Composition for preventing or treating degenerative brain diseases containing gossypetin

Abstract

The present invention relates to a composition for preventing or treating degenerative brain diseases, which comprises gossypetin or a salt thereof. The composition of the present invention has excellent effects in preventing and treating degenerative brain diseases and also has excellent effects in improving memory and cognitive function, so the composition of the present invention is useful not only for preventing and treating degenerative brain diseases but also for improving memory and cognitive function.

Description

[Technical Field]

[0001] The present invention relates to a composition for preventing or treating degenerative brain diseases, which comprises gossypetin or a salt thereof, and a method for treating degenerative brain diseases using the same. The present invention also relates to a composition for improving memory and cognitive function, which comprises gossypetin or a salt thereof. [Background technology]

[0002] Due to the development of income levels and medical and healthcare environments, the proportion of elderly populations is gradually increasing, especially in Western Europe. In Korea, the proportion of the population aged 65 or over exceeded 14% in 2017, making it an aging society, and it is predicted that Korea will enter a super-aged society by 2025. Therefore, solving geriatric diseases among the elderly population is emerging as an urgent social issue.

[0003] In particular, there is currently no effective treatment for Alzheimer's disease, which accounts for roughly half of all cases of dementia in the elderly, and no way to prevent it. There are five types of drugs approved by the US Food and Drug Administration (FDA) that are prescribed to Alzheimer's patients, including acetylcholinesterase inhibitors and glutamate receptor inhibitors, but all of these only serve to alleviate symptoms.

[0004] Degenerative brain diseases occur due to the deposition of protein aggregates in brain tissue. In Alzheimer's disease, Aβ peptides become tangled and form extracellular plaques. This generates reactive oxygen species, triggering an inflammatory response by microglia. Microglia secrete IL-1, TNFα, PGE2, NO, NOO-, O2, and H2O2, promoting the death of surrounding neurons. Reactive oxygen species are generated by oxidative stress, which induces the release of cytochrome C in mitochondria and the activation of caspase-3, promoting apoptosis. As Aβ peptide aggregation increases, the aggregation of tau protein, which stabilizes microtubules within neurons, is also induced. Microtubule disruption leads to atrophy of neuronal axons and dendrites, resulting in neuronal degeneration. Furthermore, the aggregation and intracellular accumulation of tau protein disrupts neuronal signaling, including the transport of substances within neurons, leading to apoptosis.

[0005] In both Alzheimer's disease and Huntington's disease, apoptosis occurs when the protein huntingtin aggregates and accumulates within cells. In particular, if the CAG repeat sequence is genetically generated in the huntingtin gene, more than 100 glutamine residues are attached to the N-terminus of the protein, making it difficult for the protein to form its normal structure. Furthermore, if the function of chaperone proteins is weakened, it becomes difficult for the protein to form its three-dimensional structure, and misfolded proteins become tangled and form aggregates. Similarly, in Parkinson's disease, the aggregation of α-synuclein protein increases, forming thread-like structures with an amyloid fibril structure. This increases the formation of Lewy bodies within cells, impairing cellular function and ultimately leading to neuronal death. Recently, the role of normal α-synuclein has been elucidated. When calcium ion concentrations increase within nerve cells, α-synuclein attaches to the endoplasmic reticulum, which stores neurotransmitters, ensuring normal secretion. It acts as a calcium sensor, maintaining a delicate balance between calcium and α-synuclein within the cell. Therefore, if this balance is disrupted, α-synuclein begins to aggregate, leading to Parkinson's disease.

[0006] Thus, a typical symptom of degenerative brain diseases is the increase in protein aggregates in brain tissue. Protein aggregates trigger an inflammatory immune response in brain tissue, which then spreads to surrounding tissues. This disrupts the function of intracellular organelles, leading to the progression of apoptosis and resulting in the degeneration of brain tissue.

[0007] Since degenerative brain diseases require long-term drug administration, it is essential to develop substances with little toxicity and high efficacy. Therefore, if natural edible substances with therapeutic and preventive functions are developed, side effects can be minimized.

[0008] Meanwhile, Korean Patent Publication No. 10-1424547 (Patent Document 1) and Korean Patent Publication No. 10-2015-0047687 (Patent Document 2) disclose compositions for treating degenerative brain diseases containing lactic acid bacteria or products fermented by lactic acid bacteria, but these have limited commercial application due to their limited therapeutic effect on degenerative brain diseases such as dementia. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] Republic of Korea Registered Patent Publication No. 10-1424547 [Patent Document 2] Republic of Korea Patent Publication No. 10-2015-0047687 Summary of the Invention [Problem to be solved by the invention]

[0010] As a result of extensive efforts, the present inventors have confirmed that gossypetin, a small molecule substance derived from natural products, has excellent efficacy in preventing and treating degenerative brain diseases, thereby completing the present invention. [Means for solving the problem]

[0011] An object of the present invention is to provide a pharmaceutical composition for preventing or treating degenerative brain diseases, which comprises gossypetin or a salt thereof.

