or a pharmaceutically acceptable salt thereof as an effective ingredient.

[0079] In the present application, "macular degeneration" includes Senile macular degeneration, Age-related macular degeneration (AMD), Macular degeneration in elderly, Myopic macular degeneration (MMD), and macular dystrophy.

[0080] In the present application, "Senile macular degeneration" includes dry macular degeneration and wet macular degeneration.

[0081] In one embodiment, the macular degeneration can be Senile macular degeneration.

[0082] In one embodiment, the macular degeneration can be dry macular degeneration.

[0083] In the present application, "preventing macular degeneration" means all actions of inhibiting, delaying or reducing macular degeneration.

[0084] In the present application, "improving macular degeneration" and "treating macular degeneration" mean all actions of improving or turning to a favorable direction of symptoms of individuals suspected of macular degeneration onset or already having macular degeneration.

[0085] In one embodiment, the prevention, improvement and / or treatment of macular degeneration can be achieved by reducing reactive oxygen species.

[0086] In one embodiment, the prevention, improvement and / or treatment of macular degeneration can be achieved by reducing retinal pigment epithelial cell degeneration.

[0087] The pharmaceutical composition of the present application can include an effective ingredient alone or further include one or more pharmaceutically acceptable carriers, excipients or diluents.

[0088] The carriers, excipients or diluents that can be included in the pharmaceutical composition of the present application can be, but are not limited to, lactose, dextrose, sucrose, dextrin, maltodextrin, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, hydroxybenzoate, hydroxypropylbenzoate, talc, magnesium stearate or mineral oil.

[0089] The pharmaceutical composition of the present application can be provided by administering to an individual.

[0090] In the present application, the term "administration" means introducing a predetermined substance into an individual in an appropriate method, and the term "individual" means all animals such as livestock and mice, and for example, can be mammals including humans.

[0091] The administration route of the pharmaceutical composition of the present application can be, but is not limited to, oral, intravenous, intramuscular, intra-arterial, intra- medullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual, or rectal.

[0092] In one embodiment, the composition of the present application can be administered orally or non-orally.

[0093] When the composition of the present application is administered non-orally, it is preferable, but not limited to, selected from the group consisting of external use on the skin or intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

[0094] The pharmaceutical composition of the present application can be formulated into a solid preparation for oral administration, such as a tablet, a pill, a powder, a granule, or a capsule.

[0095] The pharmaceutical composition of the present application can be formulated into a liquid preparation for oral administration, such as a suspension, a liquid for internal use, an emulsion, or a syrup.

[0096] The pharmaceutical composition of the present application can be formulated into a preparation for non-oral administration, such as a sterile aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized preparation, or a suppository.

[0097] The pharmaceutical composition of the present application can be administered to an individual in a pharmaceutically effective dose, and the term "pharmaceutically effective dose" means a dose sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment.

[0098] The pharmaceutically effective dose of the pharmaceutical composition of the present application can be determined according to factors including the severity of a disease, the activity of the pharmaceutical composition, the sensitivity of an individual or a patient to the pharmaceutical composition, the administration time, the administration route, and the excretion rate, the treatment period, and the combination of drugs, as well as other factors known in the medical field, and can be appropriately selected by one of ordinary skill in the art.

[0099] The pharmaceutical composition of the present application can be administered as an independent therapeutic agent, or in combination with other therapeutic agents of the past, and when administered in combination, can be administered sequentially or simultaneously, and can be administered once or multiple times, all of which can be easily determined by one of ordinary skill in the art.

[0100] The present application provides a health functional food for preventing or improving amyloidosis, which comprises a peptide of Chemical Formula 1 or a food-acceptable salt thereof.

[0101] In addition, the present application provides a health functional food for preventing or improving macular degeneration, which comprises the peptide of Chemical Formula 1 or a food-acceptable salt thereof.

[0102] The health functional food of the present application means a food prepared and processed using a material or ingredient having a functionality beneficial to the human body according to the relevant laws on health functional foods, and the functionality means ingestion for the purpose of regulating nutrition or obtaining an effect beneficial to health uses such as physiological action, with respect to the structure or function of the human body.

