Pharmaceutical composition for treating obesity and meibomian gland dysfunction comprising PLK1 inhibitor as active ingredient

Abstract

The present invention relates to a composition for inhibiting adipogenesis, the composition comprising a PLK1 inhibitor as an active ingredient. In addition, it was found that high-fat diet-induced obesity is alleviated and treated by administering a low concentration of the PLK1 inhibitor, and it was found that the PLK1 inhibitor has an inhibitory effect on adipogenesis in human meibomian gland epithelial cells, and thus can treat eye disorders such as dry eye syndrome. In addition, it was found that the PLK1 inhibitor interferes with adipose tissue accumulation in liver cells, and thus can alleviate or treat liver diseases including fatty liver and liver fibrosis.

Description

Pharmaceutical composition for treating obesity and meibomian gland dysfunction containing a PLK1 inhibitor as an active ingredient

[0001] The present invention relates to the technical field of regulating intracellular lipid metabolism, and specifically to a pharmaceutical composition comprising a PLK1 inhibitor as an active ingredient.

[0002] Conventional obesity treatments aim for weight loss and primarily work by suppressing appetite, interfering with fat absorption, or accelerating metabolism. While these drugs can effectively induce initial weight loss, they are often accompanied by various side effects. For instance, appetite-suppressing medications can affect the central nervous system, potentially causing neurological side effects such as an increased heart rate. Additionally, drugs that block fat absorption can impair the absorption of essential nutrients along with fat, which may lead to gastrointestinal side effects such as diarrhea.

[0003] Long-term use of these drugs initially leads to weight loss, but over time, the effect diminishes, eventually resulting in a yo-yo effect where weight returns. This implies that the effects of obesity treatments may be temporary, making it difficult for patients to maintain a consistent weight. Therefore, there was an urgent need to develop a new mechanism that could overcome the limitations of existing treatments and enable long-term weight management. To this end, researchers completed this invention by focusing on gaining a deeper understanding of the fundamental causes of obesity and exploring various biological pathways involved in weight control.

[0004] The objective of the present invention is to provide a composition for inhibiting the formation of fat cells comprising a PLK1 inhibitor as an active ingredient.

[0005] Another objective of the present invention is to provide a pharmaceutical composition for the prevention or treatment of obesity comprising a PLK1 inhibitor as an active ingredient.

[0006] Another objective of the present invention is to provide a pharmaceutical composition for the prevention or treatment of ocular disorders comprising a PLK1 inhibitor as an active ingredient.

[0007] However, the technical problems that the present invention aims to solve are not limited to those mentioned above, and other unmentioned problems will be clearly understood by those skilled in the art from the description below.

[0008] Various embodiments of the present invention are described with reference to the drawings. In the following description, for a complete understanding of the present invention, various specific details, such as specific forms, compositions, and processes, are described. However, specific embodiments may be practiced without one or more of these specific details, or in combination with other known methods and forms. In other examples, known processes and manufacturing techniques are not described as specific details so as not to make the present invention unnecessary or obscure. Reference throughout this specification to one embodiment implies that the particular features, forms, compositions, or characteristics described in association with the embodiment are included in one or more embodiments of the present invention. Accordingly, the circumstances of the embodiments expressed at various locations throughout this specification do not necessarily represent the same embodiment of the present invention. Additionally, particular features, forms, compositions, or characteristics may be combined in any suitable way in one or more embodiments. Unless otherwise defined in the specification, all scientific and technical terms used in this specification have the same meaning as commonly understood by those skilled in the art to which the present invention pertains.

[0009] The term "PLK1 inhibitor" in the present invention refers to a compound or drug that inhibits the activity of a specific protein, polo-like kinase 1 (PLK1). Such inhibitors are used in cancer treatment by interfering with the growth and division of cancer cells.

[0010] The above PLK1 inhibitors may include, but are not limited to, Volasetib, Rigosertib, GSK461364, TAK-960, ZK-thiazolidinone, and BI 2536.

[0011] The term "BI 6727" in this invention, also known as Volasetib, is a small molecule inhibitor that specifically inhibits PLK1. This compound is being studied primarily for the treatment of solid tumors and hematological cancers and is currently in the clinical trial phase. Its CAS number is 755038-65-4, and its molecular weight is 618.31. Its IUPAC name is N-((1r,4R)-4-(4-(cyclopropylmethyl)piperazine-1-yl)cyclohexyl)-4-(((R)-7-ethyl-8-isopropyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridine-2-yl)amino)-3-methoxybenzamide. The above BI 6727 is IC 50 It inhibits PLK2 and PLK3 and plays a role in inducing somatic cell division arrest and apoptosis. In addition, BI 6727, which is also a dihydropteradinone derivative, shows significant antitumor activity in various cancer models. For cancer treatment purposes, the above BI 6727 can be administered at a dose of 25 mg / kg / day or more and 50 mg / kg / day or less.

[0012] It also has the structure of the following chemical formula 2.

[0013] Chemical formula 2

[0014]

[0015] The term "BI 2536" in this invention refers to a potent inhibitor targeting PLK1 that can contribute to cancer treatment by inhibiting the division of cancer cells. Furthermore, as a Pan-PLK1 inhibitor, it is a compound that simultaneously inhibits not only PLK1 but also several members of the Polo-like kinase family, such as PLK2, PLK3, and PLK4. This type of inhibitor can exhibit broader cellular regulatory effects by regulating activity across the PLKs. The CAS number is 755038-02-9, and the molecular weight is 521.65. The IUPAC name is (R)-4-((8-cyclopentyl-7-ethyl-5-methyl-6-oxo-5,6,7,8-tetrahydropteridine-2-yl)amino)-3-methoxy-N-(1-methylpiperidine-4-yl)benzamide. Additionally, BI 2536 is IC 50 It inhibits PLK1 and BRD4 and inhibits IFNB (encoding IFN-β) gene transcription.

[0016] In addition, when BI 2536 was administered to human meibomian gland epithelial cells, it was confirmed that PPARγ, which regulates adipogenesis, and SREBP-1 and ACC, which regulate lipogenesis, were significantly reduced.

[0017] It also has the structure of the following chemical formula 3.

[0018] Chemical formula 3

[0019]

[0020] BI 6727 and BI 2536, the PLK1 inhibitors of the present invention, have a significant effect on inhibiting adipocyte formation compared to other PLK1 inhibitors.

[0021] The term "Rigosertib" in the present invention is a multikinase inhibitor that inhibits the growth of cancer cells by regulating intracellular signaling pathways. It is an investigational drug primarily used for the treatment of myelodysplastic syndrome (MDS).