[0012] Another object of the present invention is to provide a composition for inhibiting VRK2 activity, which comprises gossypetine or a salt thereof.

[0013] Another object of the present invention is to provide a functional health food composition containing gossypetin or a salt thereof for preventing or ameliorating degenerative brain diseases.

[0014] Another object of the present invention is to provide a pharmaceutical composition for improving memory or cognitive function, comprising gossypetine or a salt thereof.

[0015] Another object of the present invention is to provide a functional health food composition for improving memory or cognitive function, which contains gossypetin or a salt thereof.

[0016] Another object of the present invention is to provide a method for preventing or treating degenerative brain diseases, which comprises administering to a non-human individual a composition containing gossypetine or a salt thereof. [Effects of the Invention]

[0017] The present invention has revealed that gossypetin has excellent effects in inhibiting protein aggregation in brain cells, and that animal experiments have also shown that it has excellent effects in preventing and treating degenerative brain diseases and improving memory.Therefore, the composition containing gossypetin of the present invention is useful for preventing and treating degenerative brain diseases and improving memory and cognitive function. [Brief explanation of the drawings]

[0018] [Figure 1] These results demonstrate the inhibitory effect of gossypetin on intracellular protein aggregation. (a) shows the amount of protein aggregates that did not pass through a filter after cells were treated with gossypetin, and (b) shows the amount of protein aggregates determined by gel electrophoresis. (c) shows the formation of protein aggregates in cells and their reduction by gossypetin treatment, as indicated by the aggregation of green fluorescent protein. (d) shows a quantitative analysis of this. (e) shows a comparative analysis of the amount of insoluble aggregates compared to soluble protein aggregates, and (f) shows the amount of soluble protein aggregates. (g) shows the number of cells containing such protein aggregates, and (h) shows the total number of cells expressing fluorescent protein. [Figure 2] The results show the degree of inhibition of VRK2 activity depending on the concentration of gossypetin. [Figure 3]This is the result of re-evaluating the short-term memory of Alzheimer's-induced mice in a Y-maze test in response to gossypetin. [Figure 4] The results showed that gossypetin had an effect on improving learning function by measuring the escape latency of Alzheimer's-induced mice through the Morris water maze test. [Figure 5] The results of measuring the memory ability of Alzheimer's-induced mice for the location of the platform in the Morris water maze test with gossypetin. (a) shows the time spent at the platform after it was removed, and (b) shows the total distance traveled by the mice in the water maze test. [Figure 6] These are the results of measuring the degree to which gossypetin reduces the deposition of amyloid aggregates in the hippocampus of mouse brains. [Figure 7] These are the results of biochemical measurements of the degree of amyloid aggregate deposition in mouse brains in response to gossypetin. (a) The amount of amyloid aggregates that did not pass through a filter among protein aggregates accumulated in the brain was measured, (b) The amount of soluble amyloid aggregates was measured, (c) The amount of amyloid aggregate polymers accumulated in the brain was analyzed by gel electrophoresis, and (d) The amount of amyloid monolayers and polymers was compared. [Figure 8] This shows the results of measuring the degree of reduction in microglial gliosis in mouse brains in response to gossypetin. [Figure 9] These are the results of measuring the degree of reduction in astrocyte-mediated gliosis in mouse brains in response to gossypetin. [Figure 10] The VRK2 activity inhibitory potency of gossypetin was compared with that of compounds known to significantly inhibit Aβ and tau protein aggregation in test tubes. [Figure 11] The results show that the degree to which gossypetin inhibits protein aggregation in cells is compared with that of compounds known to significantly inhibit Aβ and tau protein aggregation in test tubes. [Figure 12]The results show that the degree of improvement in short-term memory ability in Alzheimer's disease-induced mice was compared with the effect of gossypetin in a Y-maze test using a compound known to significantly inhibit the aggregation of Aβ and tau proteins in test tubes. [Figure 13] The results show that a compound known to significantly inhibit Aβ and tau protein aggregation in test tubes was used to compare the degree of improvement in short-term memory ability in Alzheimer's disease-induced mice with the effect of gossypetin in a maze experiment. DETAILED DESCRIPTION OF THE INVENTION

[0019] This will be explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention may also be applied to each different description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention is not limited by the specific description given below.

[0020] Additionally, those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein, and such equivalents are intended to be encompassed by the present invention.

[0021] One aspect of the present invention provides a pharmaceutical composition for preventing or treating a degenerative brain disease, comprising gossypetin or a salt thereof.

