Composition for treating Alzheimer's disease containing mixed extracts of Pinellia sinensis, Licorice Root, Glycyrrhiza Root, Korean Ginseng, Chinese Rhizome, Scutellaria Root, and Coptis Rhizome as active ingredients

Abstract

The present invention relates to the use of a composition containing Hangeshashinto as an active ingredient for the treatment, amelioration or prevention of Alzheimer's disease caused by ApoE4 gene mutation. The composition and treatment method of the present invention containing Hangeshashinto as an active ingredient exhibits excellent effects of reducing amyloid β protein and inhibiting accumulation specifically in Alzheimer's disease caused by ApoE4 gene mutation, and is highly useful as a composition for prevention, amelioration or treatment specific to Alzheimer's disease caused by ApoE4 gene mutation.

Description

[Technical Field]

[0001] The present invention relates to a pharmaceutical composition for preventing or treating Alzheimer's disease, which contains as an active ingredient a mixed extract of Pinellia Sinensis, Rhizome Rhizome, Licorice Root, Korean ginseng, Chinese cabbage, Scutellaria Root, and Coptis Rhizome; a functional food composition for preventing or ameliorating Alzheimer's disease, which contains as an active ingredient a mixed extract of Pinellia Sinensis, Rhizome Rhizome, Licorice Root, Korean ginseng, Chinese cabbage, Scutellaria Root, and Coptis Rhizome; a quasi-drug composition for preventing or ameliorating Alzheimer's disease, which contains as an active ingredient a mixed extract of Pinellia Sinensis, Rhizome Rhizome, Licorice Root, Korean ginseng, Chinese cabbage, Scutellaria Root, and Coptis Rhizome; and a method for treating Alzheimer's disease, which comprises the step of administering the pharmaceutical composition to an individual. [Background technology]

[0002] Alzheimer's disease is the most common degenerative brain disease, characterized by a gradual deterioration of cognitive functions, including memory. Major symptoms of Alzheimer's disease include memory loss, impaired language ability, impaired spatial and temporal awareness, impaired judgment and daily living abilities, personality changes, agitation, depression, delusions, and other psychobehavioral symptoms, as well as physical symptoms such as urinary and fecal incontinence and behavioral disorders.

[0003] The incidence of Alzheimer's disease is increasing worldwide due to the increase in the elderly population brought about by the extension of life expectancy. Experts predict that the number of patients with Alzheimer's disease will reach 13.8 million in the United States and 2.37 million in Korea by 2050, making the disease an increasingly serious problem. As a result, research into the treatment and improvement of Alzheimer's disease is being actively conducted worldwide.

[0004] On the other hand, Alzheimer's disease is known to develop through various mechanisms, such as hyperphosphorylation of tau protein, inflammatory responses, intracerebral deposition due to excessive production of beta-amyloid, and oxidative damage. Although the exact causes of these mechanisms have not been elucidated, it has been reported that genetic causes account for approximately 40-50% of all cases of Alzheimer's disease.

[0005] Typically, mutations in the amyloid precursor protein gene and presenilin gene are known to be genetic factors that increase the risk of early-onset Alzheimer's disease, in which symptoms begin before the age of 65, while the ApoE4 allele of the apolipoprotein E gene is known to be a genetic factor that increases the risk of late-onset Alzheimer's disease, in which symptoms begin mainly after the age of 65.

[0006] As described above, Alzheimer's disease develops due to various causes. Some research has been conducted on specific therapeutic agents corresponding to these specific causes of Alzheimer's disease. For example, Patent Document 1 discloses a composition containing superoxide dismutase and S-adenosylmethionine for treating Alzheimer's disease caused by overexpression of presenilin 1 and BACE. Although some research has been conducted, there is currently no active research on specific therapeutic agents corresponding to the specific causes of Alzheimer's disease.

[0007] The ApoE gene refers to a gene encoding the sequence of apolipoprotein E (ApoE). The ApoE gene encodes a component of lipoproteins, such as high-density lipoprotein (HDL), low-density lipoprotein (LDL), and very-low-density lipoprotein (VLDL), which are normal components of plasma. The ApoE gene is located on chromosome 19 and is known to play a key role as a fat transporter, regulating fat metabolism after injury to the central and peripheral nervous systems. The ApoE gene has three alleles: E2, E3, and E4. Of these alleles, E2 and E4 are known to be associated with Alzheimer's disease dementia. The apolipoprotein E4 allele (ApoE4) is known to be a risk factor for the development of Alzheimer's disease dementia, while the apolipoprotein E2 allele (ApoE2) is known to be a protective factor for the development of Alzheimer's disease dementia. It is known that approximately 10 to 15% of all patients with Alzheimer's disease develop the disease due to the ApoE4 genetic mutation.

[0008] Thus, the ApoE4 genotype is one of the main causes of Alzheimer's disease. Therefore, the present inventors have conducted extensive research to develop a composition specifically active against Alzheimer's disease caused by ApoE4 genetic mutations. As a result, they have elucidated the amyloid beta-reducing effect of a mixed extract of Pinellia Root, Licorice Root, Ginseng Root, Radix Candida, Scutellaria Root, and Coptis Rhizome (hereinafter referred to as Hangeshashinto) specific to ApoE4-induced Alzheimer's disease, and developed the use of Hangeshashinto for the treatment of Alzheimer's disease caused by ApoE4 genetic mutations, thereby completing the present invention.

