Composition for promoting mitochondrial autophagy activity comprising isoquinoline derivative compound as active ingredient and use thereof

Abstract

The present invention relates to novel isoquinoline derivatives. The derivative has the effect of promoting mitochondrial autophagy activity, so that mitochondria with damaged structure and / or function is reduced. Specifically, the novel isoquinoline derivative according to the present invention does not induce mitochondrial dysfunction and has been determined as a safe compound that is non-toxic to cells, and thus it can be used to prevent, ameliorate and / or treat various diseases that may be caused by mitochondrial abnormality.

Description

Technical Field

[0001] The present invention relates to a composition for promoting mitochondrial autophagy activity, comprising an isoquinoline derivative compound as an active ingredient, and use thereof.

[0002] The present invention claims priority based on Korean Patent Application No. 10-2022-0056058 filed on May 6, 2022, and all contents disclosed in the specification and drawings of these applications are incorporated herein by reference. Background Art

[0003] Mitochondria are common organelles in cells, commonly known as "powerhouses of cells". Mitochondria not only generate energy through oxidative phosphorylation in cells, but also perform functions necessary for the survival of cells and organisms, including apoptosis, ion homeostasis, intermediate metabolism of sugars and lipids, pyrimidine synthesis, urea synthesis and heme synthesis.

[0004] The activity of mitochondria in vivo is maintained by the balance of mitochondrial production, fusion and fission mechanisms, as well as the production of new mitochondria and the removal of damaged or expired mitochondria through mitochondrial autophagy. Due to abnormal mitochondrial autophagy, reactive oxygen species produced by damaged mitochondria are considered to be the main cause of aging, and the accumulation of damaged mitochondria is closely related to various metabolic diseases, including diabetes, as well as degenerative diseases associated with aging (Bratic and Larsson, 2013).

[0005] Mitophagy is an intracellular degradation mechanism that removes damaged or unnecessary mitochondria. When mitochondria are damaged, they form autophagosomes by surrounding the damaged mitochondria with a membrane, which then fuses with lysosomes to selectively eliminate the damaged mitochondria. Mitophagy activity is important for regulating mitochondrial function in various cells, including neurons, and for maintaining tissue function.

[0006] Mitophagy is a selective degradation mechanism of mitochondria. Initially, it was thought to be part of autophagy, a process that breaks down various intracellular components to survive conditions of nutrient deprivation. However, it was later demonstrated to be a mechanism that effectively eliminates damaged or unnecessary mitochondria by surrounding them with a membrane, fusing with lysosomes, and completely degrading them. Mitophagy plays a protective role in mitochondria by preventing the release of pro-apoptotic proteins, the generation of reactive oxygen species, and unnecessary ATP hydrolysis in aged, damaged, and depolarized mitochondria.

[0007] According to recent studies, inhibition of mitophagy activity leads to the accumulation of damaged mitochondria, which induces motor neuron death and may lead to degenerative brain diseases such as Parkinson's disease. In addition, abnormal mitophagy activity has been reported to be associated with a variety of human diseases, including degenerative brain diseases such as Parkinson's disease, Alzheimer's disease, and Lou Gehrig's disease, as well as peripheral neuropathy, heart disease, metabolic disease, and cancer, which has aroused researchers' interest in the role of mitophagy in human diseases and its potential therapeutic applications.

[0008] Currently, experimental approaches to induce mitophagic activity involve treatment with so-called "mitochondrial toxins (e.g., CCCP, FCCP, and rotenone)" that induce mitochondrial dysfunction. However, CCCP and FCCP are mitochondrial membrane potential uncouplers that depolarize the mitochondrial membrane potential, and rotenone acts as a Complex I inhibitor. These mitochondrial toxins induce mitophagic activity by directly causing mitochondrial damage, a mechanism for removing damaged mitochondria, but their strong toxicity to cells limits their use as drugs to promote mitophagic activity. Summary of the invention Technical issues

[0009] The object of the present invention is to provide a pharmaceutical composition for promoting mitochondrial autophagy activity, which comprises an isoquinoline derivative compound or a salt thereof as an active ingredient.

[0010] Another object of the present invention is to provide a food composition for promoting mitochondrial autophagy activity, comprising an isoquinoline derivative compound or a salt thereof as an active ingredient.

[0011] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating diseases caused by mitochondrial dysfunction, comprising an isoquinoline derivative or a salt thereof as an active ingredient.

[0012] Another object of the present invention is to provide a food composition for preventing or improving diseases caused by mitochondrial dysfunction, which comprises an isoquinoline derivative or a salt thereof as an active ingredient.

[0013] Another object of the present invention is to provide a method for increasing mitophagy activity in cells, comprising the step of treating cells with an isoquinoline derivative compound in vitro.

[0014] Another object of the present invention is to provide a method for inhibiting mitochondrial dysfunction, which comprises the step of treating cells in vitro with an isoquinoline derivative compound. Technical Solution

[0015] To achieve the above object, the present invention provides a pharmaceutical composition for promoting mitochondrial autophagy activity, which comprises an isoquinoline derivative compound or a salt thereof as an active ingredient.

[0016] In addition, the present invention provides a food composition for promoting mitochondrial autophagy activity, comprising an isoquinoline derivative compound or a salt thereof as an active ingredient.

[0017] In addition, the present invention provides use of an isoquinoline derivative compound or a salt thereof in preparing a drug for promoting mitochondrial autophagy activity.

[0018] In addition, the present invention provides a method for promoting mitophagy activity, comprising the step of administering an isoquinoline derivative compound or a salt thereof to an individual in need thereof.