[0022] Another aspect of the present invention provides a health functional food composition for improving memory or cognitive function, comprising gossypetin or a salt thereof.

[0023] The composition may contain gossypetin as an active ingredient.

[0024] In the present invention, "gossypetine" may be a compound represented by the following chemical formula 1.

[0025] [ka]

[0026] The gossypetin may be a compound designated as 2-(3,4-dihydroxyphenyl)-3,5,7,8-tetrahydroxy-4H-chromen-4-one.

[0027] The gossypetine can be obtained by chemical synthesis, by extraction from plants, or by using commercially available substances, but the method of obtaining it is not particularly limited, and any method known in the art can be used.

[0028] The pharmaceutical composition of the present invention may contain not only the above-mentioned gossypetin but also its pharmaceutically acceptable salt. In the present invention, the term "pharmaceutically acceptable salt" refers to a salt form in which the above-mentioned compound is combined with another substance, which exhibits similar pharmacological activity.

[0029] The pharmaceutically acceptable salts include, but are not limited to, inorganic acid salts such as hydrochloride, hydrobromide, phosphate, or sulfate, and organic acid salts such as carboxylates and sulfonates. The carboxylates include, but are not limited to, acetate, maleate, fumarate, malate, citrate, tartrate, lactate, or benzoate. The sulfonates include, but are not limited to, methanesulfonate, ethanesulfonate, benzenesulfonate, toluenesulfonate, or naphthalenedisulfonate.

[0030] In the present invention, the term "degenerative brain disease" refers to a disease caused by brain cell damage.

[0031] The degenerative brain disease may be caused by or exhibits one or more of the following pathological features: protein aggregation, increased VRK2 activity, increased degradation of chaperone proteins, protein misfolding, Lewy body formation, polyglutamine aggregation, and glial inflammatory response. The protein aggregation may be, but is not limited to, aggregation of amyloid Aβ, tau protein, huntingtin, or α-synuclein.

[0032] Specifically, the degenerative brain disease of the present invention may be one or more diseases selected from the group consisting of dementia, Alzheimer's disease, Huntington's disease, Parkinson's disease, multiple system atrophy, multiple sclerosis, tauopathies, brain tumors, Pick's disease, and Creutzfeldt-Jakob disease, and more specifically, may be one or more diseases selected from Alzheimer's disease, Huntington's disease, and Parkinson's disease, but is not limited thereto.

[0033] In the present invention, "memory / cognitive function" refers to the ability to efficiently manipulate knowledge and information, including all processes using the brain, such as thinking, speaking, remembering, judging, and executing. Damage to brain cells can result in the loss or impairment of memory and cognitive function, which manifests as symptoms such as amnesia, memory decline, and memory impairment.

[0034] In the present invention, the term "prevention" means any action that suppresses or delays the onset of cognitive dysfunction or neuroinflammation by administering the composition according to the present invention, "treatment" means any action that improves or beneficially changes the symptoms of an individual suspected of or affected with cognitive dysfunction or neuroinflammation by administering the pharmaceutical composition, and "improvement" means any action that at least reduces a parameter related to the condition being treated by administering the composition of the present invention, for example, the severity of the symptom.

[0035] In one embodiment of the present invention, gossypetin was confirmed to inhibit VRK2 activity in vitro, and administration of gossypetin to dementia-induced mice in vivo was confirmed to improve memory and inhibit the progression of dementia. Therefore, the composition of the present invention can be used to inhibit VRK2 activity and is useful for preventing and treating degenerative brain diseases as well as improving memory and cognitive function.

[0036] The composition of the present invention may further contain suitable carriers, excipients, and diluents that are commonly used in the manufacture of pharmaceutical compositions, and may be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, external preparations, suppositories, and sterile injection solutions by conventional methods.

[0037] The pharmaceutical compositions containing gossypetin or a pharmaceutically acceptable salt thereof according to the present invention can be formulated into external preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and sterile injection solutions in the usual manner, and can have the dosage forms of creams, gels, patches, sprays, ointments, plasters, lotions, liniments, pastes, or cataplasms, but are not limited thereto.

[0038] Carriers, excipients, and diluents that can be contained in the above-mentioned gossypetin-containing composition include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. When formulated, they are prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc. These solid dosage forms are prepared by mixing the above-mentioned composition with at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid dosage forms include suspensions, oral solutions, emulsions, syrups, etc., which may contain various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Parenteral dosage forms include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin.

[0039] Another aspect of the present invention provides a functional health food composition for preventing or ameliorating degenerative brain diseases, comprising gossypetin or a salt thereof.

[0040] Another aspect of the present invention provides a health functional food composition for improving memory or cognitive function, comprising gossypetin or a salt thereof.

[0041] The composition may contain gossypetin as an active ingredient.

[0042] Gossypetin, degenerative brain diseases, memory and cognitive function, prevention, treatment, and improvement have been described above.

