Aminoaromatic compounds or pharmaceutically acceptable salts of the compounds and pharmaceutical compositions for preventing or treating neurodegenerative diseases comprising the same as an active ingredient

Abstract

Provided are a novel aminoaromatic compound or a pharmaceutically acceptable salt of the compound, a pharmaceutical composition for preventing or treating a neurodegenerative disease containing the same as an active ingredient, and a health food composition for preventing or ameliorating a neurodegenerative disease.

Description

Technical Field

[0001] This invention relates to a novel amino aromatic compound or a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising such compound as an active ingredient for the prevention or treatment of neurodegenerative diseases, and a health food composition for the prevention or improvement of neurodegenerative diseases. Background Technology

[0002] Neurodegenerative diseases are diseases caused by the weakening or disappearance of nerve cell function, leading to abnormalities in motor regulation, cognitive function, sensory function, and autonomic nervous system function. Typical examples include dementia, Alzheimer's disease (AD), Parkinson's disease (PD), and memory disorders.

[0003] One of the leading causes of neurodegenerative diseases is oxidative stress in neurons caused by the generation of reactive oxygen species (ROS). Oxidative stress, defined as an imbalance between the antioxidant and oxidative systems in an organism, is known to arise from the accumulation of intracellular ROS. This oxidative stress causes lipid peroxidation, intracellular DNA damage, and other adverse effects, thereby inducing apoptosis and neuronal cell apoptosis.

[0004] In particular, the brain has a high oxygen saturation and is rich in highly unsaturated fatty acids or metal ions that directly target oxidative stress. Furthermore, the brain's meurotransmitters undergo auto-oxidation. Under oxidative stress caused by reactive oxygen species (ROS), the brain increases the production of neurotoxic oxidative products to compensate for the decrease in unsaturated fatty acid content. Therefore, the brain is an organ that is very sensitive to oxidative stress and has limited antioxidant and recovery capabilities.

[0005] Reactive oxygen species (ROS) are chemically reactive free radicals and non-free radicals containing oxidizing capabilities. They can include superoxide anion radicals (·O2-), perhydroxyl radicals (HO2·), hydroxyl radicals (·OH), and singlet oxygen. 1 O2), hydrogen peroxide (H2O2), alkoxy radicals (·OR), superoxide radicals (·OOR), etc.

[0006] Among reactive oxygen species (ROS), hydrogen peroxide (H2O2) is the most common product of various oxidation reactions (such as oxidases, dehydrogenases, and peroxidases) that occur primarily in the mitochondria of living organisms. Hydrogen peroxide (H2O2), beyond its role as a byproduct itself, is produced simultaneously as a signaling transducer and a toxic molecule. During mitochondrial respiration, superoxide dismutase (SOD) catalyzes the conversion of superoxide anion radicals (·O2-) into hydrogen peroxide (H2O2) and oxygen (O2).

[0007] Another source of hydrogen peroxide (H2O2) is NADPH oxidase (nicotinamide adenine dinucleotide phosphate oxidase), which catalyzes the formation of superoxide anion radicals (·O2-) from oxygen (O2), ultimately generating hydrogen peroxide (H2O2). Furthermore, xanthine oxidase is responsible for generating hydrogen peroxide (H2O2) during hypoxanthine oxidation, and members of the monoamine oxidase (MAO) family oxidize monoamines (e.g., dopamine and noradrenaline) and polyamines (e.g., Nacetyl putrescine) to produce hydrogen peroxide (H2O2). In addition, many other methods exist for generating hydrogen peroxide (H2O2).

[0008] Low concentrations of reactive oxygen species (ROS) temporarily generated in specific regions by external signals can regulate cellular functions such as growth, apoptosis, and immunity. Therefore, appropriate amounts of hydrogen peroxide (H2O2) are crucial for living organisms. However, excessive amounts of hydrogen peroxide (H2O2) beyond cellular regulation can act as toxic substances within cells. In other words, hydrogen peroxide (H2O2) presents both beneficial and harmful effects on cells.

[0009] Moderate levels of hydrogen peroxide (H₂O₂) function as cell signaling molecules (CSMs), including transcription factors, protein kinases, and growth factors. However, moderate levels of H₂O₂ can damage DNA, lipids, and proteins. Base breakdown, single- or double-strand DNA breaks, protein cross-linking, and DNA modifications involving purines or pyrimidines are caused by damage from H₂O₂. H₂O₂ disrupts the lipid bilayer and subsequently affects tissue stability through lipid peroxidation; fragmented proteins, protein cross-linking, and amino acid oxidation are also produced by H₂O₂.

[0010] To balance fluctuating hydrogen peroxide (H2O2) levels in the body, a variety of antioxidant systems have been established. Catalase (CAT), glutathione peroxidase (GPx), and horseradish peroxidase (HRP) are known hydrogen peroxide (H2O2)-degrading enzymes. CAT is one of the most potent antioxidant enzymes containing a heme cofactor that breaks down hydrogen peroxide (H2O2) into harmless water and oxygen. GPx is a selenium cofactor enzyme, and the breakdown of hydrogen peroxide (H2O2) is accompanied by the oxidation of glutathione (GSH). Furthermore, HRP contains heme as a cofactor and exhibits catalase-like activity in reducing hydrogen peroxide (H2O2) to water and oxygen. All these antioxidant systems maintain equivalence of hydrogen peroxide (H2O2) levels under physiological conditions.

[0011] However, the balance of hydrogen peroxide (H2O2) levels breaks down under various pathological conditions, which are closely related to pathology. Typically, in neurodegenerative diseases, tumors, and autoimmune diseases, the balance of hydrogen peroxide (H2O2) is excessively skewed to one side. For example, the accumulation of amyloid-β induces the proliferation of monoamine oxidase B (MAO-B) and reactive astrocytes.

[0012] Therefore, drugs that prevent oxidative stress caused by hydrogen peroxide (H2O2) have become therapeutic targets. However, no effective drug targets have yet been identified for preventing damage caused by hydrogen peroxide (H2O2). Summary of the Invention

[0013] Technical issues

[0014] In their research to discover novel compounds with preventative or therapeutic effects against neurodegenerative diseases, the inventors focused on the fact that hydrogen peroxide, as a type of reactive oxygen species, is present at much higher levels in the pathological state of neurodegenerative diseases (especially Alzheimer's disease) compared to its byproduct formation from endogenous oxidative reactions such as mitochondrial respiration. They sought to develop novel compounds that could scavenge hydrogen peroxide, one of many types of reactive oxygen species. As a result, they discovered that amino-aromatic compounds with a specific structure remove hydrogen peroxide without scavenging hydroxyl radicals, thus completing this invention.

[0015] The object of this invention is to provide a novel amino-aromatic compound or a pharmaceutically acceptable salt thereof for use as a hydrogen peroxide scavenger in the blood.

[0016] Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of neurodegenerative diseases comprising a novel amino aromatic compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.

[0017] Furthermore, the present invention provides a health food composition for the prevention or improvement of neurodegenerative diseases, comprising a novel amino aromatic compound of the present invention or a food science-acceptable salt thereof as an active ingredient.

[0018] Technical solution

[0019] In order to achieve the above objectives,

[0020] According to one aspect of the invention, an amino aromatic compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof is provided:

[0021] [Chemical Formula 1]

[0022]

[0023] In the chemical formula 1,

[0024] Ar is C6-C 20 arylene, wherein the arylene group of Ar can be further selected from C1-C2. 10 Alkyl, C1-C 10 Alkoxy, amino, mono-C1-C 10 Alkylamino or di-C1-C 10 Alkylamino, Halogenated C1-C 10 Alkyl, halogenated C1-C 10 One or more of the alkoxy and hydroxyl groups are substituted;

[0025] R 1 and R2 Each independently can be either hydrogen or C1-C 10 alkyl;

[0026] R 3 Halogen, C1-C 10 Alkoxy, halogenated C1-C 10 Alkyl or halogenated C1-C 10 Alkoxy;

[0027] n is an integer of 1 or 2;

[0028] In R 3 In the case of halogens, n is an integer of 1.