[0009] Prior to this patent application, there had been no literature or technology disclosing the fact that Hangeshashinto is specifically applicable to improving Alzheimer's disease caused by ApoE4 gene mutations. [Prior art documents] [Patent documents]

[0010] [Patent Document 1] Korean Patent Publication No. 10-2010-0126326 Summary of the Invention [Problem to be solved by the invention]

[0011] The present invention aims to provide a pharmaceutical composition for preventing or treating Alzheimer's disease, which comprises a mixed extract of Pinellia Rhizome, Licorice Root, Glycyrrhiza Root, Korean Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome as an active ingredient.

[0012] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating Alzheimer's disease caused by ApoE4 gene mutations, which contains a mixed extract of Pinellia Root, Licorice Root, Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome as active ingredients.

[0013] Another object of the present invention is to provide a functional food composition for preventing or ameliorating Alzheimer's disease, which contains a mixed extract of Pinellia Root, Licorice Root, Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome as an active ingredient.

[0014] Another object of the present invention is to provide a quasi-drug composition for preventing or ameliorating Alzheimer's disease, which contains a mixed extract of Pinellia Root, Licorice Root, Ginseng Root, Radix Candida, Scutellaria Root, and Coptis Rhizome as an active ingredient.

[0015] A further object of the present invention is to provide a method for treating Alzheimer's disease, which comprises the step of administering the composition to a non-human individual. [Means for solving the problem]

[0016] Each description and embodiment disclosed in the present invention applies to other descriptions and embodiments. That is, any combination of various elements disclosed in the present invention is included in the present invention. Furthermore, the present invention is not limited to the following specific description.

[0017] 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 which equivalents are intended to be encompassed by the present invention.

[0018] Furthermore, throughout the specification of the present invention, when a part is said to "comprise" a certain component, this does not mean that it excludes other components, but that it may further include other components, unless otherwise specified.

[0019] The present invention will now be described in more detail. The present invention relates to use of a composition containing a mixed extract of Pinellia Rhizome, Licorice Root, Glycyrrhiza Root, Panax Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome as an active ingredient for the prevention, amelioration, or treatment of Alzheimer's disease.

[0020] To achieve the above object, one aspect of the present invention provides a pharmaceutical composition containing a mixed extract of Pinellia Rhizome, Rhizome Root, Glycyrrhiza Root, Korean Ginseng, Radix Scutellaria, Scutellaria Root, and Coptis Rhizome as an active ingredient.

[0021] In the present invention, "Hangeshashinto" refers to a mixed extract of Pinellia Root, Licorice Root, Glycyrrhiza Root, Korean Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome, and these are used interchangeably in the present specification. The Hangeshashinto can be prepared by a method commonly used in the art. In traditional Chinese medicine, Hangeshashinto is generally known to be used for symptoms such as gastritis, gastric dilatation, gastroduodenal ulcers, and gastroenteritis.

[0022] In one embodiment, the Hange-shashin-to is a mixed extract of Pinellia Rhizome, Rhizome Root, Radix Candida, Radix Scutellaria, Rhizome Root, Rhizome Root, Glycyrrhiza Root and Rhizome Root, or a mixture of extracts of each of the above ingredients, but is not limited thereto.

[0023] In one specific example, the ratio of Pinellia Root, Licorice Root, Glycyrrhiza Root, Panax Rhizome, Radix Candida, Scutellaria Root, and Coptis Rhizome contained in the Hange-Sha-Shi-To is approximately 1.2-2.0:0.8-1.2:0.8-1.2:0.8-1.2:0.6-1.0:0.8-1.2:0.2-0.5, but is not limited thereto.

[0024] In another specific example, the Hange-shashin-to is an extract obtained by mixing about 1.2 to 2.0 g of Pinellia Root, about 0.8 to 1.2 g of Rhizome Rhizome, about 0.8 to 1.2 g of Glycyrrhiza Root, about 0.8 to 1.2 g of Korean Ginseng, about 0.6 to 1.0 g of Radix Candida, about 0.8 to 1.2 g of Scutellaria Root, and about 0.2 to 0.5 g of Coptis Rhizome, but is not limited thereto.

[0025] In a preferred embodiment, the Hange-shashin-to is an extract of a mixture of about 1.67 g of Pinellia Root, about 1 g of Rhizome Rhizome, about 1 g of Glycyrrhiza Root, about 1 g of Korean Ginseng, about 0.83 g of Radix Candida, about 1 g of Scutellaria Root, and about 0.33 g of Coptis Rhizome, or an extract of a mixture of these ingredients in similar proportions but with different masses, but is not limited to these.

[0026] The components of Hangeshashinto will be described in more detail below. In this invention, "pinellia ternata" generally refers to the tuber of Pinellia ternata Breitenbach, a plant of the Araceae family, from which the cork layer has been removed. It is also known as Jiwen or Yangyan Pinellia. It has almost no odor, is slightly mucous, has a spicy taste, and has a warming nature. In traditional Chinese medicine, pinellia ternata is used for phlegm, cough, and asthma, as well as for symptoms such as headaches caused by stiffness, dizziness, chest tightness, bloating, vomiting, sore throat, back pain, mastitis, and vomiting during pregnancy. Its pharmacological effects have been reported to include expectorant, antitussive, anti-vomiting, silicosis prevention, and anticancer effects.

[0027] In this invention, "Daizhu (Da jujube)" refers to the fruit of the jujube tree, whose scientific name is Ziziphus jujuba var. inermis. It is also known as jujube or kumimi (lit., "lit. jujube"). It has a reddish-brown surface, is oval in shape, and reaches a length of 1.5 to 2.5 cm. It becomes sweeter when ripe. In traditional Chinese medicine, Daizhu (Da jujube) has been reported to be used as a diuretic, tonic, and emollient.