[0043] When the gossypetin of the present invention is used as a food additive, it can be added directly or in combination with other foods or food ingredients, and can be used in a conventional manner. The amount of the active ingredient to be added can be appropriately determined depending on the intended use (prevention, health, or therapeutic treatment). Generally, the gossypetin of the present invention is added in an amount of 15% by weight or less, preferably 10% by weight or less, based on the raw materials during the production of foods or beverages. However, when taking the gossypetin for long-term purposes such as health and hygiene or health regulation, the amount may be less than the above range, and since there are no safety issues, the active ingredient may also be used in an amount greater than the above range.

[0044] There are no particular limitations on the types of foods to which the substances can be added. Examples of foods to which the substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, drinking water, tea, health drinks, alcoholic beverages, and vitamin complexes, all of which include health functional foods in the usual sense.

[0045] The health drink composition of the present invention may contain various flavorings or natural carbohydrates as additional ingredients, as in conventional beverages. The natural carbohydrates mentioned above include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Examples of sweeteners that can be used include natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrates is generally about 0.01 to 0.20 g, preferably about 0.04 to 0.10 g, per 100 mL of the composition of the present invention.

[0046] In addition to the above, the compositions of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, etc. The compositions of the present invention may also contain fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These ingredients may be used independently or in combination. The proportion of these additives is not critical, but is typically selected in the range of 0.01 to 0.20 parts by weight per 100 parts by weight of the composition of the present invention.

[0047] Other ingredients that may be added include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohol, colorants, fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, etc.

[0048] Examples of the oil and fat component include ester-based oils, hydrocarbon-based oils, silicone-based oils, fluorine-based oils, animal oils, and vegetable oils.

[0049] Ester-based oils and fats include glyceryl tri-2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, butyl stearate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, diisopropyl adipate, neo Isoalkyl pentanoate, caprylic / capric triglyceride, trimethylolpropane tri-2-ethylhexanoate, trimethylolpropane triisostearate, pentaerythritol tetra-2-ethylhexanoate, cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, cetyl ricinoleate, isostearyl laurate, isotridecyl myristate, isocetyl palmitate, octyl stearate, stearin Isocetyl oleate, isodecyl oleate, octyldodecyl oleate, octyldodecyl linoleate, isopropyl isostearate, cetostearyl 2-ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, propylene glycol dicaprylate, propylene glycol dicaprylate, neopentyl glycol dicaprate, neopentyl glycol dioctanoate, glyceryl tricaprylate, tri Glyceryl undecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyldodecyl neopentanoate, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, octyldecyl isostearate, polyglycerin oleate, polyglycerin isostearate, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate,Examples of esters include cetyl lactate, octyldecyl lactate, triethyl citrate, acetyltriethyl citrate, acetyltributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearate, di-2-ethylhexyl succinate, diisobutyl adipate, diisopropyl sebacate, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dehydrocholesteryl oleate, phytosteryl isostearate, phytosteryl oleate, isocetyl 12-stearoylhydroxystearate, stearyl 12-stearoylhydroxystearate, and isostearyl 12-stearoylhydroxystearate.

[0050] Examples of hydrocarbon oils include squalene, liquid paraffin, α-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, and petrolatum.

[0051] Examples of silicone oils and fats include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane-methylcetyloxysiloxane copolymer, dimethylsiloxane-methylstearoxysiloxane copolymer, alkyl-modified silicone oil, and amino-modified silicone oil.

[0052] Examples of fluorine-based oils include perfluoropolyethers.

[0053] Examples of animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, safflower oil, soybean oil, corn oil, rapeseed oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil, cottonseed oil, palm oil, coconut oil, wheat germ oil, rice germ oil, shea butter, evening primrose oil, macadamia nut oil, meadowfoam oil, egg yolk oil, beef tallow, hemp oil, mink oil, orange roughy oil, jojoba oil, candelilla wax, carnauba wax, liquid lanolin, and hydrogenated castor oil.

[0054] Examples of the moisturizing agent include water-soluble low-molecular moisturizing agents, fat-soluble molecular moisturizing agents, water-soluble polymers, and fat-soluble polymers.

[0055] Examples of water-soluble low-molecular-weight moisturizers include serine, glutamine, sorbitol, mannitol, sodium pyrrolidone carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (degree of polymerization n=2 or more), polypropylene glycol (degree of polymerization n=2 or more), polyglycerin B (degree of polymerization n=2 or more), lactic acid, and lactate salts.

[0056] Examples of the fat-soluble low-molecular-weight moisturizing agent include cholesterol and cholesterol esters.

[0057] Examples of water-soluble polymers include carboxyvinyl polymers, polyaspartates, tragacanth, xanthan gum, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, water-soluble chitin, chitosan, and dextrin.