[0103] The food composition of the present application can comprise general food additives, and whether or not a food additive is applicable is determined according to the relevant standards and specifications of the corresponding item, in accordance with the General Principles and General Test Methods of the Food Additives Code approved by the Food and Drug Safety Department, etc., in the absence of other provisions.

[0104] The items included in the Food Additives Code, for example, can include chemical synthetics such as ketones, glycine, potassium citrate, nicotinic acid, cinnamic acid, etc., natural additives such as persimmon pigment, licorice extract, crystalline cellulose, sorghum pigment, guar gum, etc., and mixed preparations such as L-glutamate sodium preparation, flour additive alkali preparation, preservative preparation, tar pigment preparation, etc.

[0105] The health functional food of the present application can comprise the peptide of Chemical Formula 1 in an amount of 0.01 to 95% by weight, preferably 1 to 80% by weight, with respect to the total weight of the health functional food, for the purpose of preventing and / or improving diseases. In addition, it can be prepared and processed into a form such as tablets, capsules, powders, granules, liquids, pills, etc., for the purpose of preventing and / or improving diseases.

[0106] Hereinafter, examples are exemplified in detail for the purpose of specifically describing the present application. However, the following examples are provided only for the purpose of making the present application easier to understand, and the content of the present application is not limited to the following examples.

[0107] Embodiments

[0108] [Example 1] Preparation of GV2001

[0109] A peptide having the following Chemical Formula 1 and composed of 17 amino acids (hereinafter, abbreviated as GV2001) was prepared.

[0110] [Chemical Formula 1]

[0111]

[0112] GV2001 was synthesized by sequentially coupling amino acids from the C-terminal, according to the well-known Fmoc (fluorenylmethyloxycarbonyl) solid phase peptide synthesis (SPPS) method.

[0113] All amino acid starting materials used in peptide synthesis employed N-terminal protection with Fmoc or Boc (tert-butyloxycarbonyl), and residues protected with acid-removable Boc, t-Bu (t-butylester), or Pbf (2,2,4,6,7-pentamethyldihydro-benzofuran-5-sulfonyl). Examples are as follows:

[0114] Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Boc-Pyr-OH, Fmoc-Lys(Boc)-OH, Fmoc-Pro-OH, Fmoc-Leu-OH, Fmoc-Ile-OH, Fmoc-Phe-OH, Fmoc-Ser-OH, Fmoc-Thr-OH, Fmoc-Lys(Boc)-OH, Fmoc-Arg(Pbf)-OH.

[0115] The coupling reagents used were DIC (diisopropylcarbodiimide), DMF (dimethylformamide), Oxyma (ethyl 2-oxime cyanoacetate), and IPA (isopropyl alcohol). Fmoc removal employed a DMF solution containing 20% ​​piperidine (piperidine in DMF). A cleavage cocktail [95% TFA (trifluoroacetic acid) / 5% H2O / 50 mg / mL DTT (dithiothreitol)] was used to separate the synthesized peptides in the resin and remove protecting groups from residues.

[0116] The peptide was synthesized by utilizing the state of the starting amino acid with an amino acid protecting group bound to a solid-phase support, allowing each amino acid to react sequentially, followed by solvent washing and deprotection, and repeating this process. The synthesized peptide was cleaved from the resin, purified by HPLC (high performance liquid chromatography), and its synthesis was confirmed by MS (mass spectrometry), followed by freeze-drying.

[0117] In the counter ion exchange process, GV2001 dissolved in water in the form of HOAc (acetic acid) salt was replaced with AmberChrom ion exchange resin into the form of chloride. The GV2001 after replacement was filtered with PVDF (polyvinylidene fluoride) filter membrane and then freeze-dried.

[0118] The specific synthesis process of GV2001 is as follows.

[0119] (1) Coupling

[0120] In the Fmoc-Rink methylbenzhydrylamine resin, each amino acid from C-terminal to N-terminal and coupling reagent DIC / Oxyma / IPA dissolved in DMF were added to perform the coupling of amino acids. Then it was cleaned with DMF.