[0029] Furthermore, another aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of neurodegenerative diseases comprising an amino aromatic compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0030] Furthermore, another aspect of the present invention provides a health food composition for the prevention or improvement of neurodegenerative diseases, comprising an amino aromatic compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0031] Technical effect

[0032] The amino aromatic compounds according to the present invention act as scavengers to remove hydrogen peroxide, which is a type of reactive oxygen species.

[0033] The amino aromatic compounds of the present invention inhibit apoptosis induced by H2O2-induced oxidative stress by scavenging hydrogen peroxide, a reactive oxygen species (ROS) in cells.

[0034] That is, since the amino aromatic compounds according to the present invention reduce the ROS concentration by acting together with enzymes containing heme, the concentration of ROS is not excessively reduced, but rather by removing excessively generated hydrogen peroxide to achieve an appropriate concentration required in vivo.

[0035] The amino aromatic compounds of the present invention, as small molecule scavengers of hydrogen peroxide in the blood, have very high blood-brain barrier (BBB) ​​permeability and can act directly on the brain, thus showing excellent effects in the treatment of brain diseases.

[0036] The amino aromatic compounds according to the present invention exhibit hydrogen peroxide scavenging activity and improve hydrogen peroxide-induced apoptosis, thereby demonstrating antioxidant properties that can improve cognitive and memory impairments.

[0037] The amino aromatic compounds of the present invention can decompose hydrogen peroxide in the presence of heme-containing peroxidase and hemoglobin (Hb) to reduce the level of hydrogen peroxide in the blood, thereby inhibiting or treating the onset of neurodegenerative diseases.

[0038] Therefore, the amino aromatic compounds of the present invention can inhibit damage caused by harmful hydrogen peroxide, and thus can be used as active ingredients in pharmaceutical compositions for the prevention or treatment of neurodegenerative diseases and in health food compositions for the prevention or improvement of neurodegenerative diseases. Attached Figure Description

[0039] Figure 1 This is a schematic image of hydrogen peroxide (H2O2) analysis after the reaction of hydrogen peroxide (H2O2) with heme protease between hydrogen peroxide (H2O2) and 10-acetyl-3,7-dihydroxyphenoxazine (Amplex Red).

[0040] Figure 2 Example 2 illustrates the in vitro analysis of the decomposition of H2O2 by a novel compound (A, B) shows the timeline of imaging using the cell-permeable H2O2 dye H2DCFDA-AM in primary cultured astrocytes and a schematic diagram of the chemical reaction based on the hydrogen peroxide measurement principle; (C) shows a graph of fluorescence intensity of the concentration of the amino-aromatic compound KDS12008 (Example 1) according to the present invention. The fluorescence intensity represents the amount of intracellular H2O2, which is normalized by control conditions).

[0041] Figure 3 The results of the in vitro analysis of the decomposition of H2O2 by the novel compound in Experimental Example 2 are shown in Figure I. ((A) shows a graph illustrating the decomposition effect of the amino aromatic compound KDS12008 (Example 1) (10 μM) and sodium pyruvate (10 mM) on H2O2; (B) shows the cell viability test results of the amino aromatic compound KDS12008 (Example 1) (100 μM) and sodium pyruvate (10 mM). Fluorescence intensity represents the amount of intracellular H2O2, which was normalized by control conditions. **P<0.01; ***P<0.001; ns, not valid).

[0042] Figure 4 The results of the memory impairment (memory disorder) recovery experiment in APP / PS1 mice treated with the amino-aromatic compounds of the present invention in Experimental Example 3 are shown in (A) schematic timeline of drug treatment and passive avoidance test (PAT); (B) bar chart of latency in the dark room in APP / PS1 mice treated with the amino-aromatic compounds KDS12008 (Example 1), KDS12017 (Example 16), or KDS12025 (Example 21) of the present invention, respectively. Data are expressed as mean ± SEM. Unpaired two-tailed t-test. *P<0.05, ****P<0.0001).

[0043] Figure 5 The results of immunohistochemistry (IHC) of brain tissue from APP / PS1 mice treated with the amino aromatic compounds KDS12008 (Example 1), KDS12017 (Example 16), or KDS12025 (Example 21) of the present invention are shown in Experimental Example 4.

[0044] Figure 6 This is the result of the Passive Avoidance Experiment (PAT) in Experiment Example 5.

[0045] Figure 7 This is the result of immunohistochemistry (IHC) II staining of the brain tissue in Experiment Example 6.

[0046] Figure 8 This is the result of the electrophysiology experiment in Experiment Example 7.

[0047] Figure 9 This is the result of the novel place recognition experiment in Experiment Example 8.

[0048] Figure 10 This is the result of the Passive Avoidance Experiment (PAT) in Experiment Example 8.

[0049] Figures 11 to 13 This is the result of a single toxicity evaluation (lethal dose 50, LD50) for Experimental Example 9. Detailed Implementation

[0050] The novel amino-aromatic compounds or pharmaceutically acceptable salts of such compounds of the present invention will be described in detail below. In this context, unless otherwise defined, the technical and scientific terms used have the meanings commonly understood by one of ordinary skill in the art to which this invention pertains. In the following description, descriptions of well-known functions and structures that may unnecessarily obscure the essence of the invention will be omitted.

[0051] The following terms, as used in this specification, are defined as follows, but are merely illustrative and not intended to limit the invention, application, or purpose.

[0052] In this specification, the terms “substituent”, “radical”, “group”, “moiety” and “fragment” are used interchangeably.

[0053] In this specification, the term "C" A -C B "" indicates "the number of carbon atoms is A or more and B or less".

[0054] In this specification, the term "alkyl" refers to a monovalent straight-chain or branched saturated hydrocarbon group consisting only of carbon and hydrogen atoms. The alkyl group may have 1 to 10 carbon atoms, 1 to 7 carbon atoms, or 1 to 4 carbon atoms. "Lower alkyl" refers to a straight-chain or branched alkyl group having 1 to 4 carbon atoms. As examples, the alkyl group includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, ethylhexyl, etc.

[0055] In this specification, the term "arylene" refers to an organic radical derived from an aromatic hydrocarbon by removing two hydrogen atoms, comprising monocyclic or fused ring systems in which each ring suitably contains 4 to 7, preferably 5 or 6 ring atoms, and in forms in which multiple aryl groups are linked by single bonds. Specific examples include, but are not limited to, phenylene, naphthylene, biphenylene, anthracene, etc.

[0056] In this specification, the term "alkoxy" refers to an -O-alkyl radical, and "alkyl" here is the same as defined herein. Specific examples include, but are not limited to, methoxy, ethoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, etc.

[0057] In this specification, the term "halogenated" or "halogen" refers to a group of halogen elements, such as fluorine, chlorine, bromine, and iodine.

[0058] As used in this specification, the terms "haloalkyl" or "haloalkoxy" refer to an alkyl group or alkoxy group in which one or more hydrogen atoms are replaced by halogen atoms, wherein alkyl and halogen are as defined above. For example, haloalkyl can include fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, perfluoroethyl, etc., and haloalkoxy can include fluoromethoxy, difluoromethoxy, trifluoromethoxy, fluoroethoxy, difluoroethoxy, perfluoroethoxy, etc.

[0059] In this specification, the term "amino" refers to -NH2 and "hydroxyl" refers to -OH.

[0060] In this specification, the term "alkylamino" refers to an amino radical in which one or two alkyl groups are substituted, and specific examples include, but are not limited to, methylamino (-NHMe), dimethylamino (-NMe2), ethylamino (-NHEt), diethylamino (-NEt2), etc.