[0019] Furthermore, the present invention provides a pharmaceutical composition for preventing or treating a disease caused by mitochondrial dysfunction, comprising an isoquinoline derivative or a salt thereof as an active ingredient.

[0020] In addition, the present invention also provides a food composition for preventing or improving diseases caused by mitochondrial dysfunction, which comprises an isoquinoline derivative or a salt thereof as an active ingredient.

[0021] In addition, the present invention also provides use of an isoquinoline derivative compound or a salt thereof in preparing a drug for preventing, improving and / or treating diseases caused by mitochondrial dysfunction.

[0022] In addition, the present invention also provides a method for preventing or treating a disease caused by mitochondrial dysfunction, which comprises the step of administering an isoquinoline derivative compound or a salt thereof to an individual in need thereof.

[0023] In addition, the present invention also provides use of an isoquinoline derivative compound or a salt thereof in preventing, improving and / or treating a disease caused by mitochondrial dysfunction.

[0024] In addition, the present invention also provides a method for increasing mitochondrial autophagy activity in cells, comprising the step of treating cells in vitro with an isoquinoline derivative compound.

[0025] In addition, the present invention also provides a method for inhibiting mitochondrial dysfunction, comprising the step of treating cells in vitro with an isoquinoline derivative compound.

[0026] In addition, the present invention also provides use of an isoquinoline derivative compound or a salt thereof in preparing a drug for improving or inhibiting mitochondrial dysfunction.

[0027] In addition, the present invention provides a method for improving or inhibiting mitochondrial dysfunction, which comprises the step of administering an isoquinoline derivative compound or a salt thereof to an individual in need thereof.

[0028] In addition, the present invention provides use of an isoquinoline derivative compound or a salt thereof for improving or inhibiting mitochondrial dysfunction.

[0029] In an exemplary embodiment of the present invention, the isoquinoline derivative compound may be represented by the following Chemical Formula 1 to Chemical Formula 6, but is not limited thereto: Chemical formula 1: Chemical formula 2: Chemical formula 3: Chemical formula 4: Chemical formula 5: Chemical formula 6:

[0030] In another exemplary embodiment of the present invention, the isoquinoline derivative compound may be able to enhance mitophagy activity, but is not limited thereto.

[0031] In another exemplary embodiment of the present invention, the increase in mitophagy activity may involve the removal of dysfunctional mitochondria, but is not limited thereto. That is, the increase in mitophagy activity may refer to an increase in the removal of dysfunctional mitochondria or a reduction in dysfunctional mitochondria.

[0032] In another exemplary embodiment of the present invention, dysfunctional mitochondria may be able to induce neurodegenerative diseases, but are not limited thereto.

[0033] In another exemplary embodiment of the present invention, the disease caused by mitochondrial dysfunction may be a neurodegenerative disease, but is not limited thereto.

[0034] In another exemplary embodiment of the present invention, the neurodegenerative disease can be selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, multiple sclerosis, immune-mediated brain dysfunction, progressive neurodegenerative diseases, metabolic encephalopathy, Niemann-Pick disease, dementia caused by cerebral ischemia and cerebral hemorrhage, but is not limited thereto.

[0035] In yet another exemplary embodiment of the present invention, the neurodegenerative disease may be Alzheimer's disease or dementia, but is not limited thereto. Beneficial Effects

[0036] The present invention relates to novel isoquinoline derivatives. The derivatives have the effect of promoting mitochondrial autophagy activity, thereby reducing mitochondria with structural and / or functional damage. Specifically, the novel isoquinoline derivatives according to the present invention do not induce mitochondrial dysfunction and have been determined to be safe compounds that are non-toxic to cells, and can therefore be used to prevent, improve and / or treat various diseases that may be caused by mitochondrial abnormalities. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 This is a diagram confirming the structure of the isoquinoline derivative of the present invention.

[0038] Figure 2 The figure confirms the effect of the isoquinoline derivatives of the present invention in promoting mitophagy activity by flow cytometry (A: quantification of mitophagy activity of six derivatives, B: quantification of mitophagy activity according to CD1-386 and CD1-409 concentrations).

[0039] Figure 3 The figure is a graph analyzing the effect of the isoquinoline derivatives of the present invention on mitochondrial membrane potential using the TMRM assay method.

[0040] Figure 4 The diagram shows the confirmation of the cytotoxicity of the isoquinoline derivatives of the present invention using a colony forming assay. DETAILED DESCRIPTION Invention Mode

[0041] Exemplary embodiments of the present invention will now be described in detail with reference to the accompanying drawings. In the following description, detailed explanations of techniques well known to those skilled in the art may be omitted. In addition, when describing the present invention, if it is determined that detailed explanations may unnecessarily obscure the main points of the present invention, the specific details of related known functions or configurations may be omitted. In addition, the terms used in this specification are selected to appropriately describe preferred exemplary embodiments of the present invention, and may change according to the intention of the user, operator or the practice of the relevant field of the present invention.

[0042] Therefore, the definitions of these terms should be based on the entirety of the present specification. Throughout the specification, when a part is described as "comprising" a certain component, this means that, unless otherwise specified, the inclusion of other components is not excluded, and additional components may be included.

[0043] The present invention provides a pharmaceutical composition for promoting mitochondrial autophagy activity, comprising an isoquinoline derivative compound or a salt thereof as an active ingredient.

[0044] In this specification, the term "isoquinoline derivative" may be used interchangeably with "isoquinoline derivative compound".

[0045] In the present specification, the term "compound" is a concept including an isoquinoline derivative compound and a salt thereof.