[0028] The term "licorice" in the present invention refers to a perennial plant belonging to the Fabaceae family, and its scientific name is Glycyrrhiza uralensis FISCH. Licorice is also known as kokuro, mikao, mikan, mitsuso, renso, tianso, and kaeiso. The outer skin of licorice is reddish-brown or dark brown and has vertical wrinkles. In traditional Chinese medicine, licorice has been reported to be used as an ingredient that neutralizes the toxicity of drugs, improves medicinal efficacy, removes cold and heat evils from the five internal organs, opens up all blood vessels, and strengthens muscles and bones.

[0029] In the present invention, "Korean ginseng" refers to the root of Panax ginseng CA Meyer (Araliaceae). Korean ginseng is also known as Onigae (ghost lid), Kanejing Magnolia (Golden Crown), Shinso (sacred herb), Jade Spirit (jewelry), Renwei (religious vine), Renji (religious vine), Jigen (earth spirit), Kier Ginseng (child ginseng), Blood Ginseng (blood ginseng), Yellow Ginseng (wild ginseng), Wild Mountain Ginseng (wild ginseng), and Beiji Ginseng (beautiful ginseng). Korean ginseng has a unique odor, a sweet and slightly bitter taste, and a warming nature. In traditional Chinese medicine, Korean ginseng is used to replenish vitality and treat physical weakness, malaise, fatigue, loss of appetite, vomiting, and diarrhea, and has also been reported to improve kidney function.

[0030] In the present invention, "kankyo (dried ginger)" refers to the dried rhizome of ginger (Zingiber officinale Roscoe). Kankyo is also called white ginger or even ginger, and has a distinctive odor and medicinal properties of pungent and hot. Kankyo has pharmacological effects such as anti-inflammatory, analgesic, and antibacterial properties.

[0031] In the present invention, "Scutellaria root" refers to the root or its periderm removed of Scutellaria baicalensis, which belongs to the genus Scutellaria in the family Lamiaceae. Scutellaria root has almost no odor and a slightly bitter taste. Scutellaria root has been reported to have pharmacological effects such as anti-cancer, hepatoprotective, and antioxidant properties.

[0032] In the present invention, "coprinus japonica" refers to an evergreen perennial plant of the dicotyledonous Ranunculaceae family. The scientific name of coprinus japonica is Coptis chinensis, and it has a bitter and cold taste. Berberine, the main component of coprinus japonica, has been reported to pharmacologically exhibit antibacterial activity against intestinal bacteria, sedative and antispasmodic activity, anti-arteriosclerotic activity, anti-inflammatory activity, choleretic activity, and pancreatic secretion promoting activity.

[0033] The term "extract" or "mixed extract" as used herein includes extracts obtained by extracting the aforementioned Pinellia Root, Rhizome Root, Licorice Root, Korean ginseng, Chinese cabbage, Scutellaria Root, and Coptis Rhizome, as well as any other extract formulation that can be prepared using the extract, such as dilutions or concentrates of the extract, dried products obtained by drying the extract, crude or purified products of the extract, and mixtures thereof. Specifically, the extract of the present invention may be prepared in the form of a dried powder after extraction for use. In the present invention, Pinellia Root, Rhizome Root, Licorice Root, Korean ginseng, Chinese cabbage, Scutellaria Root, or Coptis Rhizome may be commercially available or cultivated or harvested in the wild.

[0034] In one specific example of the present invention, the mixed extract of Pinellia Root, Brassica Root, Licorice Root, Korean ginseng, Chinese cabbage, Scutellaria Root, and Coptis Rhizome may be prepared by extracting and mixing each plant, or by mixing all of Pinellia Root, Brassica Root, Licorice Root, Korean ginseng, Chinese cabbage, Scutellaria Root, and Coptis Rhizome and then extracting. The mixing process may be carried out before or after the extraction. Furthermore, it is preferable to extract each plant and then mix them, but the method is not limited thereto.

[0035] In the present invention, the type of extraction solvent used to extract the extract is not particularly limited, and any solvent known in the art can be used. Examples of the extraction solvent include water, lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propyl alcohol, and butyl alcohol, polyhydric alcohols such as glycerin, butylene glycol, and propylene glycol, hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane, and mixtures thereof, but are not limited to these examples. The extract is preferably a water or lower alcohol extract having 1 to 4 carbon atoms, but is not limited to these.

[0036] In the present invention, the method for extracting the extract is not particularly limited, and can be a method commonly used in the art. Examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, and reflux extraction, but are not limited thereto. These methods may be used alone or in combination of two or more.

[0037] In one embodiment of the present invention, the mixed extract of Pinellia Root, Licorice Root, Ginseng Root, Radix Scutellaria Root, and Coptis Rhizome includes a concentrate, filtrate, fraction, dried product, and pulverized product prepared by additional processes typically applied to plant extracts, such as concentration, filtration, fractionation, drying, and grinding, after extraction. Here, the "concentrate" refers to a substance obtained by removing a solvent or the like from a chemical substance to increase the concentration of a major solid component, the "filtrate" refers to a substance obtained by a method of separating a mixture of liquid and solid based on particle size, the "fraction" refers to a product obtained by a fractionation method for separating a specific component or group from a mixture containing various components, the "dried product" refers to a product obtained by removing water from a substance containing a small amount of water, and the "pulverized product" refers to a product obtained by applying force to crush or break down solid particles into smaller particles. Here, the concentrate, filtered product, fractionated product, dried product, pulverized product, etc. can be obtained by a conventional method for obtaining the resulting product, and there are no particular limitations.