[0061] In this specification, the term "pharmaceutical acceptable" means that the composition has no toxic properties to cells or individuals, such as humans, exposed to it, and is generally considered safe for use in pharmaceutical preparations. Such use is formally approved by national authorities or listed in the Korean Pharmacopoeia or the United States Pharmacopeia.

[0062] In this specification, the term "pharmaceutically acceptable salt" refers to any organic or inorganic addition salt of the compounds of the present invention, the concentration of which has a relatively non-toxic and harmless effect on the patient, and the side effects caused by the salt do not reduce the beneficial efficacy of the compounds of the present invention.

[0063] In this specification, the terms "pharmaceuticalally acceptable excipient" and "pharmaceuticalally acceptable carrier" refer to substances that facilitate the administration of the active agent and its absorption by the subject.

[0064] In this specification, the term “oxidative stress” is used in its usual sense and refers to an abnormal level of reactive oxygen species.

[0065] In this specification, the term "prevention" refers to all actions that inhibit or delay the onset, spread, and recurrence of neurodegenerative diseases.

[0066] In this specification, the term "improvement" means all actions that at least reduce parameters (e.g., the severity of symptoms) related to the condition being treated.

[0067] In this specification, the term "treatment" refers to all behaviors that result in improvement or favorable change of symptoms in neurodegenerative diseases.

[0068] In this specification, the term "individual" refers to all animals (including humans) that have developed or may develop neurodegenerative diseases. The animals may include, but are not limited to, mammals such as cattle, horses, sheep, pigs, goats, camels, antelopes, dogs, and cats that require treatment for similar symptoms.

[0069] In this specification, the term "administration" means introducing the pharmaceutical composition of the present invention into an individual by some suitable method. As for the route of administration of the composition of the present invention, it can be administered by a variety of routes, including oral or non-oral administration, as long as the target tissue can be reached.

[0070] In this specification, the term "pharmaceutical effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment without causing side effects. For effective dosage standards, the person in charge can readily determine based on factors including the patient's sex, age, weight, health status, type and severity of the disease, drug activity, drug sensitivity, method of administration, time of administration, route of administration and excretion rate, treatment duration, factors of concomitant or concurrent medications, or other factors known in the medical field.

[0071] In this specification, the term "food" includes meat, sausage, bread, chocolate, confectionery, snacks, biscuits, pizza, instant noodles, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health foods and health foods, etc., including all foods in the general sense.

[0072] In this instruction manual, the term "health food" refers to food prepared and processed using raw materials or ingredients that have functional properties beneficial to the human body, in accordance with Korean Law No. 6727 concerning health foods. "Functional" means that it is ingested for the purpose of obtaining effects beneficial to health purposes, such as regulating nutrients or physiological functions, based on the structure and function of the human body.

[0073] In this specification, the term "food-acceptable salt" refers to a dosage form of a compound that does not cause severe irritation to the organism to which the compound is administered, nor does it impair the biological activity and physical properties of the compound.

[0074] This invention provides a treatment for neurodegenerative diseases comprising a novel amino aromatic compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt of such compound, as an active ingredient, wherein the novel amino aromatic compound or the pharmaceutically acceptable salt of such compound can be used as a hydrogen peroxide scavenger in the blood.

[0075] [Chemical Formula 1]

[0076]

[0077] In the chemical formula 1,

[0078] Ar is C6-C 20 arylene, wherein the arylene group of Ar can be further selected from C1-C2. 10 Alkyl, C1-C 10 Alkoxy, amino, mono-C1-C 10 Alkylamino or di-C1-C 10 Alkylamino, Halogenated C1-C 10 Alkyl, halogenated C1-C 10 One or more of the alkoxy and hydroxyl groups are substituted;

[0079] R 1 and R 2 Each independently can be either hydrogen or C1-C 10 alkyl;

[0080] R 3 Halogen, C1-C 10 Alkoxy, halogenated C1-C 10 Alkyl or halogenated C1-C 10 Alkoxy;

[0081] n is an integer of 1 or 2;

[0082] But in R 3 In the case of halogens, n is an integer of 1.

[0083] The amino aromatic compounds of the present invention have low cytotoxicity and, as small molecular weight compounds, can act as scavengers to remove hydrogen peroxide, a type of reactive oxygen species.

[0084] The amino aromatic compounds according to the present invention do not remove hydroxyl radicals and are not MAO-B inhibitors. ROS-GLO analysis confirmed that the amino aromatic compounds according to the present invention act as catalysts in the presence of endogenous peroxidases, particularly heme-containing peroxidases and hemoglobin (Hb), to perform the reaction that decomposes hydrogen peroxide into water.

[0085] That is, because the amino-aromatic compounds according to the present invention act together with heme-containing peroxidases and hemoglobin (Hb) present in the body to remove excess hydrogen peroxide until an appropriate concentration is reached, the amino-aromatic compounds of the present invention inhibit neuronal apoptosis caused by harmful hydrogen peroxide, thereby effectively preventing, improving, or treating neurodegenerative diseases. Furthermore, the amino-aromatic compounds according to the present invention have the ability to penetrate the blood-brain barrier (BBB) ​​with excellent efficiency, thus allowing for administration at low concentrations and achieving rapid, fast, and more effective therapeutic effects.

[0086] In one embodiment of the present invention, Ar is C6-C. 12 The Ar group is preferably phenylene or biphenylene; the Ar group may be further substituted with one or more of the following: C1-C7 alkyl, C1-C7 alkoxy, amino, and hydroxyl groups.

[0087] In one embodiment of the present invention, Ar is a phenylene oxide, and preferably, nitrogen atoms are introduced at positions 1 and 4 of the phenylene oxide, respectively.

[0088] Specifically, an amino aromatic compound according to an embodiment of the present invention may be represented by the following chemical formula 2 or 3.

[0089] [Chemical Formula 2]

[0090]

[0091] [Chemical Formula 3]

[0092]

[0093] In the chemical formulas 2 and 3,

[0094] R 1 and R 2 Each is independently hydrogen or a C1-C7 alkyl group;

[0095] Hal is a halogen;

[0096] R 3 It is a C1-C7 alkoxy or a halo-C1-C7 alkyl;

[0097] R' is a C1-C7 alkyl, C1-C7 alkoxy, amino, or hydroxyl group;

[0098] a is an integer from 0 to 4;

[0099] n is an integer that is either 1 or 2.

[0100] Preferably, in chemical formulas 2 and 3 according to an embodiment of the present invention, the R1 and R 2 Each is independently hydrogen or C1-C4 alkyl; Hal is a halogen; R 3 It is a C1-C4 alkoxy or a halo-C1-C4 alkyl; a is an integer 0; n can be an integer of 1 or 2.

[0101] According to one embodiment, chemical formula 2 can be represented by the following chemical formula 4:

[0102] [Chemical Formula 4]

[0103]

[0104] In chemical formula 4,

[0105] R 1 and R 2 Each is independently hydrogen or C1-C4 alkyl;

[0106] Hal is a halogen.

[0107] Specifically, in the chemical formula 4, the R 1 and R 2 Each is independently a C1-C4 alkyl group; Hal can be a halogen.

[0108] Specifically, in the chemical formula 4, the R 1 It is hydrogen; R 2 It is a C1-C4 alkyl group; Hal can be a halogen.

[0109] Specifically, in the chemical formula 4, the R 1 and R 2 Each can be hydrogen independently; Hal can be a halogen.

[0110] According to one embodiment, chemical formula 3 can be represented by the following chemical formula 5:

[0111] [Chemical Formula 5]

[0112]

[0113] In chemical formula 5,

[0114] R 1 and R 2 Each is independently hydrogen or C1-C4 alkyl;

[0115] R 3 It is a C1-C4 alkoxy or a halo-C1-C4 alkyl;

[0116] n is an integer that is either 1 or 2.

