A ligand that acts on an adenosine receptor or a composition for preventing, improving, or treating obesity containing the same

Abstract

The present invention relates to A 2A a dual - acting ligand that acts on both adenosine receptors and / or A3 adenosine receptors, and a composition for preventing, improving, or treating obesity containing the ligand. Further, the present invention relates to a method for treating A 2A adenosine receptor (AR) and / or A3 adenosine receptor (AR) - related diseases or A 2A use for the manufacture of a medicament for treating adenosine receptor (AR) and / or A3 adenosine receptor (AR) - related diseases. The ligand LJ - 4378 or its derivatives that act on the adenosine receptors of the present invention 2A acts doubly as an agonist for AR and an antagonist for A3AR, significantly increasing mitochondrial proteins, UCP1, and COXIV respectively, and showing an excellent anti - obesity effect on high - fat diet mice. Therefore, the ligand that acts on the adenosine receptors of the present invention can be effectively utilized in the manufacture of a composition, treatment method, or medicament for preventing, improving, or treating obesity.

Description

[Technical Field]

[0001] This application claims priority from Korean Patent Application No. 10-2022-0091348, filed on July 22, 2022, the entire specification of which is incorporated herein by reference.

[0002] This invention is A 2A The present invention relates to a dual-acting ligand that acts on both adenosine receptors and / or A3 adenosine receptors, and a composition containing said ligand for preventing, ameliorating, or treating obesity.

[0003] The present invention also provides a method for producing a compound A using the ligand. 2A Methods for treating adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related disorders 2A The present invention relates to use in the manufacture of a medicament for treating adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases.

[0004] This invention was made under the support of the Ministry of Health and Welfare of the Republic of Korea under grant number HN22C0687 (1465037037). [Background technology]

[0005] Obesity is defined as excessive fat accumulation, which is a major risk factor for metabolic diseases such as type 2 diabetes. Adipose tissue can be classified into brown adipose tissue (BAT) and white adipose tissue (WAT). BAT contains numerous mitochondria and is responsible for non-shivering thermogenesis, whereas WAT is specialized for fat storage and mobilization. Therefore, activating and recruiting BAT is a strategy to increase energy expenditure and ultimately could be exploited with the potential goal of preventing obesity.

[0006] As an inducer of brown adipocytes, pharmacological stimulation of adenosine receptor signaling has received considerable attention as a potential target for improving metabolic health. Adenosine receptors are classified into four types: G-coupled receptors A1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A11, A12, A13, A14, A15, A16, A17, A18, A19, A20, A21, A22, A23, A24, A25, A26, A27, A28, A29, A30, A31, A32, A33, A34, A35, A36, A37, A38, A39, A40, A41, A42, A43, A44, A45, A46, A47, A48, A49, A51, A52, A53, A 2A , A 2B, and can be classified into A3. A1 adenosine receptor (A1AR) and A3 adenosine receptor (A3AR) bind to G i / o protein to suppress the production of cAMP and the signal transduction of cAMP-dependent protein kinase (PKA). In contrast, A 2A adenosine receptor (A 2A AR) and A 2B adenosine receptor (A 2B AR) bind to G s / olf protein to promote the activity of adenylate cyclase (AC) and the production of cAMP.

[0007] Among adenosine receptors, A 2A AR is more abundantly expressed in BAT than in WAT, and its anti-obesity effect has been supported by various studies. Activation of A 2A AR stimulates AC activity to promote PKA-dependent lipolysis, and the signal transduction of A 2A AR is necessary for the complete physiological function of BAT. Also, recent studies have shown that the expression of thermogenic genes including UCP1 is significantly improved both in vitro and in vivo by CGS21680, an agonist of A 2A AR. In contrast, A3AR binds to Gi and suggests an inhibitory effect on A 2A AR stimulation. The expression of A3AR is abundant in white adipose tissue, but the regulatory role of A3AR in adipocyte metabolism is not fully understood.

Prior Art Documents

Patent Documents

[0008]

Patent Document 1

Non-Patent Documents

[0009]

Non-Patent Document 1

[0010] The inventors of the present invention have made intensive efforts to provide a dual active ligand that acts on both the A 2A adenosine receptor (AR) and / or the A3 adenosine receptor (AR). As a result, it has been confirmed that LJ-4378 or its derivatives are agonists for the A 2A AR and act as antagonists for the A3AR, and can provide an excellent anti-obesity effect, thus completing the present invention.

[0011] Therefore, an object of the present invention is to provide a ligand that acts on the adenosine receptor and a composition for preventing, improving, or treating obesity containing the ligand.

[0012] Another object of the present invention is to provide a method for treating A 2A adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases using the ligand, or to provide a use for manufacturing a medicament for treating A 2A adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases. Means for Solving the Problems

[0013] The present invention provides pharmaceutical compositions for preventing or treating adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases, which contain a compound represented by the following chemical formula 1, its isomers, or pharmaceutically acceptable salts: 2A

Chemical formula

[0014] According to a preferred embodiment of the present invention, R1 is H, a methyl group, or an aryl-C1-C10 alkyl group, and R2 is a C1-C4 alkyl group.

[0015] …According to a preferred embodiment of the present invention, the compound represented by the chemical formula 1, its isomers, or pharmaceutically acceptable salts are: (1) 2-(6-amino-2-(hex-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (2) (2R,3R,4S)-2-(6-amino-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (3) (2R,3R,4S)-2-(6-((3-fluorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (4) (2R,3R,4S)-2-(6-((3-chlorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (5) (2R,3R,4S)-2-(6-((3-bromobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (6)(2R,3R,4S)-2-(6-((3-Iodobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (7)(2R,3R,4S)-2-(6-amino-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (8)(2R,3R,4S)-2-(2-(but-1-yn-1-yl)-6-((3-fluorobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; (9)(2R,3R,4S)-2-(2-(but-1-yn-1-yl)-6-((3-chlorobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; (10)(2R,3R,4S)-2-(6-((3-bromobenzyl)amino)-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; and (11)(2R,3R,4S)-2-(2-(but-1-yn-1-yl)-6-((3-iodobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; is any one or more selected from the group consisting of.

[0016] According to a preferred embodiment of the present invention, the compound represented by the above chemical formula 1, its isomers, or a pharmaceutically acceptable salt is an agonist for adenosine receptor (AR) and an antagonist for A3 adenosine receptor (AR). 2A

[0017] According to a preferred embodiment of the present invention, the above A 2A adenosine receptor (AR) and / or A3 adenosine receptor (AR) related disease is obesity.

[0018] In addition, the present invention provides a health functional food composition for preventing or improving obesity, comprising the compound represented by the following chemical formula 1, its isomers, or a pharmaceutically acceptable salt: [Chemical formula] Said R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group, R2 is a C1-C10 alkyl group.

[0019] Also, the present invention provides a method for treating a patient with a disease related to adenosine receptor (AR) and / or A3 adenosine receptor (AR), comprising the step of administering to the patient a compound represented by the following chemical formula 1, its isomers, or a pharmaceutically acceptable salt thereof: 2A adenosine receptor (AR) and / or A3 adenosine receptor (AR): 2A [Chemical formula] Said R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group, R2 is a C1-C10 alkyl group.

[0020] Also, the present invention provides the use of a compound represented by the following chemical formula 1, its isomers, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating a disease related to adenosine receptor (AR) and / or A3 adenosine receptor (AR): 2A [Chemical formula] Said R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group, R2 is a C1-C10 alkyl group.

[0021] Extracellular adenosine is a potent endogenous signaling molecule that regulates various physiological and pathological events through four types of adenosine receptors. Studies using genetically modified mice with altered adenosine receptor expression have further confirmed the fact that adenosine exerts diverse effects on energy metabolism. Therefore, targeting adenosine receptors may have the potential for translation in treating metabolic diseases. The present invention is about A 2A confirmed the effects of a dual-action ligand with A

[0022] G αs -coupled A 2A receptor stimulation activates the adenylate cyclase and subsequently the cAMP-PKA signaling pathway, while activation of G αi -coupled A3AR suppresses adenylate cyclase and opposes the effects of A 2A receptor signaling. Therefore, the inventors hypothesized that a dual ligand with A 2A receptor agonist and A3AR antagonist activities could synergistically activate PKA-dependent lipolysis and thermogenesis in adipocytes. The present invention has confirmed that LJ-4378 treatment increases the mitochondrial content and activity in adipocytes, and the effect of LJ-4378 is even more potent than that of an A 2A receptor agonist or an A3AR antagonist. In vivo LJ-4378 treatment increased energy consumption and the expression of the brown adipocyte marker CIDEA, as determined by non-invasive imaging of the bioluminescence signal in CIDEA reporter mice. In vivo LJ-4378 treatment decreased body weight and fat content and improved glucose tolerance in mice fed a high-fat diet, showing an anti-obesity effect.