[0022] The term "GSK461364" in this invention refers to a selective inhibitor of PLK1 that inhibits the growth of cancer cells by intervening in the regulation of the cancer cell cycle. This compound is an anticancer therapeutic agent currently under development.

[0023] The term "TAK-960" in this invention refers to a small molecule inhibitor targeting PLK1, used to block the division and growth of cancer cells. This compound is being studied as a potential therapeutic agent for various types of cancer.

[0024] The term "ZK-thiazolidinone" in the present invention refers to a compound of the thiazolidinone class and is used for certain drugs having biological activity. This compound can exhibit anti-inflammatory and anticancer effects.

[0025] The term "pyrimidine ring" in this invention refers to a six-atom heteroaromatic compound used in organic chemistry that has a ring structure containing two nitrogen atoms. This structure is utilized as the basic framework for various drugs and biologically active molecules.

[0026] The term "amide bond" in the present invention refers to a covalent bond formed by a condensation reaction between a carboxyl group and an amine group. This bond plays an important role as a peptide bond in protein structures.

[0027] The term "ether bond" in the present invention refers to a bond within an organic compound in which two carbon atoms are connected through an oxygen atom. Ether bonds serve as stable mediators in various organic synthesis reactions.

[0028] The terms “alkyl” or “alk” of the present invention refer to linear or branched-chain alkanes (hydrocarbons) radicals comprising 1 to 12 carbon atoms, preferably 1 to 6 carbon atoms. Exemplary “alkyl” groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutylpentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, etc. For example, the term “(C1-C4)alkyl” refers to linear or branched-chain alkanes (hydrocarbons) radicals comprising 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, and isobutyl. "Substituted alkyl" refers to an alkyl group substituted with one or more substituents, preferably 1 to 4 substituents, at any available attachment site.

[0029] In the present invention, the term “aryl” refers to a monocyclic or polycyclic carbon ring that is wholly or partially unsaturated and aromatic. For example, a C6-C10 aryl refers to a monocyclic or polycyclic carbon ring that is wholly or partially unsaturated and aromatic, composed of 6 to 10 carbon atoms. For example, if it includes two or more aromatic rings (bicyclic, etc.), the aromatic rings of the aryl group may be connected at a single point (e.g., biphenyl) or fused (e.g., naphthyl, phenanthrenyl, etc.). “Substituted aryl” refers to an aryl group substituted by one or more substituents, preferably 1 to 3 substituents, at any available attachment point.

[0030] In the present invention, the term “carbocycle” refers to a fully saturated or partially saturated cyclic hydrocarbon group comprising 1 to 4 rings and 3 to 8 carbons per ring, or a cyclic, aromatic hydrocarbon group having 1 to 5 aromatic rings, in particular a monocyclic or bicyclic group such as phenyl, biphenyl, or naphthyl. The term “carbocycle” encompasses cycloalkyl, cycloalkenyl, cycloalkynyl, and aryl as previously defined. The term “substituted carbocycle” refers to a carbocycle or carbocyclic group substituted with one or more substituents, preferably 1 to 4 substituents, at any available attachment site.

[0031] In the present invention, the terms “cycloalkyl” or “cycloalkyl” refer to a fully saturated cyclic hydrocarbon group comprising 1 to 4 rings and 3 to 8 carbons per ring. They include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclodecyl. The terms “C3-C8 cycloalkyl”, “C3-C7 cycloalkyl”, and “C3-C6 cycloalkyl” have similar meanings; for example, “C3-C7 cycloalkyl” refers to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl. “Substituted cycloalkyl” refers to a cycloalkyl group substituted with one or more substituents, preferably 1 to 4 substituents, at any available attachment site.

[0032] In the present invention, the terms “heterocycle” and “heterocyclic” refer to a fully saturated, or partially or fully unsaturated, comprising an aromatic (i.e., “heteroaryl”) cyclic group (e.g., a 4 to 7-membered monocyclic, a 7 to 11-membered bicyclic, or an 8 to 16-membered tricyclic ring system) having at least one heteroatom within at least one carbon atom-containing ring. Each ring of the heterocyclic group containing the heteroatom may have 1, 2, 3, or 4 heteroatoms selected from a nitrogen atom, an oxygen atom, and / or a sulfur atom, wherein the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized. (The term "heteroarylium" refers to a heteroaryl group containing a quaternary nitrogen atom and is therefore positively charged.) A heterocyclic group can be attached to any heteroatom or carbon atom of the rest of the molecule in a ring or ring system. Exemplary monocyclic and heterocyclic groups include azetidinyl, pyrrolidinyl, pyrrolyl, pyrazolyl, oxetanil, pyrazolinyl, imidazollyl, imidazollinyl, imidazollinyl, oxazolyl, oxazolidinyl, isoxazolidinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazollyl, isothiazollyl, furyl, tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperajinil, 2-oxopiperajinil, 2-oxopyrrolodinyl, 2-oxoazefinil, azefinil, hexahydrodiazefinil, 4-piperidonyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, Includes triazolyl, tetrazolyl, tetrahydropyranil, morpholinyl, thiamofolinyl, thiamofolinyl sulfoxide, thiamofolinyl sulfone, 1,3-dioxolane and tetrahydro-1,1-dioxothienyl, etc.The terms “substituted heterocycle” and “substituted heterocyclic” (e.g., “substituted heteroaryl”) refer to a heterocycle or heterocyclic group substituted with one or more substituents, preferably 1 to 4 substituents, at any available attachment site.

[0033] In the present invention, the terms “aromatic heterocyclic” or “heteroaryl” have the same meaning and refer to a heteroaromatic group comprising one or more heteroatoms. The heteroatoms referred to herein include oxygen, sulfur, and nitrogen. Examples include furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring may be condensed to an aryl, heterocyclic group, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. The heteroaryl is optionally substituted or unsubstituted.

[0034] In the present invention, the term "alkylamino" refers to a group having the structure-NHR', where R' is hydrogen, alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl as defined above.