[0117] Specifically, in the chemical formula 5, the R 1and R 2 Each is independently a C1-C4 alkyl group; R 3 It is a C1-C4 alkoxy or a halo-C1-C4 alkyl; n is an integer of 1 or 2.

[0118] Specifically, in the chemical formula 5, R 1 It is hydrogen; R 2 It is a C1-C4 alkyl group; R 3 It is a C1-C4 alkoxy or a halo-C1-C4 alkyl; n is an integer of 1 or 2.

[0119] Specifically, in the chemical formula 5, the R 1 and R 2 Each is independently hydrogen; R 3 It is a halogenated C1-C4 alkyl group; n is an integer of 1 or 2.

[0120] In any compound described in this specification, the halogen or halogen may be fluorine.

[0121] The amino aromatic compound according to one embodiment may be selected from any of the following groups of compounds, but is not limited thereto.

[0122]

[0123]

[0124] It will be apparent to those skilled in the art that the method for preparing an amino aromatic compound according to an embodiment of the present invention can be carried out by utilizing or appropriately modifying methods known in the art. Furthermore, the reaction time in the preparation method of Formula 1 according to an embodiment of the present invention can vary depending on the type and amount of reactants and solvent. As an example, the reaction is terminated after confirmation by TLC or the like that the starting material has been completely consumed. When the reaction is complete, the solvent is distilled under reduced pressure, and the target product can then be separated and purified by conventional methods such as column chromatography. As an example, it can be prepared by reacting an aryldiamine compound with a phenylalkyl bromide compound; more details will be described in the following examples.

[0125] [Reaction Formula 1]

[0126]

[0127] (In reaction 1 above, Ar, R) 1 R 2 R 3 (and n is the same as the content in the above chemical formula 1).

[0128] This invention includes not only the above-mentioned amino aromatic compounds and pharmaceutically acceptable salts of such compounds, but also all prodrugs, hydrates and solvates that can be prepared therefrom.

[0129] That is, the amino aromatic compounds of the present invention can be used in the form of prodrugs, hydrates, solvates, and pharmaceutically acceptable salts to promote absorption in vivo or increase solubility. Therefore, the prodrugs, hydrates, solvates, and pharmaceutically acceptable salts also fall within the scope of the present invention.

[0130] The amino aromatic compounds of the present invention can be used in the form of pharmaceutically acceptable salts, which are salts prepared according to conventional methods in the art, methods known to those skilled in the art. Specifically, the pharmaceutically acceptable salts include, but are not limited to, salts derived from free acids and bases that are pharmacologically or physiologically acceptable.

[0131] Acid addition salts formed from pharmaceutically acceptable free acids include inorganic acids (hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc.) and organic acids (methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutamate, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, etc.). These pharmaceutically non-toxic salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, fluorides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, decanoates, heptanoates, propynylates, oxalates, malonates, succinates, caprylates, sebacic acid, fumarates, maleates, and butynedi-1,4-dicarboxylic acid. Salts, hexane-1,6-diacidates, benzoates, chlorobenzoates, methyl benzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalates, terephthalates, benzenesulfonates, toluenesulfonates, chlorobenzenesulfonates, xylenesulfonates, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate esters, methanesulfonates, propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, mandelates, etc.

[0132] Acid addition salts can be prepared by conventional methods, such as dissolving the amino aromatic compounds of the present invention in water-miscible organic solvents such as methanol, ethanol, acetone, dichloromethane, and acetonitrile, adding organic or inorganic acids, filtering and drying the resulting precipitate, or preparing them by vacuum distillation of the solvent and excess acid followed by drying and crystallization in an organic solvent.

[0133] Furthermore, pharmaceutically acceptable metal salts can be prepared using alkalis. Alkali metal salts or alkaline earth metal salts can be obtained, for example, by dissolving the amino aromatic compound of the present invention in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the undissolved amino aromatic compound salt, and then evaporating and drying the filtrate. In this case, the preparation of sodium, potassium, or calcium salts as metal salts is pharmaceutically suitable, but not limited thereto. Additionally, the corresponding silver salts can be obtained by reacting the alkali metal salt or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0134] Preferably, the pharmaceutically acceptable salt of the amino aromatic compound according to an embodiment of the present invention can be a hydrochloride salt.

[0135] That is, the amino aromatic compound according to an embodiment of the present invention can be a compound in the form of a hydrochloride salt selected from the following structures:

[0136]

[0137] The hydrate of the amino aromatic compound of the present invention refers to an amino aromatic compound of the present invention or a pharmaceutically acceptable salt thereof containing a stoichiometric or non-stoichiometric amount of water bound by non-covalent intermolecular forces.

[0138] The solvates of the amino aromatic compounds of the present invention represent the heterocyclic compounds of the present invention or pharmaceutically acceptable salts of such compounds comprising stoichiometric or non-stoichiometric amounts of solvent bound by non-covalent intermolecular forces. Available solvents include volatile, non-toxic solvents.

[0139] The amino-aromatic compounds of the present invention can be administered in the form of a prodrug, which is decomposed in a human or animal body to provide the compounds of the present invention as the active ingredient. The prodrug can be used to modify and / or improve the physical and / or pharmacokinetic profile of the parent compound, and can be formed when the parent compound contains suitable groups or substituents that can be induced in a manner that forms a prodrug.

[0140] If a compound (prodrug) is isolated from the body to generate the amino-aromatic compound or its salt of the present invention, such compounds are also included within the scope of the present invention. As used herein, unless otherwise stated, the term "prodrug" refers to a compound of the present invention that can be hydrolyzed, oxidized, and otherwise reacted under biological conditions (in vitro or in vivo) to provide an active compound, particularly a compound of the present invention. Examples of prodrugs include compounds that generate the compounds of the present invention through biohydrolysis, said compounds comprising, but not limited to, biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogs. Preferably, prodrugs of compounds having a carboxyl functional group are lower alkyl esters of carboxylic acids. Carboxylic acid esters are typically formed by esterifying a portion of the carboxylic acid present in the molecule. The prodrug can be readily prepared using the methods described in Burger's Medicinal Chemistry and Drug Discovery, 6th edition (Donald J. Abrahamed., 2001, Wiley) and Design and Application of Prodrugs (H. Bundgaarded., 1985, Harwood Academic Publishers Gmfh).

[0141] The present invention provides a hydrogen peroxide scavenger comprising an amino aromatic compound of the chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0142] Furthermore, the present invention provides a pharmaceutical composition for the prevention or treatment of neurodegenerative diseases comprising an amino aromatic compound of the chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0143] Neurodegenerative diseases refer to diseases or conditions that impair the function of the nervous system, including motor regulation, cognitive function, sensory function, and autonomic nervous system function. They are synonymous with "degenerative brain diseases." Specifically, examples include dementia, Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease, amyotrophic lateral sclerosis (ALS), post-traumatic stress disorder, multiple sclerosis (MS), cerebral ischemic disease, and amyotrophic lateral sclerosis.

[0144] The amino aromatic compounds according to the present invention can decompose excess hydrogen peroxide in the presence of heme-containing peroxidase and hemoglobin (Hb) in the blood, thereby reducing the level of hydrogen peroxide in the blood to an appropriate level, thereby inhibiting or treating the onset of neurodegenerative diseases.

[0145] Furthermore, the amino-aromatic compounds of the present invention, as low-molecular-weight hydrogen peroxide scavengers in the blood, possess very high blood-brain barrier (BBB) ​​permeability, thus allowing them to act directly on the brain and exhibit excellent efficacy in treating brain diseases. Therefore, the amino-aromatic compounds of the present invention can be effectively used for the prevention or treatment of neurodegenerative diseases.