[0046] In the present invention, derivatives include pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically acceptable hydrates, pharmaceutically acceptable anhydrates, pharmaceutically acceptable enantiomers, pharmaceutically acceptable esters, pharmaceutically acceptable polymorphs, pharmaceutically acceptable prodrugs, pharmaceutically acceptable complexes, and the like.

[0047] In the present invention, the term "salt" includes pharmaceutically acceptable salts and food-grade acceptable salts. Pharmaceutically or food-grade acceptable salts refer to salts derived from pharmaceutically or food-grade acceptable organic acids, inorganic acids or bases.

[0048] The example of suitable acid comprises hydrochloric acid, bromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, p-toluenesulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid, benzoic acid, malonic acid, gluconic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid etc.Acid addition salt can be prepared by ordinary method, for example, compound is dissolved in excessive acid aqueous solution, and uses polar organic solvent such as methanol, ethanol, acetone or acetonitrile to precipitate salt.In addition, can be by equimolar compound and acid or alcohol are heated in water, then mixture is evaporated to dryness, or by suction filtration separation precipitated salt prepares salt.

[0049] Salts derived from suitable bases may include alkali metals (e.g., sodium and potassium), alkaline earth metals (e.g., magnesium and ammonium), but are not limited thereto. Alkali metal or alkaline earth metal salts may be obtained, for example, by dissolving the compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the insoluble compound salt, and then evaporating and drying the supernatant. In this case, from a pharmaceutical point of view, it is particularly suitable to prepare metal salts, particularly sodium, potassium or calcium salts. In addition, the corresponding silver salts may be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

[0050] The scope of the compounds of the present invention includes not only pharmaceutically acceptable salts but also all isomers, hydrates and solvates which can be prepared by conventional methods.

[0051] The term "prevention" used in the present invention refers to any action to inhibit or delay the symptoms of a specific disease by administering the composition of the present invention.

[0052] The term "treatment" as used in the present invention refers to any action to improve or beneficially alter the symptoms of a particular disease by administering the composition of the present invention.

[0053] The term "mitophagy" used in the present invention refers to an intracellular degradation mechanism that removes damaged or unnecessary mitochondria. When mitochondrial damage occurs, it can form autophagosomes, which fuse with lysosomes to selectively degrade and remove damaged mitochondria. Mitochondrial autophagy is different from autophagy. Autophagy is a mechanism that produces macromolecular precursors and generates energy by degrading and recycling unnecessary cellular components (such as old proteins, protein aggregates, organelles, and pathogens that have infiltrated cells) when cells are in a state of nutrient deficiency. Mitochondrial autophagy is regulated independently of regulatory signals (such as nutrients, energy, and pressure) that control autophagy. In the present invention, the phrase "promotes mitophagy" covers all effects that enhance or increase the activity, frequency, degree, and level of mitophagy.

[0054] The composition according to the present invention may also include a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable" in the present invention refers to a property that indicates no toxicity to cells or humans exposed to the composition. The carrier may be any carrier known in the art, including but not limited to buffers, preservatives, anesthetics, solubilizers, emulsifiers, stabilizers, excipients and lubricants.

[0055] In addition, the composition of the present invention can be prepared into various forms for use according to conventional methods, including oral dosage forms (such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc.), as well as local preparations, suppositories, sterile injections, etc. In addition, it can be used in the form of local skin preparations (such as ointments, lotions, sprays, patches, creams, powders, suspensions, gels or jellies). Excipients, carriers and diluents that can be included in the composition of the present invention include lactose, glucose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil, etc. When formulating, a commonly used diluent or excipient (such as a filler, a swelling agent, a binder, a wetting agent, a disintegrant and a surfactant) is used to prepare the composition.

[0056] Oral solid preparations include tablets, powders, granules and capsules, which are prepared by mixing one or more excipients with Psychotria rubra extracts, such as starch, calcium carbonate, sucrose, lactose or gelatin. In addition, lubricants such as magnesium stearate and talcum powder are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups, which can also contain various excipients (including wetting agents, sweeteners, flavoring agents and preservatives, etc.) in addition to commonly used simple diluents (such as water and liquid paraffin). Preparations for parenteral administration include sterile solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils (such as olive oil) and injectable esters (such as ethyl oleate, etc.). The matrix of the suppository can include Witepsol, polyethylene glycol, Tween61, cocoa butter, lauric acid and glycerin gelatin, etc.

[0057] Pharmaceutical composition according to the present invention is used in a pharmaceutically effective amount. In the present invention, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease, with a reasonable benefit / risk ratio applicable to medical treatment. The effective dose level can be determined according to the type and severity of the patient's condition, the activity of the drug, the sensitivity to the drug, the time of application, the route of administration, the excretion rate, the duration of treatment, the concomitant medication, and other factors well-known in the medical field.

[0058] The pharmaceutical composition according to the present invention can be used as a single therapeutic agent or in combination with other therapeutic agents, and can be used sequentially or simultaneously with conventional therapy, as a single dose or multiple doses. Taking all the above factors into consideration, it is important to use the amount that achieves the maximum effect with the minimum dose and has no side effects, which can be easily determined by a technician in the field to which the present invention belongs.

[0059] The pharmaceutical composition according to the present invention can be administered to an individual by various routes. All administration methods are contemplated, including, for example, oral, intravenous, intramuscular, subcutaneous and intraperitoneal injections.

[0060] The content of the compound in the composition of the present invention can be appropriately adjusted according to the symptoms of the disease, the degree of progression of the symptoms, the condition of the patient, etc. For example, the range can be 0.0001 to 99.9% by weight, or 0.001 to 50% by weight of the total weight of the composition, but is not limited thereto. The above content ratio is based on dry weight excluding solvent.