[0121] (2) Fmoc and Boc deprotection

[0122] DMF solution containing 20% piperidine was added to perform deprotection.

[0123] (3) Cleavage

[0124] The cleavage mixture was added to the synthesized peptide resin to separate the peptide from the resin.

[0125] (4) Filtration

[0126] The obtained mixture was concentrated, precipitated with MTBE (methyl tert-butyl ether) / hexane, and then filtered and dried.

[0127] (5) Purification

[0128] The obtained dry filtrate was purified with Prep-HPLC (preparative high performance liquid chromatography), the molecular weight was confirmed by LC / MS (liquid chromatograph-mass spectrometer), and then freeze-dried.

[0129] (6) Powder preparation

[0130] The AmberChrom resin was replaced with chloride, and then freeze-dried after PVDF filtration to prepare the powder.

[0131] [Example 2] Confirmation of stability of GV2001 in human plasma

[0132] GV2001 and GV1001 were added to 100% human plasma and diluted to 5 μM, respectively. The mixtures were stirred at 37°C for 10, 60, and 240 minutes, and then collected. Pretreatment was performed using methanol containing 0.1% formic acid via plasma protein precipitation. The supernatant was collected and analyzed by LC-MS (liquid chromatography-mass spectrometry) according to the conditions in Table 1 below to determine the percentage (%) of residual peptide relative to the amount of peptide before the reaction.

[0133] [Table 1]

[0134]

[0135]

[0136] like Figure 1 As shown, the ratio of residual peptide to unreacted peptide in GV2001 at each time point was higher than that in GV1001. This confirms that GV2001 is more stable in plasma than GV1001.

[0137] [Example 3] Pharmacokinetic curve confirmation of GV2001

[0138] To confirm the pharmacokinetic curve of GV2001 prepared according to Example 1, plasma from rats that had been administered GV2001 was collected, and methanol containing 0.1% formic acid was added at a volume of 9 times that of the plasma. The mixture was pretreated by protein precipitation, and after centrifugation at 16100g for 10 minutes, the supernatant was collected and analyzed by LC-MS according to the conditions in Table 2 below.

[0139] [Table 2]

[0140]

[0141] 3-1. Confirm the pharmacokinetic curve of GV2001 by intravenous administration.

[0142] GV2001 and GV1001 were administered intravenously (IV) to three SD rats (Sprague Dawley Rats) in each group. Blood samples were collected at 0.033, 0.083, 0.17, 0.25, 0.5, 1, and 2 hours post-administration. Plasma was separated from the blood samples by centrifugation, pretreated, and analyzed by LC-MS. Pharmacokinetic parameters were calculated using the non-compartmental analysis model of the Phoenix WinNonlin (Pharsight ver 6.4, USA) program.

[0143] The result, such as Figure 2 As shown, the AUC of GV2001t The area under the plasma concentration-time curve (AUC) was 0.107 ug·h / mL, which is higher than the AUC of GV1001. t The value (0.024 ug·h / mL) is more than 4 times higher than that of GV1001. This confirms that even in organisms, GV2001 is more stable than GV1001, less prone to degradation, and has a longer residence time.

[0144] 3-2. Confirm the pharmacokinetic profile of GV2001 by subcutaneous administration.

[0145] GV2001 and GV1001 were administered subcutaneously (SC) to three SD rats in each group. Blood samples were collected at 0.033, 0.083, 0.17, 0.25, 0.5, 1, and 2 hours post-administration. Plasma was separated from the blood samples by centrifugation, pretreated, and analyzed by LC-MS. Pharmacokinetic parameters were calculated using a non-compartmental analysis model in the Phoenix WinNonlin (Pharsight ver 6.4, USA) program.

[0146] The result, such as Figure 3 As shown, it was confirmed that even when administered subcutaneously, GV2001 is more stable than GV1001 in vivo, less prone to degradation, and has a longer residence time.