[0023] In the present invention, qPCR analysis shows that A 2A R and A3AR are the main subtypes of adenosine receptors expressed in adipocytes. In contrast, the expression level of A 2B R in adipocytes was lower than that of other adenosine receptors. Previous studies have shown that A 2B R is mainly expressed in muscle and BAT, and A 2BIt has been proven that the activation of AR exerts the effects of anti-aging and anti-obesity. Interestingly, A 2B AR plays a lenient role in A 2A -mediated lipolysis and forms a heterodimer of two receptors. In contrast, the A1 receptor bound to G αi suppresses the activity of adenylate cyclase and is highly expressed in human white adipocytes. Therefore, A1AR has been characterized as an antilipolytic agent that can reduce the levels of circulating FFA and triglycerides and promote insulin sensitivity in adipose tissue. In addition, A1AR signaling promotes adipogenesis and regulates inflammation as shown by studies on A1AR knockout mice.

[0024] In addition to its effect on lipolysis, the activation of A 2A AR exerts an anti-inflammatory effect in a mouse model of diet-induced obesity.

[0025] Conclusively, since the present invention can be orally administered, has a high patient preference, and its mechanism of action does not stimulate the central nervous system but promotes and consumes fat through the adenosine receptor of adipocytes, the probability of developing mental side effects is low. As an anti-obesity agent with a new mechanism, it can provide an option for patients who do not respond to conventional dosing methods.

[0026] In addition, in the case of conventional GLP-1 derivatives and central nervous system stimulant anti-obesity agents, obesity is treated by the effect of appetite reduction. In contrast, the present invention can increase patient preference by a mechanism that directly targets brown adipocytes to reduce fat, and an increase in effect and alleviation of side effects can be expected by combined administration.

[0027] Therefore, the present invention can provide a pharmaceutical composition for preventing or treating A 2A adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases, comprising a compound represented by the following Chemical Formula 1, its isomers, or a pharmaceutically acceptable salt:

Chemical Formula

[0028] According to a preferred embodiment of the present invention, said R1 is H, a methyl group, or an aryl-C1-C10 alkyl group, R2 may be a C1-C4 alkyl group.

[0029] In the present invention, said pharmaceutically acceptable salts are usefully acid addition salts formed by pharmaceutically acceptable free acids. The acid addition salts are obtained from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, or phosphorous acid, and non-toxic organic acids such as aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates and alkanedioates, aromatic acids, aliphatic and aromatic sulfonic acids. Such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, or mandelate.

[0030] The acid addition salts according to the present invention can be prepared by conventional methods, for example, by dissolving the compound represented by Chemical Formula 1 in an excess of aqueous acid solution and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone, or acetonitrile. Alternatively, the solvent and the excess acid can be evaporated from the mixture and then dried, or the precipitated salt can be collected by suction filtration.

[0031] Pharmaceutically acceptable metal salts may also be prepared using a base. Alkali metal or alkaline earth metal salts can be obtained, for example, by dissolving the compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the precipitated compound salt, and evaporating and drying the filtrate. In this case, it is pharmaceutically suitable to prepare sodium, potassium, or calcium salts as the metal salts. The corresponding silver salts can be obtained by reacting the alkali metal or alkaline earth metal salts with a suitable silver salt (e.g., silver nitrate).

[0032] According to a preferred embodiment of the present invention, the compound represented by Chemical Formula 1, its isomers, or pharmaceutically acceptable salts are (1) 2-(6-Amino-2-(hex-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (2) (2R,3R,4S)-2-(6-Amino-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (3) (2R,3R,4S)-2-(6-((3-Fluorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (4) (2R,3R,4S)-2-(6-((3-Chlorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (5)(2R,3R,4S)-2-(6-((3-Bromobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (6)(2R,3R,4S)-2-(6-((3-Iodobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (7)(2R,3R,4S)-2-(6-Amino-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (8)(2R,3R,4S)-2-(2-(but-1-yn-1-yl)-6-((3-fluorobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; (9)(2R,3R,4S)-2-(2-(but-1-yn-1-yl)-6-((3-chlorobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; (10)(2R,3R,4S)-2-(6-((3-Bromobenzyl)amino)-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; and (11)(2R,3R,4S)-2-(2-(but-1-yn-1-yl)-6-((3-iodobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; It may be any one or more selected from the group consisting of.

[0033] According to a preferred embodiment of the present invention, the compound represented by the chemical formula 1, its isomers, or a pharmaceutically acceptable salt thereof is A 2A An agonist for the adenosine receptor (AR) and may be an antagonist for the A3 adenosine receptor (AR).

[0034] According to a preferred embodiment of the present invention, the A 2A The adenosine receptor (AR) and / or the A3 adenosine receptor (AR)-related disease may be obesity.

[0035] "Prevention" in the present invention means the prevention of A by the compound represented by Chemical Formula 1 of the present invention, its isomer, or pharmaceutically acceptable salt. 2A It refers to any action that inhibits or delays the onset of adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases.

[0036] "Improvement" or "treatment" of the present invention refers to the improvement of a patient's condition by a compound of Formula 1, its isomer, or a pharmaceutically acceptable salt thereof. 2A It means any action that improves or alleviates parameters related to adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases, for example, the severity of symptoms.

[0037] The pharmaceutical compositions of the present invention can be in various oral or parenteral dosage forms. When formulating the compositions, they can be prepared using one or more buffering agents (e.g., saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, extenders, binders, adjuvants (e.g., aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrating agents or surfactants, diluents or excipients, etc.

[0038] Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations are prepared by mixing one or more compounds with at least one or more excipients, such as starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol, maltitol, cellulose, methylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, or gelatin, etc. For example, tablets or sugar-coated tablets can be obtained by blending an active ingredient with a solid excipient, pulverizing the mixture, adding suitable excipients, and then processing the mixture into a granule.

[0039] In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of liquid preparations for oral administration include suspensions, internal liquids, oils, or syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, or preservatives may be included. In some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as disintegrants, and anticoagulants, flavorings, emulsifiers, solubilizers, dispersants, flavoring agents, antioxidants, packaging agents, pigments, and preservatives may be additionally included.

[0040] Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspension solvents, oils, lyophilized preparations, or suppositories. As non-aqueous solvents and suspension solvents, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As the base of suppositories, witepsol, macrogol, tween 61, cocoa butter, laurin fat, glycerol, gelatin, etc. may be used.

[0041] The composition of the present invention can be administered orally or parenterally, and when administered parenterally, it can be formulated into a dosage form by a method known in the art in the form of topical skin application; intraperitoneal, rectal, intravenous, subcutaneous, or transdermal administration agents.

[0042] In the case of the injection, it must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. In the case of an injection, examples of suitable carriers include, but are not limited to, solvents or dispersion media containing water, ethanol, polyols (such as glycerol, propylene glycol, and liquid polyethylene glycol), mixtures thereof, and / or vegetable oils. More preferably, as a suitable carrier, Hank's solution, Ringer's solution, PBS (phosphate buffered saline) containing triethanolamine, or sterile water for injection, isotonic solutions such as 10% ethanol, 40% propylene glycol, and 5% dextrose can be used. To protect the injection from microbial contamination, it may additionally contain various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. Also, the injection may almost always additionally contain an isotonicity agent such as sugar or sodium chloride.

[0043] In the case of a transdermal administration agent, forms such as ointments, creams, lotions, topical solutions, pastes, liniments, aerosols, etc. are included. In the above, transdermal administration means that a pharmaceutical composition is locally administered to the skin so that an effective amount of the active ingredient contained in the pharmaceutical composition is transmitted into the skin.

[0044] The composition of the present invention is administered in a pharmaceutically effective amount. A pharmaceutically effective amount means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The level of the effective volume can be determined by factors such as the type and severity of the patient's disease, the activity of the drug, the sensitivity to the drug, the administration time, the administration route and excretion rate, the treatment period, factors including drugs used simultaneously, and other factors well known in the medical field. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered in a single or multiple doses. That is, the total effective amount of the composition of the present invention can be administered to a patient in a single dose or by a fractionated treatment protocol administered in multiple doses over a long period. It is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects considering all of the above factors, which can be easily determined by those skilled in the art.

[0045] The dosage of the pharmaceutical composition of the present invention can vary widely depending on the patient's weight, age, gender, health status, diet, administration time, administration method, excretion rate, and disease severity.