[0061] The dosage selection of the pharmaceutical composition according to the present invention is based on factors such as the age, weight, sex, physical condition, etc. of the individual. Obviously, the concentration of the active ingredient contained in the pharmaceutical composition can vary according to the subject, and preferably, it is contained in the pharmaceutical composition at a concentration of 0.01 to 5000 μg / ml. If the concentration is less than 0.01 μg / ml, it may not show pharmacological activity, and if the concentration exceeds 5,000 μg / ml, it may show toxicity to the human body.

[0062] In the present invention, the term "individual" refers to a subject in need of disease treatment, more specifically, a mammal, such as a human or non-human primate, mouse, rat, dog, cat, horse and cow.

[0063] In the present invention, the term "administration" refers to providing a specific composition of the present invention to an individual by any appropriate method.

[0064] In the present invention, "prevention" refers to any action that inhibits or delays the onset of a target disease, and "treatment" refers to any action that improves or favorably changes the symptoms of a target disease and associated metabolic abnormalities by administering a pharmaceutical composition according to the present invention. In addition, "improvement" refers to any action that reduces parameters associated with a target disease (e.g., the severity of symptoms) by administering a composition according to the present invention.

[0065] According to an exemplary embodiment of the present invention, the isoquinoline derivative compound may be represented by the following Chemical Formulas 1 to 6: Chemical formula 1: Chemical formula 2: Chemical formula 3: Chemical formula 4: Chemical formula 5: Chemical formula 6:

[0066] In an exemplary embodiment, the pharmaceutically acceptable salt is preferably an acid addition salt formed from a pharmaceutically acceptable organic acid. The term "pharmaceutically acceptable" as used in the present invention refers to a property indicating non-toxicity to cells or humans exposed to the compound or its acid addition salt. Acid addition salts are obtained from inorganic acids (e.g., hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or arsenic acid) and non-toxic organic acids (e.g., aliphatic monocarboxylates and dicarboxylates, phenyl-substituted alkanoyl acid salts, hydroxyalkanoyl acid salts, alkanedioates, aromatic acids, and aliphatic and aromatic sulfonic acids). Examples of such pharmacologically nontoxic salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, fluorides, acetates, propionates, decanoates, octanoates, acrylates, formates, isobutyrates, decanoates, heptanoates, propionates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyrates, 1,4-dimethoxybenzene, 1,6-tetrahydrofuran ... dioic acid salt, hexane-1,6-dioic acid salt, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, or glycolate.

[0067] The acid addition salt according to the present invention can be prepared by conventional methods, for example, dissolving the compound shown in Chemical Formula 1 to Chemical Formula 6 in a large excess of an acidic aqueous solution, and using a water-miscible organic solvent (e.g., methanol, ethanol, acetone or acetonitrile) to precipitate the salt. In addition, the mixture can be dried by evaporating the solvent or excess acid, or the precipitated salt can be prepared by suction filtration.

[0068] In addition, a base can be used to prepare a pharmaceutically acceptable metal salt. The compound is dissolved in an excess of an alkali metal hydroxide or alkaline earth metal hydroxide solution, the insoluble compound salt is filtered, and the supernatant is evaporated and dried to obtain an alkali metal or alkaline earth metal salt. At this time, it is pharmaceutically appropriate to prepare a metal salt such as a sodium, potassium or calcium salt. In addition, an alkali metal or alkaline earth metal salt is reacted with a suitable anion (e.g., silver nitrate) to obtain the corresponding silver salt.

[0069] In an exemplary embodiment, the compounds shown in the above chemical formulas 1 to 6 of the present invention may include not only their stereoisomers and pharmaceutically acceptable salts, but also any solvates, hydrates, racemates or prodrugs derived therefrom. In particular, the compounds shown in the above chemical formulas 1 to 6 may include isomers on all asymmetric carbon atoms. In addition, when there is an asymmetric carbon center in the substituent of the compound of the present invention, they may exist in the form of R or S isomers, racemates, diastereomers, isomer mixtures and individual diastereomers, and all these isomers and mixtures thereof are included within the scope of the present invention.

[0070] According to an exemplary embodiment of the present invention, the isoquinoline derivative compound can enhance mitochondrial autophagy activity.

[0071] According to an exemplary embodiment of the present invention, the enhancement of mitophagy activity may involve the removal of dysfunctional mitochondria.

[0072] According to an exemplary embodiment of the present invention, the mitochondrial dysfunction may induce a neurodegenerative disease, and the neurodegenerative disease may be selected from the group consisting of: Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, multiple sclerosis, immune-mediated brain dysfunction, progressive neurodegenerative diseases, metabolic encephalopathy, Niemann-Pick disease, dementia caused by cerebral ischemia and cerebral hemorrhage, preferably Alzheimer's disease or dementia, but not limited thereto.

[0073] In addition, another object of the present invention is to provide a food composition for promoting mitochondrial autophagy, which comprises an isoquinoline derivative compound or a salt thereof as an active ingredient. The salt may be a food grade acceptable salt. In addition, the food composition includes a health functional food composition.

[0074] The term "improvement" as used in the present invention refers to any action that at least reduces a parameter associated with the condition being treated, such as the severity of symptoms.

[0075] The food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional components in addition to the active ingredient of the present invention, similar to conventional food compositions.