[0035] In the present invention, the term "dialkylamino" refers to a group having the structure-NRR', wherein R and R' are each independently an alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl, heterocyclil or substituted heterocyclil as defined above. R and R' may have the same or different dialkylamino moiety. Examples of dialkylamino groups include, but are not limited to, dimethylamino, methyl ethylamino, diethylamino, methylpropylamino, di(n-propyl)amino, di(iso-propyl)amino, di(cyclopropyl)amino, di(n-butyl)amino, di(tert-butyl)amino, di(neopentyl)amino, di(n-pentyl)amino, di(hexyl)amino, di(cyclohexyl)amino, etc. In certain embodiments, R and R' are connected to form a cyclic structure. The resulting cyclic structure may be aromatic or non-aromatic. Examples of cyclic diaminoalkyl groups include, but are not limited to, aziridinyl, pyrrolidinyl, piperidinyl, morpholinyl, pyrrolyl, imidazolyl, 1,3,4-trianolyl, and tetrazolyl.

[0036] In the present invention, the term “substitution” means that one or more hydrogen atoms on a specified group are substituted by a specified substituent. The specified substituent is the substituent correspondingly described above or the substituent shown in each example. Unless specifically described, the specified substituted group may have one substituent selected from a specific group at any substitutable site of the group, and said substituent may be the same or different at each position. Cyclic substituents, such as heterocycloalkyl, may be connected to another ring, such as cycloalkyl, to form a spiro-double ring system, such as a two-ring system having one common carbon atom. Those skilled in the art should understand that the combination of substituents anticipated in the present invention is a stable or chemically feasible combination. The above substituents are, for example, C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C3-8 cycloalkyl, 3- to 12-membered heterocyclic groups, aryl, heteroaryl, halogen, hydroxyl, carboxyl (-COOH), C1-8 aldehyde group, C2-10 acyl, C2-10 ester group, C1-C12 alkoxycarbonyl, amino, alkoxy, C1-10 sulfonyl, etc., but are not limited thereto.

[0037] Unless otherwise determined, any heteroatom with an unfulfilled valence is assumed to have enough hydrogen atoms to satisfy the valence.

[0038] As used herein, the terms “combination” or “pharmaceutical combination” refer to a product produced from a mixture or combination of more than one active ingredient and comprising both fixed and non-fixed combinations of the active ingredient. The term “fixed combination” means that an active ingredient, e.g., a compound of the present invention, and one or more additional therapeutic agents are administered to a patient simultaneously in the form of a single substance or dosage. The term “non-fixed combination” means that an active ingredient, e.g., a compound of the present invention, and one or more additional therapeutic agents are administered to a patient simultaneously, jointly, or sequentially without specific time limits as individual substances, wherein such administration provides a therapeutically effective level of the active ingredient within the patient’s body. The latter also applies to cocktail therapy, e.g., the administration of three or more active ingredients.

[0039] As used herein, the terms “composition” or “pharmaceutical composition” refer to a mixture of at least one of the compounds of the present invention and optionally one or more other pharmaceutically acceptable chemical components, such as a carrier, stabilizer, diluent, dispersant, suspending agent, thickener and / or excipient.

[0040] As used herein, the terms "optical isomer" or "stereoisomer" refer to any of the various stereoisomer coordinations that may exist for a given compound of the present invention, including geometric isomers. Substituents are understood to be capable of attaching to the chiral center of a carbon atom. The term "chiral" refers to a molecule having the characteristic of non-superimposability on its mirror image partner, whereas the term "achiral" refers to a molecule capable of superposition on its mirror image partner. Accordingly, the present invention includes enantiomers, diastereomers, or racemics of a compound. "Enantiomers" are a pair of stereoisomers that are mirror images of each other and are non-superimposable. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The term is used to designate racemic mixtures where appropriate. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-Prelog RS system. If the compound is a pure enantiomer, the stereochemistry at each chiral carbon may be specified by R or S. Resolved compounds whose absolute coordination is unknown may be designated as (+) or (-) depending on the direction in which they rotate plane polarization at the wavelength of the sodium D line (left-handed or right-handed). The specific compounds described herein may contain one or more asymmetric centers or axes and thus produce enantiomers, diastereomers, and other stereoisomer forms that can be defined as (R)- or (S)- in terms of absolute stereochemistry.

[0041] Salts of the compounds of the present invention having at least one salt-forming group can be prepared in a manner known to those skilled in the art. For example, salts of the compounds of the present invention having an acid group can be formed, for example, by treating the compound with a metal compound, e.g., an alkali metal salt of a suitable organic carboxylic acid, e.g., a sodium salt of 2-ethylhexanoic acid; an organic alkali metal or alkaline earth metal compound, e.g., a corresponding hydroxide, carbonate, or hydrogen carbonate, e.g., sodium hydroxide or potassium hydroxide, carbonate, or hydrogen carbonate; a corresponding calcium compound; or ammonia or a suitable organic amine, preferably in stoichiometric amounts or only a slight excess of the salt-forming agent. Acid addition salts of the compounds of the present invention are obtained in a conventional manner, for example, by treating the compound with an acid or a suitable anion exchange reagent. The internal salt of the compound of the present invention containing acid and basic salt-forming groups, for example, free carboxyl groups and free amino groups, can be formed, for example, by neutralizing a salt, such as an acid addition salt, to an isoelectric point using a weak base, for example, or by treating with an ion exchanger.

[0042] Salts can be converted into free compounds according to methods known to those skilled in the art. Metal and ammonium salts can be converted, for example, by treatment using a suitable acid, and acid addition salts can be converted, for example, by treatment using a suitable basic agent.

[0043] All of the above-mentioned process steps may be carried out under reaction conditions known to a person skilled in the art, including those specifically mentioned, in the absence or ordinarily present of a solvent or diluent (e.g., a solvent or diluent that is inert to the reagent used and dissolves it), in the absence or present of a catalyst, condensing agent or neutralizing agent, e.g., an ion exchanger depending on the nature of the reaction and / or reactants, e.g., a cation exchanger (e.g., in the form of H+), at reduced, ordinary, or elevated temperatures, e.g., in a temperature range of about -100°C to about 190°C (e.g., approximately -80°C to about 150°C, e.g., -80 to -60°C, room temperature, -20 to 40°C, or reflux temperature), under atmospheric pressure or in a sealed container, under pressure where appropriate, and / or in an inert atmosphere, e.g., under an argon or nitrogen atmosphere.

[0044] The term "PPARγ" in this invention refers to a type of nuclear receptor that plays an important role in regulating lipid metabolism and glucose homostasis. This receptor is a major target in the development of treatments for diabetes, obesity, and cardiovascular disease.

[0045] The term "SREBP-1" of the present invention refers to Sterol Regulatory Element-Binding Protein-1, an important transcription factor that regulates lipid synthesis. This protein plays a crucial role in fatty acid and cholesterol metabolism by regulating the expression of genes that promote the synthesis of fatty acids and triglycerides, and is involved in the regulation of metabolic diseases. The term "FASN PI3K" of the present invention refers to the binding of Fatty Acid Synthase and Phosphoinositide 3-Kinase. This binding is important for activating signaling pathways essential for cell growth and survival.