[0046] The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

[0047] The pharmaceutical composition of the present invention can also be provided in a dosage form of an external preparation containing the compound represented by the above chemical formula 1, its isomers, or a pharmaceutically acceptable salt as an active ingredient. A of the present invention 2AWhen a pharmaceutical composition for preventing or treating adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases is used as a topical skin preparation, it may additionally contain adjuvants commonly used in the field of dermatology, such as fatty substances, organic solvents, solubilizers, thickeners, and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic emulsifiers, non-ionic emulsifiers, metal ion sequestering agents, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic activators, lipophilic activators, or lipid vesicles, etc. Also, the above components may be introduced in amounts commonly used in the field of dermatology.

[0048] A of the present invention 2A When a pharmaceutical composition for preventing or treating adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases is provided as a topical skin preparation, it may be in dosage forms such as, but not limited to, ointments, patches, gels, creams, or sprays.

[0049] In addition, the present invention may provide a health functional food composition for preventing or improving obesity, comprising a compound represented by the following Chemical Formula 1, its isomers, or a pharmaceutically acceptable salt: [Chemical Formula] R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group, R2 is a C1-C10 alkyl group.

[0050] Since the compound represented by Chemical Formula 1, its isomers, or the pharmaceutically acceptable salt is the same as that used in the above pharmaceutical composition, the description is replaced by that description.

[0051] The health - functional food composition according to the present invention can be manufactured in various forms by ordinary methods known in the art. As general foods, but not limited to these, beverages (including alcoholic beverages), fruits and their processed foods (e.g., canned fruits, bottled fruits, jams, marmalades, etc.), fish, meats, and their processed foods (e.g., hams, sausages, corned beef, etc.), breads and noodles (e.g., udon, soba, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, candies, dairy products (e.g., butter, cheese, etc.), edible vegetable oils, margarine, vegetable proteins, retort foods, frozen foods, various seasonings (e.g., miso, soy sauce, sauce, etc.), etc., can be manufactured by adding the compound represented by Chemical Formula 1 of the present invention, its isomers, or a pharmaceutically acceptable salt thereof. Also, as nutritional supplements, but not limited to these, capsules, tablets, pills, etc., can be manufactured by adding the compound represented by Chemical Formula 1 of the present invention, its isomers, or a pharmaceutically acceptable salt thereof. Further, as health - functional foods, but not limited to these, for example, the compound represented by Chemical Formula 1 of the present invention, its isomers, or a pharmaceutically acceptable salt itself can be liquefied, granulated, encapsulated, and powdered so as to be taken by being manufactured in the form of tea, juice, and drinks for drinking (health drinks). Also, the compound represented by Chemical Formula 1 of the present invention, its isomers, or a pharmaceutically acceptable salt can be manufactured and used in the form of a powder or a concentrated solution for use in the form of a food additive. Also, the compound represented by Chemical Formula 1 of the present invention, its isomers, or a pharmaceutically acceptable salt and A 2A can be mixed with a known active ingredient known to have a preventive or ameliorating effect on adenosine receptor (AR) and / or A3 adenosine receptor (AR) - related diseases and manufactured in the form of a composition.

[0052] When using the compound represented by Chemical Formula 1 of the present invention, its isomers, or a pharmaceutically acceptable salt as a health beverage, the health beverage composition may contain various flavoring agents or natural carbohydrates, etc. as additional components like ordinary beverages. The above-mentioned carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetening agent may be a natural sweetening agent such as thaumatin and stevia extract; a synthetic sweetening agent such as saccharin and aspartame, etc.

[0053] In addition, the compound represented by Chemical Formula 1 of the present invention, its isomers, or a pharmaceutically acceptable salt may be contained as an active ingredient in a health functional food composition for preventing or improving adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases, and the amount thereof is 2A not particularly limited as long as it is an amount effective for achieving the preventive or ameliorating effect on adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases, but is preferably 0.01 to 100% by weight based on the total weight of the whole composition. Further, the health functional food composition of the present invention may be produced by mixing together the compound represented by Chemical Formula 1, its isomers, or a pharmaceutically acceptable salt with 2A other active ingredients known to be effective for adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases. 2A In addition to the above, the health functional food of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonating agents, etc. Further, the health functional food of the present invention may contain the pulp for producing natural fruit juice, fruit juice beverage, or vegetable beverage. Such components may be used independently or in combination.

[0054] In addition, the health functional food of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonating agents, etc. Further, the health functional food of the present invention may contain the pulp for producing natural fruit juice, fruit juice beverage, or vegetable beverage. Such components may be used independently or in combination.

[0055] In addition, the present invention 2AA method for treating adenosine receptor (AR) and / or A3 adenosine receptor (AR)-associated diseases, comprising administering a compound represented by the following formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof to a patient: 2A Methods for treating adenosine receptor (AR) and / or A3 adenosine receptor (AR)-associated disorders may be provided: [ka] R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group; R2 is a C1-C10 alkyl group.

[0056] The compound represented by Chemical Formula 1, its isomer, or pharmaceutically acceptable salt is the same as that used in the pharmaceutical composition, and therefore the description therefor is substituted therefor.

[0057] According to a preferred embodiment of the present invention, 2A The adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related disorder can be obesity.

[0058] The present invention also provides 2A The present invention provides a use of a compound represented by the following chemical formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof for use in the manufacture of a medicament for treating an adenosine receptor (AR) and / or A3 adenosine receptor (AR)-associated disease: [ka] R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group; R2 is a C1-C10 alkyl group.

[0059] The compound represented by Chemical Formula 1, its isomer, or pharmaceutically acceptable salt is the same as that used in the pharmaceutical composition, and therefore the description therefor is substituted therefor.

[0060] According to a preferred embodiment of the present invention,2A Adenosine receptor (AR) and / or A3 adenosine receptor (AR)-related diseases may be obesity.

Advantages of the Invention

[0061] LJ-4378 or its derivatives, which are ligands acting on the adenosine receptor of the present invention, 2A act doubly as an agent acting on AR and an antagonist acting on A3AR, significantly increase mitochondrial proteins, UCP1 and COXIV, respectively, and showed excellent anti-obesity effects on high-fat diet mice. Therefore, the ligand acting on the adenosine receptor of the present invention can be effectively utilized for the production of compositions, treatment methods or drugs for the prevention, improvement or treatment of obesity.

Brief Description of the Drawings

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Figure 1a

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Figure 1b

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Figure 1c

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Figure 2

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Figure 3

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Figure 4

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Figure 5

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Figure 6

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Figure 7

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Figure 8

Mode for Carrying Out the Invention

[0072] Hereinafter, the present invention will be described in more detail through examples. It is self-evident to those with ordinary knowledge in the relevant technical field that these examples are solely for illustrative purposes of the present invention and should not be construed as limiting the scope of the present invention by these examples.

[0073] [Example 1] <1-1>Cell culture Adipocyte progenitor cells capable of differentiating into brown adipocytes were isolated from the brown adipose tissue of mice. The cells were cultured in Dulbecco's modified Eagle's medium (Welgene, LM001-07) containing 10% fetal bovine serum (FBS, Gibco, 16000044) and 1% Penicillin Streptomycin (Welgene, LS202-02) in humidified air at 37 °C and 5% CO2. When the cells reached approximately 100% confluency, the cells were exposed to a differentiation medium supplemented with 2.5 mM isobutylmethylxanthine (IBMX, Cayman, I5879), 0.125 mM indomethacin (Cayman, 70270), 1 μM dexamethasone (Cayman), 1 μg / mL insulin (Sigma, I9278), and 1 nM triiodothyronine (T3, Cayman, 6028) for 3 days for adipocyte differentiation. Next, the cells were maintained in a growth medium containing 1 μg / mL insulin and 1 nM T3 (triiodothyronine) for 3 days.

[0074] <1-2>Animal Six-week-old C57BL / 6 male mice and CIDEA reporter mice were used according to a protocol approved by the Seoul National University Institutional Animal Care and Use Committee (SNU-201107-1, SNU-201221-3) for in vivo experiments. The mice were housed under a 12h-light / 12h-dark cycle and allowed free access to normal chow diet (NCD, Purina Lab, 38057, protein: 24.52% calories, carbohydrates: 63.07% calories, fat: 12.41% calories) and water at 22 ± 1 °C. LJ-4378 (1 mg·kg−1 day−1) dissolved in 0.2% DMSO (diluted in PBS) was intraperitoneally administered for 10 days.

[0075] In the case of the diet-induced obesity model, mice were fed a high-fat diet (HFD) (Research Diets, D12492, protein: 20% kcal, carbohydrates: 20% kcal, fat: 60% kcal) for 8 weeks. Next, mice were injected with LJ-4378 (1 mg·kg−1) once a day for 10 days.