[0038] The pharmaceutical composition of the present invention may contain Hangeshashinto as an active ingredient, as described above. Furthermore, the Hangeshaxin Tang is contained in the composition at a concentration of 50 to 800 μg / ml, but is not limited thereto. For example, the Hangeshaxin Tang is contained in the composition at a concentration of 100 to 700 μg / ml, but is not limited thereto. The Hangeshaxin Tang is preferably contained in the composition at a concentration of 200 to 500 μg / ml, but is not limited thereto. The Hangeshaxin Tang is more preferably contained in the composition at a concentration of about 300 μg / ml.

[0039] The pharmaceutical composition of the present invention may contain one or more additional active ingredients in addition to Hange Shashin Tang. The pharmaceutical composition of the present invention may further contain a pharmaceutically acceptable carrier, excipient, or diluent commonly used in the manufacture of pharmaceutical compositions, and the carrier may be a non-naturally occurring carrier.

[0040] In the present invention, "pharmaceutical acceptable" means the property of not being toxic to cells or humans that come into contact with the composition. Specific examples of the carrier, excipient, and diluent include, but are not limited to, lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, etc. These may be used alone or in combination of two or more.

[0041] The pharmaceutical compositions may be formulated into any of the following forms, including tablets, pills, powders, granules, capsules, suspensions, oral solutions, emulsions, syrups, sterile aqueous solutions, non-aqueous solvents, lyophilized preparations, and suppositories, using conventional methods. These compositions may be used in a variety of oral or parenteral dosage forms. When formulated, they are prepared using commonly used diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Oral solid formulations include tablets, pills, powders, granules, and capsules, and these solid formulations contain at least one excipient, such as starch, calcium carbonate, sucrose or lactose, or gelatin. In addition to conventional excipients, lubricants, such as magnesium stearate and talc, may also be used. Oral liquid preparations include suspensions, oral solutions, emulsions, syrups, etc., and in addition to the commonly used diluents of water and liquid paraffin, various excipients such as humectants, sweeteners, flavorings, and preservatives are used. Parenteral preparations include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, etc. 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, but are not limited to, witepsol, macrogol, Tween 61, cocoa butter, lauric butter, and glycerogelatin.

[0042] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating Alzheimer's disease caused by ApoE4 gene mutation, which comprises Hangeshashinto as an active ingredient. In the present invention, "Alzheimer's disease" refers to a degenerative brain disease that develops gradually and causes gradual deterioration of cognitive functions, including memory.

[0043] Mutations in the apolipoprotein E4 allele (ApoE4) gene are known to be one of the causes of Alzheimer's disease. The ApoE gene refers to a gene encoding the sequence of apolipoprotein E (ApoE). The ApoE4 genotype is known to be a risk factor for the development of Alzheimer's disease dementia. Previous studies have reported that the incidence of Alzheimer's disease in individuals with one APOE4 allele (heterozygous) is approximately 50%, while the incidence of Alzheimer's disease in individuals with two APOE4 alleles (homozygous) is approximately 91%. Furthermore, it is known that individuals with the APOE4 allele are at an earlier age at onset of Alzheimer's disease.

[0044] The Alzheimer's disease in the present invention may be Alzheimer's disease caused by an ApoE4 gene mutation. Here, "Alzheimer's disease caused by an ApoE4 gene mutation" in the present invention refers to, but is not limited to, Alzheimer's disease caused by the expression of ApoE4 in an individual having the ApoE4 genotype.

[0045] The Alzheimer's disease caused by the ApoE4 gene mutation is, but is not limited to, Alzheimer's disease dementia caused by the ApoE4 gene mutation. Here, "Alzheimer's disease dementia" in the present invention means a dementia caused by Alzheimer's disease.

[0046] The Alzheimer's disease may be, for example, late onset Alzheimer's disease. In the present invention, "treatment" refers to any action that improves or favorably changes the symptoms of Alzheimer's disease caused by ApoE4 gene mutation by administering a composition.

[0047] In the present invention, "prevention" refers to any action of suppressing or delaying the onset, progression, and recurrence of Alzheimer's disease caused by ApoE4 gene mutations using the composition of the present invention.

[0048] Specifically, as an example of the treatment, in one embodiment of the present invention, it was confirmed that the treatment activates the phagocytic action of amyloid beta, inhibits the accumulation of amyloid beta, and removes amyloid beta in individuals with Alzheimer's disease caused by ApoE4 gene mutations.

[0049] In another specific example, the composition of the present invention may suppress hyperphosphorylation of tau protein in individuals with Alzheimer's disease caused by ApoE4 gene mutation.

[0050] In yet another specific example, the composition of the present invention may improve cognitive function in an individual with Alzheimer's disease caused by an ApoE4 gene mutation by promoting phagocytosis of amyloid beta or suppressing hyperphosphorylation of tau protein.

[0051] Thus, it has been confirmed that the composition of the present invention containing Hangeshashinto as an active ingredient is highly useful as a therapeutic composition specific to Alzheimer's disease caused by ApoE4 gene mutations.

[0052] Yet another aspect of the present invention provides a method for treating Alzheimer's disease caused by ApoE4 gene mutations. The treatment method may include a step of testing whether the individual has the ApoE4 genotype, and determining an individual who is ApoE4 positive and has Alzheimer's disease as a treatment target.

[0053] The treatment method may also include a step of administering to an individual the pharmaceutical composition containing Hange Shashin Tang as an active ingredient. The "Hangeshashinto", "treatment", "Alzheimer's disease caused by ApoE4 gene mutation", "prevention" and "administration" are as described above.