[0076] Examples of the above-mentioned natural carbohydrates include monosaccharides (e.g., glucose and fructose); disaccharides (e.g., maltose and sucrose); and polysaccharides (e.g., dextrin and cyclodextrin), as well as conventional sugars and sugar alcohols (e.g., xylitol, sorbitol, and erythritol). The above-mentioned flavoring agents may advantageously include natural flavoring agents (taumatine), stevia extracts (e.g., stevioside A, glycyrrhizin, etc.), and synthetic flavoring agents (e.g., saccharin and aspartame). The food composition of the present invention can be formulated in the same manner as the pharmaceutical composition to be used as a functional food, or can be added to various foods. Examples of foods to which the composition of the present invention can be added include, but are not limited to, beverages, meats, chocolates, foods, snacks, pizzas, noodles, other pasta products, chewing gum, candy, ice cream, alcoholic beverages, vitamin complexes, and health products.

[0077] In addition, except active ingredient extract, food composition can also contain various nutrients, vitamins, mineral substances (electrolytes), synthetic flavoring agents and natural flavoring agents, coloring agents and emulsifiers (such as cheese and chocolate), pectin and its salt, alginic acid and its salt, organic acid, protective colloid thickener, pH adjusting agent, stabilizer, preservative, glycerine, alcohol and the carbonating agent used in carbonated beverage. In addition, food composition of the present invention can contain the pulp used to produce natural fruit juice, fruit juice beverage and vegetable beverage.

[0078] The functional food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids and lozenges for the purpose of promoting mitochondrial autophagy activity. In the present invention, the term "health functional food composition" refers to a food manufactured and processed using raw materials or ingredients having beneficial functions to the human body as defined in Health Functional Food Act No. 6727, and is intended to be eaten to achieve a health-promoting effect, such as regulating nutrients related to human body structure and function or for physiological effects. The health functional food of the present invention may contain conventional food additives, and unless otherwise specified, the applicability of these additives is determined according to the general provisions and general test methods outlined in the food additive regulations approved by the Food and Drug Administration, according to the specifications and standards related to each project. The items listed in the "Food Additive Regulations" include, for example, chemically synthesized substances (such as ketones, glycine, calcium citrate, niacin and cinnamic acid, etc.); natural additives (such as colorants, licorice extracts, crystalline cellulose, caramel color and guar gum); and mixed preparations (such as sodium glutamate preparations, alkaline agents for noodle additives) and preservative preparations and tartrazine preparations, etc. For example, a health functional food in tablet form can be prepared by mixing the active ingredient of the present invention with excipients, binders, disintegrants and other additives using conventional methods to form granules, and then pressing them using a lubricant, or the mixture can be directly pressed into tablets. In addition, the tablet form of the health functional food can contain a glidant or other similar agents as needed. The hard capsule form of the health functional food can be prepared by filling a conventional hard capsule with a mixture of the active ingredient of the present invention and additives such as excipients. The soft capsule form can be prepared by filling gelatin or similar capsule materials with a mixture of the active ingredient and additives such as excipients. The soft capsule form can contain a plasticizer such as glycerol or sorbitol, as well as a colorant and a preservative as needed. The health functional food in tablet form can be prepared by molding the mixture of the active ingredient of the present invention with excipients, binders, disintegrants, etc. by methods known in the art. In addition, they can be coated with powdered sugar or other coating agents as needed, or coated with substances such as starch or talcum powder on the surface. The health functional food in granule form can be prepared by forming granules from the mixture of the active ingredient of the present invention with excipients, binders, disintegrants, etc. by methods known in the art. In addition, they can contain flavoring agents, glidants or other additives as needed.

[0079] When the compound of the present invention, its derivative or food-grade acceptable salt is used as a food additive, it can be directly added or used in combination with other foods or food ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to the intended use (prevention, health care or treatment). Generally, when manufacturing food or beverages, the addition amount of the compound of the present invention can be 15% by weight or less, or 10% by weight or less of the total weight of the raw materials. However, for long-term intake for the purpose of health and hygiene or health regulation, the addition amount can be lower than the specified range, and since there is no safety problem, the active ingredient can also be used beyond this range.

[0080] There is no specific restriction on the types of food. Examples of foods that may contain the above substances include meat, sausages, bread, chocolate, candy, snacks, biscuits, pizza, noodles, other pasta products, chewing gum, dairy products (such as ice cream), various soups, beverages, teas, drink mixes, alcoholic beverages and vitamin complexes, covering all forms of health functional foods in the traditional sense.

[0081] The health drink composition according to the present invention can contain various flavoring agents or natural carbohydrates as additional ingredients, similar to conventional beverages. The above-mentioned natural carbohydrates include monosaccharides (such as glucose and fructose); disaccharides (such as maltose and sucrose); polysaccharides (such as dextrin and cyclodextrin); and sugar alcohols (such as xylitol, sorbitol and erythritol). Sweeteners can include natural sweeteners (such as stevioside and stevia extract), and synthetic sweeteners (such as saccharin and aspartame). The ratio of natural carbohydrates is generally about 0.01-0.20 grams per 100 milliliters of the composition of the present invention, or about 0.04-0.10 grams.

[0082] In addition, the composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectin and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerine, alcohol and the carbonating agent used in carbonated beverages. In addition, the composition of the present invention may contain the pulp used to produce natural fruit juice, fruit juice beverages and vegetable beverages. These ingredients can be used alone or in combination. The ratio of these additives is not important, but is usually selected in the range of 0.01 to 0.20 weight parts in every 100 weight parts of the composition of the present invention.

[0083] In this specification, "health functional food" refers to the same term as food for special health use (FoSHU), that is, food that has been processed to effectively play a biological regulatory function and provide nutrition at the same time, with significant medical and health benefits. Such food can be made into various forms, including tablets, capsules, powders, granules, liquids and lozenges, to achieve the beneficial effect of preventing or improving peripheral neuropathy.