[0147] [Example 4] Efficacy evaluation of GV2001 based on an animal model of Alzheimer's disease

[0148] According to Table 3 below, the efficacy evaluation of GV2001 based on an animal model of Alzheimer's disease was carried out.

[0149] [Table 3]

[0150]

[0151] The 5xFAD mouse is a mouse model of Alzheimer's disease. Its neurons produce β-amyloid protein, which begins to accumulate from 2 months of age. Neurological damage occurs from 6 months of age, and spatial learning ability begins to be impaired from 9 months of age. It is purchased from Jackson Lab. and 7-month-old mice are obtained through mating for experiments.

[0152] 4-1. Evaluate the improvement in spatial learning ability using the Morris water maze test.

[0153] The water maze test showed that the WT group exhibited a gradual rapid finding of the platform during the 10-day training period, and showed a difference from the 5xFAD group. The 5xFAD group did not significantly reduce the latency during the 10-day training period, but the GV1001 group and the GV2001 group exhibited an improved situation Figure 4a ). The probe test was performed on the 11th day and the zone cross was analyzed, and the results showed that the 5xFAD group had a reduced number of crossings compared to the WT group, and the GV1001 group and the GV2001 group were improved Figure 4b ). The GV1001 group and the GV2001 group also exhibited an improved effect in terms of target quadrant residence time Figure 4c .

[0154] 4-2. Evaluation of reduction of beta amyloid by immunoblotting and enzyme-linked immunosorbent assay (ELISA)

[0155] To confirm the beta amyloid levels in the brains of the mice in each group, immunoblotting analysis was performed using mouse hippocampal protein samples and an anti-amyloid antibody (6E10). The results showed that the GV2001 group exhibited a decreasing tendency (Fig. 5). In addition, the beta amyloid levels were measured in the mouse cerebral cortex protein samples by Aβ1-42 enzyme-linked immunosorbent assay (ELISA). The results showed that the beta amyloid was reduced in the GV2001 group Figure 6 .

[0156] 4-3. Evaluation of reduction of inflammatory cytokines

[0157] The mRNA levels of cytokines (IL-1β, IL-6, and TNF-α) involved in inflammation were analyzed by qRT-PCR (quantitative reverse transcription polymerase chain reaction) using mouse hippocampal samples. For all cytokines, the 5xFAD group exhibited an increased cytokine expression compared to the WT group, and the GV2001 group exhibited a more significantly decreased cytokine expression compared to the 5xFAD group (Fig. 7).

[0158] 4-4. Evaluation of reduction of beta amyloid plaques and beta amyloid by histological analysis

[0159] Using mouse brain tissue sections, β-amyloid plaques and β-amyloid were stained by thioflavin-S staining and anti-amyloid antibody (6E10) staining, respectively, and analyzed. The results of thioflavin-S staining showed that both GV1001 and GV2001 groups exhibited a reduced degree of β-amyloid plaque deposition in the cerebral cortex (Figure 8). In the hippocampus, the GV2001 group exhibited a significantly reduced degree of plaque deposition (Figure 9).

[0160] The results of 6E10 antibody staining to confirm the β-amyloid level showed that the GV2001 group exhibited a more significant reduction in β-amyloid level in the cerebral cortex and hippocampus (Figures 10 and 11).

[0161] 4-5. Evaluation of reduction in glial proliferation

[0162] Mouse brain tissue sections were stained using anti-Iba-1 antibody, which can stain microglia, and the level of mouse brain glial proliferation was analyzed.

[0163] The results of microglia staining showed that the GV2001 group exhibited a more significant reduction in glial proliferation level (Figures 12 and 13).

[0164] [Example 5] Evaluation of GV2001 efficacy based on a cell model of age-related macular degeneration

[0165] A retinal pigment epithelial cell line, ARPE-19, was treated with hydrogen peroxide (H2O2) to generate reactive oxygen species, establishing a cell model of age-related macular degeneration.

[0166] The experimental groups were set according to Table 4 below, and the efficacy of GV2001 was evaluated.

[0167] [Table 4]

[0168]

[0169] * GV2001 pre-treatment is a 2-hour treatment with GV2001 followed by a 24-hour treatment with H2O2.

