WO2025116596A1 - Novel compound, and composition for preventing or treating eye diseases, comprising same - Google Patents

Abstract

The present invention relates to a novel compound, and a composition for preventing or treating eye diseases, comprising same, and, more specifically, to: a series of phenyl-triazole derivative compound or a pharmaceutically acceptable salt thereof; a pharmaceutical composition for preventing or treating eye disease, comprising the compound; a method for preventing or treating eye diseases, comprising administering the pharmaceutical composition; an autophagy activation composition comprising the compound or a pharmaceutically acceptable salt thereof; a food composition for preventing or alleviating eye diseases; and a feed composition.

Description

Novel compound and composition containing same for preventing or treating eye disease

The present invention relates to a novel compound and a composition for preventing or treating ocular diseases comprising the same, and more particularly, to a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof; a pharmaceutical composition for preventing or treating ocular diseases comprising the compound; a method for preventing or treating ocular diseases comprising administering the pharmaceutical composition; an autophagy activating composition comprising the compound or a pharmaceutically acceptable salt thereof; a food composition for preventing or improving ocular diseases; and a feed composition:

[Chemical Formula 1]

Light can be divided into ultraviolet rays, visible light, and infrared rays according to wavelength. Visible light is light that can be commonly seen with the human eye, and its wavelength corresponds to the range of 380–780 nm. Among these, the blue series that exists between 380 and 500 nm can be classified as blue light, and this short-wavelength blue light has high energy and passes through the lens to affect retinal epithelial cells.

Humans are constantly exposed to blue light because they have to adapt to the constant changes in light both indoors and outdoors, which causes eye fatigue to increase and can lead to a vicious cycle of macular degeneration and decreased eyesight. In addition, as the use of electronic devices has increased due to recent changes in the living environment, research is being conducted on the harmful effects on the human body due to unprotected exposure to blue light emitted from displays of smart devices such as TVs, computers, and smartphones as well as LED lighting devices.

Moreover, the smartphone usage rate among adults in our country was 97% as of 2022, the highest among the countries surveyed, and the smartphone distribution and usage age is also decreasing every year.

The nerve tissue located in the center of the retina inside the eye is called the macula, and since most of the photoreceptor cells are gathered here and the image of an object is formed in the center of the macula, it plays a very important role in vision. The macula can cause vision impairment due to various causes such as aging, genetic factors, toxicity, and inflammation, which is called macular degeneration, and in severe cases, it is a disease that can lead to blindness.

In addition, it is known that the accumulation of lipofuscin in the central part of the retina is largest in the retinal pigment epithelial cells beneath the central part of the retina. N-retinyl N-retinylidene ethanolamine (A2E) is the main chromophore of lipofuscin, and causes the production of reactive oxygen species when exposed to blue light. In addition, visual impairment due to retinal cell dysfunction is closely related to the oxidation of A2E, and therefore can be a major cause of eye diseases including macular degeneration. Meanwhile, autophagy is a self-digestive system in which cellular components are decomposed and recycled as nutrients and energy sources. Aged or dysfunctional cell organelles and damaged or improperly folded proteins are the targets of autophagy, and it regulates the protein degradation process that is essential to maintain cellular homeostasis and genetic stability. It plays a central role in maintaining cellular homeostasis by regulating chemical metabolism, mitigating the production of reactive oxygen species (ROS) by regenerating damaged or long-lived mitochondria, and protecting cells from intracellular and extracellular stimuli such as nutrient deficiency, oxygen depletion, invasion of pathogens including bacteria and viruses, and UV exposure. Consequently, if autophagy is not properly regulated, various diseases related to cellular homeostasis can occur.

As a prior study, Korean Patent Publication No. 10-2023-0140410 disclosed a 3-phenylisoxazole derivative and a pharmaceutical composition containing the same as an active ingredient for preventing or treating ocular diseases.

Against this backdrop, as a result of research efforts on compounds that are effective in treating diseases related to autophagy, especially ophthalmic diseases such as dry macular degeneration, dry eye, eye fatigue, and decreased vision caused by aging and blue light, the present invention was completed by confirming that a 4-phenyl-triazole derivative compound represented by chemical formula 1 inhibits death of human retinal pigment epithelial cells (ARPE-19) induced by A2E and blue light, removes A2E fluorescent marker and A2E in ARPE-19, and activates autophagy.

One object of the present invention is to provide a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

.

Another object of the present invention is to provide a method for producing the compound or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating ocular diseases comprising the compound or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a method for preventing or treating an ocular disease, comprising a step of administering the pharmaceutical composition.

Another object of the present invention is to provide an autophagy activating composition comprising the compound or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a food composition for preventing or improving ocular diseases, comprising the compound or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a feed composition for preventing or treating ocular diseases comprising the compound or a pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a use of the compound or a pharmaceutically acceptable salt thereof for preventing, improving or treating ocular diseases.

This is explained specifically as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. That is, all combinations of various elements disclosed in the present invention fall within the scope of the present invention. In addition, the scope of the present invention cannot be considered limited by the specific description described below.

One aspect of the present invention to achieve the above object provides a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

In the above chemical formula 1,

R ₁ is hydrogen, C _1-4 alkyl, carbamoyl-C _1-4 alkyl, carboxy-C _1-4 alkyl, C _1-4 alkylcarbonyl, C _1-4 alkoxycarbonyl-C _1-4 alkyl, C _6-10 arylcarbonyl-C _1-4 alkyl, 5-10 membered heteroaryl-C _1-4 alkyl, 3 to 10 membered heterocyclyl-C _1-4 alkyl, 3 to 10 membered heterocyclylcarbonyl, C _3-10 cycloalkylcarbamoyl-C _1-4 alkyl, or (C _1-4 alkoxycarbonyl)(C _6-10 aryl-C _1-4 alkyl)carbamoyl-C _1-4 alkyl,

R ₂ is halogen, C _1-4 alkyl, C _2-4 alkenyl, C _3-10 cycloalkyl, C _3-10 cycloalkenyl, C _6-10 aryl, or 5-10 membered heteroaryl,

R ₃ is C _1-4 alkyl, C _2-4 alkenyl, C _3-10 cycloalkyl-C _1-4 alkyl, or C _6-10 aryl-C _1-4 alkyl,

The above C _3-10 cycloalkyl, C _6-10 aryl, 3 to 10 membered heterocyclyl and 5 to 10 membered heteroaryl may be unsubstituted or substituted with one or more selected from the group consisting of hydroxy, halogen, C _1-4 haloalkyl, C _1-4 alkylsulfonyl, and C _1-4 alkoxy.

For example, in the above chemical formula 1, R ₁ can be, but is not limited to, hydrogen, methyl, carbamoylmethyl, carboxymethyl, methylcarbonyl, ethoxycarbonylmethyl, ethoxycarbonylisopropyl, tert-butoxycarbonyl-methyl, methylsulfonyl-substituted phenylcarbonylmethyl, pyridinylmethyl, piperidinylmethyl, isopropylsulfonyl-substituted piperidinylmethyl, tert-butoxycarbonyl-substituted piperidinylmethyl, morpholinylcarbonyl, cyclohexylcarbamoylmethyl, hydroxy-substituted cyclohexylcarbamoylmethyl or (methoxycarbonyl)(phenylethyl)carbamoylmethyl.

For example, in the above chemical formula 1, R ₂ can be, but is not limited to, bromo, propyl, isobutyl, propenyl, cyclopentyl, cyclopentenyl, fluoro-substituted phenyl, pyridinyl or trifluoromethyl-substituted pyrazolyl.

For example, in the chemical formula 1, R ₃ can be, but is not limited to, propyl, propenyl, cyclobutylmethyl, or benzyl.

Specifically, the compound is

1-Benzyl-4-(5-bromo-2-methoxyphenyl)-1H-1,2,3-triazole

4-(5-allyl-2-methoxyphenyl)-1-benzyl-1H-1,2,3-triazole

4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenol

Ethyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate

4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenyl acetate

(R)-methyl 2-(2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetamido)-3-phenylpropanoate

(S)-methyl 2-(2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetamido)-3-phenylpropanoate

tert-butyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate

2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-1-(4-(methylsulfonyl)phenyl)ethanone),

2-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)pyridine),

3-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)pyridine),

4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)pyridine),

4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenyl morpholine-4-carboxylate

Ethyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-2-methylpropanoate

4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)piperidine),

2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetic acid

4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)-1-(isopropylsulfonyl)piperidine),

2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetamide,

2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-N-cyclohexylacetamide),

2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-N-(4-hydroxycyclohexyl)acetamide

tert-butyl 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)piperidine-1-carboxylate),

2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-bromophenol,

Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-bromophenoxy)acetate

Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-isobutylphenoxy)acetate

Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-cyclopentenylphenoxy)acetate

Ethyl 2-(3-(1-benzyl-1H-1,2,3-triazol-4-yl)-4'-fluorobiphenyl-4-yloxy)acetate

Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-(1-(trifluoromethyl)-1H-pyrazol-4-yl)phenoxy)acetate

Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-(pyridin-4-yl)phenoxy)acetate

Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-cyclopentylphenoxy)acetate

1-benzyl-4-(2-methoxy-5-propylphenyl)-1H-1,2,3-triazole

2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-propylphenol,

2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-propylphenyl acetate

Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-propylphenoxy)acetate

4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenyl acetate

Ethyl 2-(4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate

4-(2-methoxy-5-propylphenyl)-1-propyl-1H-1,2,3-triazole

4-propyl-2-(1-propyl-1H-1,2,3-triazol-4-yl)phenyl acetate

Ethyl 2-(4-propyl-2-(1-propyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate

4-(5-bromo-2-methoxyphenyl)-1-(cyclobutylmethyl)-1H-1,2,3-triazole

4-(5-allyl-2-methoxyphenyl)-1-(cyclobutylmethyl)-1H-1,2,3-triazole

4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenol

4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenyl acetate

Ethyl 2-(4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenoxy)acetate

1-(cyclobutylmethyl)-4-(2-methoxy-5-propylphenyl)-1H-1,2,3-triazole, or

Ethyl 2-(2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)-4-propylphenoxy)acetate may be, but is not limited to.

In the present invention, the term "alkyl" includes a straight-chain or branched-chain saturated hydrocarbon residue, unless otherwise specified. For example, "C1-6 alkyl" means an alkyl having a skeleton of 1 to 6 carbons. Specifically, C1-6 alkyl includes methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, t-pentyl, sec-pentyl, neopentyl, hexyl, and the like.

"Alkoxy" means a radical in which alkyl is substituted through an oxygen atom. Here, alkyl means a saturated hydrocarbon including both straight-chain or branched chain. For example, "C1-6 alkyl" means an alkyl having a skeleton of 1 to 6 carbons. Specifically, C1-6 alkyl includes methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, i-pentyl, t-pentyl, sec-pentyl, neopentyl, hexyl, and the like, and C6 alkyl is a fully saturated hydrocarbon having 6 carbons, including all structural isomers that can be formed as a saturated hydrocarbon having 6 carbons. C1-10 alkoxy includes C1-6 alkoxy, C1-3 alkoxy, or methoxy.

The term “cycloalkyl”, unless otherwise specified, means a cyclic hydrocarbon residue which is a saturated hydrocarbon residue forming a ring, and “3-6 membered cycloalkyl” means a cyclic hydrocarbon residue containing 3 to 6 carbon atoms as ring atoms.

The term “heterocyclyl”, unless otherwise specified, means a monovalent saturated moiety consisting of 1 to 3 rings containing one or more (e.g., 1-5, 1-4, 1-3, 1-2, 1) heteroatoms selected from N, O or S. It may be bicyclic or tricyclic containing 2 or 3 rings, wherein these 2 or 3 rings may be bridged, fused or spiro-shaped heterocycloalkyl. When consisting of multiple rings, the heteroatoms may be present in all or only some of the rings.

The term “cycloalkenyl” means a monovalent monocyclic group having at least one unsaturated double bond in the ring and lacking aromaticity.

The term “aryl” refers to an aromatic radical having a single ring or two or three fused hydrocarbon aromatic rings, and C6-10 aryl refers to an aromatic ring compound containing 6 to 10 carbons, including phenyl or naphthyl.

The term “heteroaryl”, unless otherwise specified, means an aromatic radical having a single ring or two or three fused rings (the remaining ring atoms being C) containing one or more (e.g., 1-5, 1-4, 1-3, 1-2, 1) heteroatoms selected from N, O or S as ring atoms. When composed of multiple rings, the heteroatoms may be present in all or only some of the rings.