[0054] The steps of the treatment method will now be described in detail. The treatment method of the present invention may include a step of testing whether or not an individual has the ApoE4 genotype. In the present invention, "testing whether or not an individual has the ApoE4 genotype" refers to the act of testing whether or not an individual has the ApoE4 allele. "Testing whether or not an individual has the ApoE4 genotype" is understood to be synonymous with "testing whether or not an individual is ApoE4 positive." Since the composition provided by the present invention exhibits specific activity against Alzheimer's disease caused by ApoE4 gene mutations, ApoE4-positive individuals are selected as treatment subjects through the above step.

[0055] In the present invention, "ApoE4 positive" means that an individual has at least one ApoE4 allele. The method for detecting the presence or absence of the ApoE4 allele may be, but is not limited to, a method commonly used in the art. The detection may be performed by, but is not limited to, isoelectric focusing, immunological methods, immunochemical methods, sequence analysis, etc. For example, the detection may be performed by polymerase chain reaction (PCR).

[0056] The treatment method of the present invention may also include a step of determining, as a treatment target, an individual who is ApoE4-positive and has Alzheimer's disease. That is, an individual who has tested ApoE4-positive in the above-described testing process and who has Alzheimer's disease may be determined as a treatment target. The treatment target may be a person who has developed Alzheimer's disease caused by an ApoE4 gene mutation. The treatment target may also be a person who has developed Alzheimer's disease caused by expression of the ApoE4 gene.

[0057] Here, the term "individual" in the present invention refers to any animal, including humans, rats, mice, livestock, etc. The animals include not only humans but also mammals such as cows, horses, sheep, pigs, goats, camels, serows, dogs, and cats, but are not limited to these.

[0058] Here, the subject of treatment may be an individual with a heterozygous ApoE4 genotype who has developed Alzheimer's disease, or an individual with a homozygous ApoE4 genotype who has developed Alzheimer's disease, or a patient with Alzheimer's disease caused by expression of the ApoE4 gene.

[0059] Specifically, the subject of treatment may be an individual who has developed Alzheimer's disease dementia caused by an ApoE4 gene mutation. The treatment method of the present invention may include a step of administering to the subject a pharmaceutical composition containing Hange Shashin Tang as an active ingredient.

[0060] Here, the pharmaceutical composition is understood to be synonymous with the aforementioned "pharmaceutical composition for preventing or treating Alzheimer's disease caused by ApoE4 gene mutation, comprising Hangeshashinto as an active ingredient."

[0061] The composition of the present invention may be administered orally or parenterally depending on the intended method. For parenteral administration, topical application to the skin or intraperitoneal, rectal, subcutaneous, intravenous, intramuscular, or intrathoracic injection may be selected. Here, "administration" in the present invention means providing a predetermined substance to a patient by any appropriate method, and the administration route of the composition of the present invention can be any common route that can deliver the substance to the target tissue. Examples of administration routes include, but are not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, pulmonary, and rectal administration.

[0062] The dosage of the composition varies depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, severity of disease, and the like. Furthermore, the compositions of the present invention may be used alone or in combination with surgery, hormone therapy, drug therapy, methods using biological response modifiers, etc. for the prevention and treatment of cognitive impairment. For purposes of the present invention, the compositions of the present invention may be administered in combination with one or more additional drugs commonly used to treat Alzheimer's disease.

[0063] In yet another aspect, the present invention provides a functional food composition for preventing or ameliorating Alzheimer's disease, comprising Hangeshashinto as an active ingredient. Here, a specific aspect of the present invention provides a functional food composition for preventing or ameliorating Alzheimer's disease caused by ApoE4 gene mutation, which contains Hangeshashinto as an active ingredient.

[0064] In this regard, the terms "Hangeshashinto," "Alzheimer's disease," and "caused by ApoE4 gene mutation" are as described above. The weight ratio or concentration of Hangeshashinto in the functional food composition is the same as that defined in the pharmaceutical composition.

[0065] The "prevention" in the present invention is the same as that explained in the pharmaceutical composition, and "amelioration" means any action of improving or favorably changing the symptoms of an individual who has developed or is suspected of having the disease using the composition of the present invention.

[0066] The term "functional food" is synonymous with "food for special health use (FoSHU)" and refers to a food manufactured and / or processed using ingredients or components with beneficial functions for the human body. It refers to a food with high medical and therapeutic value that is processed to efficiently exhibit bioregulatory functions in addition to providing nutrients. Here, "functionality" refers to regulating nutrients for the structure and function of the human body or providing beneficial health effects, such as physiological effects. Functional foods are foods that contain ingredients beneficial to the human body and whose functions and stability have been recognized by the Ministry of Food and Drug Safety. Other foods that have not been approved by the Ministry of Food and Drug Safety but are considered to have beneficial effects on the human body are called health (supplemental) foods. The food of the present invention can be manufactured using methods commonly used in the art and can be manufactured using ingredients and components commonly added in the art. The food may be in any dosage form recognized as a food.

[0067] The health food composition of the present invention may contain any food-related acceptable adjuvants, additives, and other ingredients in addition to Hange Shashin Tang. Here, the food composition of the present invention may contain herbal extracts, food supplementary additives, natural carbohydrates, and the like as additional ingredients in addition to Hange Shashin Tang, which is an essential ingredient.

[0068] The functional food composition may further contain a food-scientifically acceptable carrier, which can be appropriately selected and used by those skilled in the art. Examples include, but are not limited to, various nutrients, vitamins, minerals (electrolytes), flavorings such as synthetic flavorings and natural flavorings, colorants and fillers, pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloids, thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonation agents used in carbonated drinks.