[0076] [Example 2] Synthesis and Profile of LJ-4378 and its Derivatives <2-1>Synthesis of LJ-4378 D-Mannose was converted to a glycosyl donor (compound 1) according to Non-Patent Document 1. The glycosyl donor (compound 1) is a Lewis acid which, in the presence of TMSOTf, was condensed with 2-amino-6-chloropurine to afford the β-anomer (compound 2, 30%) as a single stereoisomer (Figure 1a). The anomeric assignment was readily achieved by the nuclear Overhauser effect between H-8 and 3'-H 1 in the 1H NMR experiment. In the presence of CuI, the 2-amino-6-chloro derivative (compound 2) was treated with isoamyl nitrite, iodine, and methylene iodide to give the 2-iodo-6-chloro derivative (compound 3), which was converted to the 2-iodo-6-amino derivative (compound 4) by treatment with methanolic ammonia. In the presence of bis(triphenylphosphine)palladium dichloride, the Sonogashira coupling reaction of compound 4 with 1-hexyne gave the 2-hexynyl derivative (compound 5). Finally, the isopropylidene of compound 5 was removed with 1N HCl to synthesize the final 2-hexynyl-4'-thioadenosine derivative (LJ-4378 (2-(6-amino-2-(hex-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol, compound 6).

[0077] <2-2>Synthesis of LJ-4378 derivatives D-Mannose was converted to a glycosyl donor (compound 1) according to Non-Patent Document 1. The glycosyl donor (compound 1) is a Lewis acid which, in the presence of TMSOTf, was Vorbruggen condensed with silylated 2-iodo-6-chloropurine to afford the β-anomer (compound 3, 60%) together with a trace amount of the α-anomer (Figure 1b). The anomeric sequence of compound 3 was 1It was confirmed by 1H NMR NOE experiment. A strong NOE effect was observed between H-8 and 3'-H of compound 3 to confirm the β configuration. On the contrary, the corresponding α-anomer did not show such a NOE effect.

[0078] Next, compound 3 was subjected to a Sonogashira coupling reaction using propyne and butyne in the presence of tetrakis(triphenylphosphine)palladium and cesium carbonate to obtain C2-propynyl derivative (compound 7a) and C2-butynyl derivative (compound 7b), respectively. Compounds 7a and 7b were treated with 1N HCl to produce 2-propynyl-6-chloro derivative (compound 8a) and 2-butynyl-6-chloro derivative (compound 8b), respectively. The 2-propynyl-6-chloro derivative (compound 8a) was treated with ammonia in tert-butanol and 3-halobenzylamine to obtain 2-propynyl-4'-thioadenosine derivative (compound 9a) and 2-propynyl-N6-3-halobenzyl-4'-thioadenosine derivatives (compounds 9b-e), respectively. Similarly, the 2-butynyl-6-chloro intermediate (compound 8b) was converted to 2-butynyl-4'-thioadenosine derivative (compound 9f) and 2-butynyl-N6-3-halobenzyl-4'-thioadenosine derivatives (compounds 9g-j). The specific production methods are as follows in Production Methods 1 to 12.

[0079] [Production Method 1] (2R,3R,4S)-2-(6-Chloro-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol ((2R,3R,4S)-2-(6-Chloro-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; compound 8a). In anhydrous N,N-dimethylformamide (25 mL), to a stirred solution of compound 3 (943 mg, 2.14 mmol) were added tetrakis(triphenylphosphine)palladium (248 mg, 0.21 mmol), copper(I) iodide (49 mg, 0.25 mmol), and cesium carbonate (700 mg, 2.14 mmol). The stirred mixture was bubbled with propyne gas at room temperature for 10 minutes and stirred for an additional 1 hour at room temperature. The reaction mixture was evaporated under reduced pressure, and the crude product compound 7a was applied to the next reaction without further purification. To a stirred ice-cooled solution of the crude product compound 7a in tetrahydrofuran (5 mL) was added 1 N HCl (5 mL), and the mixture was stirred at room temperature for 15 hours. The reaction mixture was neutralized with 1 N NaOH solution and carefully evaporated under reduced pressure. The residue was purified by silica gel column chromatography (methylene chloride / methanol = 24:1) to give compound 8a (0.301 g, 45% from compound 3) as a white solid: mp 101 - 103 °C; UV (MeOH) λ max 284 nm; 1 H NMR (500 MHz, CD3OD): δ 8.85 (s, 1H), 6.08 (d, J = 6.6 Hz, 1H), 4.71 - 4.69 (m, 1H), 4.46 (q, J = 3.6 Hz, 1H), 3.55 (dd, J = 4.4, 11.0 Hz, 1H), 2.96 (dd, J = 3.4, 11.0 Hz, 1H), 2.11 (s, 3H); [α] 20 D = -45.0 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 12 H 12 ClN4O2S (M + H) + 311.0369, Found: 311.0374. Anal. Calcd for C 12 H 12 ClN4O2S: C, 46.38; H, 3.57; N, 18.03. Found: C, 46.78; H, 3.24; N, 18.04.

[0080] [Preparation 2] (2R,3R,4S)-2-(2-(But-1-yn-1-yl)-6-chloro-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; Compound 8b). To a stirred solution of Compound 3 (1.63 g, 3.71 mmol) in anhydrous N,N-dimethylformamide (25 mL) were added tetrakis(triphenylphosphine)palladium (429 mg, 0.37 mmol), copper(I) iodide (85 mg, 0.44 mmol), and cesium carbonate (1.21 g, 3.71 mmol). The stirred mixture was bubbled with butyne gas at room temperature for 10 minutes and stirred for an additional 1 hour at room temperature. The reaction mixture was evaporated under reduced pressure and used in the next reaction without further purification. To a stirred ice-cooled solution of the crude product Compound 7b in tetrahydrofuran (5 mL) was added 1N HCl (5 mL), and the mixture was stirred at room temperature for 15 hours. The reaction mixture was neutralized with 1N NaOH solution and evaporated under reduced pressure. The residue was purified by silica gel column chromatography (methylene chloride / methanol = 24:1) to give Compound 8b (530 mg, 44% from Compound 3) as a white solid: mp 105-107 °C; UV (MeOH) λ max 284 nm; 1 H NMR (500 MHz, CD3OD): δ 8.86 (s, 1H), 6.08 (d, J = 6.5 Hz, 1H), 4.69 (m, 1H), 3.54 (dd, J = 4.4, 11.0 Hz, 1H), 3.30 (bs, 1H), 2.96 (dd, J = 3.5, 11.0 Hz, 1H), 2.50 (q, J = 7.5 Hz, 2H), 1.27 (t, J = 7.5 Hz, 3H); 13C NMR (125 MHz, CD3OD): δ 154.2, 151.7, 148.7, 147.8, 132.6, 93.4, 81.6, 80.5, 75.1, 65.3, 36.1, 14.3, 14.2; [α] 20 D = -38.20 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 13 H 14 ClN4O2S (M + H)+ 325.0526, Found: 325.0528. Anal. Calcd for C 13 H 14 ClN4O2S: C, 48.08; H, 4.03; N, 17.25. Found: C, 48.09; H, 4.23; N, 17.01.

[0081] [Preparation Method 3] (2R,3R,4S)-2-(6-Amino-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol ((2R,3R,4S)-2-(6-Amino-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; Compound 9a) A solution of Compound 8a (50 mg, 0.16 mmol) in NH3 / tBuOH (5 mL) was stirred at 100 °C for 12 h in a steel bomb. The reaction mixture was evaporated and the residue was purified by silica gel column chromatography (methylene chloride / methanol = 24:1) to give 9a (45 mg, 85%) as a white solid: mp 247 - 249 °C; UV (MeOH) λ max 271 nm; 1 1H NMR (500 MHz, DMSO-d6): δ 8.46 (s, 1H), 7.35 (bs, 2H), 5.84 (d, J = 7.2 Hz, 1H), 5.54 (d, J = 6.2 Hz, 1H), 5.35 (d, J = 4.1 Hz, 1H), 4.60 (dt, J = 3.2, 6.8 Hz, 1H), 4.34 - 4.32 (m, 1H), 3.40 (dd, J = 4.1, 10.8 Hz, 1H), 2.78 (dd, J = 2.8, 10.8 Hz, 1H), 2.01 (s, 3H); 13C NMR (125 MHz, DMSO-d6): δ 155.4, 149.9, 145.3, 140.5, 118.3, 82.0, 80.2, 78.4, 72.1, 61.3, 34.3, 3.40; [α] 20 D = -38.0 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 12 H 14 N5O2S (M + H) +292.0868, Found: 292.0867. Anal. Calcd for C 12 H 13 N5O2S: C, 49.47; H, 4.50; N, 24.04. Found: C, 49.29; H, 4.21; N, 23.98.