[0084] The health functional food of the present invention can be manufactured using methods commonly used in the industry, and can include raw materials and ingredients commonly added in the industry during the production process. In addition, unlike traditional drugs, health functional foods are made from food ingredients, which have the advantage of not having the side effects that may occur with long-term use of drugs, and they can also provide excellent portability.

[0085] Furthermore, the present invention provides a pharmaceutical composition for preventing or treating a disease caused by mitochondrial dysfunction, comprising an isoquinoline derivative or a salt thereof as an active ingredient.

[0086] According to an exemplary embodiment of the present invention, the disease caused by mitochondrial dysfunction may be a neurodegenerative disease.

[0087] In addition, the present invention also provides a food composition for preventing or improving diseases caused by mitochondrial dysfunction, which comprises an isoquinoline derivative or a salt thereof as an active ingredient.

[0088] In addition, the present invention also provides a method for improving mitochondrial autophagy activity in cells, which comprises the step of treating cells in vitro with an isoquinoline derivative compound.

[0089] The cell of the present invention can be a cell expressing mt-Keima, and "mt-Keima" refers to a coral-derived fluorescent protein that exhibits pH-dependent changes in fluorescence properties (Katayama H et al., 2011, Chem Biol). Due to changes in protein structure, mt-Keima exhibits a significantly enhanced fluorescence signal at 458nm under neutral conditions (pH 7) and exhibits strong fluorescence at 561nm under acidic conditions (pH 4). Utilizing these characteristics, mt-Keima can be specifically expressed in mitochondria, and by comprehensively analyzing the fluorescence signals at 458nm and 561nm, the fluorescence signal when the mitochondria in a neutral environment (pH = 7.4-7.8) are converted to an acidic environment (pH = 4.5) by the lysosomes delivered by mitophagy can be analyzed, thereby quantitatively measuring the mitophagy activity.

[0090] In addition, the present invention also provides a method for inhibiting mitochondrial dysfunction, comprising the step of treating cells in vitro with an isoquinoline derivative compound.

[0091] Unless otherwise stated, all numbers, values ​​and / or expressions used in this specification to represent components, reaction conditions and ingredient contents should be understood as approximate values, reflecting the various measurement uncertainties generated when these values ​​are obtained from fundamentally different values, and should be qualified in all cases using the term "about". In addition, when a numerical range is disclosed in this specification, the range is continuous and, unless otherwise stated, includes all values ​​from the minimum value to the maximum value, including the maximum value. In addition, when the range refers to an integer, unless otherwise stated, it includes all integers from the minimum value to the maximum value, including the maximum value.

[0092] For example, in this specification, when describing the range of a variable, it is understood that the variable covers all values ​​within the range, including the specified endpoints of the range. For example, the range of "5 to 10" includes not only the values ​​5, 6, 7, 8, 9 and 10, but also any sub-ranges, such as 6 to 10, 7 to 10, 6 to 9, 7 to 9, etc. It is also understood to include any valid integer value between the ranges, such as 5.5, 6.5, 7.5, and sub-ranges such as 5.5 to 8.5 and 6.5 to 9. In addition, for example, the range of "10% to 30%" includes not only the integer values ​​of 10%, 11%, 12%, 13% to 30%, but also any sub-ranges, such as 10% to 15%, 12% to 18% and 20% to 30%. It is also understood to include any valid value within the range, such as 10.5%, 15.5% and 25.5%.

[0093] The present invention will be described in more detail below by way of examples. These examples are provided only to illustrate the present invention in more detail, and those skilled in the art will appreciate that the scope of the present invention is not limited to these examples. <Example 1> Synthesis of isoquinoline derivatives

[0094] The isoquinoline derivatives CD1-271, CD1-363, CD1-367, CD1-386, CD1-404 and CD1-409 of the present invention were synthesized. <1-1> Synthesis of CD1-271

[0095] Precursor CD1-012 (1 g, 2.6 mmol) was dissolved in methanol (30 ml) in a dry round-bottom flask, and 0.5 M aqueous hydrochloric acid solution (8 ml) was added at 0°C, stirred for 6 hours, and then concentrated under reduced pressure to obtain CD1-271 with a yield of 99%. <1-2> Synthesis of CD1-363

[0096] The precursor berberine (0.5 g, 1.2 mmol) was dissolved in methanol (20 ml) in a dry round-bottom flask, and potassium carbonate (0.5 g, 3.6 mmol) was then added. After that, a 2.2 M sodium borohydride aqueous solution (0.6 ml) containing 5% sodium hydroxide was added at 0°C and stirred for 3 hours. The obtained solid was filtered, washed with distilled water, and vacuum dried to obtain compound CD1-363 with a yield of 84%. <1-3> Synthesis of CD1-367

[0097] CD363 was used as a precursor and synthesized in the same manner as in Example 1-1 to obtain CD1-367 with a yield of 96%. <1-4> Synthesis of CD1-386, CD1-404 and CD1-409

[0098] The precursor berberine (0.5g, 1.3mmol) was dissolved in a 5N sodium hydroxide aqueous solution (20ml) in a dry round-bottom flask, and then acetone (5ml) was added at 0°C and stirred for 6 hours. After concentration under reduced pressure, cold water (30ml) was added and the resulting solid was filtered. The filtered solid was washed with distilled water and then dried under vacuum. The dried solid (0.4g, 1mmol) was dissolved in anhydrous CH2CL2 (10ml) in a pressure tube, and then allyl iodide (0.4g, 1mmol) was added and stirred at 100°C for 4 hours. It was then concentrated under reduced pressure, and the resulting mixture was adsorbed on silica gel and separated by silica gel chromatography purification (CH2Cl2: acetone = 50: 1 → 15: 1v / v) to obtain compound CD1-386 with a yield of 35%. In addition, CD1-404 was obtained with a yield of 14%, and CD1-409 was obtained with a yield of 47%. <1-5> NMR analysis