[0046] The term “metabolic disease” in the present invention refers to a disease resulting from abnormalities in energy metabolism, lipid metabolism, or glucose metabolism, and is a concept that encompasses a group of diseases including the disruption of the body’s energy balance, abnormal fat accumulation, insulin resistance, persistence of inflammatory responses, or tissue dysfunction resulting therefrom. The metabolic diseases include, but are not limited to, obesity, fatty liver, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and cirrhosis, and these diseases may occur alone or in combination. In particular, fatty liver and liver fibrosis can be understood as a mode of metabolic disease associated with lipid accumulation in the liver, inflammatory responses, and progression of fibrosis caused by obesity or metabolic abnormalities.

[0047] The term “metabolic disease” in this invention refers to a concept that can occur regardless of weight gain or loss, and encompasses pathological conditions associated with the long-term disruption of metabolic homeostasis, extending beyond simple weight control disorders.

[0048] The term "p-AKT(Ser473) / AKT" of the present invention refers to the phosphorylated form of protein kinase B (AKT) at the serine 473 position. This form indicates the activation of AKT and is involved in regulating various cellular functions such as cell survival, growth, and division.

[0049] The term "obesity" in this invention refers to a condition in which excessive fat accumulates in the body and has a negative impact on health. This increases the risk of chronic diseases and is associated with various health problems, such as cardiovascular disease and diabetes.

[0050] The term "ocular disorder" in this invention is a general term for various diseases occurring on the surface of the eye, including tear film instability, eyelid abnormalities, or conjunctival inflammation.

[0051] The above ocular disorders may include, but are not limited to, dry eye syndrome, blepharitis, conjunctivitis, squamous cell dysplasia, and bullous keratoconjunctivitis, and these diseases may be caused by the excessive accumulation of epithelial fat cells.

[0052] The term "dry eye syndrome" in this invention refers to dryness of the eyes caused by insufficient tear secretion or a deterioration in tear quality. This condition may result in eye discomfort and vision problems.

[0053] The term "blepharitis" in the present invention refers to an inflammatory condition of the eyelids, which may be caused by bacteria or an allergic reaction. This may include itching, redness, and dandruff formation of the eyelids.

[0054] The term "conjunctivitis" in this invention refers to a condition in which the conjunctiva becomes inflamed, caused by various factors such as viruses, bacteria, and allergies. It is accompanied by redness of the eye, increased discharge, and itching.

[0055] The term "squamous cell dysplasia" in this invention refers to a disease characterized by abnormal growth or changes in squamous epithelial cells. This results in abnormal cell growth and sometimes increases the risk of progression to cancer.

[0056] The term "bullous keratoconjunctivitis" in this invention refers to a disease in which inflammation occurs in the cornea and conjunctiva, which are primarily located in the upper part of the eye, causing eye pain, redness, and discomfort. This disease impairs the stability of the eye and requires appropriate treatment.

[0057] The term "meibomian gland" in this invention refers to a small oil gland located in the eyelid that functions to secrete the lubricating oil of the tear film. The normal function of this gland is important for preventing dry eyes and maintaining eye health.

[0058] The term "epithelial cell" in this invention refers to a cell that covers the surface of the body or the interior of an organ, performing functions such as protection, secretion, and absorption. These cells play a very important role in the body and constitute various tissues and organs.

[0059] The term “HMGECs (Human Meibomian Gland Epithelial Cells)” in this invention refers to human meibomian gland epithelial cells. Meibomian glands are located in the eyelids and play a role in reducing tear evaporation by producing an oily layer of the eye. Therefore, HMGECs are a type of cell primarily used in research related to meibomian gland function.

[0060] The term "fat cell" in this invention refers to a cell that primarily stores fat and plays roles such as energy storage, heat retention, and shock absorption. These cells are important for maintaining energy balance within the body.

[0061] Gene expression regulation refers to the ability to control the expression of genes related to adipocyte differentiation and lipid synthesis by regulating the activity of specific transcription factors, such as PPARγ (peroxisome proliferator-activated receptor gamma). By regulating adipocyte maturation or lipid storage through the activation or inhibition of factors like PPARγ, the production of lipids within adipocytes can be suppressed.

[0062] The term "pre-adipocyte" in this invention refers to a cell that has not yet differentiated into a full adipocyte and possesses the potential to transform into an adipocyte as needed. These cells are important for the growth and regeneration of adipose tissue.

[0063] The term "adipocyte" in the present invention refers to a cell that stores or accumulates fat as a major component, and includes cells involved in energy storage, metabolic regulation, and hormone secretion within the body. Such adipocytes may include typical white adipocytes and brown adipocytes. Furthermore, the term "adipocyte" is not limited to cells present in typical adipose tissue but may be used to include cells that excessively accumulate fat or have activated fat synthesis pathways, such as steatotic hepatocytes or cells in a pathological state associated with fat accumulation.

[0064] Accordingly, in the present invention, inhibition of adipocyte formation may include not only inhibiting differentiation from preadipocytes into adipocytes, but also inhibiting the formation or functional activation of cells with increased lipid accumulation or lipidization.

[0065] The term “inhibition of adipocyte formation” in the present invention refers to a state in which biological processes related to the formation of adipocytes or fat accumulation are reduced or inhibited, which may include the inhibition of the formation, synthesis, or accumulation of fat within adipocytes. Such inhibition of adipocyte formation may be related to the prevention or treatment of obesity, metabolic disorders, or pathological conditions associated with fat accumulation (e.g., fibrosis). Although not limited thereto, said inhibition of adipocyte formation may be achieved through the inhibition of fatty acid synthesis, the promotion of fatty acid oxidation, the regulation of insulin signaling pathways, or the regulation of gene expression related to adipogenesis.

[0066] In one embodiment, inhibition of fatty acid synthesis can be achieved by inhibiting the activity or expression of key enzymes involved in fatty acid synthesis, which may be induced directly or indirectly by specific compounds or drugs. Additionally, the promotion of fatty acid oxidation may act to reduce fat accumulation by activating pathways in which fatty acids are converted into energy within cells. Furthermore, regulation of the insulin signaling pathway refers to influencing fat production through the regulation of insulin signals involved in glucose uptake and fat synthesis within fat cells, and fat accumulation within fat cells may be reduced through such regulation.