[0058] Lipid-soluble polymers include polyvinylpyrrolidone-eicosene copolymer, polyvinylpyrrolidone-hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and polymeric silicone.

[0059] Emollients include long-chain acyl glutamic acid cholesteryl ester, cholesteryl hydroxystearate, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl ester, and the like.

[0060] Examples of the surfactant include a nonionic surfactant, an anionic surfactant, a cationic surfactant, and an amphoteric surfactant.

[0061] Nonionic surfactants include self-emulsifying glycerin monostearate, propylene glycol fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, POE (polyoxyethylene) sorbitan fatty acid esters, POE sorbit fatty acid esters, POE glycerin fatty acid esters, POE alkyl ethers, POE fatty acid esters, POE hydrogenated castor oil, POE castor oil, POE·POP (polyoxyethylene·polyoxypropylene) copolymers, POE·POP alkyl ethers, polyether-modified silicones, lauric acid alkanolamide, alkylamine oxide, and hydrogenated soybean phospholipids.

[0062] Examples of anionic surfactants include fatty acid soaps, α-acylsulfonates, alkylsulfonates, alkylarylsulfonates, alkylnaphthalenesulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, alkylamide phosphates, alkyloylalkyl taurine salts, N-acylamino acid salts, POE alkyl ether carboxylates, alkyl sulfosuccinates, sodium alkylsulfoacetates, acylated hydrolyzed collagen peptide salts, and perfluoroalkyl phosphate esters.

[0063] Examples of cationic surfactants include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, stearic acid diethylaminoethylamide, stearic acid dimethylaminopropylamide, and lanolin derivative quaternary ammonium salts.

[0064] Examples of amphoteric surfactants include carboxybetaine type, amidobetaine type, sulfobetaine type, hydroxysulfobetaine type, amidosulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, and amidoamine type amphoteric surfactants.

[0065] Organic and inorganic pigments include inorganic pigments such as silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, red iron oxide, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, chromium oxide, chromium hydroxide, calamine, and composites thereof; organic pigments such as polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenolic resin, fluororesin, silicone resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene-styrene copolymer, silk powder, cellulose, CI Pigment Yellow, and CI Pigment Orange, as well as composite pigments of these inorganic and organic pigments.

[0066] Organic powders include metal soaps such as calcium stearate; alkyl phosphate metal salts such as zinc sodium cetyl phosphate, zinc lauryl phosphate, and calcium lauryl phosphate; polyvalent metal salts of acyl amino acids such as calcium N-lauroyl-β-alanine, zinc N-lauroyl-β-alanine, and calcium N-lauroylglycine; polyvalent metal salts of amidosulfonic acids such as calcium N-lauroyl-taurate and calcium N-palmitoyl-taurate; N-acyl basic amino acids such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoyl lysine, N-α-palmitoylornithine, N-α-lauroylarginine, and N-α-hardened tallow fatty acid acylarginine; N-acyl polypeptides such as N-lauroylglycylglycine; α-amino fatty acids such as α-aminocaprylic acid and α-aminolauric acid; polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene-styrene copolymer, and tetrafluoroethylene.

[0067] UV absorbers include para-aminobenzoic acid, ethyl para-aminobenzoate, amyl para-aminobenzoate, octyl para-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomenthyl salicylate, benzyl cinnamate, 2-ethoxyethyl para-methoxycinnamate, octyl para-methoxycinnamate, mono-2-ethylhexaneglyceryl di-para-methoxycinnamate, isopropyl para-methoxycinnamate, diisopropyl and diisopropyl cinnamate ester mixture, uroca Examples of such benzotriazoles include hydroxybenzoic acid, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and its salts, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone sodium disulfonate, dihydroxybenzophenone, tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2'-ethylhexyl-1'-oxy)-1,3,5-triazine, and 2-(2-hydroxy-5-methylphenyl)benzotriazole.

[0068] Examples of disinfectants include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcinol, isopropylmethylphenol, azulene, salicylic acid, zinc pyrithione, benzalkonium chloride, photosensitizer No. 301, mononitroguaiacol sodium, and undecylenic acid.

[0069] Antioxidants include butylhydroxyanisole, propyl gallate, erythorbic acid, and the like.

[0070] Examples of pH adjusters include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, and sodium monohydrogen phosphate.

[0071] Examples of alcohol include ethyl alcohol and higher alcohols.

[0072] Furthermore, other ingredients that may be added are not limited to these, and any of the above ingredients can be added within a range that does not impair the objects and effects of the present invention. However, they are preferably added in an amount of 0.01 to 5% by weight, more preferably 0.01 to 3% by weight, based on the total weight.

[0073] When the dosage form of the present invention is a lotion, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or the like may be used as a carrier component.