The term “halogen” means a halogen atom such as F, Cl, Br, I, and when it is said to be substituted with “halogen”, it means substituted with one or more halogen atoms. In this case, the halogen atoms to be substituted can be selected within the range of 1 to 5.

The compound of the present invention may exist in the form of a pharmaceutically acceptable salt. As a salt, an acid salt formed by a pharmaceutically acceptable free acid is useful. The term "pharmaceutically acceptable salt" of the present invention means a salt form which is relatively nontoxic and harmless at a concentration that has an effective effect on a patient, and means any organic or inorganic addition salt of the compound represented by the chemical formula 1, the side effects due to which the salt does not reduce the beneficial effects of the compound represented by the chemical formula 1.

Acid addition salts are prepared by conventional methods, for example, by dissolving the compound in an excess of an aqueous acid solution and precipitating the salt using a water-miscible organic solvent, for example, methanol, ethanol, acetone or acetonitrile. Equimolar amounts of the compound and an acid or alcohol (e.g., glycol monomethyl ether) in water can be heated, and the mixture can then be evaporated to dryness, or the precipitated salt can be filtered off with suction.

At this time, organic acids and inorganic acids can be used as the free acid, and inorganic acids such as hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, and tartaric acid can be used, and organic acids such as methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, and hydroiodic acid can be used, but are not limited thereto.

In addition, a pharmaceutically acceptable metal salt can be prepared using a base. An alkali metal salt or an alkaline earth metal salt is obtained, for example, by dissolving a compound in an excess of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering out the undissolved compound salt, and then evaporating and drying the filtrate. At this time, sodium, potassium, or calcium salts are particularly suitable for preparing the metal salt, but are not limited thereto. In addition, a corresponding silver salt can be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

Pharmaceutically acceptable salts of the compounds of the present invention include salts of acidic or basic groups which may be present in the compounds of Formula 1, unless otherwise indicated. For example, pharmaceutically acceptable salts may include sodium, calcium and potassium salts of hydroxy groups, and other pharmaceutically acceptable salts of amino groups include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts, and the like, and can be prepared by methods for preparing salts known in the art.

The chemical formula 1 of the present invention is represented by The salt of the compound is a pharmaceutically acceptable salt, represented by the chemical formula 1. Any salt that exhibits pharmacological activity equivalent to that of the compound may be used without restriction.

In addition, the compound represented by the chemical formula 1 according to the present invention includes, without limitation, not only pharmaceutically acceptable salts thereof, but also solvates such as possible hydrates that can be prepared therefrom, and all possible stereoisomers. The solvates and stereoisomers of the compound represented by the chemical formula 1 can be prepared from the compound represented by the chemical formula 1 using methods known in the art.

Furthermore, the compound represented by the chemical formula 1 according to the present invention can be prepared in a crystalline or amorphous form, and when prepared in a crystalline form, can be optionally hydrated or solvated. The present invention may include not only a stoichiometric hydrate of the compound represented by the chemical formula 1 but also compounds containing various amounts of water. The solvate of the compound represented by the chemical formula 1 according to the present invention includes both a stoichiometric solvate and a non-stoichiometric solvate.

Another aspect of the present invention for achieving the above object provides a method for preparing a compound represented by the chemical formula 1 or a pharmaceutically acceptable salt thereof, comprising a first step of reacting 4-bromo- ₂ -ethynyl-1-methoxybenzene with a mixture of R ₃ '-X ₁ and NaN ₃ in the presence of CuBr and PMDETA (N,N,N',N",N"-pentamethyldiethylenetriamine) to prepare a phenyl-triazole derivative having R 3 ' substituted on the triazole ring.

At this time, the R ₃ ' may be R ₃ or a precursor thereof, a halogen X ₁ .

For example, in the manufacturing method of the present invention, the first step may be performed by adding a reactant other than the above to a mixture of R ₃ '-X ₁ and NaN ₃ mixed at room temperature and cooled to 0°C, reacting the mixture at the above temperature, and then heating the mixture to room temperature to perform additional reaction, but is not limited thereto.

For example, the manufacturing method of the present invention

Step a, wherein Br on the phenyl ring of the product obtained from the previous step is replaced with R ₂ ';

Step b of replacing OMe on the phenyl ring of the product obtained from the previous step with OH; and

It may further include at least one step selected from the group consisting of step c of replacing OH on the phenyl ring of the product obtained from step b with R ₁ ',

The above R ₁ ' and R ₂ ' may be R ₁ or a precursor thereof, and R ₂ or a precursor thereof, respectively, but are not limited thereto.

Specifically,

Step a is carried out by reacting i) R ₂ '-SnBu ₃ or ii) R ₂ '-boronic acid and Cs ₂ CO ₃ in the presence of Pd(PPh ₃ ) ₄ ,

Step b is performed by reacting B(X ₂ ) ₃ at -60°C to 0°C.

Step c is carried out i) in the presence of R ₁ '-X ₃ and K ₂ CO ₃ , or ii) in the presence of acetic anhydride or R ₁ '-X ₃ and Et ₃ N (triethylamine) and optionally DMAP (4-dimethylaminopyridine),

The above step a can be performed after step 1 or after step c, but is not limited thereto.

At this time, X ₂ and X ₃ can each independently be halogen.

For example, X ₁ , X ₂ , and X ₃ may each independently be bromo or chloro, but are not limited thereto.

For example, the step a may be performed by mixing the reactants, bubbling with nitrogen, and reacting at 80° C. to 110° C. under a nitrogen atmosphere, but is not limited thereto. At this time, a microwave may be used to achieve the reaction temperature, but is not limited thereto.

For example, in the step b, B(X ₂ ) ₃ may be BCl ₃ or BBr ₃ , and the reaction may be performed under a nitrogen atmosphere, but is not limited thereto.

For example, the step c above can be performed at 50°C to 70°C in the case of i), at 0°C to 40°C in the case of ii), and i), ii) or both can be performed under a nitrogen atmosphere, but is not limited thereto.

In addition, the manufacturing method of the present invention may additionally include a step d of reducing R ₂ ', R ₃ ' or both to alkyl when they contain alkenyl.

For example, the above step d can be performed in the presence of a Pd/C catalyst under an H ₂ atmosphere, but is not limited thereto.

Furthermore, in the manufacturing method of the present invention, the precursors of R ₁ , R ₂ , and/or R ₃ may mean functional groups that can be converted or substituted with R ₁ , R ₂ , and/or R ₃ by a known method. Therefore, the manufacturing method of the present invention may further include a step for additionally modifying R ₁ , R ₂ , R ₃ or one or more of their precursors. The additional step may be performed without limitation using a method known in the art.

Meanwhile, each step of the manufacturing method of the present invention can be independently performed in an organic solvent. The organic solvent can be any solvent used in organic synthesis in the art without limitation. For example, the organic solvent may include DMF (dimethylformamide), toluene, dichloromethane, acetone, acetonitrile, THF (tetrahydrofuran), EtOAc (ethyl acetate), methanol, etc., which may be used alone or in a mixture of two or more thereof. Specifically, as the organic solvent, DMF may be used in the first step, DMF or toluene may be used in the a-step, dichloromethane may be used in the b-step, and EtOAc or methanol may be used in the c-step and d-steps, but is not limited thereto.

Another aspect of the present invention for achieving the above object provides a pharmaceutical composition for preventing or treating an ocular disease comprising the compound or a pharmaceutically acceptable salt thereof.

The term "eye disease" of the present invention refers to a disease related to the eye, also called an eye disease, an ophthalmic disease, etc., and for the purpose of the present invention, may refer to an eye disease related to autophagy induced by A2E and blue light. Specifically, the eye disease may refer to dry macular degeneration, xerophthalmia, eye fatigue, or decreased vision, but is not limited thereto.

The above term, "A2E (N-retinyl N-retinylidene ethanolamine)" is the main chromophore of lipofuscin in the central part of the retina, and is a substance that causes the production of reactive oxygen species when exposed to blue light. If A2E accumulates excessively inside cells, retinal pigment epithelial cells may die.

The above term, "blue light", refers to the blue light that exists between 380 and 500 nm in the visible light range, and is emitted in large quantities from displays of smart devices such as TVs and computers and LED lighting devices. Blue light with a short wavelength, known to have the highest energy among the light that the human eye can distinguish, passes through the lens and affects retinal epithelial cells.

The term "prevention" of the present invention means any act of inhibiting or delaying the worsening or progression of an eye disease by administering a pharmaceutical composition containing the compound as an active ingredient.

The term "treatment" of the present invention means any act in which symptoms of an ocular disease are improved or beneficially changed by administering a pharmaceutical composition containing the compound as an active ingredient.

The term "pharmaceutical composition" of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent commonly used in the manufacture thereof, and the carrier may include a non-naturally occurring carrier. Specific examples of the carrier, excipient and diluent include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

In addition, the pharmaceutical composition may have any one dosage form selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, oral solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilizers, and suppositories, respectively, according to a conventional method, and may be various oral or parenteral dosage forms. When formulated, it is prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Tablets, pills, powders, granules, capsules, etc. may be used as solid preparations for oral administration, and the solid preparations may use at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives can be used. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, or suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include, but are not limited to, witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerogelatin.

The content of the compound in the pharmaceutical composition of the present invention can be appropriately adjusted depending on the symptoms and progression of the disease, the patient's condition, etc., and may be, for example, 0.0001 to 99.9 wt% or 0.001 to 50 wt% based on the total composition weight, but is not limited thereto.

In one embodiment of the present invention, in order to evaluate the autophagy activity by treatment with the compound represented by the chemical formula 1 and its derivative compound (novel compound), human retinal pigment epithelial cells, ARPE-19, were used to confirm whether LC3-II, one of the major indicator proteins of autophagy activation, increased, and as a result, the autophagy activation effect by treatment with the novel compound was confirmed (Figs. 1a to 1b and Tables 2 to 3).

In addition, in one embodiment of the present invention, as a result of confirming the A2E-fluorescent labeling substance (A2E-BDP) and A2E removal ability by the novel compound treatment, it was confirmed that A2E-BDP and A2E accumulated in cells were removed by the compound treatment (Figs. 2a to 6c).

In addition, in one embodiment of the present invention, improvement in retinal damage and recovery of ocular tissue by treatment with the novel compound was confirmed using a retinal damage animal model induced by blue light (Figs. 7a to 7c).

These results suggest that the novel compound of the present invention can be useful for preventing or treating eye diseases.

Another aspect of the present invention for achieving the above purpose provides a method for preventing or treating an ocular disease comprising administering the pharmaceutical composition.

The pharmaceutical composition, ocular disease, prevention or treatment is as described above.

The term "subject" of the present invention means all animals including rats, livestock, etc., including humans, who have developed or may develop eye diseases. Specifically, it may be mammals such as cows, horses, sheep, pigs, goats, camels, antelopes, dogs, cats, etc., as well as humans who require prevention or treatment of symptoms similar to the above diseases, but is not limited thereto.

The term "administration" of the present invention means introducing the composition of the present invention into a patient by any appropriate method, and the administration route of the composition may be through any common route as long as it can reach the target tissue.

The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount.

The above term, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dosage level can be determined according to the type and severity of the individual, age, sex, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. For example, an effective amount of 0.001 mg/kg to 1000 mg/kg, 0.01 mg/kg to 100 mg/kg, or 0.1 to 20 mg/kg or 0.1 to 500 mg/kg is included. The amount of the pharmaceutical composition of the present invention can be selected and implemented within an appropriate range by a person skilled in the art.

The above pharmaceutical composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. In addition, it may be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects by considering all of the above-mentioned factors, and this can be easily determined by those skilled in the art.

In addition, the pharmaceutical composition may be administered orally or parenterally (for example, intravenously, subcutaneously, intraperitoneally, or topically) depending on the intended method, and the dosage may vary depending on the patient's condition and weight, the extent of the disease, the drug form, the administration route, and the time, but may be appropriately selected by those skilled in the art. As a specific example, the dosage may generally be administered once a day or several times a day, but a preferred dosage may be appropriately selected by those skilled in the art depending on the patient's condition and weight, the extent of the disease, the drug form, the administration route, and the time.

Another aspect of the present invention for achieving the above purpose provides a food composition for preventing or improving ocular disease comprising the compound or a pharmaceutically acceptable salt thereof.

The compounds, pharmaceutically acceptable salts, ocular diseases and prevention thereof are as described above.