[0067] The term "adipogenesis" in this invention refers to the process in which adipocytes differentiate into mature adipocytes. This process is important for the formation and maintenance of adipose tissue in the body.

[0068] The term "lipogenesis" in this invention refers to the process of synthesizing fatty acids and triglycerides. This process is one of the primary methods of storing energy in the body.

[0069] The term “chronic liver disease” in the present invention refers to a pathological condition in which structural or functional abnormalities of the liver persist for a long period, and includes fatty liver, liver fibrosis, and diseases corresponding to the advanced stages thereof. Such liver disease may be characterized by one or more of hepatocyte damage, inflammatory response, fat accumulation, and fibrosis.

[0070] The above liver diseases are not limited to but may include fatty liver, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and cirrhosis.

[0071] The term “hepatic steatosis” in the present invention refers to a condition in which triglycerides are excessively accumulated within the liver due to an abnormal increase in the production of adipocytes or the accumulation of fat in liver cells, and may include both alcoholic and non-alcoholic causes. The hepatic steatosis includes non-alcoholic fatty liver disease (NAFLD) that occurs regardless of alcohol consumption, and includes an early pathological state that can progress from simple hepatic steatosis to steatohepatitis and fibrosis.

[0072] In one embodiment of the present invention, fatty liver may occur in association with the excessive production of adipocytes in liver tissue, fat accumulation, or activation of fat synthesis pathways, and such fat accumulation may contribute to the development and progression of fatty liver by inducing an increase in fat droplets within liver cells, disruption of metabolic homeostasis, and liver dysfunction.

[0073] The above fatty liver includes, but is not limited to, simple fatty liver and may include an early metabolic abnormality state prior to progression to inflammation or fibrosis.

[0074] Accordingly, in the present invention, “prevention or treatment of fatty liver” may include the alleviation or inhibition of fat synthesis and accumulation within liver tissue associated with the inhibition of fat cell production or the reduction of fat accumulation.

[0075] The term “fibrosis” in this invention refers to a pathological condition in which fibroblasts or equivalent cells are activated due to chronic tissue damage, metabolic abnormalities, or inflammatory responses, and an excessive accumulation of extracellular matrix containing collagen leads to structural and functional deformation of the tissue. Furthermore, this invention can alleviate inflammatory responses within the liver by inhibiting the production or reducing the accumulation of hepatic fat cells, and thereby inhibit the activation of hepatic stellate cells, thereby suppressing the occurrence or progression of hepatic fibrosis.

[0076] In one embodiment of the present invention, fibrosis may occur in association with the excessive production of adipocytes in tissues, fat accumulation, or activation of fat synthesis pathways, and such fat accumulation may contribute to the progression of fibrosis by inducing oxidative stress, secretion of inflammatory cytokines, and activation of fibrosis signaling pathways.

[0077] Fibrosis according to the present invention includes, but is not limited to, liver fibrosis, and may include, for example, a pathological condition in which increased expression of fibrosis-related genes (TGF-β1, α-SMA, Col1a1, etc.) is observed along with increased expression of genes or proteins related to fat synthesis in liver tissue induced by a high-fat diet. Although not limited thereto, the fibrosis may include pulmonary fibrosis, liver fibrosis, renal fibrosis, pancreatic fibrosis, or cutaneous fibrosis.

[0078] Accordingly, in the present invention, “prevention or treatment of fibrosis” can be interpreted as a concept that includes the alleviation or inhibition of the activation of fibrosis-related signals associated with the inhibition of adipocyte production or the reduction of fat accumulation.

[0079] In one embodiment of the present invention, fatty liver and liver fibrosis can be understood as different stages of phenotypes existing on a pathological continuum with fat accumulation in liver tissue as a common starting point. Specifically, fatty liver may be induced when triglycerides accumulate due to the excessive production of adipocytes or the activation of lipid synthesis pathways in liver tissue; if such fat accumulation persists, oxidative stress and inflammatory responses are induced, which can lead to liver fibrosis through the activation of hepatic stellate cells and fibrosis signaling pathways.

[0080] Therefore, the approach of the present invention, which alleviates fat accumulation in liver tissue by inhibiting the generation of liver adipocytes or reducing fat accumulation, can be usefully applied to prevent or alleviate the occurrence or progression of liver fibrosis by blocking the activation of inflammatory and fibrotic signals induced by fat accumulation, while simultaneously inhibiting the occurrence or progression of fatty liver.

[0081] The term "diet" in this invention refers to the sum of all foods and beverages consumed. This has a significant impact on an individual's health, nutritional status, and weight management.

[0082] The term "high-fat diet" in the present invention refers to a diet high in fat content. Such a diet may increase the risk of weight gain, cardiovascular disease, and metabolic disease.

[0083] The term "CDC25C" in this invention refers to a kinase associated with cell cycle regulation that is involved in key regulatory steps of cell division. This enzyme plays a critical role at the G2 / M checkpoint of the cell cycle and is considered important in the study of diseases such as cancer.

[0084] In the present invention, "prevention" may include, without limitation, any act of blocking disease symptoms or suppressing or delaying disease symptoms using the pharmaceutical composition of the present invention. Specifically, prevention of obesity refers to strategies and activities to prevent weight gain and excessive accumulation of body fat, and may include administering the composition of the present invention. Additionally, prevention of ocular disorders refers to any measures to maintain the surface health of the eyeball and prevent potential damage or disease, and may include administering the composition of the present invention.

[0085] In the present invention, "treatment" and "improvement" may be included without limitation as long as they are any act of improving or benefiting from disease symptoms by irradiating the pharmaceutical composition of the present invention.

[0086] Specifically, the treatment and improvement of obesity includes all medical and behavioral measures to reduce body fat and restore a healthy weight. This may include personalized dietary adjustments, regular physical activity, and sometimes drug treatment and surgical approaches, and may include the administration of the composition of the present invention.

[0087] A PLK1 inhibitor may be included in a pharmaceutical composition for the treatment and improvement of obesity at an amount of 0.01 mg / kg / day or more and 1.0 mg / kg / day or less, specifically at an amount of 0.1 mg / kg / day or more and 0.5 mg / kg / day or less, more specifically at an amount of 0.2 mg / kg / day or more and 0.4 mg / kg / day or less, and even more specifically at an amount of 0.25 mg / kg / day or more and 0.35 mg / kg / day or less.

[0088] Treatment and improvement of ocular disorders refers to a medical approach to manage disorders or diseases occurring on the surface of the eye and to alleviate symptoms, and may include administering the composition of the present invention.