[0074] When the dosage form of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly when it is a spray, it may further contain a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether.

[0075] When the dosage form of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan fatty acid esters.

[0076] When the dosage form of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth may be used as a carrier component.

[0077] When the dosage form of the present invention is a surfactant-containing cleanser, the carrier component may be a fatty alcohol sulfate, a fatty alcohol ether sulfate, a sulfosuccinic acid monoester, isethionic acid, an imidazolinium derivative, methyl taurate, sarcosinate, a fatty acid amide ether sulfate, an alkylamidobetaine, a fatty alcohol, a fatty acid glyceride, a fatty acid diethanolamide, a vegetable oil, a lanolin derivative, or an ethoxylated glycerol fatty acid ester.

[0078] Yet another aspect of the present invention provides a method for preventing or treating a degenerative brain disease, which comprises administering the above pharmaceutical composition to an individual.

[0079] In the present invention, the term "individual" refers to a subject requiring treatment for a disease. Specifically, it may refer to mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cows, and more specifically, it may refer to animals other than humans or humans, but is not limited thereto.

[0080] The pharmaceutical compositions of the present invention may be administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to any medical treatment, and the effective dose level can be determined by factors including the type and severity of the patient's disease, drug activity, drug sensitivity, administration time, administration route and excretion rate, treatment duration, concurrently used drugs, and other factors well known in the medical field. In one embodiment, the concentration of gossypetin administered may be 0.1 mg / kg to 500 mg / kg based on the body weight of the individual to be administered, specifically, but not limited to, 0.1 mg / kg to 100 mg / kg, 0.5 mg / kg to 100 mg / kg, 0.5 mg / kg to 50 mg / kg, 0.5 mg / kg to 30 mg / kg, 0.5 mg / kg to 15 mg / kg, 7 mg / kg to 13 mg / kg, and more specifically, 10 mg / kg. The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, sequentially or simultaneously with conventional therapeutic agents, or in single or multiple administrations. Taking all of the above factors into consideration, it is important to administer an amount that will achieve maximum efficacy at the minimum dose without side effects, which can be easily determined by one of ordinary skill in the art to which the present invention pertains.

[0081] The pharmaceutical composition of the present invention can be administered to an individual via various routes. All administration methods are conceivable, and may be administered, for example, by oral administration, subcutaneous, intravenous, intramuscular, intrauterine dura or intracerebrovascular injection. The pharmaceutical composition of the present invention is determined depending on the type of active ingredient drug, as well as various related factors such as the disease to be treated, the administration route, the patient's age, sex, weight and disease severity. [Example]

[0082] The present invention will be described in more detail below through examples and experimental examples. However, these examples and experimental examples are for illustrative purposes only and the scope of the present invention is not limited to these examples and experimental examples.

[0083] Example 1: Confirmation of the in vitro effects of gossypetin An in vitro experiment was carried out to determine whether gossypetin, a naturally occurring compound, is effective in preventing degenerative brain diseases.

[0084] Example 1-1: Confirmation of the concentration-dependent inhibitory effect of gossypetin on intracellular polyglutamine protein aggregation Polyglutamine protein aggregation is a cause of degenerative brain diseases. To confirm the inhibitory effect of gossypetin on polyglutamine protein aggregation, we expressed polyglutamine protein bound to green fluorescent protein in U2OS cells expressing VRK2, then treated them with various concentrations of gossypetin and confirmed the effect using a dot blot assay. In addition, protein aggregates formed within the cells were observed by electrophoresis, and the level of intracellular protein aggregate formation was quantitatively evaluated using a fluorescence microscope.

[0085] As a result, we confirmed that polyglutamine protein aggregates were reduced in a concentration-dependent manner (Fig. 1A, 1B). Furthermore, insoluble protein aggregates were significantly reduced (Fig. 1C-E), and the number of cells bearing protein aggregates was also reduced (Fig. 1F-1H).

[0086] Example 1-2: Confirmation of the mutual binding between gossypetin (GSP) and VRK2, and confirmation of gossypetin's concentration-dependent inhibition of VRK2 activity and conservation of chaperone proteins The kinase VRK2 is known to regulate the TRiC chaperone protein in cells and prevent the removal of polyglutamine aggregates. Therefore, we predicted that inhibiting VRK2 would inhibit polyglutamine aggregation. To confirm whether gossypetin has an inhibitory effect on VRK2 activity, we treated cells with various concentrations of gossypetin and measured the degree of VRK2 activity inhibition.