[0082] * General procedure for the synthesis of compounds 9b - e To a stirred solution of diol compound 8a (50 mg, 0.16 mmol) in ethanol (5 mL) was added triethylamine (3 equiv) and 3 - halobenzylamine (1.5 equiv) at room temperature. The mixture was stirred at room temperature for 24 h. The solvent was evaporated over 48 h and the residue was purified by silica gel column chromatography (methylene chloride / methanol = 35:1) to give compounds 9b - e as white solids.

[0083] [Production Method 4] (2R,3R,4S)-2-(6-((3-Fluorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol ((2R,3R,4S)-2-(6-((3-Fluorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydro thiophene-3,4-diol; Compound 9b). Yield: 81%; white solid; mp 237 - 239 °C; UV (MeOH) λ max 275 nm; 11H NMR (500 MHz, DMSO-d6): δ 8.52 - 8.45 (m, 2H), 7.34 (q, J = 7.7 Hz, 1H), 7.15 - 7.03 (m, 3H), 5.86 (d, J = 7.2 Hz, 1H), 5.55 (d, J = 6.2 Hz, 1H), 5.36 (d, J = 4.0 Hz, 1H), 4.68 - 4.61 (m, 3H), 4.34 (bs, 1H), 3.39 (dd, J = 3.9, 10.7 Hz, 1H), 2.80 (dd, J = 2.6, 10.8 Hz, 1H), 2.01 (s, 3H); 13C NMR (125 MHz, DMSO-d6): δ 163.1, 161.1, 153.9, 149.3, 145.5, 142.9, 140.6, 130.2, 122.9, 118.7, 113.5 (CF), 81.9, 80.7, 78.4, 72.1, 61.3, 42.3, 34.3, 3.4; [α] 20 D = -32.0 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 19 H 19 FN5O2S (M + H) + 400.1244, Found: 400.1249. Anal. Calcd for C 19 H 18 FN5O2S: C, 57.13; H, 4.54; N, 17.53. Found: C, 57.24; H, 4.94; N, 17.13.

[0084] [Production Method 5] (2R, 3R, 4S)-2-(6-((3-Chlorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol ((2R, 3R, 4S)-2-(6-((3-Chlorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydro thiophene-3,4-diol; Compound 9c). Yield: 78%; white solid; mp 224 - 226 °C; UV (MeOH) λ max 275 nm; 11H NMR (500 MHz, DMSO-d6): δ 8.51 (s, 1H), 8.47 (bs, 1H), 7.37 - 7.28 (m, 4H), 5.86 (d, J = 7.2 Hz, 1H), 5.54 (d, J = 6.2 Hz, 1H), 5.34 (d, J = 4.0 Hz, 1H), 4.67 (bs, 2H), 4.61 (s, 1H), 4.34 (s, 1H), 3.40 (dd, J = 3.9, 10.7 Hz, 1H), 2.80 (dd, J = 2.6, 10.8 Hz, 1H), 2.02 (s, 3H); 13C NMR (125 MHz, DMSO-d6): δ 153.9, 149.3, 145.5, 142.4, 140.6, 132.8, 130.1, 126.8, 126.5, 125.6, 118.7, 81.9, 80.7, 78.4, 72.1, 61.3, 42.2, 34.3, 3.47; [α] 20 D = -42.0 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 19 H 19 ClN5O2S (M + H) + 416.0948, Found: 416.0952. Anal. Calcd for C 19 H 18 ClN5O2S: C, 54.87; H, 4.36; N, 16.84. Found: C, 54.76; H, 4.46; N, 16.45.

[0085] [Production Method 6] (2R, 3R, 4S)-2-(6-((3-Bromobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol ((2R, 3R, 4S)-2-(6-((3-Bromobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydro thiophene-3,4-diol; Compound 9d). Yield: 79%; white solid; mp 240 - 242 °C; UV (MeOH) λ max 275 nm; 11H NMR (500 MHz, DMSO-d6): δ 8.51 (s, 1H), 8.47 (bs, 1H), 7.52 (s, 1H), 7.42 (d, J = 7.6 Hz, 1H), 7.32 - 7.25 (m, 2H), 5.86 (d, J = 7.2 Hz, 1H), 5.53 (d, J = 6.2 Hz, 1H), 5.35 (d, J = 4.0 Hz, 1H), 4.67 (bs, 2H), 4.61 (bs, 1H), 4.34 (s, 1H), 3.40 (dd, J = 3.9, 10.7 Hz, 1H), 2.80 (dd, J = 2.6, 10.8 Hz, 1H), 2.02 (s, 3H); 13C NMR (125 MHz, DMSO-d6): δ 153.9, 149.3, 145.5, 142.7, 140.6, 130.4, 129.7, 129.5, 126.1, 121.5, 118.7, 81.9, 80.7, 78.4, 72.1, 61.3, 42.1, 34.3, 3.45; [α] 20 D = 1.40 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 19 H 19 BrN5O2S (M + H) + 462.0424, Found: 462.0446. Anal. Calcd for C 19 H 18 BrN5O2S: C, 49.57; H, 3.94; N, 15.21. Found: C, 49.17; H, 4.12; N, 15.43.

[0086] [Production Method 7] (2R, 3R, 4S)-2-(6-((3-Iodobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol ((2R, 3R, 4S)-2-(6-((3-Iodobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydro thiophene-3,4-diol; Compound 9e). Yield: 77%; white solid; mp 253 - 255 °C; UV (MeOH) λ max 275 nm; 11H NMR (500 MHz, DMSO-d6): δ 8.50 (s, 1H), 8.45 (bs, 1H), 7.71 (s, 1H), 7.58 (d, J = 7.2 Hz, 1H), 7.33 (d, J = 7.6 Hz, 1H), 7.11 (t, J = 7.7 Hz, 1H), 5.86 (d, J = 7.1 Hz, 1H), 5.54 (d, J = 6.1 Hz, 1H), 5.34 (d, J = 3.8 Hz, 1H), 4.63 (bs, 3H), 4.34 (s, 1H), 3.41 (dd, J = 3.8, 10.6 Hz, 1H), 2.80 (dd, J = 2.6, 10.8 Hz, 1H), 2.02 (s, 3H); 13C NMR (125 MHz, DMSO-d6): δ 153.9, 149.3, 145.5, 142.5, 140.6, 135.7, 135.4, 130.4, 126.5, 118.7, 94.7, 81.9, 80.7, 78.4, 72.1, 61.3, 40.0, 34.3, 3.4; [α] 20 D = 107.0 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 19 H 19 IN5O2S (M + H) + 508.0304, Found: 508.0312. Anal. Calcd for C 19 H 18 IN5O2S: C, 44.98; H, 3.58; N, 13.80. Found: C, 45.13; H, 3.18; N, 14.12.

[0087] [Production Method 8] (2R, 3R, 4S)-2-(6-Amino-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol ((2R, 3R, 4S)-2-(6-Amino-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; Compound 9f) Compound 8b (50 mg, 0.15 mmol) was converted to compound 9f (40 mg, 84%) as a white solid according to the same procedure used for the production of compound 9a: mp 202 - 204 °C; UV (MeOH) λ max 275 nm; 11H NMR (500 MHz, DMSO-d6): δ 8.47 (s, 1H), 7.36 (s, 1H), 5.86 (d, J = 7.2 Hz, 1H), 5.55 (d, J = 5.2 Hz, 1H), 5.37 (bs, 1H), 4.59 - 4.60 (m, 1H), 4.34 (bs, 1H), 3.41 - 3.35 (m, 2H; merged with water peak), 2.79 (dd, J = 2.4, 10.0 Hz, 1H), 2.40 (q, J = 7.5 Hz, 2H), 1.15 (t, J = 7.5 Hz, 3H); 13C NMR (125 MHz, DMSO-d6): δ 155.6, 149.9, 145.7, 140.4, 118.2, 86.7, 80.6, 78.5, 72.1, 61.1, 34.3, 13.3, 11.9; [α] 20 D = 90.00 (c 0.12, MeOH); HRMS (FAB+): m / z calculated for C 13 H 16 N5O2S (M + H) + 306.1025, Found: 306.1011. Anal. Calcd for C 13 H 15 N5O2S: C, 51.13; H, 4.95; N, 22.94. Found: C, 51.42; H, 4.55; N, 23.10.

[0088] * General procedure for the synthesis of compounds 9g - j. To a stirred solution of diol compound 8b (50 mg, 0.15 mmol) in ethanol (5 mL) were added triethylamine (3 equiv) and 3 - halobenzylamine (1.5 equiv) at room temperature, and the mixture was stirred at room temperature for 24 h. The solvent was evaporated in 48 h, and the residue was purified by silica gel column chromatography (methylene chloride / methanol = 20:1) to afford compounds 9g - j as white solids.