The term "improvement" of the present invention means any action that at least reduces a parameter associated with a condition being treated, for example, the severity of a symptom, by administration of a composition comprising the novel compound.

The term "food" of the present invention includes meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, and health foods, and includes all foods in the conventional sense.

The above health functional food is the same term as food for special health use (FoSHU), and refers to food with high medical and healthcare effects that is processed to effectively exhibit bioregulatory functions in addition to providing nutrition.

Here, 'function(s)' means regulating nutrients for the structure and function of the human body or obtaining a useful effect for health purposes such as physiological functions. The health food above means a food that has a more active health maintenance or promotion effect than general foods, and health supplement food means a food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and health supplement food may be used interchangeably. Specifically, the health functional food above means a food that is prepared by adding the novel compound of the present invention to food materials such as beverages, teas, spices, gums, and confectionery, or by encapsulating, powdering, or manufacturing it in the form of a suspension, and may mean that when consumed, it brings about a specific health effect.

The food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art.

In addition, the food composition can be manufactured without limitation into various forms of formulations as long as it is a formulation recognized as a food.

In addition, the food composition may additionally include a food-wise acceptable carrier. The type of the carrier is not particularly limited, and any carrier commonly used in the relevant technical field may be used.

In addition, the food composition may include additional ingredients commonly used in food compositions to improve odor, taste, and sight. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, and the like. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr); and amino acids such as lysine, tryptophan, cysteine, and valine.

In addition, the food composition may include food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), bactericides (bleaching powder and highly bleaching powder, sodium hypochlorite, etc.), antioxidants (butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), etc.), colorants (tar color, etc.), color developers (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), reinforcing agents, emulsifiers, thickeners (glutinating agents), film agents, gum bases, foam suppressants, solvents, and improvers. The above additives can be selected according to the type of food and used in an appropriate amount.

Another aspect of the present invention for achieving the above purpose provides a feed composition for preventing or improving ocular diseases, comprising the compound or a pharmaceutically acceptable salt thereof.

The compounds, pharmaceutically acceptable salts, ocular diseases, prevention and improvement thereof are as described above.

The above term, "feed" means any natural or artificial diet, meal, etc. or ingredients of said meal, intended for eating, ingesting and digesting by animals or suitable therefor.

The type of the above feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of the above feed include plant feed such as grains, nuts, food processing by-products, algae, fiber, pharmaceutical by-products, fats, starches, meal, or grain by-products; animal feed such as proteins, inorganic substances, fats, mineral substances, fats, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

Another aspect of the present invention for achieving the above object provides a use or purpose of the compound or a pharmaceutically acceptable salt thereof for preventing, improving or treating ocular diseases.

The compounds, pharmaceutically acceptable salts, ocular diseases, prevention, improvement and treatment are as described above.

The novel compound according to the present invention and its derivative compounds can be utilized to treat, prevent and improve autophagy-related diseases, particularly eye diseases, by inhibiting death of human retinal pigment epithelial cells (ARPE-19) induced by A2E and blue light, removing A2E-fluorescent markers and A2E in ARPE-19, and activating autophagy.

Figures 1a and 1b illustrate the evaluation of autophagy activation by treatment with a compound represented by chemical formula 1.

Figures 2a to 2c show the ability to remove intracellular accumulated A2E-fluorescent labeling substance (A2E-BDP) by treatment with a compound represented by chemical formula 1 or the compound of Comparative Example 1.

Figure 3 shows the results of measuring the amount of A2E present in cells by treatment with a compound represented by chemical formula 1 or the compound of Comparative Example 1.

Figure 4 shows the ability of the compound represented by chemical formula 1 to remove accumulated A2E-BDP within cells by treatment with various derivative compounds.

Figures 5a to 5b and 6a to 6c show the removal ability of intracellular A2E by treatment with the derivative compound.

Figures 7a to 7d show the improvement in retinal damage and recovery of ocular tissue by treatment with the derivative compound or the compound of Comparative Example 1 using a blue light-induced retinal damage animal model.

Hereinafter, the present invention will be described in more detail through examples. These examples are intended to explain the present invention more specifically, and the scope of the present invention is not limited by these examples.

Comparative Example 1: Preparation of 3-(5-allyl-2-methoxyphenyl)-5-((allyloxy)methyl)isoxazole

The title compound was obtained in the same manner as in Korean Patent Publication No. 10-2023-0140410.

Manufacturing Example 1: Manufacturing of 4-bromo-2-ethynyl-1-methoxybenzene (b)

5-Bromo-ortho-anisaldehyde ( a , 2.0 g, 1 eq) and K ₂ CO ₃ (1.67 g, 1.3 eq) were dissolved in methanol (40 mL). The mixture was cooled to 0 °C and Bestmann-Ohira reagent (0.1 M in MeOH, 1.3 eq) was slowly added at 0 °C. The mixture was warmed to ambient temperature and stirred for 6 h. After the reaction was complete, the solution was quenched with water and the solvent was removed in vacuo . The residue was diluted with EtOAc (100 mL) and washed with water and brine. The resulting organic layers were combined, dried over MgSO ₄ and concentrated in vacuo. The residue was purified by flash column chromatography (EtOAc/hexane = 1:20) to obtain the title compound ( b , 1.8 g, 92%) as a white solid.

Example 1: Preparation of 1-benzyl-4-(5-bromo-2-methoxyphenyl)-1H-1,2,3-triazole (GS27)

Benzyl bromide (0.26 mL, 1 eq) and NaN ₃ (142 mg, 1 eq) were dissolved in DMF (40 mL). The mixture was stirred at room temperature for 18 h. The mixture was cooled to 0 °C, and 4-bromo-2-ethynyl-1-methoxybenzene ( b , 460 mg, 1 eq), Cu(I)Br (63 mg, 2 eq), and PMDETA (N,N,N',N",N"-pentamethyldiethylenetriamine, 0.89 mL, 2 eq) dissolved in DMF were added at 0 °C. After maintaining the mixture at the same temperature for 30 min, the mixture was warmed to room temperature and stirred for 3 h. After the reaction was completed, the mixture was diluted with EtOAc (100 mL) and washed with water and brine. The obtained organic layers were combined, dried over Na ₂ SO ₄ , and concentrated in vacuo. The residue was purified by flash column chromatography (EtOAc/hexane = 1:5) to obtain the title compound ( GS27 , 663 mg, 88%).

^1H NMR (600 MHz, Chloroform -d ) δ 8.49 (d, J = 2.6 Hz, 1H), 7.96 (s, 1H), 7.37 (m, 4H), 7.29 (ddt, J = 7.3, 1.5, 0.7 Hz, 2H), 6.81 (d, J = 8.8) Hz, 1H), 5.59 (s, 2H), 3.86 (s, 3H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 154.8, 142.5, 135.1, 131.5, 130.3, 129.2, 128.8, 127.9, 123.5, 121.4, 113.7, 112.6, 55.8, 54.2.

Example 2: Preparation of 4-(5-allyl-2-methoxyphenyl)-1-benzyl-1H-1,2,3-triazole (GS28)

To a solution of 1-benzyl-4-(5-bromo-2-methoxyphenyl)-1H-1,2,3-triazole ( GS27 , 660 mg, 1 eq) in DMF (0.8 M) was added Pd(PPh ₃ ) ₄ (222 mg, 0.1 eq) and allyltributylstannane (0.89 mL, 1.5 eq). After bubbling with nitrogen, the mixture was stirred at 90 °C for 5 h under _{N 2 atmosphere} . The mixture was diluted with EtOAc and washed with water and brine. The combined organic layers were dried over Na ₂ SO ₄ and concentrated in vacuo . The residue was purified by flash column chromatography (EtOAc/hexane = 1:5) to obtain the title compound ( GS28 , 548 mg, 94%).

^1H NMR (600 MHz, Chloroform -d ) δ 8.19 (d, J = 2.3 Hz, 1H), 7.97 (s, 1H), 7.36 (m, 3H), 7.28 (m, 2H), 7.11 (dd, J = 8.4, 2.4 Hz, 1H), 6.88 (d, J = 8.4 Hz, 1H), 5.99 (ddt, J = 16.8, 10.0, 6.7 Hz, 1H), 5.59 (s, 2H), 5.07 (m, 2H), 3.86 (s, 3H), 3.41 (d, J = 6.7 Hz, 2H).

Example 3: Preparation of 4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenol (GS30)

4-(5-Allyl-2-methoxyphenyl)-1-benzyl-1H-1,2,3-triazole ( GS28 , 428 mg, 1 eq) was added boron tribromide (4.2 mL, 3 eq, 1.0 M solution in _CH2Cl2 ) to a _CH2Cl2 solution ₍ 6 mL) under _{N2 atmosphere} at -20 °C. After the reaction was completed, the solution was quenched with saturated _NH4Cl . The reaction mixture was _diluted with _CH2Cl2 , washed with water, dried over _Na2SO4 and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane = 1: ₄ ) to give the title compound ( GS30 , 370 mg, 90%) as a white solid.

^1H NMR (600 MHz, Chloroform -d ) δ 7.74 (s, 1H), 7.40 (m, 3H), 7.32 (m, 2H), 7.14 (m, 1H), 7.04 (dd, J = 8.4, 2.0 Hz, 1H), 6.98 (d, J = 8.4 Hz, 1H), 5.93 (ddt, J = 15.9, 10.8, 6.6 Hz, 1H), 5.60 (s, 2H), 5.05 (m, 1H), 5.03 (t, J = 1.5 Hz, 1H), 3.30 (d, J = 6.8 Hz, 1H);

¹³ C NMR (150 MHz, CDCl ₃ ) δ 154.4, 148.4, 137.9, 134.3, 130.9, 130.3, 129.4, 129.2, 128.3, 125.8, 118.8, 117.8, 115.8, 113.7, 54.7, 39.4.

Example 4: Preparation of ethyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate (GS32)

To an acetone solution of 4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl) _phenol ( GS30 , 400 mg, 1.0 eq) and _K2CO3 (641 mg, 3 eq) was added ethyl 2-bromoacetate (0.26 mL, 1.5 eq). The mixture was stirred at 60 °C for 3 h under _N2 atmosphere. The reaction mixture was cooled to room temperature and then quenched with saturated _NH4Cl . The mixture was diluted with EtOAc, washed with water, dried over _Na2SO4 and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane = 1:4) _to give the title compound ( GS32 , 458 mg, 88%) was obtained.

¹ H-NMR (400 MHz, Chloroform -d ) δ 8.58 (d, J = 2.5 Hz, 1H), 8.26 (d, J = 2.2 Hz, 1H), 7.34 (d, J = 3.3 Hz, 5H), 7.09 (d, J = 8.4 Hz, 1H), 6.76 (d, J = 8.4 Hz, 1H), 5.98 (m, 1H), 5.61 (s, 2H), 5.08 (m, 2H), 4.64 (s, 2H), 4.26 (q, J = 7.3 Hz, 2H), 3.41 (d, J = 6.7 Hz, 2H), 1.29 (t, J = 7.2 Hz, 3H).

Example 5: Preparation of 4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenyl acetate (GS33)

To a CH ₂ Cl ₂ solution of GS30 (30 mg, 1.0 eq) and acetic anhydride (0.01 mL, 1 eq) were added DMAP (4-dimethylaminopyridine, 1.3 mg, 0.1 eq) and Et ₃ N (triethylamine, 0.14 mL, 10 eq) under N ₂ atmosphere at 0 °C. The mixture was stirred under N ₂ atmosphere at 0 °C for 18 h. After the reaction was completed, the reaction mixture was quenched with water. The mixture was diluted with CH ₂ Cl ₂ , washed with water, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane = 1:4) to give the title compound ( GS33 , ) as a white solid. 26 mg, 73%) was obtained.

^1H -NMR (400 MHz, Chloroform -d ) δ 7.42 (d, 1H), 7.40 (d, 1H), 7.38 (m, 3H), 7.31 (m, 2H), 7.17 (m, 1H), 7.06 (s, 1H), 5.97 (m, 1H), 5.58 (s, 2H), 5.10 (m, 2H), 3.42 (d, 2H), 2.12 (s, 3H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 169.0, 145.4, 143.5, 138.4, 136.9, 134.6, 129.2, 129.2, 128.9, 128.6, 128.2, 123.1, 122.9, 121.6, 116.3, 54.2, 39.6, 29.7, 21.1.