[0087] First, we chemically conjugated gossypetin to Sepharose 4B (Fig. 2A) and then examined whether gossypetin and VRK2 interacted with each other. VRK2 did not bind to Sepharose 4B, but only to gossypetin-Sepharose 4B, demonstrating the interaction between VRK2 and gossypetin (Fig. 2B). Furthermore, we confirmed that the interaction between gossypetin and VRK2 was specific, as we confirmed that the addition of gossypetin disrupted the binding between VRK2 and gossypetin-Sepharose 4B (Fig. 2C). We also demonstrated that gossypetin concentration-dependently inhibited VRK2 activity (Fig. 2D), and confirmed that the addition of gossypetin increased, but not decreased, the amount of the chaperone protein CCT-4 (Fig. 2E).

[0088] Experimental results confirmed that gossypetin inhibits VRK2 activity in vitro, suggesting that it may be effective in preventing and treating degenerative brain diseases.

[0089] Example 2: Confirmation of the in vivo effects of gossypetin Based on the results of Example 1, gossypetin was predicted to be effective in preventing and treating degenerative brain diseases. Animal experiments were conducted to confirm whether gossypetin is effective in actual animals.

[0090] For this experiment, 7-week-old 5xFAD Alzheimer's-induced mice were orally administered gossypetin at a concentration of 10 mg / kg for 14 weeks (Figures 3A and 3B). Subsequently, the mice were subjected to the Y-maze and Morris water maze tests, which are widely used tests to assess cognitive function. Specifically, the Y-maze test measured short-term memory. One week after the Y-maze test, the Morris water maze test was performed. The mice were trained to memorize the location of the platform for 5 days, and the time it took to find the platform was measured. Additionally, on the 6th day after platform training, the platform was removed from the tank, and the frequency with which the mice remained in the platform zone was measured.

[0091] The Y-maze experiment confirmed that the short-term memory of 5xFAD Alzheimer's-induced mice treated with gossypetin was improved to the same extent as that of normal mice (Figure 3C). Through training Platform Location Check whether you want to remember , compared with a normal control group. result, 5xFAD mice took longer to find the platform than normal controls. Gossypetine administration 5xFAD mice is a normal mouse At the level of We confirmed that the platform was found within a short period of time (Figure 4).

[0092] In addition, after the platform was removed, the frequency with which the mice stayed in the area where the platform was located was checked. Although the total distance traveled was similar, the group of Alzheimer's-induced mice fed gossypetin showed an increased frequency of staying in the area where the platform was located, confirming that the memory of the Alzheimer's-induced mice had improved (Figure 5).

[0093] Through this, it was proven that gossypetine has the effect of preventing and treating Alzheimer's disease, and it was also confirmed that it has an excellent effect in improving cognitive function.

[0094] Example 3: Analysis of brain tissue from Alzheimer-induced mice treated with gossypetin Example 3-1: Analysis of brain tissue from Alzheimer-induced mice treated with gossypetin The brains of Alzheimer's disease-induced mice (5xFAD) were extracted from groups fed gossypetin and those not fed gossypetin, and then sliced. immunohistochemistry The level of Aβ amyloid aggregate formation was analyzed using staining methods. When observing the brain tissues of the hippocampus (Figures 6A-6C) and frontal lobe (Figures 6D-6F), it was confirmed that Aβ amyloid aggregates were significantly reduced in the gossypetin-fed mice.

[0095] Example 3-2: Biochemical analysis of hippocampal regions in Alzheimer-induced mouse brains treated with gossypetin Brain tissue from Alzheimer's-induced mice fed gossypetin was crushed and analyzed using biochemical methods such as protein quantification and Western blot. Dot blot and Western blot results confirmed a statistically significant decrease in the amount of soluble Aβ amyloid protein, as well as both its monomeric and polymeric forms (Figures 7A-E). Aβ amyloid protein is a protein produced by enzymatic cleavage of the APP amyloid precursor protein. To determine whether the decrease in Aβ amyloid protein was due to a decrease in the amount of APP amyloid precursor protein, Western blot analysis confirmed that the amount of APP amyloid precursor protein remained unchanged (Figures 7F and 7G).

[0096] Example 3-3: Analysis of the degree of gliosis due to hyperactivity of microglia in the brains of Alzheimer-induced mice treated with gossypetin The brains of Alzheimer's disease-induced mice (5xFAD) were extracted and sectioned from groups fed gossypetin and unfed, and the distribution and proliferation of microglia and astrocytes were analyzed using antibodies against IB-1, a microglia-specific protein, and GFAP, an astrocyte-specific protein.

[0097] When we examined the tissues of the hippocampus (Figures 8A-8B, Figures 9A-9B) and frontal lobe (Figures 8C-8D, Figures 9C-9D) of the brain, we found that the hyperactivity of microglia and astrocytes, i.e., gliosis, was significantly reduced in the mice fed gossypetin.

[0098] Example 4: Comparison of VRK2 activity inhibitory effects of gossypetin with other compounds The level of VRK2 activity inhibition by gossypetin was compared with that of other compounds known to inhibit protein aggregation, which causes degenerative brain diseases, in vitro.