[0146] In addition to the aforementioned active ingredient, a pharmaceutical composition according to one embodiment also contains a generally non-toxic, pharmaceutically acceptable carrier and / or excipient, thereby enabling it to be dosage formulated into a pharmaceutically acceptable formulation, i.e., an oral or non-oral formulation. Furthermore, it may further include fillers, extenders, binders, wetting agents, disintegrants, surfactants, and other diluents.

[0147] Examples of pharmaceutically acceptable carriers, excipients, or diluents may include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, or mineral oil.

[0148] The pharmaceutical compositions of the present invention can be formulated into various forms according to conventional methods, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, and other oral dosage forms, as well as injectable sterile injection solutions, according to their intended use. They can also be administered via various routes, including oral, intravenous, intraperitoneal, subcutaneous, rectal, and local administration.

[0149] Furthermore, the pharmaceutical compositions of the present invention may additionally include fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc.

[0150] Oral dosage forms include tablets, pills, hard / soft capsules, liquids, suspensions, emulsifiers, syrups, granules, and elixirs. In addition to the active ingredient, these dosage forms may use one or more commonly used fillers, expanders, wetting agents, disintegrants, lubricants, binders, surfactants, or other diluents or excipients. Disintegrants may include agar, starch, alginate or its sodium salt, anhydrous calcium monohydrogen phosphate, etc.; lubricants may include silica, talc, stearic acid or its magnesium or calcium salts, polyethylene glycol, etc.; and binders may include magnesium aluminum silicate, starch paste, gelatin, tragacanth gum, methylcellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidone, and low-substituted hydroxypropyl cellulose, etc. In addition, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc. can be used as diluents, and depending on the situation, commonly known boiling mixtures, absorbents, colorants, flavorings, sweeteners, etc. can be used together.

[0151] Examples of formulations for non-oral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Non-aqueous solvents and suspensions can include vegetable oils such as propylene glycol, polyethylene glycol, olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, synthetic fatty acid esters (witepsol), polyethylene glycol, Tween 61, cocoa butter, glyceryl laurate, glycerin, and gelatin can be used. Furthermore, injections can include conventional additives such as solvents, isotonic agents, suspending agents, emulsifiers, stabilizers, and preservatives. To formulate an injection, the amino-aromatic compound of the present invention or a pharmaceutically acceptable salt thereof can be mixed with a stabilizer or buffer in water to prepare a solution or suspension, which is then prepared into a unit-dose form in ampoules or vials.

[0152] The pharmaceutical compositions of the present invention may be sterile, or may contain preservatives, stabilizers, thickeners, wettable powders or emulsification promoters, salts and / or buffers for adjusting osmotic pressure, and may also contain other therapeutically useful substances, and may be formulated by conventional methods such as dissolving, dispersing, mixing, granulating, gelling or coating.

[0153] The pharmaceutically effective amount of the amino aromatic compounds of the present invention depends on factors including the patient's health status, the type and severity of the disease, the activity of the drug, the sensitivity to the drug, the method of administration, the time of administration, the route of administration, the excretion rate, the duration of treatment, the drugs used in combination or concurrently, and other factors known in the medical field. Specifically, the effective amount of the compound in the pharmaceutical composition of the present invention can vary depending on the patient's age, sex, and weight, and is generally from about 0.01 mg / kg / day to 500 mg / kg / day, preferably from 0.1 mg / kg / day to 100 mg / kg / day, and can be administered daily or every other day, or once daily or divided into several doses. However, since the dosage can be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, etc., the dosage is not intended to limit the scope of the present invention in any way.

[0154] The pharmaceutical composition of the present invention can be administered orally or non-orally, preferably non-oral via subcutaneous injection, intravenous injection, intramuscular injection or intraperitoneal injection.

[0155] The pharmaceutical compositions of the present invention can be administered as a single therapeutic agent or in combination with other therapeutic agents, can be administered sequentially or simultaneously with existing therapeutic agents, and can be administered once or multiple times. Considering all the above factors, it is particularly important to administer the drug in the amount that achieves the maximum effect with the minimum amount without side effects, and this can be readily determined by those skilled in the art.

[0156] Furthermore, the present invention provides a method for preventing or treating neurodegenerative diseases, comprising the step of administering the amino aromatic compound or a pharmaceutically acceptable salt of the compound or the pharmaceutical composition to a subject suffering from or at risk of developing neurodegenerative diseases.

[0157] Furthermore, the present invention provides a health food composition for the prevention or improvement of neurodegenerative diseases, comprising the amino aromatic compound or a food-grade acceptable salt thereof as an active ingredient.

[0158] The food-grade acceptable salts may include those obtained by reacting the amino aromatic compounds of the present invention with inorganic acids (hydrochloric acid, bromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.), sulfonic acids (methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, etc.), organic carbonic acids (such as tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, decanoic acid, isobutyric acid, malonic acid, succinic acid, phthalic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid, etc.). Furthermore, the compounds of the present invention may react with bases to obtain alkali metal salts (ammonium salts, sodium salts, or potassium salts, etc.), alkaline earth metal salts (calcium salts or magnesium salts, etc.), salts of organic bases (e.g., dicyclohexylamine, N-methyl-D-glucosamine, tris(hydroxymethyl)methylamine, etc.), and amino acid salts (arginine, lysine, etc.), but are not limited thereto.

[0159] The health food composition may be provided in the form of powder, granules, tablets, capsules, syrup, or beverage. In addition to the amino aromatic compound as the active ingredient, the health food may be used with other foods or food additives, and may be used appropriately according to conventional methods. The amount of active ingredient mixed may be appropriately determined according to its intended use (e.g., preventative, health-promoting, or therapeutic treatment).

[0160] The health food composition may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents (synthetic and natural flavoring agents, etc.), coloring agents and fillers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, and carbonating agents for carbonated beverages. In addition, it may contain fruit pulp used in the preparation of natural fruit juices, fruit juice beverages, and vegetable beverages. These ingredients may be used alone or in combination.

[0161] Furthermore, the health food may also contain food additives. As for the suitability of "food additives", unless otherwise specified, it shall be determined based on the general principles and general test methods of the Food Additives Code approved by the Ministry of Food and Drug Safety, according to the specifications and standards related to the corresponding items.

[0162] Items included in the aforementioned "Codex Alimentarius" may include, for example, chemically synthesized products (ketones, glycine, potassium citrate, niacin, cinnamic acid, etc.), natural additives (persimmon pigment, licorice extract, crystalline cellulose, guar gum, etc.), and mixed preparations (L-glutamate preparations, alkali additives for flour products, preservative preparations, tar pigment preparations, etc.).

[0163] The amino aromatic compounds contained in the health food composition may be used based on the effective dosage of the pharmaceutical composition. However, in the case of long-term intake for health and hygiene purposes or for health regulation purposes, the dosage may be below the above range. Since there are no safety issues with the active ingredients, it is obvious that the active ingredients may be used in amounts above the above range.

[0164] The health food composition can be formulated into various dosage forms such as meat, sausage, bread, chocolate, candy, snacks, biscuits, pizza, instant noodles, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes.

[0165] The present invention will now be described in more detail through preferred embodiments. However, this is merely illustrative of the invention, and the scope of the invention is not limited thereto in any sense, but is defined only by the scope of the claims.