Example 6: Preparation of (R)-methyl 2-(2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetamido)-3-phenylpropanoate (GT01)

To a CH ₂ Cl ₂ solution of 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetic acid ( GT11 , 30 mg, 1.0 eq) at 0 °C were added methyl D-phenylalaninate (15.4 mg, 1.0 eq), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC, 25 mg, 1.5 eq), 1-hydroxybenzotriazole (HOBt, 5.8 mg, 0.5 eq), and Et ₃ N (36 μL, 3 eq). The mixture was stirred at room temperature for 4 h, and the reaction mixture was quenched with saturated NH ₄ Cl. The mixture was diluted with CH ₂ Cl ₂ , washed with water and brine, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane = 1:3) to give the title compound ( GT01 , 38 mg, 81%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 7.90 (m, 2H), 7.73 (d, J = 8.5 Hz, 1H), 7.33 (m, 5H), 7.16 (m, 4H), 7.03 (dd, J = 7.0, 2.5 Hz, 2H), 6.83 (d, J = 8.3 Hz, 1H), 5.99 (ddt, J = 16.8, 9.8, 6.7 Hz, 1H), 5.53 (d, J = 14.9 Hz, 1H), 5.40 (d, J = 14.9 Hz, 1H), 5.11 (m, 2H), 5.01 (dt, J = 8.0, 6.0 Hz, 1H), 4.58 (d, J = 2.6 Hz, 2H), 3.77 (s, 3H), 3.41 (d, J = 6.7 Hz, 2H), 3.16 (m, 2H).

Example 7: Preparation of (S)-methyl 2-(2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetamido)-3-phenylpropanoate (GT02)

Except that methyl L-phenylalaninate (15.4 mg, 1.0 eq) was used instead of methyl D-phenylalaninate, the reaction was carried out in the same manner as in Example 6, and the title compound ( GT02 , ) was obtained as a white solid by purification. 35 mg, 79%) was obtained.

^1H NMR (400 MHz, Chloroform- d ) δ 7.89 (m, 2H), 7.74 (d, J = 8.5 Hz, 1H), 7.33 (ddd, J = 19.8, 8.6, 5.1 Hz, 5H), 7.15 (m, 4H), 7.03 (dd, J = 19.8, 8.6, 5.1 Hz, 5H) 6.9, 2.4 Hz, 2H), 6.83 (d, J = 8.4 Hz, 1H), 5.99 (ddt, J = 16.8, 9.8, 6.7 Hz, 1H), 5.53 (d, J = 14.8 Hz, 1H), 5.40 (d, J = 14.9 Hz, 1H), 5.11 (m, 2H), 5.01 (dt, J = 8.0, 6.0 Hz, 1H), 4.58 (d, J = 2.6 Hz, 2H), 3.77 (s, 2H), 3.41 (d, J = 6.7 Hz, 2H), 3.18 (m, 2H).

Example 8: Preparation of tert-butyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate (GT03)

The reaction was carried out in a similar manner as in Example 4, except that t - butyl-2-bromoacetate was used instead of ethyl 2-bromoacetate, and the title compound ( GT03 , ) was obtained as a white solid by purification. 170 mg, 72%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 8.62 (s, 1H), 8.25 (s, 1H), 7.35 (d, J = 1.1 Hz, 2H), 7.07 (dd, J = 8.4, 2.2 Hz, 1H), 6.74 (d, J = 8.4 Hz, 1H), 5.98 (m, 1H), 5.60 (s, 2H), 5.08 (m, 2H), 4.52 (d, J = 1.2 Hz, 2H), 3.40 (d, J = 6.7 Hz, 2H), 1.49 (d, J = 1.2 Hz, 9H).

Example 9: Preparation of 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-1-(4-(methylsulfonyl)phenyl)ethanone (GT04)

The reaction and purification were carried out in a similar manner as in Example 4, except that 2-bromo-1-(4-(methylsulfonyl)phenyl)ethanone was used instead of ethyl 2-bromoacetate, to give the title compound ( GT04 , ) as a white solid. 30 mg, 90%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 8.63 (s, 1H), 8.23 (s, 1H), 8.06 (m, 4H), 7.37 (m, 5H), 7.11 (d, J = 8.3 Hz, 1H), 6.86 (d, J = 8.3 Hz, 1H), 5.98 (ddt, J = 16.7, 9.4, 6.6 Hz, 1H), 5.60 (s, 1H), 5.32 (s, 2H), 5.09 (m, 2H), 3.41 (d, J = 6.7 Hz, 2H), 3.10 (s, 3H).

Example 10: Preparation of 2-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)pyridine (GT05)

The reaction and purification were carried out in a similar manner as in Example 4, except that 2-(bromomethyl)pyridine was used instead of ethyl 2-bromoacetate, to obtain the title compound ( GT05 , ) as a pale yellow solid. 19.6 mg, 75%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 8.58 (d, J = 4.8 Hz, 1H), 8.22 (s, 1H), 8.13 (s, 1H), 7.58 (t, J = 7.7 Hz, 1H), 7.37 (m, 3H), 7.28 (m, 4H), 7.10 (d, J = 8.3 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 6.01 (ddt, J = 16.8, 9.4, 6.9 Hz, 1H), 5.58 (s, 2H), 5.27 (s, 2H), 5.10 (m, 2H), 3.43 (d, J = 6.7 Hz, 2H).

Example 11: Preparation of 3-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)pyridine (GT06)

The reaction was carried out in a similar manner as in Example 4, except that 3-(bromomethyl)pyridine was used instead of ethyl 2-bromoacetate, and the title compound ( GT06 , ) was obtained as a white solid by purification. 21 mg, 79%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 8.64 (s, 1H), 8.61 (d, J = 4.6 Hz, 1H), 8.20 (s, 1H), 7.74 (s, 1H), 7.61 (d, J = 7.8 Hz, 1H), 7.33 (m, 3H), 7.21 (m, 3H), 7.12 (d, J = 8.4 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 5.99 (ddt, J = 16.7, 9.7, 6.7 Hz, 1H), 5.49 (s, 2H), 5.09 (s, 2H), 5.08 (m, 2H), 3.41 (d, J = 6.7 Hz, 2H).

Example 12: Preparation of 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)pyridine (GT07)

The reaction was carried out in a similar manner as in Example 4, except that 4-(bromomethyl)pyridine was used instead of ethyl 2-bromoacetate, and the title compound ( GT07 , ) was obtained as a white solid by purification. 17 mg, 63%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 8.53 (d, J = 5.1 Hz, 2H), 8.20 (s, 1H), 7.83 (s, 1H), 7.37 (m, 3H), 7.23 (m, 4H), 7.10 (d, J = 8.4 Hz, 1H), 6.87 (d, J = 8.4 Hz, 1H), 5.99 (ddt, J = 16.8, 9.6, 6.8 Hz, 1H), 5.53 (s, 2H), 5.09 (s, 2H), 5.08 (m, 2H), 3.41 (d, J = 6.7 Hz, 2H).

Example 13: Preparation of 4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenyl morpholine-4-carboxylate (GT08)

To a CH ₂ Cl ₂ solution of GS30 (20 mg, 1.0 eq) and DMAP (2 mg, 0.2 eq) were added morpholine-4-carbonyl chloride (0.12 mL, 1.5 eq) and Et ₃ N (0.02 mL, 2 eq). The mixture was stirred at room temperature for 3 h under N ₂ atmosphere, and the reaction mixture was quenched with saturated NH ₄ Cl. The mixture was diluted with EtOAc, washed with water, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane = 1:3) to give the title compound ( GT08 , 20 mg, 72%) was obtained.

^1H NMR (600 MHz, Chloroform -d ) δ 7.87 (d, J = 2.2 Hz, 1H), 7.50 (s, 1H), 7.40 (m, 3H), 7.30 (m, 2H), 7.16 (dd, J = 8.3, 2.2 Hz, 1H), 7.06 (d, J = 8.3 Hz, 1H), 5.97 (ddt, J = 16.8, 10.0, 6.7 Hz, 1H), 5.56 (s, 2H), 5.09 (m, 2H), 3.57 (d, J = 50.0 Hz, 4H), 3.42 (dd, J = 8.8, 3.9 Hz, 6H).

Example 14: Preparation of ethyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-2-methylpropanoate (GT09)

The reaction was carried out in a similar manner as in Example 4, except that ethyl 2-bromo-2-methylpropanoate was used instead of ethyl 2-bromoacetate and DMF was used as a solvent, to obtain the title compound ( GT09 , ) as a white solid. 18 mg, 64%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 8.12 (s, 1H), 7.98 (s, 1H), 7.34 (m, 5H), 6.98 (m, 1H), 6.67 (d, J = 8.4 Hz, 1H), 5.97 (ddt, J = 16.8, 10.0, 6.8 Hz, 1H), 5.57 (s, 2H), 5.07 (m, 2H), 4.20 (q, J = 7.1 Hz, 2H), 3.38 (d, J = 6.7 Hz, 2H), 1.46 (s, 9H), 1.21 (t, J = 7.1 Hz, 5H).

Example 15: Preparation of 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)piperidine (GT10)

To a CH ₂ Cl ₂ solution of tert-butyl 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)piperidine-1-carboxylate ( GT16 , 30 mg) was added trifluoroacetic acid (1 mL) under N ₂ atmosphere at 0°C. The mixture was stirred at room temperature for 4 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (CH ₂ Cl ₂ /MeOH = 15:1) to give the title compound ( GT10-TFA salt , ) as a white solid TFA salt. 29 mg, 97%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 8.06 (s, 1H), 7.76 (s, 1H), 7.32 (m, 5H), 7.09 (d, J = 8.2 Hz, 1H), 6.80 (d, J = 8.5 Hz, 1H), 5.96 (ddt, J = 16.8, 9.8, 6.8 Hz, 2H), 5.57 (s, 3H), 5.06 (m, 3H), 4.80 (s, 8H), 3.84 (d, J = 6.3 Hz, 3H), 3.37 (d, J = 7.2 Hz, 6H), 2.72 (t, J = 12.7 Hz, 3H), 1.90 (s, 2H), 1.78 (d, J = 14.0 Hz, 3H), 1.54 (q, J = 11.3 Hz, 3H).

Example 16: Preparation of 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetic acid (GT11)

Except that tert-butyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate ( GT03 ) was used instead of tert-butyl 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)piperidine-1-carboxylate ( GT16 ), the reaction and purification were similar to Example 15, to give the title compound (GT11-TFA salt , ) as a white solid TFA salt. 130 mg, 99%) was obtained.

^1H NMR (400 MHz, Methanol- d ₄ ) δ 8.76 (s, 1H), 7.98 (s, 1H), 7.35 (m, 5H), 7.11 (d, J = 8.4 Hz, 1H), 6.91 (d, J = 8.3 Hz, 1H), 5.97 (ddt, J = 16.8, 9.5, 6.7 Hz, 1H), 5.61 (s, 2H), 5.05 (m, 2H), 4.71 (s, 2H), 3.37 (d, J = 6.7 Hz, 2H).

Example 17: Preparation of 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)-1-(isopropylsulfonyl)piperidine (GT12)

To an acetonitrile solution of 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)piperidine ( GT10 , 27 mg, 1.0 eq) and propane-2-sulfonyl chloride (0.01 mL, 1.2 eq) was added Et ₃ N (0.03 mL, 3 eq). The mixture was stirred overnight at room temperature under N ₂ atmosphere, and the reaction mixture was quenched with saturated NH ₄ Cl. The mixture was diluted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane/MeOH = 1:2:0.1) to give the title compound ( GT12 , 13 mg, 38%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 8.15 (s, 1H), 7.87 (s, 1H), 7.35 (m, 5H), 7.08 (d, J = 8.4 Hz, 1H), 6.83 (d, J = 8.4 Hz, 1H), 5.98 (ddt, J = 16.8, 9.9, 6.7 Hz, 1H), 5.60 (s, 2H), 5.07 (m, 2H), 3.88 (d, J = 5.9 Hz, 2H), 3.80 (d, J = 13.1 Hz, 2H), 3.39 (d, J = 6.7 Hz, 2H), 3.18 (p, J = 6.8 Hz, 1H), 2.79 (t, J = 12.1 Hz, 2H), 1.85 (m, 1H), 1.65 (m, 2H), 1.42 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H).