[0099] Specifically, the VRK2 inhibitory activity of rosmarinic acid and lacmoid, which are known to significantly suppress the aggregation of Aβ and tau proteins, the causes of degenerative brain diseases, in vitro, was measured using an in vitro kinase assay. In addition, cells were induced to undergo polyglutamine protein aggregation, and each compound was treated. The degree of reduction in intracellular protein aggregation was then observed using a fluorescence microscope. The total area of ​​protein aggregation in the cells, the average size of protein aggregates, and the degree of protein aggregation inhibition compared to the untreated control group were then tabulated. The cytotoxicity of each compound was also confirmed.

[0100] Measurement of VRK2 autophosphorylation activity showed that at the same concentration of 25 μM, gossypetin most potently inhibited VRK2 autophosphorylation activity, rosmarinic acid had a very weak inhibitory effect, and lacmoid had only a slight inhibition of VRK2 activity (Figure 10).

[0101] Furthermore, each compound showed almost no cytotoxicity (Fig. 11C), but gossypetin inhibited intracellular protein aggregation by 43%, whereas the other compounds did not inhibit intracellular protein aggregation (Figs. 11A-11B).

[0102] Through this, they confirmed that gossypetin is not toxic within cells and has the best effect in inhibiting protein aggregation. Furthermore, they confirmed that even if gossypetin has the activity to inhibit some of the causative substances of Alzheimer's in vitro, that is, at the test tube level, it does not have the activity against all of the causative substances of Alzheimer's, and therefore predicted that the effect would be different in in vivo experiments.

[0103] Example 5: Comparison of the in vivo effects of gossypetine with other compounds The compound of Example 4 was administered to an Alzheimer's mouse model to confirm its effect on improving actual cognitive function.

[0104] Example 5-1: Effect of prevention and treatment of Alzheimer's disease confirmed through Y-maze experiment Specifically, we used 5xFAD, an Alzheimer's disease-induced mouse model, and orally administered gossypetin (GSP), rosmarinic acid, and lacmoid at 10 mg / kg each for 15 weeks starting from the time the 5xFAD mice reached 8 weeks of age, and then performed a Y-maze experiment to examine cognitive function.

[0105] The experimental results showed that the performance of 5xFAD mice was significantly impaired compared to normal mice (Fig. 12A). Treatment with lacmoid (Fig. 12B) or rosmarinic acid (Fig. 12C), compounds known to inhibit Aβ and tau protein aggregation in vitro, failed to improve cognitive function. However, administration of gossypetin restored the behavioral changes to near-normal levels (Fig. 12D), confirming that gossypetin treatment improved cognitive function.

[0106] Example 5-2: Effects of Alzheimer's prevention and treatment confirmed through water maze experiments Specifically, 5xFAD mice with induced dementia were orally administered 10 mg / kg of gossypetin (GSP), rosmarinic acid, or lacmoid for 16 weeks, starting from the age of 8 weeks, and then subjected to a water maze experiment to examine cognitive function. The water maze experiment involved training the mice to find a platform placed under the water surface in a tank for 5 days, and then removing the platform on the 6th day to assess how well they could remember the location of the platform.

[0107] The results of the experiment showed that the memory ability of 5xFAD mice was significantly impaired compared to that of normal mice (Figure 13A). In addition, the addition of lacmoid and rosmarin, compounds known to inhibit Aβ and tau protein aggregation in vitro, significantly reduced the memory ability of 5xFAD mice. 5xFAD mice When treated with gossypetin, cognitive function was found to be worse than when no treatment was administered (Fig. 13B) and was not significantly improved (Fig. 13C). However, when treated with gossypetin, the mice exhibited better memory performance than normal mice (Fig. 13D).

[0108] Through this, it was confirmed that even compounds known to be effective in vitro show different results in in vivo experiments, and that gossypetine has an extremely effective effect in improving cognitive function.

[0109] From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are merely illustrative and not limiting. The scope of the present invention should be interpreted as including within the meaning and scope of the claims below, and any modifications or variations derived from the equivalent concepts thereof, rather than the above detailed description.

Claims

1. A pharmaceutical composition for preventing or treating Alzheimer's disease, comprising gossypetin or a salt thereof as an active ingredient, wherein the gossypetin is represented by the following chemical formula 1: 【Chemical 1】

2. 2. The pharmaceutical composition for preventing or treating Alzheimer's disease according to claim 1, wherein Alzheimer's disease is caused by one or more causes selected from the aggregation of beta-amyloid (Aβ) protein and the aggregation of tau protein.

3. A functional health food composition for preventing or ameliorating Alzheimer's disease, comprising gossypetin or a salt thereof as an active ingredient, wherein the gossypetin is represented by the following chemical formula 2: 【Chemistry 2】

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