[0089] [Production Method 9] (2R,3R,4S)-2-(2-(But-1-yn-1-yl)-6-((3-fluorobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; Compound 9g Yield: 80%; white solid; mp 230 - 231 °C; UV (MeOH) λ max 275 nm; 1 1H NMR (500 MHz, DMSO-d6): δ 8.52 (s, 1H), 8.46 (bs, 1H), 7.35 (q, J = 7.7 Hz, 1H), 7.16 - 7.11 (m, 2H), 7.06 - 7.02 (m, 1H), 5.88 (d, J = 7.3 Hz, 1H), 5.55 (d, J = 6.2 Hz, 1H), 5.36 (d, J = 3.8 Hz, 1H), 4.69 (bs, 2H), 4.60 (bs, 1H), 4.34 (s, 1H), 3.41 (dd, J = 3.9, 10.7 Hz, 1H), 2.80 (dd, J = 2.6, 10.8 Hz, 1H), 2.40 (q, J = 7.5 Hz, 2H), 1.15 (t, J = 7.5 Hz, 3H); 13C NMR (125 MHz, DMSO-d6): δ 163.1, 161.2, 153.9, 149.4, 145.6, 142.9, 140.5, 130.1, 123.0, 118.6, 113.5 (CF), 86.9, 80.9, 78.5, 72.1, 61.1, 42.3, 34.3, 13.2, 11.9; [α] 20 D = -15.10 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 20 H 21 FN5O2S (M + H) + At 414.1400, Found: 414.1396. Anal. Calcd for C 20 H 20 FN5O2S: C, 58.10; H, 4.88; N, 16.94. Found: C, 57.99; H, 5.12; N, 16.44.

[0090] [Preparation Method 10] (2R,3R,4S)-2-(2-(But-1-yn-1-yl)-6-((3-chlorobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol (2R,3R,4S-2-(2-(But-1-yn-1-yl)-6-((3-chlorobenzyl)amino)-9H-purin-9-yl)tetrahydrothio -phene-3,4-diol; Compound 9h). Yield: 78%; white solid; mp 236 - 238 °C; UV (MeOH) λ max 275 nm; 1 1H NMR (500 MHz, DMSO-d6): δ 8.52 (s, 1H), 8.47 (bs, 1H), 7.38 - 7.27 (m, 4H), 5.88 (d, J = 7.2 Hz, 1H), 5.56 (d, J = 6.1 Hz, 1H), 5.37 (d, J = 3.5 Hz, 1H), 4.68 (bs, 2H), 4.60 (s, 1H), 4.34 (bs, 1H), 3.41 (dd, J = 3.8, 10.6 Hz, 1H), 2.80 (dd, J = 2.4, 10.6 Hz, 1H), 2.40 (q, J = 7.5 Hz, 2H), 1.15 (t, J = 7.5 Hz, 3H); 13C NMR (125 MHz, DMSO-d6): δ 153.9, 149.4, 145.6, 142.4, 140.5, 132.8, 130.1, 126.9, 126.6, 125.7, 118.6, 86.9, 80.8, 78.5, 72.1, 61.1, 42.2, 34.3, 13.2, 11.9; [α] 20 D = -28.0 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 20 H 21 ClN5O2S (M + H) + 430.1104, Found: 430.1111. Anal. Calcd for C 20 H 20 ClN5O2S: C, 55.88; H, 4.69; N, 16.29. Found: C, 55.54 H, 4.25; N, 16.29.

[0091] [Preparation 11] (2R,3R,4S)-2-(6-((3-Bromobenzyl)amino)-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol ((2R,3R,4S)-2-(6-((3-Bromobenzyl)amino)-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothio -phene-3,4-diol; Compound 9i). Yield: 77%; white solid; mp 222 - 224 °C; UV (MeOH) λ max 275 nm; 1 H NMR (500 MHz, DMSO-d6): δ 8.52 (s, 1H), 8.48 (bs, 1H), 7.53 (s, 1H), 7.42 (d, J = 7.6 Hz, 1H), 7.33 - 7.25 (m, 2H), 5.87 (d, J = 5.2 Hz, 1H), 5.54 (d, J = 5.2 Hz, 1H), 5.36 (bs, 1H), 4.67 (bs, 1H), 4.60 (bs, 2H), 4.34 (bs, 1H), 3.40 (dd, J = 3.9, 10.7 Hz, 1H), 2.80 (dd, J = 2.5, 10.7 Hz, 1H), 2.41 (q, J = 7.4 Hz, 2H), 1.16 (t, J = 7.5 Hz, 3H); 13C NMR (125 MHz, DMSO-d6): δ 153.9, 149.4, 145.6, 142.7, 140.5, 130.4, 129.8, 129.5, 126.1, 121.5, 118.6, 87.0, 80.8, 78.5, 72.1, 61.1, 42.2, 34.3, 13.2, 11.9; [α] 20 D = -9.90 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 20 H 21 BrN5O2S (M + H) + 476.0580, Found: 476.0594. Anal. Calcd for C 20 H 20 BrN5O2S: C, 50.64; H, 4.25; N, 14.76. Found: C, 50.76; H, 4.32; N, 14.98.

[0092] [Preparation 12] (2R,3R,4S)-2-(2-(But-1-yn-1-yl)-6-((3-Iodobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-phene-3,4-diol; Compound 9j). Yield: 77%; white solid; mp 239 - 241 °C; UV (MeOH) λ max 275 nm; 1 1H NMR (500 MHz, DMSO-d6): δ 8.52 (s, 1H), 8.45 (bs, 1H), 7.72 (s, 1H), 7.58 (d, J = 7.6 Hz, 1H), 7.33 (d, J = 7.3 Hz, 1H), 7.11 (t, J = 7.7 Hz, 1H), 5.88 (d, J = 7.3 Hz, 1H), 5.55 (d, J = 6.1 Hz, 1H), 5.36 (d, J = 3.9 Hz, 1H), 4.63 - 4.60 (m, 3H), 4.30 (bs, 1H), 3.41 (dd, J = 3.9, 10.7 Hz, 1H), 2.81 (dd, J = 2.7, 10.8 Hz, 1H), 2.41 (q, J = 7.4 Hz, 2H), 1.16 (t, J = 7.5 Hz, 3H); 13C NMR (125 MHz, DMSO-d6): δ 153.8, 149.3, 145.6, 142.5, 140.5, 135.8, 135.3, 130.4, 126.5, 118.6, 94.6, 86.9, 80.8, 78.5, 72.1, 61.1, 42.1, 34.3, 13.2, 11.9; [α] 20 D = -13.60 (c 0.10, MeOH); HRMS (FAB+): m / z calculated for C 20 H 21 IN5O2S (M + H) + 522.0461, Found: 522.0468. Anal. Calcd for C 20 H 20 IN5O2S: C, 46.07; H, 3.87; N, 13.43. Found: C, 46.23; H, 3.47; N, 13.95.

[0093] <2-1>Profile of LJ-4378 or its derivatives LJ-4378 is a substance with excellent binding ability to A 2A AR and A3AR, and showed very high subtype selectivity. Also, the most important A 2A Evaluation results of the cAMP activation ability of AR, showing an EC similar to that of the full agonist CGS21680 50 and efficacy. In the case of A3AR, as a result of experiments with Cl-IB-MECA, which is an A3AR full agonist, high concentrations of LJ-4378 act as a full antagonist that can perfectly inhibit cAMP production, and the K B value showed high efficacy at the nM level (Figure 1c). That is, LJ-4378, which acts strongly as an agonist and an antagonist on A 2A AR and A3AR respectively, could effectively increase cAMP production.

[0094] The binding affinities of LJ-4378 or its derivatives for A 2A AR and A3AR are as shown in Table 1 below. All binding affinity experiments were performed using adherent HEK293 cells stably transfected with cDNA encoding appropriate hAR binding, and were performed using 1 nM [3H]17, 10 nM [3H]18, or 0.2 nM [3H]16 as radioactive ligands for A 2A and A3AR respectively. The values were normalized against the non-specific binder NECA (5’-N-ethylcarboxamidoadenosine, 10 μM).

[0095]

Table 1

[0096] [Example 3] Volume-dependent effects of LJ-4378 on lipolysis and mitochondrial content <3-1>Cytotoxicity LJ-4378 is A 2AIt is a dual - acting ligand with the activities of an AR agonist and an A3AR antagonist. EZ - CYTOX (DoGEN, EZ - 3000) was used to evaluate cell viability according to the manufacturer's guidelines. Absorbance was measured at 450 nm using a microplate reader (Thermo MULTISKAN GO, 8816 - 2015).

[0097] Analysis of the cytotoxicity of LJ - 4378 showed that concentrations below 100 μM did not affect the cell viability of brown adipocytes (Figure 2(a)).