Example 18: Preparation of 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetamide (GT13)

A THF solution of 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetic acid (GT11 , 26 mg, 1.0 eq) and CDI (carbonyldiimidazole, 24 mg, 2 eq) was stirred at room temperature for 4 h under N ₂ atmosphere. To the mixture was added NH ₄ OH aqueous solution (0.5 mL, excess) and stirred for another 3 h at the same temperature. The reaction mixture was quenched with water. The mixture was diluted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane = 1:4) to give the title compound ( GT13 , 18 mg, 70%) was obtained.

^h1H NMR (400 MHz, Chloroform -d ) δ 7.79 (s, 1H), 7.59 (s, 1H), 7.35 (m, low 5H), 7.14 (d, J = 8.4 Hz, 1H), 6.86 (d, J = 8.5 Hz, 1H), 5.94 (ddt, J = 17.5, 8.5, 6.6 Hz, 1H), 5.57 (s, 2H), 5.07 (m, 2H), 4.57 (s, 2H), 3.36 (d, J = 6.6 Hz, 2H).

Example 19: Preparation of 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-N-cyclohexylacetamide (GT14)

A DMF solution of GT11 (31 mg, 1.0 eq), cyclohexylamine (13 mg, 1.5 eq), EDC (41 mg, 3 eq), HOBt (14 mg, 1 eq), DMAP (11 mg, 0.1 eq), and DIPEA (N,N-diisopropylethylamine, 0.15 mL, 10 eq) was stirred overnight at 60 °C under N ₂ atmosphere. The reaction mixture was cooled to room temperature and then quenched with water. The mixture was diluted with EtOAc, washed with water and brine, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane = 1:2) to give the title compound ( GT14 , 23 mg, 60%) was obtained.

^1H NMR (400 MHz, Chloroform -d ) δ 7.78 (s, 1H, -NH), 7.74 (d, J = 8.4 Hz, 1H), 7.59 (s, 1H), 7.35 (m, 5H), 7.12 (d, J = 8.3 Hz, 1H), 6.85 (d, J) = 8.3 Hz, 1H), 5.94 (m, 1H), 5.60 (s, 2H), 5.06 (m, 2H), 4.56 (s, 2H), 3.84 (m, 1H), 3.36 (d, J = 6.6 Hz, 2H), 1.89 (m, 2H), 1.73 (m, 2H), 1.62 (m, 1H), 1.28 (m, 4H).

Example 20: Preparation of 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-N-(4-hydroxycyclohexyl)acetamide (GT15)

The reaction and purification were carried out in a similar manner as in Example 19, except that 4-aminocyclohexanol was used instead of cyclohexylamine, to give the title compound ( GT15 , ) as a white solid. 25 mg, 53%) was obtained.

¹ H NMR (400 MHz, Chloroform- d ) δ 8.29 (d, J = 8.2 Hz, 1H), 7.74 (s, 1H), 7.36 (m, 6H, including -NH), 7.12 (d, J = 8.4 Hz, 1H), 6.85 (d, J = 8.4 Hz, 1H), 5.92 (ddd, J = 16.4, 11.5, 6.8 Hz, 1H), 5.59 (s, 2H), 5.04 (m, 2H), 4.56 (s, 2H), 3.83 (m, 1H), 3.66 (m, 1H), 3.34 (d, J = 6.6 Hz, 2H), 1.99 (m, 4H), 1.78 (s, 1H, -OH), 1.41 (t, J = 9.5 Hz, 4H).

Example 21: Preparation of tert-butyl 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)piperidine-1-carboxylate (GT16)

The reaction and purification were carried out in a similar manner as in Example 4, except that t - butyl 4-(bromomethyl)piperidine-1-carboxylate was used instead of ethyl 2-bromoacetate, to give the title compound ( GT16 , ) as a white solid. 43 mg, 51%) was obtained.

^1H NMR (600 MHz, Chloroform -d ) δ 8.16 (d, J = 2.3 Hz, 1H), 7.84 (s, 1H), 7.39 (m, 3H), 7.30 (m, 2H), 7.08 (dd, J = 8.4, 2.3 Hz, 1H), 6.83 (d, J = 8.4 Hz, 1H), 5.98 (ddt, J = 16.8, 10.0, 6.7 Hz, 1H), 5.57 (s, 2H), 5.06 (m, 2H), 4.07 (s, 2H), 3.83 (d, J = 6.5 Hz, 2H), 3.39 (d, J = 6.8 Hz, 2H), 2.62 (s, 2H), 1.85 (m, 1H), 1.61 (m, 2H), 1.48 (s, 9H), 1.15 (qd, J = 12.4, 4.4 Hz, 2H).

Example 22: Preparation of 2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-bromophenol (c)

To a CH ₂ Cl ₂ solution (20 mL) of GS27 (857 mg, 1 eq) was added boron trichloride (7.47 mL, 3 eq, 1.0 M solution in CH ₂ Cl ₂ ) under N ₂ atmosphere at 0 °C. After the reaction was completed, the solution was quenched with saturated NH ₄ Cl. The reaction mixture was diluted with CH ₂ Cl ₂ , washed with water, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane = 1:4) to give the title compound ( c , 821 mg, 100%) as a white solid.

^1H NMR (600 MHz, DMSO _-d6 ) δ 10.52 (bs, 1H), 8.51 (s, 1H), 8.13 (d, J = 2.6 Hz, 1H), 7.36 (m, 5H), 7.31 (d, J = 2.6 Hz, 1H), 6.93 (d, J = 8.7 Hz, 1H), 5.68 (s, 2H).

Example 23: Preparation of ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-bromophenoxy)acetate (d)

The reaction and purification were carried out in a similar manner as in Example 4, except that a DMF solution of 2-(1 - benzyl-1H-1,2,3-triazol-4-yl)-4-bromophenol ( c ) was used instead of the acetone solution of GS30, to afford the title compound ( d ,) as a white solid. 753 mg, 71%) was obtained.

^1H NMR (600 MHz, Chloroform -d ) δ 8.59 (s, 1H), 8.55 (d, J = 2.3 Hz, 1H), 7.35 (m, 6H), 6.68 (d, J = 8.6 Hz, 1H), 5.60 (s, 2H), 4.62 (s, 2H), 4.27 (q, J = 7.0 Hz, 2H), 1.30 (t, J = 7.1 Hz, 3H).

Example 24: Preparation of ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-isobutylphenoxy)acetate (GT17)

To a solution of ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-bromophenoxy)acetate ( d , 30 mg, 1 eq) in toluene (0.14 M) were added Pd(PPh ₃ ) ₄ (8.4 mg, 0.1 eq), isobutylboronic acid (14.7 mg, 2 eq) and Cs ₂ CO ₃ (71 mg, 3 eq). After bubbling with nitrogen, the reaction was performed using a microwave at 100 °C for 1.5 h. The reaction mixture was cooled to room temperature and then quenched with saturated NH ₄ Cl. The mixture was diluted with EtOAc and washed with water and brine. The resulting organic layers were combined, dried over Na ₂ SO ₄ and concentrated in vacuo. The residue was purified by flash column chromatography (EtOAc/hexane = 1:5) to obtain the title compound ( GT17 , 15.6 mg, 55%).

^1H NMR (600 MHz, Chloroform -d ) δ 8.58 (s, 1H), 8.22 (d, J = 2.3 Hz, 1H), 7.34 (m, 5H), 7.03 (dd, J = 8.3, 2.3 Hz, 1H), 6.73 (d, J = 8.4 Hz, 1H), 5.61 (s, 2H), 4.64 (s, 2H), 4.26 (q, J = 7.1 Hz, 2H), 2.49 (d, J = 7.2 Hz, 2H), 1.90 (m, 1H), 1.29 (t, J = 7.1 Hz, 3H), 0.90 (d, J = 6.6 Hz, 6H).

Example 25: Preparation of ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-cyclopentenylphenoxy)acetate (GT18)

The reaction and purification were carried out in a similar manner as in Example 24, except that cyclopent-1-en-1-ylboronic acid was used instead of isobutylboronic acid, to give the title compound ( GT18 , 22.3 mg, 76%) was obtained.

^1H NMR (600 MHz, Chloroform -d ) δ 8.59 (s, 1H), 8.50 (d, J = 2.3 Hz, 1H), 7.35 (m, 6H), 6.77 (d, J = 8.5 Hz, 1H), 6.19 (m, 1H), 5.61 (s, 2H), 4.66 (s, 2H), 4.26 (q, J = 7.2 Hz, 2H), 2.75 (m, 2H), 2.53 (ddd, J = 10.2, 4.8, 2.4 Hz, 2H), 2.03 (m, 2H), 1.29 (t, J = 7.1 Hz, 3H).

Example 26: Preparation of ethyl 2-(3-(1-benzyl-1H-1,2,3-triazol-4-yl)-4'-fluorobiphenyl-4-yloxy)acetate (GT19)

The title compound ( GT19 , ) was obtained by reaction and purification in a similar manner as in Example 24, except that 4-fluorophenylboronic acid was used instead of isobutylboronic acid. 29.3 mg, 94%) was obtained.

^1H NMR (600 MHz, Chloroform -d ) δ 8.64 (d, J = 2.4 Hz, 1H), 8.62 (s, 1H), 7.61 (m, 2H), 7.45 (dd, J = 8.5, 2.4 Hz, 1H), 7.36 (m, 5H), 7.11 (t, J = 8.7 Hz, 2H), 6.88 (d, J = 8.5 Hz, 1H), 5.62 (s, 2H), 4.70 (s, 2H), 4.28 (q, J = 7.1 Hz, 2H), 1.31 (t, J = 7.2 Hz, 3H).

Example 27: Preparation of ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-(1-(trifluoromethyl)-1H-pyrazol-4-yl)phenoxy)acetate (GT20)

The reaction and purification were carried out in a similar manner as in Example 24, except that 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(trifluoromethyl)-1H-pyrazole was used instead of isobutylboronic acid, to give the title compound ( GT20 , 31 mg, 91%) was obtained.

^1H NMR (600 MHz, Chloroform -d ) δ 8.82 (d, J = 2.3 Hz, 1H), 8.65 (s, 1H), 7.91 (dd, J = 8.6, 2.3 Hz, 1H), 7.84 (d, J = 2.7 Hz, 1H), 7.36 (m, 5H), 6.89 (d, J = 8.6 Hz, 1H), 6.86 (d, J = 2.6 Hz, 1H), 5.63 (s, 2H), 4.71 (s, 2H), 4.28 (q, J = 7.1 Hz, 2H), 1.31 (t, J = 7.1 Hz, 3H).

Example 28: Preparation of ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-(pyridin-4-yl)phenoxy)acetate (GT21)

To a solution of d (60 mg, 1 eq) in DMF (0.08 M) were added Pd(PPh ₃ ) ₄ (16.7 mg, 0.1 eq) and 4-(tributylstannyl)pyridine (80 mg, 1.5 eq). After bubbling with nitrogen, the reaction was carried out overnight at 90 °C using a shield tube. The reaction mixture was cooled to room temperature and quenched with saturated NH ₄ Cl. The mixture was diluted with EtOAc and washed with water and brine. The obtained organic layers were combined, dried over Na ₂ SO ₄ and concentrated in vacuo. The residue was purified by flash column chromatography (EtOAc/hexane = 1:3) to give the title compound ( GT21 , 6.9 mg, 11.5%).

^1H NMR (600 MHz, Chloroform -d ) δ 8.98 (d, J = 2.3 Hz, 1H), 8.67 (ddd, J = 4.8, 1.8, 0.9 Hz, 1H), 8.64 (s, 1H), 8.09 (dd, J = 8.6, 2.4 Hz, 1H), 7.87 (dt, J = 8.0, 1.0 Hz, 1H), 7.75 (ddd, J = 8.0, 7.5, 1.8 Hz, 1H), 7.36 (m, 5H), 7.20 (ddd, J = 7.5, 4.8, 1.1 Hz, 1H), 6.94 (d, J = 8.6 Hz, 1H), 5.63 (s, 2H), 4.73 (s, 2H), 4.28 (q, J = 7.1 Hz, 2H), 1.31 (t, J = 7.1 Hz, 3H).

Example 29: Preparation of ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-cyclopentylphenoxy)acetate (GT22)

A solution of ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-cyclopentenylphenoxy)acetate ( GT18 , 10 mg, 1 eq) in EtOAc and Pd/C (5% wet, catalytic amounts) were stirred at room temperature under a H ₂ atmosphere. After the reaction was complete, the reaction mixture was filtered through a pad of Celite. The filtered solution was concentrated in vacuo, and the residue was purified by flash column chromatography (EtOAc/hexane = 1:5) to obtain the title compound ( GT22 , 6.3 mg, 63%).