[0069] The functional food composition may also contain additional ingredients commonly used in food compositions to improve aroma, taste, visual appearance, etc. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. Minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), and copper (Cu) may also be included. Amino acids such as lysine, tryptophan, cysteine, and valine may also be included.

[0070] Furthermore, food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder and highly bleached powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), etc.), colorants (tar dyes, etc.), color formers (sodium nitrite, sodium acetate, etc.), bleaching agents (sodium sulfite), seasonings (monosodium glutamate (MSG), etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), fragrances (vanillin, lactones, etc.), leavening agents (alum, potassium D-bitartrate, etc.), strengtheners, emulsifiers, thickeners (thickening agents), coating agents, gum bases, foam inhibitors, solvents, and improvers may also be added.

[0071] Yet another aspect of the present invention provides a quasi-drug composition for preventing or ameliorating Alzheimer's disease, which comprises Hangeshashinto as an active ingredient. Here, a specific aspect of the present invention provides a quasi-drug composition for preventing or ameliorating Alzheimer's disease caused by ApoE4 gene mutation, which contains Hangeshashinto as an active ingredient.

[0072] In this regard, the terms "Hangeshashinto," "Alzheimer's disease," "disease caused by ApoE4 gene mutation," "prevention," and "amelioration" are as described above. The weight ratio or concentration of Hangeshashinto in the quasi-drug composition is the same as that defined in the pharmaceutical composition.

[0073] In the present invention, "quasi-drugs" refer to products that are used for the purpose of diagnosing, treating, ameliorating, mitigating, managing, or preventing diseases in humans or animals, and have a milder effect than pharmaceuticals. For example, according to the Pharmaceutical Affairs Law, quasi-drugs exclude products used for pharmaceutical purposes, and include products used for treating or preventing diseases in humans / animals, and products that have a mild effect on the human body or do not have a direct effect.

[0074] The quasi-drug composition may be prepared by adding the composition containing Hangeshashinto as an active ingredient according to the present invention as it is, or may be used in combination with other quasi-drugs or quasi-drug ingredients, and may be used appropriately in a conventional manner. The amount of the active ingredient to be mixed is determined appropriately depending on the purpose of use (prevention, health, or therapeutic treatment).

[0075] The quasi-drug composition includes, but is not limited to, a personal hygiene product, a hand sanitizer, or an ointment. The personal hygiene product may be, but is not limited to, a toothpaste, a mouthwash, a cleaning gel, or a storage agent for a prosthesis such as a denture.

[0076] Furthermore, the quasi-drug composition is prepared and used in the form of, but not limited to, an ointment, a lotion, a spray, a patch, a cream, a powder, a suspension, a gel, or a gel.

[0077] Yet another aspect of the present invention provides use of a composition containing a mixed extract of Pinellia Rhizome, Rhizome Root, Licorice Root, Korean Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome as an active ingredient for the prevention or treatment of Alzheimer's disease. The Alzheimer's disease may be Alzheimer's disease caused by ApoE4 gene mutation. The Alzheimer's disease may also be late-onset Alzheimer's disease. In this regard, the terms "Hangeshashinto," "Alzheimer's disease," "caused by ApoE4 gene mutation," "prevention," and "treatment" are as described above.

[0078] Another aspect of the present invention provides use of a functional food composition containing a mixed extract of Pinellia Root, Licorice Root, Licorice Root, Korean Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome as an active ingredient for the prevention or treatment of Alzheimer's disease. The Alzheimer's disease may be Alzheimer's disease caused by ApoE4 gene mutation. The Alzheimer's disease may also be late-onset Alzheimer's disease.

[0079] Another aspect of the present invention provides use of a quasi-drug composition containing a mixed extract of Pinellia Root, Licorice Root, Ginseng Root, Radix Candida, Scutellaria Root, and Coptis Rhizome as an active ingredient for the prevention or treatment of Alzheimer's disease. The Alzheimer's disease may be Alzheimer's disease caused by an ApoE4 gene mutation. The Alzheimer's disease may also be late-onset Alzheimer's disease. [Effects of the Invention]

[0080] The composition and treatment method of the present invention containing Hangeshashinto as an active ingredient exhibit excellent effects of reducing and inhibiting the accumulation of amyloid beta protein specifically in Alzheimer's disease caused by ApoE4 gene mutations, and are highly useful as a therapeutic composition specific to Alzheimer's disease caused by ApoE4 gene mutations. In the following examples, this specification confirms the ApoE4-specific amyloid beta-reducing effect of the composition of the present invention using ApoE4 human cerebral astrocytes and cerebral organoids, and clearly elucidates the results.

[0081] Thus, the compositions and treatment methods of the present invention may provide more effective customized treatment for patients with Alzheimer's disease due to their ApoE4 genotype. [Brief explanation of the drawings]