[0166] [Preparation Example 1] Preparation of (4-aminophenyl)(methyl)carbamate tert-butyl ester

[0167]

[0168] Preparation of N-methyl-4-nitroaniline

[0169] Fluoro-4-nitrobenzene (1.0 g, 7.1 mmol) and methylamine (3.41 g, 109.9 mmol) were dissolved in ethanol, and the solution was refluxed for 24 hours. The solution was then cooled to room temperature and the solvent was removed under reduced pressure. The organic layer was separated from the residue by ethyl acetate and brine and washed. The separated organic layer was dried over sodium sulfate, and then ethyl acetate was removed under reduced pressure to give 913.3 mg (84.6%) of the target compound as a yellow solid. (400 MHz) 1 H NMR(DMSO-d6)δ8.01(d,2H,J=9.28),7.31(d,1H,J=4.28),6.61(d,2H,J=9.36),2.80(d,2H,J=5.00),2.33(s,1H)

[0170] Preparation of (methyl)(4-nitrophenyl)carbamate tert-butyl ester

[0171] N-methyl-4-nitroaniline (800 mg, 5.26 mmol), di-tert-butyl dicarbonate (1.72 g, 7.89 mmol), and 4-dimethylaminopyridine (32.1 mg, 0.26 mmol) were dissolved in tetrahydrofuran, and the solution was refluxed for 12 hours. The solution was then cooled to room temperature and the solvent was removed under reduced pressure. The organic layer was separated from the residue by ethyl acetate and brine and washed. The separated organic layer was dried over sodium sulfate, and then ethyl acetate was removed under reduced pressure to give 1.29 g (97.3%) of the target compound as a yellow oil. (400 MHz) 1 H NMR(DMSO-d6)δ8.20(d,2H,J=9.20),7.60(d,2H,J=9.16),3.29(s,3H),1.45(s,9H)

[0172] Preparation of (4-aminophenyl)(methyl)carbamate tert-butyl ester

[0173] 1.29 g (5.11 mmol) of tert-butylmethyl (nitrophenyl)carbamate was dissolved in a mixed solvent of distilled water (12 mL), methanol (25 mL), and tetrahydrofuran (6 mL). Iron powder (1.36 g, 26.1 mmol) and ammonium chloride (2.80 g, 52.4 mmol) were then added, and the mixture was stirred at 50 °C for 3 hours. The mixture was then cooled to room temperature and filtered through diatomaceous earth (Celite). The solvent was removed under reduced pressure after filtration. The organic layer was separated from the residue by ethyl acetate and brine and washed. The separated organic layer was dried over sodium sulfate, and then ethyl acetate was removed under reduced pressure to give 890 mg (78.1%) of the target compound as a yellow solid. (400 MHz) 1 H NMR(DMSO-d6)δ6.86(d,2H,J=8.52),6.50(d,2H,J=8.60),5.01(s,2H),3.06(s,3H),1.35(s,9H)

[0174] Example I: Preparation of amino aromatic compounds

[0175]

[0176] Potassium carbonate (3.0 equivalents), potassium iodide (0.1 equivalents), and a phenylalkyl bromide compound (b, 1.0 equivalents) were added to an acetonitrile solution containing a p-phenylenediamine compound (a, 1.2 equivalents). The mixture was then stirred at 110°C for 36 hours. Afterward, the mixture was cooled to room temperature, diluted with ethyl acetate, and washed with brine. The remaining organic layer was dried over Na₂SO₄ and then the solvent was removed under reduced pressure. The residue was purified by column chromatography to obtain compound P1. The purified compound P1 was dissolved in dichloromethane (DCM), and then 4.0 M hydrogen chloride solution was added. The mixture was then stirred at room temperature for 48 hours, and the resulting precipitate was filtered to obtain compound P2 in its hydrochloride form. Several amino aromatic compounds listed in Table 1 were prepared using the above method.

[0177] [Table 1]

[0178]

[0179]

[0180]

[0181]

[0182]

[0183] [Experimental Example]

[0184] Primary cultured astrocytes

[0185] Primary cortical astrocytes were prepared from C57BL / 6 mice from birth to day 3. The cerebral cortex was dissected and homogenized without adherent meninges, and then triturated into single-cell suspensions. Cells were grown in DMEM medium (Dulbecco's modified Eagle's medium) (Invitrogen) supplemented with 25 mM glucose, 10% heat-inactivated horse serum, 10% heat-inactivated fetal bovine serum (FCS), 2 mM glutamine, and 1000 U / ml penicillin-streptomycin. The cells were incubated at 37°C in a humidified 5% CO2 incubator. On day 3, cells were vigorously washed by repeated pipetting and the medium was changed to remove remnants and other suspension cell types.

[0186] [Experimental Example 1] Evaluating hydrogen peroxide scavenging capacity using enzymes present in vivo

[0187] To understand the hydrogen peroxide scavenging capacity of the proposed enzyme and the amino aromatic compounds of the present invention, the following experiments were performed.

[0188] Amplex Red is a fluorescent dye that transforms into a resorufin when horseradish peroxidase (HRP) and H2O2 are present, enabling the measurement of changes in H2O2 (i.e., the H2O2 assay). However, HRP is an enzyme not present in the body. Considering that hemoglobin has the same heme group as HRP, experiments were conducted based on the idea that it would act in conjunction with the amino-aromatic compound of this invention. The experiment involved adding hemoglobin instead of HRP to the H2O2 measurement via Amplex Red. Figure 1 The reaction was carried out in a manner described above. The final concentrations of each substance were 10 μM H2O2 and 80 μg / ml hemoglobin, and the amino aromatic compounds of this invention were treated according to concentration. After reacting at 37°C for approximately 30 minutes, fluorescence (excitation: 540 nm, emission: 580 nm) was measured using a microplate reader. At this point, the fluorescence intensity can be considered proportional to the amount of H2O2. Based on the drug-free fluorescence value, after normalization according to each concentration, the fluorescence value was statistically processed and the program was used to calculate the 50% value, i.e., the half-maximum effect concentration (EC5). 50 (half maximal effective concentration). The results are shown in Table 2 below, with AAD-2004 used as a control group.

[0189] [Table 2]

[0190]

[0191]

[0192]

[0193] As shown in Table 2 above, it can be seen that the amino-aromatic compounds according to the present invention effectively react with hemoglobin (Hb) to reduce hydrogen peroxide. Therefore, it can be seen that the amino-aromatic compounds according to the present invention can be used as hydrogen peroxide scavengers, which work together with hemoglobin present in the body to reduce the excess hydrogen peroxide generated, thereby achieving an appropriate level.

[0194] [Experimental Example 2] Evaluation of H2O2 decomposition experiment I and cell viability conducted in vitro.

[0195] To verify the H2O2 scavenging effect at the cellular level, the amount of intracellular H2O2 was measured by fluorescence in astrocytes using the cell-permeant 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA-AM) as a hydrogen peroxide probe. Figure 2 (A, B)). Using the amino-aromatic compound KDS12008 (Example 1) of the present invention as the test substance, sodium pyruvate, known to remove H2O2, was used as the control substance. When glial cells were treated with various concentrations of the amino-aromatic compound KDS12008 (Example 1) of the present invention, it was confirmed that H2O2 decreased with increasing concentration (…). Figure 2 (C)).

[0196] Figure 2 (A) shows an experimental timeline for understanding the effect of the amino-aromatic compound KDS12008 (Example 1) (10 μM) and sodium pyruvate (10 mM) of the present invention on the degradation of naturally occurring H2O2 in cultured astrocytes. The cultured astrocytes were allowed to stabilize for three days before being treated with the amino-aromatic compound KDS12008 (Example 1) and sodium pyruvate of the present invention. Two days later, a cell-permeable H2O2 dye (H2DCFDA-AM) was used as a method for measuring intracellular H2O2. Measurements were performed using 10 μM of H2DCFDA-AM as a sample that reacts with H2O2 to produce green fluorescence. The results are shown in… Figure 3 (A). From Figure 3 (A) confirms that when using the amino aromatic compound KDS12008 (Example 1) (10 μM) of the present invention, there is a statistically significant reduction in H2O2, and the reduction trend is greater than that of high concentrations of 10 mM sodium pyruvate.