^1H NMR (600 MHz, Chloroform -d ) δ 8.57 (s, 1H), 8.31 (d, J = 2.3 Hz, 1H), 7.34 (m, 5H), 7.13 (ddd, J = 8.4, 2.3, 0.6 Hz, 1H), 6.74 (d, J = 8.4) Hz, 1H), 5.60 (s, 2H), 4.63 (s, 2H), 4.26 (q, J = 7.1 Hz, 2H), 3.02 (m, 1H), 2.07 (m, 2H), 1.82 (m, 2H), 1.65 (m, 4H), 1.29 (t, J = 7.1 Hz, 3H).

Example 30: Preparation of 1-benzyl-4-(2-methoxy-5-propylphenyl)-1H-1,2,3-triazole (GS29)

The reaction and purification were carried out in a similar manner as in Example 29, except that GS28 was used instead of GT18 and methanol was used as a solvent, to give the title compound ( GS29 , 9 mg, 90%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 8.18 (d, 1H), 7.97 (s, 1H), 7.36 (m, 3H), 7.30 (m, 2H), 7.10 (m, 1H), 6.87 (d, 1H), 5.60 (s, 2H), 3.86 (s, 3H), 2.60 (t, 2H), 1.66 (m, 2H), 0.94 (t, 3H);

¹³ C-NMR (150 MHz, Chloroform -d ) δ 153.8, 143.8, 135.2, 135.2, 129.0, 128.8, 128.5, 127.7, 127.6, 123.0, 118.9, 110.6, 55.4, 53.9, 37.2, 24.8, 13.8.

Example 31: Preparation of 2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-propylphenol (GS31)

The reaction and purification were carried out in a similar manner as in Example 29, except that GS30 was used instead of GT18 and methanol was used as a solvent, to give the title compound ( GS31 , 7.8 mg, 77%) was obtained.

^1H -NMR (400 MHz, Chloroform -d ) δ 10.64 (s, 1H), 7.74 (s, 1H), 7.41 (m, 3H), 7.33 (m, 2H), 7.14 (d, 1H), 7.04 (m, 1H), 6.97 (m, 1H), 5.60 (s, 2H), 2.50 (t, 2H), 1.59 (m, 2H), 0.91 (t, 3H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 153.9, 148.3, 134.2, 133.5, 130.0, 129.3, 129.1, 128.1, 125.5, 118.7, 117.4, 113.4, 54.6, 37.2, 24.8, 13.8.

Example 32: Preparation of 2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-propylphenyl acetate (GS34)

The reaction and purification were carried out in a similar manner as in Example 29, except that 4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenyl acetate ( GS33 ) was used instead of GT18 and EtOAc was used as a solvent, to give the title compound ( GS34 , 8.3 mg, 82%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 7.89 (s, 1H), 7.59 (s, 1H), 7.39 (m, 3H), 7.31 (m, 2H), 7.15 (m, 1H), 7.03 (d, 1H), 5.58 (s, 2H), 2.62 (t, 2H), 2.12 (s, 3H), 1.66 (m, 2H), 0.95 (t, 3H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 169.1, 145.1, 143.6, 140.9, 134.6, 129.2, 129.1, 128.9, 128.4, 128.4, 128.2, 122.8, 122.7, 121.5, 54.2, 37.4, 24.5, 21.1, 13.8.

Example 33: Preparation of ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-propylphenoxy)acetate (GS35)

The reaction and purification were carried out in a similar manner as in Example 29, except that ethyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl) phenoxy )acetate ( GS32 ) was used instead of GT18 and EtOAc was used as a solvent, to give the title compound ( GS35 , 8.3 mg, 82%) was obtained.

^1H -NMR (400 MHz, Chloroform -d ) δ 8.58 (s, 1H), 8.25 (d, 1H), 7.34 (m, 5H), 7.07 (m, 1H), 6.74 (d, 1H), 5.61 (s, 2H), 4.64 (s, 2H), 4.26 (m, 2H), 2.61 (t, 2H), 1.66 (m, 2H), 1.28 (t, 3H), 0.92 (t, 3H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 168.6, 151.7, 143.3, 136.3, 135.3, 128.9, 128.6, 128.4, 127.9, 127.6, 124.1, 119.7, 111.3, 65.6, 61.4, 54.0, 37.2, 24.7, 14.2, 13.8.

Example 34: Preparation of 4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenyl acetate (e)

4-Allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenol (4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenol, 20 mg, 1.0 eq) and acetic anhydride (7 μL, 1.2 eq) were added to a CH ₂ Cl ₂ solution at ₀ °C under N ₂ atmosphere. The mixture was stirred at room temperature for 3 h under N ₂ atmosphere. After the reaction was completed, the reaction mixture solution was quenched with water. The mixture was diluted with CH ₂ Cl ₂ , washed with water, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash column chromatography (EtOAc/hexane = 1:4) to give the title compound ( e , 20.5 mg, 87%) was obtained.

Example 35: Preparation of ethyl 2-(4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate (GS50)

The reaction and purification were carried out in a similar manner as in Example 4, except that 4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenol was used instead of GS30 , to give the title compound ( GS50 , ) as a white solid. 35 mg, 88%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 8.58 (s, 1H), 8.26 (d, 1H), 7.10 (m, 1H), 6.78 (d, 1H), 6.10 (m, 1H), 5.98 (m, 1H), 5.35 (m, 2H), 5.07 (m, 4H), 4.68 (s, 2H), 4.31 (q, 2H), 3.42 (d, 2H), 1.34 (t, 3H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 168.6, 152.0, 143.0, 137.5, 133.7, 131.7, 128.6, 127.9, 124.0, 119.9, 119.6, 115.8, 111.5, 65.6, 61.4, 52.6, 39.5, 14.2.

Example 36: Preparation of 4-(2-methoxy-5-propylphenyl)-1-propyl-1H-1,2,3-triazole (GS48)

The reaction and purification were carried out in a similar manner as in Example 29, except that 1-allyl-4-(5-allyl-2-methoxyphenyl)-1H-1,2,3-triazole was used instead of GT18 , to give the title compound ( GS48 , 13.2 mg, 87%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 8.18 (d, 1H), 8.02 (s, 1H), 7.11 (m, 1H), 6.89 (d, 1H), 4.37 (t, 2H), 3.92 (s, 3H), 2.61 (t, 2H), 1.99 (m, 2H), 1.67 (m, 2H), 0.99 (t, 3H), 0.94 (t, 3H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 153.8, 143.2, 135.3, 128.6, 127.6, 122.9, 119.1, 110.7, 55.5, 51.8, 37.2, 24.8, 23.8, 13.8, 11.1.

Example 37: Preparation of 4-propyl-2-(1-propyl-1H-1,2,3-triazol-4-yl)phenyl acetate (GS51)

The reaction and purification were carried out in a similar manner as in Example 29, except that 4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenyl acetate ( e ) was used instead of GT18 , to give the title compound ( GS51 , 13.7 mg, 90%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 7.89 (d, 1H), 7.73 (s, 1H), 7.17 (m, 1H), 7.07 (d, 1H), 4.38 (t, 2H), 2.63 (t, 2H), 2.33 (s, 3H), 1.97 (m, 2H), 1.68 (m, 2H), 0.99 (t, 3H), 0.96 (t, 3H);

¹³ C-NMR (150 MHz, DMSO) δ 169.4, 145.1, 143.3, 141.0, 129.1, 128.5, 122.9, 122.7, 121.5, 52.0, 37.5, 24.6, 23.9, 21.5, 13.9, 11.2.

Example 38: Preparation of ethyl 2-(4-propyl-2-(1-propyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate (GS52)

The reaction and purification were carried out in a similar manner as in Example 29, except that ethyl 2-(4-allyl-2- ( 1-allyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate ( GS50 ) was used instead of GT18, to give the title compound ( GS52 , 12.8 mg, 84%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 8.63 (s, 1H), 8.25 (d, 1H), 7.09 (m, 1H), 6.77 (d, 1H), 4.69 (s, 2H), 4.40 (t, 2H), 4.33 (q, 2H), 2.61 (t, 2H), 2.02 (m, 2H), 1.67 (m, 2H), 1.35 (t, 3H), 0.99 (t, 3H), 0.93 (t, 3H);

¹³ C-NMR (150 MHz, DMSO) δ 168.8, 151.6, 142.8, 136.3, 128.5, 127.5, 124.0, 119.7, 111.2, 65.5, 61.5, 51.9, 37.2, 24.8, 23.9, 14.3, 13.9, 11.2.

Example 39: Preparation of 4-(5-bromo-2-methoxyphenyl)-1-(cyclobutylmethyl)-1H-1,2,3-triazole (GS36)

The title compound ( GS36 ,) was obtained by reaction and purification in a similar manner as in Example 1, except that (bromomethyl)cyclobutane was used instead of benzyl bromide. 407 mg, 89%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 8.48 (d, 1H), 7.97 (s, 1H), 7.38 (m, 1H), 6.84 (d, 1H), 4.41 (d, 2H), 3.93 (s, 3H), 2.89 (m, 1H), 2.12 (m, 2H), 1.90 (m, 4H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 154.6, 141.8, 131.2, 130.2, 123.1, 121.5, 113.6, 112.5, 55.7, 55.2, 35.6, 25.8, 18.1.

Example 40: Preparation of 4-(5-allyl-2-methoxyphenyl)-1-(cyclobutylmethyl)-1H-1,2,3-triazole (GS37)

The reaction and purification were carried out in a similar manner as in Example 2, except that 4-(5-bromo-2-methoxyphenyl)-1-(cyclobutylmethyl)-1H-1,2,3-triazole ( GS36 ) was used instead of GS27 , to give the title compound ( GS37 , 302 mg, 86%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 8.18 (d, 1H), 7.98 (s, 1H), 7.12 (m, 1H), 6.91 (d, 1H), 6.91 (m, 1H), 5.08 (m, 2H), 4.41 (d, 2H), 3.92 (s, 3H), 3.41 (d, 2H), 2.89 (m, 1H), 2.12 (m, 2H), 1.90 (m, 4H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 154.1, 143.0, 137.8, 132.7, 128.7, 127.8, 122.8, 119.4, 115.6, 110.9, 55.5, 55.1, 39.4, 35.6, 25.8, 18.1.

Example 41: Preparation of 4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenol (GS38)

The reaction was carried out in a similar manner as in Example 3, except that 4-(5-allyl-2-methoxyphenyl)-1-(cyclobutylmethyl)-1H-1,2,3-triazole ( GS37 ) was used instead of GS28 and the reaction was carried out at 0°C, to obtain the title compound ( GS38 , ) as a white solid. 257 mg, 90%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 10.76 (s, 1H), 7.80 (s, 1H), 7.221 (d, 1H), 7.05 (m, 1H), 6.99 (d, 1H), 5.97 (m, 1H), 5.06 (m, 2H), 4.45 (d, 2H), 3.34 (d, 1H), 2.91 (m, 1H), 2.15 (m, 2H), 1.91 (m, 4H) ;

¹³ C-NMR (150 MHz, Chloroform- d ) δ 154.3, 147.7, 137.8, 130.7, 130.0, 125.6, 118.5, 117.7, 115.6, 113.8, 55.6, 39.3, 35.5, 25.8, 18.1.

Example 42: Preparation of 4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenyl acetate (GS39)

Except that 4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenol ( GS38 ) was used instead of 4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenol, the reaction and purification were carried out in a similar manner as in Example 34, to give the title compound ( GS39 ,) as a white solid. 28 mg, 81%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 7.90 (d, 1H), 7.67 (s, 1H), 7.19 (m, 1H), 7.09 (d, 1H), 6.00 (m, 1H), 5.11 (m, 2H), 4.42 (d, 2H), 3.44 (d, 2H), 2.87 (m, 1H), 2.33 (s, 3H), 2.14 (m, 2H), 1.93 (m, 4H);

¹³ C-NMR (150 MHz, Chloroform -d ) δ 169.1, 145.4, 143.1, 138.3, 136.9, 129.1, 128.7, 123.2, 122.9, 121.3, 116.3, 55.3, 39.6, 35.6, 25.9, 21.3, 18.1.