[0089] In particular, regarding the mechanism of dry eye syndrome, excessive accumulation of lipids in the meibomian glands disrupts their normal function. This leads to a decrease in secretory function, which in turn prevents the tear film from receiving sufficient oil, potentially resulting in dry eye syndrome. If the lipid layer of the tear film fails to function properly due to reduced secretion or changes in its composition, the tear film evaporates more easily, increasing its instability and further exacerbating dryness of the eyes.

[0090] The "pharmaceutical composition" of the present invention is not limited to these, but may be formulated and used in the form of oral formulations such as powders, granules, capsules, tablets, and aqueous suspensions, as well as topical preparations, suppositories, and sterile injectable solutions, according to conventional methods. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. For oral administration, the pharmaceutically acceptable carrier may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, colorants, flavorings, etc. For injectable preparations, it may include buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, etc., in combination; and for topical administration, a base, excipients, lubricants, preservatives, etc. may be used. The formulations of the pharmaceutical composition of the present invention may be prepared in various ways by mixing with the pharmaceutically acceptable carriers described above. For example, for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injectables, it can be manufactured in the form of unit dosing ampoules or multiple dosing ampoules. In addition, it can be formulated as a solution, suspension, tablet, capsule, sustained-release formulation, etc. Meanwhile, examples of carriers, excipients, and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil. In addition, it may additionally include fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, etc.

[0091] In the present invention, "routes of administration" for a pharmaceutical composition include, but are not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual, or rectal. Oral or parenteral administration is preferred.

[0092] In the present invention, "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intradural, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

[0093] The "use and dosage" of the composition of the present invention may vary depending on several factors including the activity of the specific compound used, age, body weight, general health, gender, diet, time of administration, route of administration, elimination rate, drug combination, and the severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may be appropriately selected by a person skilled in the art, depending on the patient's condition, body weight, degree of disease, form of medication, route of administration, and duration, and may be administered at a dose of 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. Administration may be administered once a day or divided into several doses. The dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as a pill, coated tablet, capsule, liquid, gel, syrup, slurry, or suspension.

[0094] In one embodiment of the present invention, the first aspect provides a pharmaceutical composition for preventing or treating obesity that inhibits the formation of fat cells and comprises a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

[0095] Chemical formula 1

[0096]

[0097] In the above chemical formula 1,

[0098] R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0099] In the first embodiment, the second embodiment provides a pharmaceutical composition in which R1 in Formula 1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group or a C5-C6 heterocyclic group.

[0100] In the first or second embodiment, the third embodiment provides a pharmaceutical composition in which R3 further comprises cyclopropane.

[0101] In any one of the first to third embodiments, the fourth embodiment provides a pharmaceutical composition in which the compound is one or more of BI 6727 and BI 2536.

[0102] In any one of the first to fourth embodiments, the fifth embodiment provides a pharmaceutical composition in which the compound has a concentration of 0.1 mg / kg / day or more and 0.5 mg / kg / day or less.

[0103] In any one of the first to fifth embodiments, the sixth embodiment provides a pharmaceutical composition in which the compound downregulates one or more selected from the group consisting of PPARγ, SREBP-1, FASN PI3K, p-AKT(Ser473) / AKT and ACC genes.

[0104] In any one of the first to sixth embodiments, the seventh embodiment provides a pharmaceutical composition in which the compound is administered orally, or by intravenous injection, intramuscular injection, intra-articular injection, intra-synovial injection, intraretinal injection, intrahepatic injection, intralesional injection, or intracranial injection.

[0105] In one embodiment of the present invention, the eighth aspect provides a pharmaceutical composition for preventing or treating eye disorders, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient and inhibiting the formation of fat cells.

[0106] Chemical formula 1

[0107]

[0108] In the above chemical formula, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocyclic group, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocyclic group.

[0109] In the eighth embodiment, the ninth embodiment provides a pharmaceutical composition in which, in the above formula 1, R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group or a C5-C6 heterocyclic group.

[0110] In the eighth or ninth embodiment, the tenth embodiment provides a pharmaceutical composition in which R3 further comprises cyclopropane.

[0111] In any one of the eighth to tenth embodiments, the eleventh embodiment provides a pharmaceutical composition in which the compound is one or more of BI 6727 and BI 2536.

[0112] In any one of the eighth to eleventh embodiments, the twelfth embodiment provides a pharmaceutical composition in which the concentration of the compound is 0.1 mg / kg / day or more and 0.5 mg / kg / day or less.

[0113] In any one of the eighth to twelve embodiments, the thirteenth embodiment provides a pharmaceutical composition in which the compound downregulates one or more selected from the group consisting of PPARγ, SREBP-1, FASN PI3K, p-AKT(Ser473) / AKT and ACC genes.

[0114] In any one of the eighth to thirteenth embodiments, the 14th embodiment provides a pharmaceutical composition in which the composition inhibits the generation of fat cells in myosemia epithelial cells.

[0115] In any one of the eighth to fourteenth embodiments, the fifteenth embodiment provides a pharmaceutical composition in which the ocular disorder is any one selected from the group consisting of dry eye, blepharitis, conjunctivitis, squamous dysplasia, and bullous keratoconjunctivitis.

[0116] In any one of the eighth to fifteenth embodiments, the 16th embodiment provides a pharmaceutical composition in which the compound is administered orally, or by intravenous injection, intramuscular injection, intra-articular injection, intra-synovial injection, intraretinal injection, intrahepatic injection, intralesional injection, or intracranial injection.

[0117] In one embodiment of the present invention, the 17th aspect provides a pharmaceutical composition for preventing or treating liver disease, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, which inhibits the formation of fat cells.

[0118] Chemical formula 1

[0119]

[0120] In the above chemical formula, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocyclic group, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocyclic group.

[0121] In the 17th embodiment, the 18th embodiment provides a pharmaceutical composition in which the liver disease is any one selected from the group consisting of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and liver cirrhosis.

[0122] In the 17th or 18th embodiment, the 19th embodiment provides a pharmaceutical composition in which, in the above formula 1, R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group or a C5-C6 heterocyclic group.

[0123] In any one of the 17th to 19th embodiments, the 20th embodiment provides a pharmaceutical composition in which R3 further comprises cyclopropane.

[0124] In one embodiment of the present invention, the 21st aspect provides a method for preventing or treating obesity, comprising the step of administering an effective amount of a compound represented by the following formula 1, or a pharmaceutically acceptable salt thereof, to a target individual.