[0197] In addition, experiments were performed using QuantiMax samples to determine cell viability. Cell viability was confirmed by the degree of luminescence observed by the QuantiMax samples, and the results are presented below. Figure 3 (B). From Figure 3 As can be seen from (B), sodium pyruvate reduces cell viability, while the amino aromatic compound KDS12008 of the present invention (Example 1), even at a concentration of 100 μM higher than the 10 μM concentration used to measure the decomposition effect of H2O2, has no effect on cell viability. That is, it is evident that it exhibits far superior efficiency compared to existing known substances.

[0198] [Experimental Example 3] Passive Avoidance Test (PAT)

[0199] The passive avoidance experiment, used to assess an animal's memory, involves applying a weak electrical stimulus to the animal in a dark room to evaluate whether the animal remembers the stimulus.

[0200] Because rats prefer dark rooms, they exhibit a tendency to move quickly from bright to dark rooms. However, if a weak electrical stimulus is applied to a rat in a dark room, it will not move from the bright room to the dark room because it develops a memory that associates the dark room with the electrical stimulus. The memory associated with the electrical stimulus and the dark room can be measured by measuring the time it takes to move from the bright room to the dark room (latency to dark room, seconds). Passive avoidance experiments were performed using APP / PS1 mice, an animal model of Alzheimer's disease. Figure 4 Wild-type (WT) mice were used as a control group.

[0201] On the first day of PAT (Physical Anesthesia), designated as the acquisition session, rats were placed in a bright room for a predetermined time. After this time, the door to a dark room was opened, and the time it took for the rats to enter the dark room was measured. If a rat entered the dark room, it was given an electric shock (0.5 mA, 2 seconds) via a foot pedal. On the second day, designated as the recovery session, rats were placed in a bright room, and the time it took for them to enter the dark room was measured. The door to the dark room was open from the beginning. Rats with good memory would not enter the dark room due to their memory of the electric stimulation, while those with poor memory would quickly enter the dark room again. WT mice, having a memory of the electric stimulation received the previous day (the acquisition session), did not want to enter the dark room. However, APP / PS1 dementia mice did not remember the electric stimulation received in the dark room the previous day and were therefore more likely to enter the dark room.

[0202] The amino aromatic compounds of the present invention (KDS12008 (Example 1), KDS12017 (Example 16), and KDS12025 (Example 21)) were injected intraperitoneally into APP / PS1 dementia mice for 16 days (KDS12008 (Example 1) 30 mg / kg / day (30 mpk); KDS12017 (Example 16) 30 mg / kg / day (30 mpk); KDS12025 (Example 21) 3 mg / kg / day (3 mpk)). Significant recovery of memory impairment in APP / PS1 dementia mice was confirmed during the passive avoidance test performed on day 26. Figure 4 ).

[0203] Specifically, it was confirmed that the Alzheimer's disease model APP / PS1 mice had poorer memory compared to wild-type mice. However, APP / PS1 mice administered the amino-aromatic compounds of the present invention (KDS12008 (Example 1), KDS12017 (Example 16), or KDS12025 (Example 21)) showed improved memory for passive avoidance, resulting in a significant increase in dwell time. Therefore, it can be concluded that the amino-aromatic compounds of the present invention are effective against Alzheimer's disease.

[0204] [Experimental Example 4] Brain tissue staining method (immunohistochemistry, IHC)

[0205] The Alzheimer's disease animal model from Experiment 3 had its brain fixed in formalin and then sliced ​​into thin sections. The amount of cells or specific proteins in the sliced ​​brain was measured in image form using tissue staining labeling. In this experiment, changes in glial cells associated with dementia were measured using antibodies that labeled glial cells. The results are shown in… Figure 5In this experimental example, tissue staining of the hippocampus was performed to observe changes in glial cells surrounding amyloid-β, a substance associated with dementia. Glial fibrillary acidic protein (GFAP) represents the staining of glial cells, and 4',6-diamidino-2-phenylindole (DAPI) represents the cell nucleus and amyloid-β.

[0206] Existing research has confirmed the accumulation of amyloid-β in the hippocampus of animal models of Alzheimer's disease, and the transformation of glial cells into reactive astrocytes in pathological conditions such as Alzheimer's. At this stage, astrocytes produce GABA and hydrogen peroxide, which act as inhibitory neurotransmitters, inhibiting nerve activity and inducing apoptosis of brain cells (neurons), thus worsening dementia. However, existing research has also confirmed a tendency for drugs that inhibit dementia to reduce the number of astrocytes.

[0207] from Figure 5 As can be seen, GFAP is increased in animal models of Alzheimer's disease, thus exhibiting symptoms of astrocytes, while GFAP is reduced in APP / PS1 mice treated with the amino-aromatic compounds KDS12008 (Example 1), KDS12017 (Example 16), and KDS12025 (Example 21) of the present invention, respectively. Therefore, it is confirmed that the astrocytes, the cause of Alzheimer's disease, are effectively reduced, and that this effect is expressed through the removal of hydrogen peroxide.

[0208] [Experimental Example 5] Passive Avoidance Test (PAT) II

[0209] APP / PS1 mice, used as an animal model of Alzheimer's disease, were injected intraperitoneally for one week with 3 mg / kg / day (3 mpk) and 10 mg / kg / day (10 mpk), respectively, and the passive avoidance test (PAT) was performed in the same manner as in Experimental Example 3. The results are shown in... Figure 6 .

[0210] Healthy, normal rats (wild-type, WT) remembered the electrical stimulation well during the retrieval process, thus spending a longer time in the dark room that had just been stimulated. Although it was an Alzheimer's animal model, the control group (WT + saline) that did not receive drug treatment immediately entered the dark room that had just been stimulated because they forgot the electrical stimulation.

[0211] Conversely, in Alzheimer's disease model APP / PS1 mice treated with the amino aromatic compound KDS12025 of the present invention (Example 21) at doses of 3 mg / kg / day (3 mpk) and 10 mg / kg / day (10 mpk), respectively, memory was maintained similarly to that of healthy normal rats, even when the administration period was reduced from the previous 16 days to one week, thus showing a statistically significant difference compared to the Alzheimer's disease model APP / PS1 mice. Figure 6 Therefore, it can be seen that the amino aromatic compounds of the present invention are effective against Alzheimer's disease.

[0212] [Experimental Example 6] Brain tissue staining method (immunohistochemistry, IHC) II

[0213] Using the animal model of Alzheimer's disease in Example 5, tissue staining of brain tissue was performed using the same method as in Example 4, and the results are presented below. Figure 7 . Figure 7 This is an image of the hippocampus tissue of an animal model, stained with hematologic staining (IHC). GFAP represents the staining results of glial cells, and Aβ represents the staining results of amyloid-β.

[0214] from Figure 7 It was confirmed that glial cells increased in the animal model of Alzheimer's disease, but in APP / PS1 mice treated with the amino-aromatic compound KDS21025 of the present invention (Example 21) at 3 mg / kg / day and 10 mg / kg / day, respectively, the glial cells returned to healthy normal levels. This result, consistent with the behavioral experiment results of Example 5, indicates that treatment with the amino-aromatic compound of the present invention can restore excessively generated H2O2 to normal levels.

[0215] [Experiment Example 7] Electrophysiology Experiment

[0216] Using the animal model of Alzheimer's disease in Experiment 5, electrophysiological experiments were conducted to confirm the mechanism by which glial cells are induced and the memory impairment caused by dementia.

[0217] Existing research has demonstrated the mechanism by which memory and cognitive impairments are induced in Alzheimer's disease patients through the continuous secretion of an inhibitory neurotransmitter called tonic gamma-aminobutyric acid (or tonic current) by reactive astrocytes, and these changes can be confirmed by electrophysiology.