Example 43: Preparation of ethyl 2-(4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenoxy)acetate (GS40)

The reaction and purification were carried out in a similar manner as in Example 4, except that GS38 was used instead of GS30 , to give the title compound ( GS40 , 36 mg, 90%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 8.58 (s, 1H), 8.26 (d, 1H), 7.09 (m, 1H), 6.78 (d, 1H), 5.98 (m, 1H), 5.07 (m, 2H), 4.68 (s, 2H), 4.43 (d, 2H), 4.33 (q, 2H), 3.41 (d, 2H), 2.91 (m, 1H), 2.12 (m, 2H), 1.90 (m, 4H), 1.35 (t, 3H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 168.6, 151.9, 142.6, 137.6, 133.7, 128.5, 127.8, 123.9, 120.11, 115.7, 111.5, 65.5, 61.4, 55.2, 39.5, 35.6, 25.8, 18.1, 14.2.

Example 44: Preparation of 1-(cyclobutylmethyl)-4-(2-methoxy-5-propylphenyl)-1H-1,2,3-triazole (GS41)

The reaction and purification were performed in a similar manner as in Example 29, except that GS37 was used instead of GT18 , to give the title compound ( GS41 , 9 mg, 90%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 8.17 (d, 1H), 7.97 (s, 1H), 7.10 (m, 1H), 6.89 (d, 1H), 4.41 (d, 2H), 3.91 (s, 3H), 2.90 (m, 1H), 2.60 (t, 1H), 2.11 (m, 2H), 1.91 (m, 4H), 1.66 (m, 2H), 0.94 (t, 1H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 153.8, 143.1, 135.3, 128.6, 127.6, 122.8, 119.1, 110.7, 55.5, 55.1, 37.2, 35.6, 25.8, 24.8, 18.1, 13.8.

Example 45: Preparation of ethyl 2-(2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)-4-propylphenoxy)acetate (GS44)

The reaction and purification were carried out in a similar manner as in Example 29, except that ethyl 2-(4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenoxy)acetate ( GS40 ) was used instead of GT18, to give the title compound ( GS44 , 8.7 mg, 87%) was obtained.

¹ H-NMR (400 MHz, Chloroform- d ) δ 8.58 (s, 1H), 8.24 (d, 1H), 7.08 (m, 1H), 6.76 (d, 1H), 4.68 (s, 2H), 4.44 (d, 2H), 4.33 (q, 2H), 2.91 (m, 1H), 2.60 (t, 2H), 2.12 (m, 2H), 1.91 (m, 4H), 1.67 (m, 2H), 1.35 (t, 3H), 0.94 (t, 3H);

¹³ C-NMR (150 MHz, Chloroform- d ) δ 168.7, 151.6, 142.7, 136.3, 128.4, 127.6, 123.9, 119.9, 111.3, 65.6, 61.4, 55.2, 37.2, 35.6, 25.8, 24.7, 18.1, 14.2, 13.8.

The structural formulas of the compounds manufactured in Example 1-45 are summarized in Table 1 below.

Experimental Example 1. Activation of autophagy by compound treatment in human retinal pigment epithelial cells

To evaluate autophagy activity by treatment with the compound represented by the above chemical formula 1, Western immunoblotting was performed using human retinal pigment epithelial cells, ARPE-19, to confirm whether there was an increase in LC3-II, one of the major indicator proteins of autophagy activation.

Specifically, ARPE-19 cells were seeded in 6-well plates at a concentration of 1 × 10 ⁵ cells/well and cultured for 24 hours, and then treated with the compound (GS-32) at concentrations of 25 and 50 μM for 6 hours. Afterwards, a cell lysis buffer containing protease inhibitors (50 mM Tris-chloride, pH 8.0, 150 mM sodium chloride, 1% NP-40, 1% sodium deoxycholate, 1% sodium dodecyl sulfate, and 2 mM ethylenediamine tetraacetic acid) was added to prepare a cell lysate. The LC3-II level was measured using an anti-LC3-II antibody (Cell Signaling Technology, MA, USA), and an antibody against β-actin (Cell Signaling Technology) was used as an internal control.

As a result, as shown in Fig. 1a, it was confirmed that the level of LC3-II, an autophagy marker, increased in a concentration-dependent manner.

In addition, when autophagy is activated, LC3-Ⅱ is involved in autophagosome formation and forms puncta in the cytoplasm, so intracellular autophagosome formation was observed.

Specifically, cells were transfected into ARPE-19 cells with a plasmid expressing RFP-GFP-LC3 (Addgene Inc. MA, USA) using Lipofectamine™ 2000 reagent (Invitrogen, Carlsbad, CA, USA). After 48 h of incubation, the cells were treated with the compound (GS-32) at concentrations of 10, 25, and 50 μM. After additional 12 h of incubation, the cells were fixed using 4% paraformaldehyde for 10 min, and then the cells were washed three times with PBS every 5 min. Finally, the puncta formation of RFP-GFP-LC3 was observed using a confocal laser scanning microscope. The nuclei were stained with 1 μg/mL of Hoechst 33342 (Thermo Scientific, Rockford, IL, USA).

As a result, as shown in Fig. 1b, not only was there an increase in intracellular GFP-LC3-II puncta (GFP) observed by the compound treatment, but a more marked increase in RFP-LC3-II puncta was also observed. When these two images were merged, RFP-LC3-II puncta were observed to be significantly more predominant.

The above results indicate that the compound of the present invention activates the autophagic flux.

Experimental Example 2. Confirmation of removal of A2E-fluorescent labeling substance by compound

Fluorescence spectrophotometry

To evaluate the intracellular A2E removal ability by treatment with the compound represented by the above chemical formula 1, an easily detectable A2E-fluorescent labeling substance (A2E-BDP) was used.

Specifically, ARPE-19 cells were seeded at a concentration of 5×10 ³ cells/well in a 96-well plate and cultured for 24 hours. Then, A2E-BDP was treated at a concentration of 10 μM for 24 hours to induce A2E-BDP accumulation in the ARPE-19 cell line. The A2E-BDP-containing ARPE-19 cells were treated once with the compound (GS-32) or the compound of Comparative Example 1 (2, 10, 25 μM) and cultured for 24 hours. A cell lysis buffer containing a protease inhibitor was added to each 96-well to lyse the cells, and the cell lysate was transferred to a white 96-well plate. The fluorescence intensity was measured (excitation 485 nm, emission 535 nm) using a fluorescence microplate reader (VICTOR™ X3, PerkinElmer, USA).

As a result, as shown in Figs. 2a and 2b, it was confirmed that the removal rate of A2E-fluorescent labeling substance (A2E-BDP) accumulated in ARPE-19 cells was increased in a concentration-dependent manner by the compound treatment. In addition, it was confirmed that it exhibited better A2E removal ability than the compound of Comparative Example 1.

Experimental Example 2-1. Confirmation of A2E removal by compound treatment

HPLC method

A2E-BDP, confirmed in the above Experimental Example 2, is known to have the same intracellular dynamics and characteristics as A2E. However, since A2E is the actual substance existing in the body, the quantitative change in intracellular A2E was directly measured using the HPLC method.

Specifically, ARPE-19 cell lines were seeded at a concentration of 4 × 10 ³ cells/well in 6-well plates and cultured for 24 hours. Then, ARPE-19 cell lines were treated with A2E at a concentration of 50 μM for 48 hours to cause intracellular accumulation of A2E. To evaluate the A2E removal ability, A2E-containing ARPE-19 cells were treated with the compound (GS-32) or the compound of Comparative Example 1 at 50 μM for 24 hours. Thereafter, ARPE-19 cells were washed with phosphate-buffered saline (PBS), and a cell lysis buffer containing a protease inhibitor was added to prepare a cell lysate. Chloroform and methanol (2:1, v/v) were added to the cell lysate, shaken to extract A2E, and then centrifuged at 10,000 g for 10 minutes. The organic phase was dried under vacuum and redissolved in 100 mL of acetonitrile. LC-MS (Shimadzu LC-MS 2010EV) analysis was performed using a C18 column (2'50 mm) with acetonitrile and 0.1% formic acid (gradient, 0-100%, 0-10 min; flow rate 1.0 ml/min; monitored at 503 nm). All operations during the extraction of A2E present in the cytoplasm of ARPE-19 cells were performed under weak red light to prevent the destruction of A2E.

As a result, as shown in Fig. 3, it was confirmed that the amount of A2E in ARPE-19 cells accumulated in A2E was significantly reduced by treatment with the compound of the present invention. In addition, it was confirmed that it exhibited superior A2E removal ability than the compound of Comparative Example 1.

The above results suggest that the compound represented by the chemical formula 1 directly removes A2E present in ARPE-19 cells.

Experimental Example 3. Autophagy-inducing activity of various derivative compounds

The effect of activating autophagy in human retinal pigment epithelial cells (ARPE-19) by various derivative compounds of the compound represented by the above chemical formula 1 was evaluated. The experimental method was the same as Experimental Example 1, and the effect of activating autophagy through the increase of LC3-Ⅱ was evaluated and shown in Tables 2 and 3 below. The results of the DMSO treatment group were used as a control.

derivative Concentration (μM) LC3-Ⅱ±β-actin DMSO 50 1.00 GS-27 25 1.32 50 1.36 GS-28 25 2.79 50 3.87 GS-29 25 1.11 50 1.19 GS-30 25 1.56 50 1.67 GS-31 25 1.25 50 1.58 GS-32 25 1.62 50 2.47 GS-33 25 1.30 50 1.47 GS-35 25 1.26 50 1.56 GS-37 25 1.37 50 1.49 GS-38 25 1.20 50 1.09 GS-39 25 0.88 50 1.07 GS-40 25 1.40 50 1.29 GS-41 25 1.14 50 1.09 GS-43 25 1.20 50 1.00 GS-44 25 1.37 50 1.25 GS-45 25 1.08 50 1.02 GS-46 25 0.81 50 1.25 GS-47 25 1.16 50 1.55 GS-48 25 1.90 50 3.00 GS-50 25 1.48 50 1.57 GS-51 25 0.97 50 1.44 GS-52 25 1.07 50 1.32

derivative Concentration (μM) LC3-Ⅱ±β-actin DMSO 50 1.00 GT-04 25 1.35 50 1.92 GT-05 25 3.64 50 4.98 GT-06 25 2.71 50 3.76 GT-07 25 2.98 50 3.42 GT-09 25 2.17 50 2.52 GT-10 25 19.00 50 17.32 GT-11 25 2.37 50 3.38 GT-12 25 6.16 50 5.89 GT-13 25 3.15 50 3.43 GT-14 25 4.07 50 3.49 GT-15 25 2.15 50 2.87 GT-16 25 1.94 50 1.57 GT-17 25 1.66 50 1.81 GT-18 25 1.80 50 2.09 GT-19 25 2.66 50 2.54 GT-20 25 1.80 50 2.81 GT-21 25 1.63 50 3.80 GT-22 25 2.52 50 2.10

As shown in Tables 2 and 3, the autophagy activation effect of derivative compounds of the compound represented by Chemical Formula 1 according to the present invention was confirmed.

Experimental Example 4. Confirmation of removal of A2E-fluorescent labeling material by treatment with derivative compound

Fluorescence spectrophotometry

The intracellular A2E removal ability by treatment with a compound represented by chemical formula 1 (GS-32 derivative compound) was evaluated using a fluorescence spectrophotometric method.

The experimental method was performed in the same manner as Experimental Example 2, treating the compound at a concentration of 50 μM.

As a result, as shown in Fig. 4, it was confirmed that the level of A2E was significantly reduced in ARPE-19 cells accumulating A2E-BDP by treatment with the compound.

Experimental Example 5. Confirmation of A2E removal ability by treatment with derivative compound

The intracellular A2E removal ability by treatment with a compound represented by chemical formula 1 (GS-32 derivative compound) was evaluated.

Specifically, ARPE-19 cells were seeded in 6-well plates at a density of 2 × 10 ⁴ cells/well and cultured for 24 h. Then, 5 μM A2E was treated three times at 48-h intervals, and 24 h after the final A2E treatment, 25 μM of each derivative was treated twice at 24-h intervals. The cells treated with the derivatives were fixed using 4% paraformaldehyde for 10 min and then washed three times with PBS every 5 min. Finally, intracellular accumulated A2E was observed using a confocal laser scanning microscope (Nikon, Minato, Tokyo, Japan). Nuclei were stained with 1 μg/mL Hoechst 33342 (Thermo Scientific, Rockford, IL, USA).

As a result, as shown in FIGS. 5a to 5d and 6a to 6c, the compounds of the present invention exhibited the effect of removing A2E accumulated in human retinal pigment epithelial cells (ARPE-19).