[0125] Chemical formula 1

[0126]

[0127] In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0128] In the 21st embodiment, the 22nd embodiment provides a method for preventing or treating obesity, wherein in the above formula 1, R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group or a C5-C6 heterocyclic group.

[0129] In the 21st or 22nd embodiment, the 23rd embodiment provides a method for preventing or treating obesity, wherein R3 further comprises cyclopropane.

[0130] In any one of the 21st to 23rd embodiments, the 24th embodiment provides a method for preventing or treating obesity, wherein the compound is one or more of BI 6727 and BI 2536.

[0131] In any one of the 21st to 24th embodiments, the 25th embodiment provides a method for preventing or treating obesity in which the compound has a concentration of 0.1 mg / kg / day or more and 0.5 mg / kg / day or less.

[0132] In one embodiment of the present invention, the 26th aspect provides a method for preventing or treating ocular disorders, comprising the step of administering an effective amount of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, to a target individual.

[0133] Chemical formula 1

[0134]

[0135] In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0136] In the 26th embodiment, the 27th embodiment provides a method for preventing or treating ocular disorders, wherein R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocycle, or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocycle.

[0137] In the 26th or 27th embodiment, the 28th embodiment provides a method for preventing or treating an eye disorder, wherein the eye disorder is any one selected from the group consisting of dry eye, blepharitis, conjunctivitis, squamous dysplasia, and bullous keratoconjunctivitis.

[0138] In one embodiment of the present invention, the 29th aspect provides a method for preventing or treating liver disease, comprising the step of administering an effective amount of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, to a target individual.

[0139] Chemical formula 1

[0140]

[0141] In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0142] In the 29th embodiment, the 30th embodiment provides a method for preventing or treating a liver disease, wherein the liver disease is any one selected from the group consisting of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and liver cirrhosis.

[0143] In one embodiment of the present invention, the 31st aspect provides a use for the prevention or treatment of obesity of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof.

[0144] Chemical formula 1

[0145]

[0146] In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0147] In the 31st embodiment, the 32nd embodiment provides an obesity prevention or treatment use in which, in the above formula 1, R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group or a C5-C6 heterocyclic group.

[0148] In the 31st or 32nd embodiment, the 33rd embodiment provides an obesity prevention or treatment use in which R3 further comprises cyclopropane.

[0149] In one embodiment of the present invention, the 34th aspect provides a use for the prevention or treatment of ocular disorders of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof.

[0150] Chemical formula 1

[0151]

[0152] In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0153] In the 34th embodiment, the 35th embodiment provides a use for preventing or treating ocular disorders, wherein in the above formula 1, R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocycle, or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocycle.

[0154] In the 34th or 35th embodiment, the 36th embodiment provides a use for preventing or treating an eye disorder, wherein the eye disorder is any one selected from the group consisting of dry eye, blepharitis, conjunctivitis, squamous dysplasia, and bullous keratoconjunctivitis.

[0155] In one embodiment of the present invention, the 37th aspect provides a use for the prevention or treatment of liver disease of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof.

[0156] Chemical formula 1

[0157]

[0158] In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0159] In the 37th embodiment, the 38th embodiment provides a use for preventing or treating liver disease, wherein in the above formula 1, R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocycle, or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocycle.

[0160] In the 37th or 38th embodiment, the 39th embodiment provides a liver disease prevention or treatment use in which R3 further comprises cyclopropane.

[0161] In any one of the 37th to 39th embodiments, the 40th embodiment provides a use for the prevention or treatment of liver disease, wherein the liver disease is any one selected from the group consisting of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and liver cirrhosis.

[0162] In one embodiment of the present invention, the 41st aspect provides a pharmaceutical composition for the prevention or treatment of metabolic diseases, comprising as an active ingredient a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof.

[0163] Chemical formula 1

[0164]

[0165] In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0166] In the 41st embodiment, the 42nd embodiment provides a pharmaceutical composition in which, in the above formula 1, R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group or a C5-C6 heterocyclic group.

[0167] In the 41st or 42nd embodiment, the 43rd embodiment provides a pharmaceutical composition in which R3 further comprises cyclopropane.

[0168] In any one of the 41st to 43rd embodiments, the 44th embodiment provides a pharmaceutical composition in which the compound is one or more of BI 6727 and BI 2536.

[0169] In any one of the 41st to 44th embodiments, the 45th embodiment provides a pharmaceutical composition in which the compound has a concentration of 0.1 mg / kg / day or more and 0.5 mg / kg / day or less.

[0170] In any one of the 41st to 45th embodiments, the 46th embodiment provides a pharmaceutical composition in which the compound downregulates one or more selected from the group consisting of PPARγ, SREBP-1, FASN PI3K, p-AKT(Ser473) / AKT and ACC genes.

[0171] In any one of the 41st to 46th embodiments, the 47th embodiment provides a pharmaceutical composition in which the compound is administered orally, or by intravenous injection, intramuscular injection, intra-articular injection, intra-synovial injection, intraretinal injection, intrahepatic injection, intralesional injection, or intracranial injection.

[0172] In any one of the 41st to 47th embodiments, the 48th embodiment provides a pharmaceutical composition in which the metabolic disease is obesity.

[0173] In any one of the 41st to 47th embodiments, the 49th embodiment provides a pharmaceutical composition in which the metabolic disease is selected from the group consisting of fatty liver, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and liver cirrhosis.

[0174] In any one of the 41st to 49th embodiments, the 50th embodiment provides a pharmaceutical composition in which the composition inhibits the formation of fat cells.

[0175] In one embodiment of the present invention, the 51st aspect provides a method for preventing or treating a metabolic disease, comprising the step of administering an effective amount of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, to a target individual.

[0176] Chemical formula 1

[0177]

[0178] In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0179] In one embodiment of the present invention, the 52nd aspect provides a use for the prevention or treatment of metabolic diseases of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof.

[0180] Chemical formula 1

[0181]

[0182] In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocycle, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

[0183] The composition of the present invention can improve obesity caused by a high-fat diet due to its effect of inhibiting fat production in fat cells.

[0184] In addition, the composition of the present invention can treat dry eye syndrome by confirming the effect of inhibiting lipid production in human meibomian gland epithelial cells.

[0185] The present invention provides an effect that inhibits the accumulation of fat in the liver and the progression of fibrosis by regulating abnormalities in energy and lipid metabolism in metabolic diseases including obesity, fatty liver, and liver fibrosis, and contributes to the overall improvement of metabolic homeostasis.