[0082] [Figure 1] Figure 1 shows an ApoE4 Alzheimer's disease dementia human induced pluripotent stem cell model created by CRISPR / Cas9 genome editing. [Figure 2a] FIG. 1 shows the astrocyte differentiation process. [Figure 2b] FIG. 1 shows the results of verifying the successful differentiation of human induced pluripotent stem cells into astrocytes via neural progenitor cells using a fluorescent staining method. [Figure 3] This figure shows a protocol for exploring the recovery of amyloid-β phagocytosis in astrocytes in Alzheimer's disease dementia using traditional Chinese medicine-based therapeutic materials. [Figure 4a] FIG. 1 shows the results of measuring Aβ42 phagocytosis in ApoE3 and ApoE4 astrocytes. [Figure 4b] FIG. 1 shows the results of the ApoE3 control group. [Figure 4c] FIG. 1 shows the restorative effect of treatment with Hangeshashinto on ApoE4 astrocytes with reduced degradative function. [Figure 5] Figure 1 shows the process of creating a dementia human induced pluripotent stem cell (APPswe) model using CRISPR / Cas9 genetic scissors technology. [Figure 6a]FIG. 1 shows a timeline of an experiment measuring the effect of Hangeshashinto on the phagocytosis of amyloid beta in APPswe astrocytes. [Figure 6b] FIG. 1 shows the results of measuring the effect of Hangeshashinto on the phagocytic activity of amyloid β in APPswe astrocytes. [Figure 7] FIG. 1 shows the progress of an experiment exploring the effects of Hangeshashinto on ApoE4 cerebral organoids. [Figure 8a] 1 shows an increase in amyloid β accumulation in ApoE4 cerebral organoids and the reducing effect of Hangeshashinto, specifically, the results of Western blot analysis to confirm amyloid β. [Figure 8b] 8A and 8B are graphs showing the increase in amyloid β accumulation in ApoE4 cerebral organoids and the reducing effect of Hangeshashinto, specifically the results of FIG. 8A, quantified in a graph. DETAILED DESCRIPTION OF THE INVENTION

[0083] The following examples and experimental examples will explain the configuration and effects of the present invention in more detail. These examples and experimental examples are merely illustrative of the present invention, and the present invention is not limited to these examples and experimental examples.

[0084] Preparation Example 1: Preparation of a composition containing Hangeshashinto as an active ingredient 1.00g of intact or periderm-removed roots and rhizomes of Glycyrrhiza uralensis Fischer, Glycyrrhiza glabra Linne, or Glycyrrhiza inflata Batal. (Fabaceae, Leguminosae)), 0.83g of dried ginger (Zingiber officinale Roscoe) (Zingiberaceae, Zingiberaceae), 1.00g of mature fruit of jujube (Zizyphus jujuba Miller var. inermis Rehder) or Boon jujube (Zizyphus jujuba Miller var. hoonensis TB Lee) (Rhamnaceae, Rhamnaceae), 1.00g of pinecone (Pine / lia ternata We prepared 1.67 g of tubers of Breitenbach (Araceae, Araceae) with the periderm completely removed, 1.00 g of roots of Korean ginseng (Panax ginseng CA Meyer (Araliaceae, Araliaceae) either intact or with the rootlets and cork layer removed), 1.00 g of roots of Scutellaria baicalensis Georgi (Labiatae, Labiatae) either intact or with the periderm removed, and 0.33 g of rhizomes of Coptis japonica Makino, Coptis chinensis Franchet, Coptis deltoidea CY Cheng et Hsiao, or Coptis teeta Wallich (Ranunculaceae) with the roots removed). The above medicinal herbs were selected, and each medicinal herb was boiled according to the amount of raw material medicine and placed in an extractor. 8 to 10 times the amount of plain water (KP) or purified water (KP) was added, and the extract was carried out at 80 to 100°C for 2 to 3 hours to produce Hange Shashin Tang. [Example]

[0085] Human induced pluripotent stem cell culture A human induced pluripotent stem cell model of Alzheimer's disease dementia carrying the ApoE4 mutation, the main genetic cause of Alzheimer's disease dementia, was created using CRISPR / Cas9 genome editing (Figure 1). To use this human induced pluripotent stem cell model for this study, ApoE3 / ApoE4 isochromosomal induced pluripotent stem cells were cultured in TeSR1 media in Matrigel-coated 6-well plates.

[0086] As a result, an ApoE4 Alzheimer's disease dementia human induced pluripotent stem cell model was created using CRISPR / Cas9 genome editing. [Example]

[0087] Astrocyte differentiation To differentiate neural progenitor cells (astrocyte precursors) from human induced pluripotent stem cells, human induced pluripotent stem cells at 100% confluence were treated with neuronal induction media [DMEM / F-12 GlutaMAX (Gibco), Neurobasal (Gibco), 0.5x N-2 (Gibco), 0.5x B27 (Gibco), 0.5x GlutaMAX (ThermoFisher Scientific), 5 μg / ml insulin (Sigma-Aldrich), 0.5x NEAA (ThermoFisher Scientific), 100 μM 2-mercaptoethanol (Sigma-Aldrich), 1x Penicillin / Streptomycin (Gibco)] together with SMAD signaling inhibitors 1 μM Dorsomorphin (Tocris) and 10 μM SB431542 (Tocris) for 11 days. The cells were then transferred to a matrigel-coated plate and the culture medium was then treated with 20 ng / ml FGF2 (Peprotech) until rosette structures were formed.

[0088] The generated neural progenitor cells were 1.5 × 10 5The cells were plated at a density of 100 μg / ml and then cultured in Astrocyte Media (Sciencell). The culture medium was changed every two days. After 1 month of culture, the cells were detached with TrypLE (Gibco) and stained with the astrocyte marker GLAST-PE antibody (MiltenylBiotec). GLAST-positive cells were then isolated using a flow cytometer (Sony SH800) and cultured (Figure 2a). To verify astrocyte differentiation, fluorescent staining was performed with antibodies to other astrocyte markers, GFAP and AQP4 (Figure 2b).