[0099] The compounds obtained in Examples 1-1 to 1-4 were subjected to NMR analysis to confirm their structures. The results are shown below. Figure 1 shown. CD1-271: 1 HNMR (DMSO-d6, 400MHz) δ9.72 (s, 1H), 8.55 (s, 1H), 7.75 (d, J = 8.8Hz, 1H), 7.47 (s, 1H), 4.83 (t, J = 6.0Hz, 2H), 3.09 (t, J = 6.4Hz, 2H) CD1-363: 1 HNMR (DMSO-d6, 400MHz) δ7.29 (s, 1H), 6.83 (d, J=8.4Hz, 1H), 6.75(s, 1H), 6.70(d, J=8.4Hz, 1H), 6.04(s, H), 6.00(s, 2H), 4.20(s, 2H), 3.75 (s, 3H), 3.71 (s, 3H), 3.36 (t, J=6.0Hz, 2H), 2.80 (t, J=6.0Hz, 2H) CD1-367: 1 HNMR (MeOH-d4, 400MHz) δ7.48 (s, 1H), 6.83-6.91 (m, 3H), 6.74 (d, J=8.4Hz, 1H), 4.12 (t, J=8.0Hz, 2H), 3.12 (t, J=8.0Hz, 2H) CD1-386: 1 HNMR (CDCl3, 400MHz) δ10.39 (s, 1H), 7.81 (s, 2H), 7.36 (s, 1H), 6.87 (s, 1H), 6.41-6.34 (m, 1H), 6.08 (s, 2H), 5.45 (d , J=10.8Hz, 1H), 5.15 (br, 2H), 4.96 (d, J=17.6Hz, 1H), 4.40 (s, 3H), 4.06 (s, 3H), 4.00 (br, 2H), 3.27 (t, J=5.6Hz, 2H) CD1-404: 1 HNMR (MeOH-d4, 400MHz) δ10.02 (s, 1H), 8.09 (d, J = 9.2Hz, 1H), 7.79 (d, J = 9.6Hz, 1H), 7.37 (t, J = 7.6Hz, 2H), 7.28 (d, J = 6.4Hz, 1H), 7.15-7.14 (m, 3H), 6.94 (s, 1H), 6.06 (s, 2H), 4.85 (br, 2H), 4.73 (s, 2H), 4.01 (s, 3H), 4.00 (s, 3H), 3.15 (t, J=5.6Hz, 2H) CD1-409: 1 HNMR (DMSO-d6, 400MHz) δ9.90 (s, 1H), 8.25-8.18 (m, 2H), 7.30 (s, 1H), 7.16 (s, 1H), 6.19 (s, 2H), 4.80 (br, 2H), 4.09 (s, 6H), 3.38 (m, 2H), 3.10 (t, J=5.2Hz, 2H), 1.48-1.43 (m, 3H) <Example 2> Analysis of the activity of isoquinoline derivatives in promoting mitochondrial autophagy

[0100] Whether the isoquinoline derivatives of the present invention promote mitochondrial autophagy activity is analyzed. Specifically, the mitochondrial autophagy fluorescent protein quantification method is used, which can quantitatively measure the mitochondrial autophagy activity in living cells. The isoquinoline derivatives prepared in Example 1 were treated with the human normal lung cell line BEAS-2B at a concentration of 20 μM for 24 hours. After treatment, the mitochondrial autophagy activity was measured using a flow cytometer. A control group (Control, con) that was not treated with the compound was used as a negative control, and CCCP (10 μM) that induced mitochondrial autophagy activity in the experiment, and CD1-012, a precursor of the compound of the present invention known to induce mitochondrial autophagy activity, were used as positive controls.

[0101] The results are as follows Figure 2 As shown, compared with the control group, the isoquinoline derivatives of the present invention significantly increased the mitochondrial autophagy activity ( Figure 2 A). In addition, when CD1-386 and CD1-409, which have the best mitophagy activity promoting effect, were compared with CD1-012, CD1-012 showed the greatest mitophagy activity at a concentration of 20 μM, while CD1-386 showed the greatest mitophagy activity at 5 μM, and CD1-409 showed the greatest mitophagy activity at 10 μM. Therefore, it was confirmed that the activity of CD1-386 was 4 times higher than that of CD1-012, a precursor of the isoquinoline derivative, and the activity of CD1-409 was 2 times higher than that of CD1-012, a precursor of the isoquinoline derivative. <Example 3> Confirmation of mitochondrial toxicity of isoquinoline derivatives

[0102] CCCP promotes mitophagic activity in experiments, it induces mitophagic activity, but also causes mitochondrial dysfunction and toxicity. Therefore, the novel isoquinoline derivatives of the present invention were tested to determine whether they induce mitochondrial dysfunction and toxicity. Specifically, mitochondrial membrane potential (known as a mitochondrial function indicator) was analyzed using the TMRM (tetramethylrhodamine methyl ester) assay.

[0103] The results are as follows Figure 3 As shown, the mitochondrial membrane potential in mitochondria treated with 10 μM CCCP was completely reduced, while the mitochondrial membrane potential in mitochondria treated with 20 μM of the isoquinoline derivative of the present invention was not significantly reduced. Therefore, it was confirmed that the novel isoquinoline derivative of the present invention does not cause mitochondrial dysfunction or toxicity. <Example 4> Confirmation of cytotoxicity of isoquinoline derivatives

[0104] A colony formation test was performed to confirm the cytotoxicity of the novel isoquinoline derivatives of the present invention. Specifically, the human normal lung cell line BEAS-2B was treated with the isoquinoline derivatives for 24 hours and then cultured for 2 weeks to analyze the colonies formed. As controls, a control group (con) not treated with any substance and a positive control treated with CCCP were used, which induces mitophagy activity but also causes mitochondrial dysfunction and toxicity.