Experimental Example 6. Inhibition of retinal damage in an animal model

Exposure to blue light is a factor that increases oxidative stress in the retina, and the improvement effect of the compound of the present invention was evaluated through a blue light-induced retinal damage animal model (Balb/c mice). All tests were conducted at the Non-Clinical Center of the Bio Headquarters (Songdo, Korea) of the Korea Conformity Laboratories (KCL) and were performed after receiving approval from the Animal Experiment Ethics Committee (Approval No.: IA20-04287).

Specifically, 80 specific pathogen free (SPF) male BALB/c mice (Orient Bio, Korea) (5 weeks old) were acclimatized for 1 week, and then weighed the day before dosing. Only animals within ±20% of the average body weight were used in the test. After group separation, the animals were maintained in a dark room for 24 hours, and then maintained in a dark room throughout the blue light irradiation period.

After anesthetizing the experimental animals with isoflurane, blue light of 10,000 lux was irradiated for a total of 14 times (1 hr/day) for 2 weeks using a blue light irradiation device. The test substance was prepared by dissolving HP-beta-cyclodextrin in a 5% CMC Na solution to make a 10% concentration, and the compound GS-32 of the present invention was dissolved therein. The administration amount was 10 ml/kg/day, and 25 mg/kg/day was administered to each experimental animal. The test substance was administered orally forcibly using an oral administration sonde (feeding needle) after grasping and correcting the skin on the dorsum of the test animal, once/day, for 7 days/week, for 4 weeks, in the morning on the day of administration. The initial 2 weeks overlapped with the blue light irradiation period, and administration was performed for a total of 4 weeks including 2 weeks after the end of blue light irradiation. On the day of administration, the test substance was administered before blue light irradiation. Before the start of administration and once a week, mydriatic agents were instilled into both eyes of all animals to dilate the pupils, and the fundus of the eyes was observed using a fundus camera (Genesis-D, Kowa Co. Ltd., Japan). Photographs were taken of each animal the week after the end of test substance administration.

After the end of the experiment, both eyes were enucleated for all animals, fixed in Davidson's solution, and stained with H&E. The average thickness of the ONL (outer nuclear layer) and IS/OS (inner segments/outer segments layer) was measured at a location 140 μm (left/right) from the retinal nerve center using the Axio Vision SE644 (ZEISS) program. The ONL and IS/OS thickness of the contralateral eye were measured in a similar way, and the average value of both eyes was used as the final thickness. One-way ANOVA test was used between each group, and it was considered statistically significant when p<0.05 or less. The SPSS 12.0 K program (SPSS, Chicago, IL, U.S.A.), a commercially widely used statistical package, was used for statistical analysis.

As a result, as shown in Figs. 7a to 7c, retinal damage was confirmed in the blue light irradiation control group compared to the normal control group, but it was confirmed that retinal damage was improved in the 25 mg/kg/day oral administration group of the compound as shown in the fundus examination (Fig. 7a). In addition, the results of histomorphological analysis of ocular tissue showed that ONL and IS/OS thickness were reduced in the blue light irradiation control group compared to the normal control group, but it was confirmed that statistically significant (p<0.01) recovery was achieved by compound administration (Figs. 7b and 7c). In addition, it was confirmed that the effect of improving retinal damage and recovering ocular tissue was superior to that of the compound of Comparative Example 1.

From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without changing the technical idea or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are exemplary in all respects and not restrictive. The scope of the present invention should be interpreted as including all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts rather than the above detailed description.

Claims (21)

A compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof: [Chemical Formula 1]

In the above chemical formula 1, R ₁ is hydrogen, C _1-4 alkyl, carbamoyl-C _1-4 alkyl, carboxy-C _1-4 alkyl, C _1-4 alkylcarbonyl, C _1-4 alkoxycarbonyl-C _1-4 alkyl, C _6-10 arylcarbonyl-C _1-4 alkyl, 5-10 membered heteroaryl-C _1-4 alkyl, 3 to 10 membered heterocyclyl-C _1-4 alkyl, 3 to 10 membered heterocyclylcarbonyl, C _3-10 cycloalkylcarbamoyl-C _1-4 alkyl, or substituted C _6-10 aryl-C _1-4 alkyl-carbamoyl-C _1-4 alkyl, wherein the substituted C _6-10 aryl-C _1-4 alkyl-carbamoyl-C _1-4 alkyl is C _6-10 C _1-4 alkoxycarbonyl is substituted on the C _1-4 alkyl of aryl-C _1-4 alkyl, R ₂ is halogen, C _1-4 alkyl, C _2-4 alkenyl, C _3-10 cycloalkyl, C _3-10 cycloalkenyl, C _6-10 aryl, or 5-10 membered heteroaryl, R ₃ is C _1-4 alkyl, C _2-4 alkenyl, C _3-10 cycloalkyl-C _1-4 alkyl, or C _6-10 aryl-C _1-4 alkyl, The above C _3-10 cycloalkyl, C _6-10 aryl, 3 to 10 membered heterocyclyl and 5 to 10 membered heteroaryl may be unsubstituted or substituted with one or more selected from the group consisting of hydroxy, halogen, C _1-4 haloalkyl, C _1-4 alkylsulfonyl, and C _1-4 alkoxycarbonyl. In the first paragraph, A compound or a pharmaceutically acceptable salt thereof, wherein R ₁ is hydrogen, methyl, carbamoylmethyl, carboxymethyl, methylcarbonyl, ethoxycarbonylmethyl, ethoxycarbonylisopropyl, tert-butoxycarbonyl-methyl, methylsulfonyl-substituted phenylcarbonylmethyl, pyridinylmethyl, piperidinylmethyl, isopropylsulfonyl-substituted piperidinylmethyl, tert-butoxycarbonyl-substituted piperidinylmethyl, morpholinylcarbonyl, cyclohexylcarbamoylmethyl, hydroxy-substituted cyclohexylcarbamoylmethyl, or N-(1-methoxycarbonyl-2-phenylethyl)carbamoylmethyl. In the first paragraph, A compound or a pharmaceutically acceptable salt thereof, wherein R ₂ is bromo, propyl, isobutyl, propenyl, cyclopentyl, cyclopentenyl, fluoro-substituted phenyl, pyridinyl or trifluoromethyl substituted pyrazolyl. In the first paragraph, A compound or a pharmaceutically acceptable salt thereof, wherein R ₃ is propyl, propenyl, cyclobutylmethyl, or benzyl. In the first paragraph, 1-Benzyl-4-(5-bromo-2-methoxyphenyl)-1H-1,2,3-triazole 4-(5-allyl-2-methoxyphenyl)-1-benzyl-1H-1,2,3-triazole 4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenol Ethyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate 4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenyl acetate (R)-methyl 2-(2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetamido)-3-phenylpropanoate (S)-methyl 2-(2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetamido)-3-phenylpropanoate tert-butyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-1-(4-(methylsulfonyl)phenyl)ethanone), 2-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)pyridine), 3-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)pyridine), 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)pyridine), 4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenyl morpholine-4-carboxylate Ethyl 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-2-methylpropanoate 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)piperidine), 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetic acid 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)-1-(isopropylsulfonyl)piperidine), 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)acetamide, 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-N-cyclohexylacetamide), 2-(4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)-N-(4-hydroxycyclohexyl)acetamide tert-butyl 4-((4-allyl-2-(1-benzyl-1H-1,2,3-triazol-4-yl)phenoxy)methyl)piperidine-1-carboxylate), 2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-bromophenol, Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-bromophenoxy)acetate Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-isobutylphenoxy)acetate Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-cyclopentenylphenoxy)acetate Ethyl 2-(3-(1-benzyl-1H-1,2,3-triazol-4-yl)-4'-fluorobiphenyl-4-yloxy)acetate Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-(1-(trifluoromethyl)-1H-pyrazol-4-yl)phenoxy)acetate Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-(pyridin-4-yl)phenoxy)acetate Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-cyclopentylphenoxy)acetate 1-benzyl-4-(2-methoxy-5-propylphenyl)-1H-1,2,3-triazole 2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-propylphenol, 2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-propylphenyl acetate Ethyl 2-(2-(1-benzyl-1H-1,2,3-triazol-4-yl)-4-propylphenoxy)acetate 4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenyl acetate Ethyl 2-(4-allyl-2-(1-allyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate 4-(2-methoxy-5-propylphenyl)-1-propyl-1H-1,2,3-triazole 4-propyl-2-(1-propyl-1H-1,2,3-triazol-4-yl)phenyl acetate Ethyl 2-(4-propyl-2-(1-propyl-1H-1,2,3-triazol-4-yl)phenoxy)acetate 4-(5-bromo-2-methoxyphenyl)-1-(cyclobutylmethyl)-1H-1,2,3-triazole 4-(5-allyl-2-methoxyphenyl)-1-(cyclobutylmethyl)-1H-1,2,3-triazole 4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenol 4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenyl acetate Ethyl 2-(4-allyl-2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)phenoxy)acetate 1-(cyclobutylmethyl)-4-(2-methoxy-5-propylphenyl)-1H-1,2,3-triazole, or A compound, which is ethyl 2-(2-(1-(cyclobutylmethyl)-1H-1,2,3-triazol-4-yl)-4-propylphenoxy)acetate, or a pharmaceutically acceptable salt thereof. A method for producing a phenyl-triazole derivative having R _{3 '} substituted on the triazole ring, comprising a first step of reacting 4-bromo-2-ethynyl-1-methoxybenzene with a mixture of R ₃ '-X ₁ and NaN ₃ in the presence of CuBr and PMDETA (N,N,N',N",N"-pentamethyldiethylenetriamine). A method for producing a compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, A manufacturing method, wherein the above R ₃ 'is R ₃ or a precursor thereof, and X ₁ is a halogen. In Article 6, Step a of replacing Br on the phenyl ring of the product obtained from the previous step with R ₂ '; Step b of replacing OMe on the phenyl ring of the product obtained from the previous step with OH; and It further comprises at least one step selected from the group consisting of step c of replacing OH on the phenyl ring of the product obtained from step b with R ₁ ', A manufacturing method, wherein the above R ₁ ' and R ₂ ' are R ₁ or a precursor thereof, and R ₂ or a precursor thereof, respectively. In Article 7, Step a is carried out by reacting i) R ₂ '-SnBu ₃ or ii) R ₂ '-boronic acid and Cs ₂ CO ₃ in the presence of Pd(PPh ₃ ) ₄ , Step b is performed by reacting B(X ₂ ) ₃ at -60°C to 0°C. Step c is carried out i) in the presence of R ₁ '-X ₃ and K ₂ CO ₃ , or ii) in the presence of acetic anhydride or R ₁ '-X ₃ and Et ₃ N (triethylamine) and optionally DMAP (4-dimethylaminopyridine), The above step a is performed after step 1 or after step c. A manufacturing method, wherein the above X ₂ and X ₃ are each independently a halogen. In Article 6, A method for producing a compound, wherein when R ₂ ', R ₃ ' or both contain alkenyl, the method further comprises a step of reducing the compound to alkyl. In Article 9, A manufacturing method, wherein the above step d is performed in the presence of a Pd/C catalyst under an H ₂ atmosphere. A pharmaceutical composition for preventing or treating an ocular disease, comprising a compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof. A pharmaceutical composition according to claim 11, wherein the eye disease is at least one selected from the group consisting of dry macular degeneration, dry eye, eye fatigue, and decreased vision. In Article 11, The composition is a pharmaceutical composition that exhibits an effect of inhibiting or removing A2E (N-retinyl-N-retinylidene ethanolamine) accumulation in human retinal pigment epithelium cells (ARPE-19) and an effect of activating autophagy in human retinal pigment epithelium cells. In Article 11, A pharmaceutical composition, wherein the composition additionally comprises a pharmaceutically acceptable carrier or excipient. In Article 11, A pharmaceutical composition characterized in that the composition suppresses the worsening or progression of an eye disease. A method for preventing or treating an eye disease, comprising administering the pharmaceutical composition of Article 11 to a subject other than a human. An autophagy-activating composition comprising a compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof. A food composition for preventing or improving ocular disease, comprising a compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof. A feed composition for preventing or improving eye disease, comprising a compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof. A method for preventing or treating an eye disease, comprising a step of administering a pharmaceutically effective amount of a compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof to a subject in need thereof. Use of a compound of any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing or treating eye diseases.

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