[0089] The differentiation of human induced pluripotent stem cells into astrocytes via neural progenitor cells, and the differentiation of astrocytes were examined using fluorescent staining. [Example]

[0090] Confirmation of amyloid-β phagocytosis in ApoE4 astrocytes induced by Hangeshashinto Synthetic amyloid beta (Aβ 42 ApoE4 astrocytes and control ApoE3 astrocytes were treated with ) and Hangeshashinto and cultured for 2 days, after which the amount of amyloid beta remaining in the culture medium was measured by ELISA (Figure 3).

[0091] In ApoE4 astrocytes, the culture medium was significantly higher in Aβ than in ApoE3. 42 When treated with Hangeshashinto, the ability to remove Aβ was significantly reduced (Fig. 4a). 42 The phagocytic capacity was confirmed to be improved to the same level or higher than that of the normal ApoE3 group. In particular, the clearance capacity under the 300 μg / ml treatment condition was significantly improved to a level higher than that of the normal group (Figure 4c). In contrast, no statistically significant difference was observed in ApoE3 astrocytes after treatment with Hangeshashinto (Figure 4b). [Example]

[0092] Hangeshashinto induces amyloid-β phagocytosis in APPswe (APP swedish) astrocytes Next, we created an APPswe astrocyte model to confirm whether Hangeshashinto also has therapeutic activity against Alzheimer's disease caused by APPswe gene mutations. APP is an amyloid precursor protein gene and is known to be a major causative gene for familial Alzheimer's disease.

[0093] Specifically, the present applicants used CRISPR / Cas9 gene scissors technology to introduce an Alzheimer's disease-inducing mutation, APPswe (KM670 / 671NL double mutation in the APP gene), into normal human induced pluripotent stem cells to create an "Alzheimer's disease-inducing human induced pluripotent stem cell model (APPswe)" (Figure 5). The sequences used in this experiment are as follows:

[0094] [Table 1]

[0095] The differentiation process of astrocytes was carried out in the same manner as in Example 2. 42 APPswe astrocytes were treated with α-amyloid-β-binding protein (Aβ) and Hangeshashinto (Hangeshashinto) and cultured for 2 days, after which the amount of amyloid-β remaining in the culture medium was measured by ELISA. The schematic timeline of this experiment is shown in Figure 6a, and the results are shown in Figure 6b.

[0096] As can be seen from the results in Figure 6b, Hangeshashinto treatment significantly reduced amyloid β (Aβ) levels in astrocytes of APPswe, a familiar Alzheimer's disease line. 42 ) No change in removal capacity was observed.

[0097] These results of Examples 3 and 4 confirm that Hangeshashinto has a specific effect of reducing amyloid β in ApoE4-containing cells. [Example]

[0098] Confirmation that Hange-shashin-to treatment alleviates Alzheimer's disease-related pathology in brain organoids ApoE4 brain organoids cultured for four months were treated with Hangeshashinto at a concentration of 300 μg / ml for seven days, and changes in amyloid beta accumulation were then observed (Figure 7).

[0099] In cultured ApoE4 cerebral organoids, a significant increase in both sub- and super-tetrameric forms of amyloid beta was observed compared to ApoE3 cerebral organoids. Amyloid beta forms tetrameric oligomers, and Aβ 40 It is known that the structure of ApoE4 is different from that of tetramers, making it more susceptible to the formation of toxic nuclei, and the formation of oligomers of tetramers or higher is the main factor affecting neurotoxicity. As shown in Figure 8b, a decrease in the amount of polymers of tetramers and higher was observed in all groups of ApoE4 brain organoids treated with Hangeshashinto (Figure 8b).

[0100] These results suggest that the present invention has excellent effects on amyloid beta suppression and phagocytosis in ApoE4-induced Alzheimer's disease, and that the present invention is useful as a specific therapeutic agent for ApoE4-induced Alzheimer's disease or late-onset Alzheimer's disease.

[0101] From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be embodied in other specific forms without changing the technical spirit or essential features thereof. It should be understood that the above examples are merely illustrative and not limiting. The present invention should be construed as including all modifications and variations derived from the meaning and scope of the claims, rather than the specification, and their equivalents.

Claims

1. A pharmaceutical composition for preventing or treating Alzheimer's disease, comprising a mixed extract of Pinellia Root, Licorice Root, Korean Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome as active ingredients, wherein the Alzheimer's disease is caused by ApoE4 gene mutation.

2. 2. The pharmaceutical composition for preventing or treating Alzheimer's disease according to claim 1, wherein the Alzheimer's disease is late-onset Alzheimer's disease.

3. The pharmaceutical composition for preventing or treating Alzheimer's disease according to claim 1, wherein the composition exerts an amyloid β reducing effect.

4. The pharmaceutical composition for preventing or treating Alzheimer's disease according to claim 1, wherein the composition is for preventing or treating Alzheimer's disease in an ApoE4-positive individual.

5. A functional food composition for preventing or ameliorating Alzheimer's disease caused by ApoE4 gene mutation, comprising a mixed extract of Pinellia Root, Licorice Root, Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome as active ingredients.

6. A quasi-drug composition for preventing or ameliorating Alzheimer's disease caused by ApoE4 gene mutation, comprising a mixed extract of Pinellia Root, Licorice Root, Ginseng, Radix Candida, Scutellaria Root, and Coptis Rhizome as active ingredients.

7. A method for treating Alzheimer's disease caused by ApoE4 gene mutations, comprising administering the pharmaceutical composition of claim 1 to a non-human individual.

8. Use of a composition containing as an active ingredient a mixed extract of Pinellia Root, Licorice Root, Licorice Root, Korean Ginseng, Radix Candida, Scutellaria Root and Coptis Rhizome for the prevention or treatment of Alzheimer's disease caused by ApoE4 gene mutations in non-human individuals.

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