[0098] <3-2>Lipolytic effect of LJ-4378 A 2A Since AR increases the level of cAMP in adipocytes and activates protein kinase A signaling, the lipolytic effect of LJ - 4378 was examined in a dose - response relationship.

[0099]

Table 2

[0100] In the case of Western blot, the primary antibodies used in the Western blot are summarized in Table 2 above. Anti - rabbit horseradish peroxidase (1:3000, Thermo Fisher, 31460) and anti - mouse horseradish peroxidase (1:3000, Jackson, 115 - 035 - 174) were used as secondary antibodies. All antibodies were diluted in blocking buffer (5% non - fat milk or 5% bovine serum albumin in TBST). Western blot images were acquired using a Fusion Solo chemiluminescence imaging system (Vilber Lourmat) and analyzed with EvolutionCapt software (version 17.03). NIH ImageJ software was used for quantification.

[0101] The level of free fatty acid (FFA) in the medium was measured using a NEFA reagent (WAKO, 436 - 91693) according to the manufacturer's product protocol.

[0102] Immunoblot analysis showed that LJ-4378 upregulated the expression level of P-HSL (phosphorylated hormone-sensitive lipase) at serine 660 in a dose-dependent manner (Figure 2(c): (10 -11 M) 1.55 ± 0.11-fold; (10 -9 M) 3.03 ± 0.21-fold, (10 -7 ) 6.64 ± 0.34-fold, (10 -5 ) 6.87 ± 0.69-fold increase). Consistently, the level of free fatty acid (FFA) was also increased by LJ-4378 (Figure 2(d): 1.92 ± 0.03-fold increase).

[0103] In addition, mitochondrial proteins were increased by the dose of LJ-4378 (Figure 2(e): ATP5A (ATP synthase alpha-subunit): (10 -11 M) 2.20 ± 0.08-fold, (10 -9 M) 3.51 ± 0.05-fold, (10 -7 ) 3.94 ± 0.06, (10 -5 ) 3.72 ± 0.04-fold increase, UQCRC2 (ubiquinol-cytochrome c reductase core protein 2): (10 -11 M) 2.15 ± 0.03-fold, (10 -9 M) 3.96 ± 0. (10 -7 ) 4.21 ± 0.13-fold, (10 -5 ) 4.02 ± 0.08-fold increase, SDHB (succinate dehydrogenase complex iron sulfur subunit B): (10 -11 M) 1.87 ± 0.28-fold, (10 -9 M) 3.47 ± 0.18-fold, (10 -7 ) 3.41 ± 0.47-fold, (10 -5 ) 3.71 ± 0.25-fold increase, MCAD: (10 -11 M) 1.53 ± 0.03-fold, (10 -9 M) 1.79 ± 0.03-fold, (10 -7 ) 1.79 ± 0.09-fold, (10 -5)An increase of 2.13 ± 0.03 times. Based on such capacity-response evaluation, a concentration of 0.1 μM was used in subsequent experiments.

[0104] [Example 4] LJ-4378 and A 2A Comparison of an AR agonist or an A3AR antagonist <4-1>Confirmation of the expression level of adenosine derivatives qPCR analysis was performed to examine the expression levels of adenosine derivatives in brown adipocytes differentiated from immortalized brown adipose progenitor cells. The primers used for qPCR analysis are listed in Table 3 below.

[0105]

Table 3

[0106] A2AAR showed the highest expression (Figure 3a: 343.7 ± 68.04), and A3AR (Figure 3a: 0.19 ± 0.03) was the second highest among the adenosine derivatives examined.

[0107] <4-2>LJ-4378, A 2A AR agonist or A 3 Comparison of the lipolytic effects of AR antagonists The lipolytic effect of LJ-4378 was compared with that of A 2A an AR agonist and the A3AR antagonist GS21680 (Cayman, 124431-80-7) and LJ-4433, respectively.

[0108] The levels of glycerol and free fatty acids (FFA) in the medium were measured using a glycerol reagent (Sigma Aldrich, F6428) and a NEFA reagent (WAKO, 436-91693) according to the product protocol of the glycerol manufacturing company.

[0109] In both glycerol and FFA analyses, the treatment with LJ-4378 showed the largest fold changes of 1.61±0.07 and 1.59±0.1, respectively, compared to CGS21680 (Figure 3b: 1.51±0.08-fold; Figure 3c: 1.29±0.09-fold) and LJ-4433 (Figure 3b: 1.22±0.07-fold; Figure 3c: 0.92±0.09-fold) (Figure 3b, c). Immunoblot analysis further confirmed the additive effect of LJ-4378, as shown by the upregulated levels of PKA downstream proteins including phosphorylated cAMP response element-binding protein (P-CREB) (increased 2.32±0.05-fold by LJ-4378; increased 2.00±0.11-fold by CGS21680; increased 1.40±0.11-fold by LJ-4433), and P-HSL (increased 2.10±0.12-fold by LJ-4378, increased 1.54±0.05-fold by CGS21680, increased 1.22±0.07-fold by LJ-4433) (Figure 3d).

[0110] [Example 5] Effect of LJ-4378 on brown adipocytes The effect of LJ-4378 on the content and metabolic activity capacity of mitochondria in brown adipocytes was evaluated.

[0111] Oxygen consumption rate (OCR) was measured with an XFp analyzer. Cells were cultured in XF DMEM basal medium (Agilent, 103575-100, pH 7.4) supplemented with 4 mM L-glutamine (Sigma, G8540) and 25 mM D-glucose (Sigma, G7021) at 37°C. The XFp Cell Mito Stress Test Kit (Agilent, 103010-100) was prepared and calculated sequentially with optimal final concentrations of 2.5 μM oligomycin, 0.5 μM FCCP, and 0.5 μM rotenone / antimycin A, and basal, maximal, and proton leak. Next, OCR was normalized by protein concentration.

[0112] Cells were stained with MitoTracker TMAfter staining mitochondria by exposing them to Red CMXRos (1:3000, Invitrogen, M7512) at 37°C for 15 minutes, they were fixed with 4% paraformaldehyde (PFA, Sigma, 158127). Images were obtained using an LSM800 confocal microscope (Zeiss, Germany) and analyzed with Zen software (version 3.0).

[0113] The expression levels of the brown adipose cell marker UCP1 and the mitochondrial protein COXIV were significantly increased by 2.93 ± 0.23-fold and 2.18 ± 0.15-fold, respectively, by the treatment with LJ-4378 (Figure 4a). MitoTracker staining also showed that LJ-4378 increased the mitochondrial membrane potential by 2.12 ± 0.06-fold in brown adipose cells (Figure 4b). In addition, the inventors compared such an effect of LJ-4378 on mitochondrial content with that of the A2AAR agonist (CGS21680) and the A3AR antagonist (LJ-4433). Both CGS21680 and LJ-4433 increased UCP1, COXIV, and MCAD (Figure 4d). However, a greater effect was observed with LJ-4378 (Figure 4d: 2.67 ± 0.38-fold increase in UCP1, 2.53 ± 0.08-fold increase in COXIV, 4.91 ± 0.02-fold increase in MCAD), suggesting an additional effect.

[0114] As a result of examining the oxygen consumption rate for functional analysis, it was shown that LJ-4378 increased the OCR associated with basal (30.5%), maximal (49.1%), and ATP production (29.3%) (Figure 4c).

[0115] [Example 6] Effect of LJ-4378 on Mitochondrial Activity in Adipose Tissue and In Vivo Energy Consumption The in vitro electron transfer activity associated with mitochondrial oxidative phosphorylation was evaluated by monitoring the reduction of 0.1% triphenyltetrazolium chloride (TTC, Sigma, T8877).

[0116] Indirect calorimetry was performed using a PhenoMaster (TSE Systems) to measure energy expenditure (EE), VO2, VCO2, activity, and food intake. Body composition was measured by nuclear magnetic resonance scanning on an EchoMRI-700 (Echo Medical Systems).

[0117] In situ staining using triphenyltetrazolium chloride (TTC) indicated that LJ-4378 upregulated mitochondrial activity in all adipose tissue depots as a redox indicator (Figure 5a: 2.86 ± 0.24-fold increase in BAT; 2.07 ± 0.17-fold increase in iWAT; 1.72 ± 0.16-fold increase in gWAT).

[0118] Indirect calorimetry analysis showed that LJ-4378 treatment increased oxygen consumption by 17.8% and energy consumption by 17.0% while activity and food intake were not affected (Figure 5b).

[0119] [Example 7] Effect of LJ-4378 on Browning of In Vivo Adipose Tissue The synergistic effect of LJ-4378 on browning of adipose tissue was examined in CIDEA reporter mice that reflect the expression of CIDEA, a brown adipocyte marker, via fluorescence and luminescence signals.