[0218] Therefore, in this experimental example, brain tissue was obtained from APP / PS1 mice (used as an animal model of Alzheimer's disease), APP / PS1 mice treated with 3 mg / kg / day and 10 mg / kg / day of the amino-aromatic compound KDS12025 of the present invention (Example 21), respectively, and healthy normal rats (wild-type, WT), and the electrical signals of living hippocampus cells were measured. Tonic Gaba, which inhibits neurotransmission in brain cells, was thus measured. The results are shown in... Figure 8 .

[0219] In animal models of Alzheimer's disease (APP / PS1, TG), Tonic Gabs were increased compared to healthy normal rats (WT), while in animal models of Alzheimer's disease treated with the amino-aromatic compound KDS12025 of the present invention (Example 21), Tonic Gabs were decreased. In this experimental example, the change in Tonic Gabs was confirmed electrophysiologically, thereby verifying the efficacy of the amino-aromatic compound of the present invention, namely, its inhibitory ability on Tonic Gabs that inhibit neurotransmission. Therefore, it can be concluded that the amino-aromatic compound of the present invention is effective against Alzheimer's disease.

[0220] [Experiment Example 8] Novel Place Recognition Experiment and Passive Avoidance Test (PAT)

[0221] The novel place recognition experiment was used to evaluate cognitive functions related to the hippocampus, thus confirming cognitive impairment and recovery. Identical objects were interacted with for a period of time, and after one hour, only the position of one of the objects was changed. In cases of normal cognitive function, past memories were retained, thus increasing attention to new objects; however, in cases of cognitive impairment, the new location could not be identified. As an animal model, the APP / PS1+GiD Alzheimer's disease model rat was used, which was developed in a manner more closely resembling the phenomena observed in Alzheimer's disease patients. The APP / PS1+GiD Alzheimer's disease model rat was prepared with reference to Nature Neuroscience 23, 1555-1566 (2020).

[0222] In this experimental example, APP / PS1+GiD rats were prepared by injecting AAV-GFAP104-DTR-GFP virus into the hippocampus of APP / PS1 rats (hereinafter referred to as "APP+DTR"). Furthermore, rats prepared by injecting AAV-GFAP104-GFP virus into the hippocampus of APP / PS1 rats were referred to as "APP+GFP". Additionally, rats to which the amino-aromatic compound KDS12025 (Example 21) of the present invention was administered were referred to as "APP+DTR+KDS12025". Furthermore, using APP / PS1 and littermate (litter cubs) as WT rats, rats prepared by injecting AAV-GFAP104-GFP virus into the hippocampus of WT rats were referred to as "WT+GFP", and rats prepared by injecting AAV-GFAP104-DTR-GFP virus into the hippocampus of WT rats were referred to as "WT+DTR".

[0223] Normal rats (WT+GFP, WT+DTR) exhibited normal cognitive function. APP+GFP showed a level of cognitive function similar to that of normal rats, while APP+DTR showed cognitive impairment similar to that of Alzheimer's disease patients. Cognitive recovery was confirmed when the amino-aromatic compound KDS12025 (Example 21) of the present invention was administered intraperitoneally (ip) at a dose of 3 mg / kg / day to APP / PS1+GiD Alzheimer's disease model rats (i.e., APP+DTR+KDS12025) exhibiting cognitive impairment. Figure 9 ).

[0224] Furthermore, the results of PAT confirmed that normal rats (WT+GFP) remembered the electrically stimulated dark room very well, thus taking a longer time to enter the dark room. Conversely, APP / PS1+GiD Alzheimer's disease model rats (i.e., APP+DTR) exhibited memory problems, but after administration of the amino-aromatic compound KDS12025 of the present invention (Example 21) (i.e., APP+DTR+KDS12025), memory was restored. Figure 10 ).

[0225] That is, the hydrogen peroxide scavenging effect of the amino aromatic compounds of the present invention in APP / PS1+GiD Alzheimer's disease model rats (which were developed in a manner closer to the phenomena observed in Alzheimer's disease patients) was confirmed, and therefore it can be concluded that the amino aromatic compounds of the present invention are effective against Alzheimer's disease.

[0226] [Experimental Example 9] Single toxicity evaluation (ethanol dose 50, LD50) 50 )

[0227] WT rats (C57BL / 6 mice) of approximately 8 weeks of age were intraperitoneally injected with the amino-aromatic compounds KDS12008 (Example 1), KDS12017 (Example 16), and KDS12025 (Example 21) of the present invention at daily doses of 100 mg / kg, 300 mg / kg, and 1000 mg / kg, respectively, and the animals were assessed for mortality. The results are shown in... Figures 11 to 13 In the study, all three drugs showed toxicity at a concentration of 1000 mg / kg. Conversely, at 300 mg / kg, all three drugs showed toxicity with only about 50% mortality in animals. At a concentration of 100 mg / kg, below 300 mg / kg, it was confirmed that none of the three drugs were fatally toxic to mice.

[0228] [Experimental Example 10] Blood-brain barrier (BBB) ​​penetration analysis

[0229] Based on the literature J Med Chem. 2001 Mar 15; 44(6): 923-30, an artificial blood-brain barrier (BBB) ​​was prepared by parallel artificial membrane permeability assay (PAMPA) and the drug permeability was evaluated.

[0230] The results confirm that the amino aromatic compound KDS12025 (Example 21) of the present invention, at a concentration of 50 μM, exhibits high permeability (KDS12025 (Example 21) 67.43 × 10⁻⁶). -6 The BBB permeability is 3.56 × 10 cm / s. In contrast, the existing drug AAD-2004, which is expected to remove reactive oxygen species, has a BBB permeability of 3.56 × 10 cm / s at the same concentration. -6 The flow rate is very low, approximately cm / s. Therefore, it can be confirmed that the amino aromatic compounds of the present invention have high permeability.

[0231] As described above, the present invention has been illustrated by specific details, limited embodiments, and accompanying drawings. However, this is provided only to facilitate a more comprehensive understanding of the invention, and the invention is not limited to the described embodiments. Those skilled in the art to which this invention pertains will be able to make various modifications and variations based on these descriptions.

[0232] Therefore, the concept of the present invention is not limited to the illustrated embodiments, nor is it limited to the scope of the claims. All contents that are equivalent or have equivalent variations to the scope of the claims are within the scope of the present invention.

Claims

1. An amino aromatic compound represented by the following Chemical Formula 5 or a pharmaceutically acceptable salt of the compound: [Chemical Formula 5] ###0001### in the Chemical Formula 5, n is an integer of 1 or 2.

2. The amino aromatic compound or the pharmaceutically acceptable salt of the compound according to claim 1, wherein the amino aromatic compound is selected from any one of the following compound group: ， 3. An amino aromatic compound represented by the following Chemical Formula 5 or a pharmaceutically acceptable salt of the compound: [Chemical Formula 5] ###0002### in the Chemical Formula 5, n is an integer of 1 or 2. R 1 and R 2 are each independently C1-C4alkyl; R 3 halo C1-C4 alkyl; 4. The amino aromatic compound or the pharmaceutically acceptable salt of the compound according to claim 3, wherein the amino aromatic compound is selected from any one of the following compound group:

5. The amino aromatic compound or the pharmaceutically acceptable salt of the compound according to any one of claims 1 to 4, wherein the pharmaceutically acceptable salt is a hydrochloride.

6. A pharmaceutical composition comprising the amino aromatic compound or the pharmaceutically acceptable salt of the compound according to any one of claims 1 to 4 as an active ingredient. 。 7. The pharmaceutical composition according to claim 6, wherein the pharmaceutical composition further comprises an excipient and a carrier. ​ ， ​ R 1 is hydrogen; R 2 is C1-C4alkyl; R 3 Ci-C4-alkyl or halogen; and R is hydrogen or Ci-C4-alkyl. ​ ​ ​ 。 ​ ​ 6. A pharmaceutical composition for the treatment and prevention of Alzheimer's disease, wherein, ​ ​ ​

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