[0105] Results, such as Figure 4 As shown, cells treated with CCCP showed a decrease in colony size and number. However, the isoquinoline derivatives of the present invention (except CD1-367) showed colony size and number similar to those of the con group, confirming that the derivatives other than CD1-367 did not show cytotoxicity.

[0106] Therefore, the present invention synthesized novel isoquinoline derivatives CD1-271, CD1-363, CD1-367, CD1-386, CD1-404 and CD1-409, and it has been confirmed that these novel isoquinoline derivatives have the effect of promoting mitochondrial autophagy activity. In addition, it has been confirmed that these derivatives will not cause mitochondrial dysfunction and are safe derivatives that are not toxic to cells. Industrial Applicability

[0107] The present invention relates to novel isoquinoline derivatives. The derivatives have the effect of promoting mitochondrial autophagy activity, thereby reducing mitochondria with structural and / or functional damage. Specifically, the novel isoquinoline derivatives according to the present invention do not induce mitochondrial dysfunction and have been determined to be safe compounds that are non-toxic to cells, and can therefore be used to prevent, improve and / or treat various diseases that may be caused by mitochondrial abnormalities.

Claims

1. A pharmaceutical composition for promoting mitochondrial autophagy activity, comprising an isoquinoline derivative compound or a salt thereof as an active ingredient.

2. The pharmaceutical composition according to claim 1, wherein the isoquinoline derivative compound is represented by the following Chemical Formula 1 to Chemical Formula 6: Chemical formula 1: Chemical formula 2: Chemical formula 3: Chemical formula 4: Chemical formula 5: Chemical formula 6:

3. The pharmaceutical composition according to claim 1, wherein the isoquinoline derivative compound increases mitophagy activity.

4. The pharmaceutical composition according to claim 3, wherein the increase in mitophagy activity removes dysfunctional mitochondria.

5. The pharmaceutical composition according to claim 4, wherein the dysfunctional mitochondria induce neurodegenerative diseases.

6. The pharmaceutical composition according to claim 5, wherein the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, multiple sclerosis, immune-mediated brain dysfunction, progressive neurodegenerative diseases, metabolic encephalopathy, Niemann-Pick disease, dementia caused by cerebral ischemia and cerebral hemorrhage.

7. The pharmaceutical composition according to claim 6, wherein the neurodegenerative disease is Alzheimer's disease or dementia.

8. A food composition for promoting mitochondrial autophagy activity, comprising an isoquinoline derivative compound or a salt thereof as an active ingredient.

9. The food composition according to claim 8, wherein the isoquinoline derivative compound is represented by the following Chemical Formula 1 to Chemical Formula 6: Chemical formula 1: Chemical formula 2: Chemical formula 3: Chemical formula 4: Chemical formula 5: Chemical formula 6:

10. A pharmaceutical composition for preventing or treating a disease caused by mitochondrial dysfunction, comprising an isoquinoline derivative compound or a salt thereof as an active ingredient.

11. The pharmaceutical composition according to claim 10, wherein the isoquinoline derivative compound is represented by the following Chemical Formula 1 to Chemical Formula 6: Chemical formula 1: Chemical formula 2: Chemical formula 3: Chemical formula 4: Chemical formula 5: Chemical formula 6:

12. The pharmaceutical composition according to claim 10, wherein the isoquinoline derivative compound increases mitophagy activity.

13. The pharmaceutical composition according to claim 10, wherein the disease caused by mitochondrial dysfunction is a neurodegenerative disease.

14. The pharmaceutical composition according to claim 13, wherein the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, multiple sclerosis, immune-mediated brain dysfunction, progressive neurodegenerative diseases, metabolic encephalopathy, Niemann-Pick disease, dementia caused by cerebral ischemia and cerebral hemorrhage.

15. A food composition for preventing or ameliorating a disease caused by mitochondrial dysfunction, comprising an isoquinoline derivative compound or a salt thereof as an active ingredient.

16. The food composition according to claim 15, wherein the isoquinoline derivative compound is represented by the following Chemical Formula 1 to Chemical Formula 6: Chemical formula 1: Chemical formula 2: Chemical formula 3: Chemical formula 4: Chemical formula 5: Chemical formula 6:

17. A method for increasing mitophagy activity in cells, comprising the step of treating the cells with an isoquinoline derivative compound.

18. The method according to claim 17, wherein the isoquinoline derivative compound is represented by the following Chemical Formula 1 to Chemical Formula 6: Chemical formula 1: Chemical formula 2: Chemical formula 3: Chemical formula 4: Chemical formula 5: Chemical formula 6:

19. A method for inhibiting mitochondrial dysfunction, comprising the step of treating cells with an isoquinoline derivative compound.

20. The method according to claim 19, wherein the isoquinoline derivative compound is represented by the following Chemical Formula 1 to Chemical Formula 6: Chemical formula 1: Chemical formula 2: Chemical formula 3: Chemical formula 4: Chemical formula 5: Chemical formula 6:

21. A method for preventing or treating a disease caused by mitochondrial dysfunction, comprising the step of administering an isoquinoline derivative or a salt thereof to an individual in need thereof.

22. Use of an isoquinoline derivative or a salt thereof in the preparation of a medicament for preventing or treating a disease caused by mitochondrial dysfunction.

23. Use of an isoquinoline derivative or a salt thereof for preventing or treating a disease caused by mitochondrial dysfunction.

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