[0120] In vivo bioluminescence was detected using an optical imaging device (Ami-X, Spectral Instruments Imaging). To detect the in vivo bioluminescence signal, mice were intraperitoneally injected with D-luciferin (150 mg kg-1, Goldbio). Quantification was performed using Aura Software (Spectral Instruments Imaging, version 2.2.1.1). In addition, ex vivo imaging tissues were collected from CIDEA reporter mice treated with D-luciferin (150 mg kg-1, Goldbio, i.p.). During imaging, the isolated tissues were maintained in 12-well plates containing D-luciferin in PBS (300 μg / ml).

[0121] LJ-4378 significantly increased the bioluminescence intensity in BAT and iWAT (Figures 6a, b). In particular, iWAT increased significantly compared to CGS21680 and LJ-4433, indicating that LJ-4378 has an additional effect on browning in white adipose tissue. The results of in vivo bioluminescence showed that LJ-4378 treatment resulted in the most significant increase in the ex vivo luminescence signal in BAT and iWAT (Figure 6c).

[0122] The mRNA levels of brown adipocyte markers were also examined, and consistent results showed that LJ-4378 produced the largest increase in gene expression related to browning, including Ucp1 (increased 2.13 ± 0.21-fold by LJ-4378, 1.72 ± 0.17-fold by CGS21680, 1.74 ± 0.08-fold by LJ-4433), and Cidea (increased 1.72 ± 0.06-fold by LJ-4378, 1.29 ± 0.12-fold by CGS21680, 1.12 ± 0.04-fold by LJ-4433, increased by LJ-4433).

[0123] [Example 8] The anti-obesity effect of LJ-4378 It was investigated whether the lipolytic and browning effects of LJ-4378 have an anti-obesity effect.

[0124] Mice fed a high-fat diet (HFD) were used as a model of diet-induced obesity. During the treatment, changes in body weight were monitored, and after the treatment, the fat and body composition of the mice were measured.

[0125] For histological analysis, adipose tissue was fixed with 10% formalin and then embedded in paraffin blocks. Paraffin sections were stained with hematoxylin / eosin (H&E).

[0126] For the glucose tolerance test (GTT), 20% D-Glucose (1 g kg-1, Sigma Aldrich, G7021) was injected intraperitoneally into the mice. Blood glucose levels were measured using a blood glucose meter (Gluco Dr.Top, allmedicus, AGM-4100).

[0127] Mice fed the HFD treated with LJ-4378 were shown to have an 8.24% decrease in body weight and a 24.18% decrease in fat mass (Figures 7a, b). Also, the weights of iWAT and gWAT were significantly decreased by LJ-4378 treatment in HFD-fed mice (Figure 7c: 30.1% decrease in iWAT, 18.6% decrease in gWAT). The decreased adipocyte size in iWAT and gWAT was further supported by H&E staining (Figures 7d, e). Additionally, LJ-4378 improved glucose tolerance in HFD-fed mice (Figure 7f).

[0128] Next, the inventors examined the expression levels of mitochondrial proteins and PKA downstream markers to confirm whether such anti-obesity and anti-diabetic effects were induced by increased mitochondrial content and activation of the PKA signaling pathway. As a result, LJ-4378 upregulated COXIV (BAT: 5.91 ± 0.23-fold, iWAT: 1.24 ± 0.04-fold), UCP1 (BAT: 1.33 ± 0.06-fold, iWAT: 3.34 ± 0.36-fold), P-CREB (BAT: 1.54 ± 0.03-fold; iWAT: 3.86 ± 0.12-fold), and P-HSL (BAT: 2.31 ± 0.18-fold, iWAT: 1.72 ± 0.04-fold) in BAT and iWAT of diet-induced obese mice (Figure 8).

[0129] [Data and Statistical Analysis] Statistical analysis was performed using Prism 7 software (GraphPad Software, USA). Data were expressed as mean ± SEM. An unpaired t-test was used to measure statistical significance between two groups. [Industrial Applicability]

[0130] LJ-4378 or its derivatives, which are ligands acting on the adenosine receptor of the present invention, 2A act doubly as an agent acting on A [Sequence Listing Free-Text]

[0131] SEQ ID NO: 1 shows the forward primer used for qPCR analysis of Ucp1. SEQ ID NO: 2 shows the reverse primer used for qPCR analysis of Ucp1. SEQ ID NO: 3 shows the forward primer used for qPCR analysis of Cidea. SEQ ID NO: 4 shows the reverse primer used for qPCR analysis of Cidea. SEQ ID NO: 5 shows the forward primer used for qPCR analysis of Ppia. SEQ ID NO: 6 shows the reverse primer used for qPCR analysis of Ppia. SEQ ID NO: 7 shows the forward primer used for qPCR analysis of Adora1. SEQ ID NO: 8 shows the reverse primer used for qPCR analysis of Adora1. SEQ ID NO: 9 represents the forward primer used in the qPCR analysis for Adora2a. SEQ ID NO: 10 represents the reverse primer used in the qPCR analysis for Adora2a. SEQ ID NO: 11 represents the forward primer used in the qPCR analysis for Adora2b. SEQ ID NO: 12 represents the reverse primer used in the qPCR analysis for Adora2b. SEQ ID NO: 13 represents the forward primer used in the qPCR analysis for Adora3. SEQ ID NO: 14 represents the reverse primer used in the qPCR analysis for Adora3.

Claims

1. A comprising a compound represented by the following chemical formula 1, an isomer thereof, or a pharmaceutically acceptable salt ２Ａ adenosine receptor (AR) and / or A ３ Pharmaceutical composition for preventing or treating adenosine receptor (AR)-related diseases: 【Chemical 1】 R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group, and R2 is a C1-C10 alkyl group.

2. R1 is H, a methyl group, or an aryl-C1-C10 alkyl group, and R2 is a C1-C4 alkyl group, and the pharmaceutical composition according to Claim 1.

3. The compound represented by the chemical formula 1, its isomers, or a pharmaceutically acceptable salt thereof is (1) 2-(6-amino-2-(hex-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (2) (2R,3R,4S)-2-(6-amino-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (3) (2R,3R,4S)-2-(6-((3-fluorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (4) (2R,3R,4S)-2-(6-((3-chlorobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (5) (2R,3R,4S)-2-(6-((3-bromobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (6) (2R,3R,4S)-2-(6-((3-iodobenzyl)amino)-2-(prop-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (7) (2R,3R,4S)-2-(6-amino-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; (8) (2R,3R,4S)-2-(2-(but-1-yn-1-yl)-6-((3-fluorobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-furan-3,4-diol; (9) (2R,3R,4S)-2-(2-(but-1-yn-1-yl)-6-((3-chlorobenzyl)amino)-9H-purin-9-yl)tetrahydrothio-furan-3,4-diol; (10) (2R,3R,4S)-2-(6-((3-bromobenzyl)amino)-2-(but-1-yn-1-yl)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; and (11) (2R,3R,4S)-2-(2-(but-1-yn-1-yl)-6-((3-iodobenzyl)amino)-9H-purin-9-yl)tetrahydrothiophene-3,4-diol; The pharmaceutical composition according to claim 1, characterized in that it is any one or more selected from the group consisting of.

4. The compound represented by the above chemical formula 1, its isomers, or a pharmaceutically acceptable salt thereof is A ２Ａ an agonist for an adenosine receptor (AR), and A ３ The pharmaceutical composition according to claim 1, which is an antagonist for an adenosine receptor (AR).

5. Said A ２Ａ An adenosine receptor (AR) and / or A ３ The pharmaceutical composition according to claim 1, characterized in that the adenosine receptor (AR)-related disease is obesity.

6. A health functional food composition for preventing or improving obesity, comprising a compound represented by the following Chemical Formula 1, an isomer thereof, or a pharmaceutically acceptable salt: 【Chemical 2】 wherein R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group, and R2 is a C1-C10 alkyl group.

7. A ２Ａ Adenosine receptor (AR) and / or A ３ Administering to a patient suffering from an adenosine receptor (AR) and / or A ２Ａ Adenosine receptor (AR) related disease a compound represented by the following chemical formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof, the method for treating an A ３ Adenosine receptor (AR) related disease: 【Chemical 3】 wherein R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group, and R2 is a C1-C10 alkyl group.

8. A ２Ａ Adenosine receptor (AR) and / or A ３ Use of a compound represented by the following Chemical Formula 1, an isomer thereof, or a pharmaceutically acceptable salt thereof for use in the manufacture of a medicament for treating adenosine receptor (AR) - related diseases: 【Chemical Formula 4】 wherein R1 is H, a C1-C10 alkyl group, or an aryl-C1-C10 alkyl group, and R2 is a C1-C10 alkyl group.

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