[0170] * GV2001 post-treatment is a 2-hour treatment with H2O2 followed by a 24-hour treatment with GV2001.

[0171] * The positive control group of GV2001 used 1mM metformin (Met), which has been shown to improve oxidative stress in H2O2-treated retinal pigment epithelial cells.

[0172] * All medications should be diluted in 0.9% saline solution before use.

[0173] 5-1. Evaluate the decrease in reactive oxygen species

[0174] The changes in reactive oxygen species (ROS) levels in GV2001-induced retinal pigment epithelial cells (ARPE-19) were confirmed by fluorescence microscopy using 2',7'-Dichlorofluorescin Diacetate (DCF-DA) as a reactive oxygen species staining reagent.

[0175] like Figure 14 As shown, DCF-DA fluorescence expression increased after H2O2-induced oxidative stress, but decreased under metformin treatment (as a positive control). Under oxidative stress conditions, DCF-DA fluorescence expression decreased in the low-concentration treatment group with GV2001 pretreatment and in the high-concentration treatment group with GV2001 posttreatment, demonstrating an effect of reducing reactive oxygen species.

[0176] 5-2. Evaluation of the reduction in retinal pigment epithelial cell degeneration

[0177] Changes in cadherin 1, a marker of epithelial cells, and vimentin, a marker of mesenchymal cells, were observed using confocal microscopy to confirm the degree of epithelial-mesenchymal transition, which represents the degeneration of retinal pigment epithelial cells.

[0178] like Figure 15 As shown, after H2O2-induced oxidative stress, the expression of epithelial cell markers decreased, while the expression of mesenchymal cell markers increased, confirming the occurrence of epithelial-mesenchymal transition (EMT). In the positive control group, metformin treatment reduced EMT. GV2001 pretreatment reduced EMT in the low-concentration treatment group and posttreatment reduced it in the high-concentration treatment group.

Claims

1. A peptide of the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] 2. A pharmaceutical composition for preventing or treating amyloidosis comprising the peptide of claim 1 or a pharmaceutically acceptable salt thereof. 。 3. The pharmaceutical composition for preventing or treating amyloidosis according to claim 2, wherein, the amyloidosis is Alzheimer's disease or cerebral amyloid angiopathy.

4. The pharmaceutical composition for preventing or treating amyloidosis according to claim 2, wherein, the prevention or treatment of amyloidosis is achieved by reducing β-amyloid protein deposition.

5. A pharmaceutical composition for preventing or treating macular degeneration comprising the peptide of claim 1 or a pharmaceutically acceptable salt thereof.

6. The pharmaceutical composition for preventing or treating macular degeneration according to claim 5, wherein, the macular degeneration is age-related macular degeneration.

7. The pharmaceutical composition for preventing or treating macular degeneration according to claim 5, wherein, the prevention or treatment of macular degeneration is achieved by reducing retinal pigment epithelial cell degeneration.

8. A health functional food for preventing or improving amyloidosis comprising the peptide of the following Chemical Formula 1 or a food-acceptable salt thereof: [Chemical Formula 1] 9. The health functional food for preventing or improving amyloidosis according to claim 8, wherein, the amyloidosis is Alzheimer's disease or cerebral amyloid angiopathy.

10. The health functional food for preventing or improving amyloidosis according to claim 8, wherein, the prevention or improvement of amyloidosis is achieved by reducing β-amyloid protein deposition.

11. A health functional food for preventing or improving macular degeneration comprising the peptide of the following Chemical Formula 1 or a food-acceptable salt thereof: [Chemical Formula 1] 12. The health functional food for preventing or improving macular degeneration according to claim 11, wherein, the macular degeneration is age-related macular degeneration.

13. The health functional food for preventing or improving macular degeneration according to claim 11, wherein, the prevention or improvement of macular degeneration is achieved by reducing retinal pigment epithelial cell degeneration. ​ 。 ​ ​ ​ ​ ​ ​ 。 ​ ​ ​ ​

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