[0186] Furthermore, the effects of the present invention are not limited to the effects described above, and should be understood to include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

[0187] Figure 1 illustrates the confirmation of fat accumulation after treating 3T3-L1 pre-adipocytes with 100 nM BI6727 during differentiation into adipocytes for 10 days. In Figure 1a, the areas stained red using Oil Red O represent triglycerides, and experimental results confirmed that the stained area decreased in the BI6727-treated group compared to the control group. In Figure 1b, the cell viability was 69% after 10 days of BI6727 treatment, and the final result was determined by dividing the daily Adiporeid analysis values ​​by the cell viability. As a result, it was confirmed that the BI6727-treated group showed a decrease compared to the control group.

[0188] Figure 2 shows the accumulation of fat after treating 3T3-L1 pre-adipocytes with 100 nM of BI2536 while differentiating them into adipocytes for 10 days. The red stained area represents neutral fat, and the experimental results confirmed that the stained area decreased in the BI2536-treated group compared to the control group.

[0189] Figure 3 shows the mRNA and protein expression of genes involved in adipogenesis and lipogenesis over time after treating 3T3-L1 preadipocytes with 100 nM BI6727 during 10 days of adipogenesis. Figures 3a and 3b confirm that treatment with BI6727 reduced the expression of PPARγ, C / EBPα, SREBP-1, FASN, and ACC genes compared to the control group as adipogenesis progressed, and also reduced p-AKT(Ser473) / AKT, a downstream regulator of SREBP-1. However, a significant decrease in PLK1 was observed only in D2.

[0190] Figure 4 shows the mRNA and protein expression of genes involved in adipogenesis and lipogenesis over time after treating 3T3-L1 adipocyte precursor cells with 100 nM of BI2536 during 10 days of differentiation into adipocytes, and it was confirmed that treatment with BI2536 reduced the expression of PPARγ, SREBP-1, and FASN genes as adipogenesis progressed.

[0191] Figure 5 illustrates the adipocyte differentiation stages affected by BI6727. Figure 5a schematically illustrates the progress of the experiment to identify the adipocyte differentiation stages affected by BI6727. Figures 4b and 4c confirmed that treatment with BI6727 during the early differentiation stage reduced PPARγ and C / EBPα, which regulate adipogenesis, as well as SREBP-1, FASN, and ACC, which regulate lipogenesis, whereas treatment with BI6727 during the late differentiation stage reduced only SREBP-1, FASN, and ACC, which regulate lipogenesis. Figure 4d confirmed that treatment with BI6727 during the early differentiation stage also showed a significant reduction in fat accumulation, and treatment with BI6727 during the late differentiation stage showed a tendency for fat accumulation to decrease compared to the control group, but it was less reduced than when the drug was administered during the early differentiation stage.

[0192] Figure 6 shows the results of confirming that PLK1 inhibitors improve obesity caused by a high-fat diet in animal experiments as well. Figure 6a shows the simultaneous administration of diet and drug using a diet-induced obesity (DIO) model to confirm the efficacy of the drug as an obesity treatment in a mouse model. The drug was orally administered for 8 weeks at a concentration of 0.3 mg / kg / day, which is about 100 times lower than the concentration of 25-50 mg / kg / day typically used as an anticancer drug. Figure 6b is a graph showing the experimental results confirming that obesity caused by a high-fat diet was improved by the administration of a low concentration of BI 6727.

[0193] Figure 7 illustrates the day-by-day cell viability and fat accumulation after treatment with BI2536 and BI6727 during adipogenesis in HMGECs. Cell viability was tested using Presto Blue reagent, and the final result was determined by dividing the Adipored assay value by the cell viability. As a result, Figures 7c and 7f confirmed that fat accumulation was reduced in the BI2536 and BI6727 treatment groups compared to the control group.

[0194] Figure 8 shows the mRNA expression of genes involved in adipogenesis and lipogenesis after treatment with BI2536 and BI6727 during adipogenesis in HMGECs. Figures 8a and 8b confirm that as differentiation progressed in the BI2536 and BI6727 treatment groups, the expression of PPARγ, SREBP-1, FASN, and ACC genes decreased compared to the control group.

[0195] Figure 9 shows the protein expression of PLK1 and CDC25C after treatment with BI2536 during adipogenesis in HMGECs. After treatment with BI2536, a Pan-PLK1 inhibitor, the protein expression levels of PLK1 and CDC25C were examined. The results showed that PLK1 decreased only in D2, while CDC25C decreased during the differentiation process.

[0196] Figure 10 shows the expression of proteins regulating adipogenesis and lipogenesis after treatment with BI 2536 during adipogenesis in HMGECs. In Figure 10a, treatment with BI 2536 reduced the expression of PPARγ, SREBP-1, FASN, and ACC genes compared to the control group as adipogenesis progressed, and in Figure 10b, it was confirmed that PI3K, an upstream regulator of SREBP-1, and p-AKT(Ser473) / AKT, a downstream regulator, were also reduced.

[0197] Figure 11 shows the protein expression of PLK1 and CDC25C after treatment with BI6727 during adipogenesis in HMGECs. As a result of measuring the protein expression levels of PLK1 and CDC25C after treatment with BI6727, a selective PLK1 inhibitor, it was confirmed that PLK1 showed a decrease only in D2, and CDC25C decreased during the differentiation process.

[0198] Figure 12 shows the expression of proteins regulating adipogenesis and lipogenesis after treatment with BI6727 during adipogenesis in HMGECs. In Figure 12a, treatment with BI6727 reduced the expression of PPARγ, SREBP-1, FASN, and ACC genes compared to the control group as adipogenesis progressed, and in Figure 12b, it was confirmed that PI3K, an upstream regulator of SREBP-1, and p-AKT(Ser473) / AKT, a downstream regulator, were also reduced.

[0199] Figure 13 confirms that drug treatment with HFD + BI6727 inhibits body weight gain and reduces visceral fat weight. Figure 13a shows the results of tracking body weight changes for 8 weeks in animals fed a normal diet (NCD) or a high-fat diet (HFD) while administering vehicle or BI6727 (0.3 mg / kg / day). It shows that while significant body weight gain was observed in the HFD + vehicle group, body weight gain was significantly inhibited in the HFD + BI6727 treatment group. Figure 13b shows the results of comparing the final body weight at the end of the experiment, indicating that the body weight of the BI6727 treatment group under HFD conditions was significantly reduced compared to the vehicle treatment group. Figure 13c is a graph showing the weekly body weight gain relative to the body weight at the start of the experiment, confirming that the rate of body weight gain was significantly reduced in the HFD + BI6727 treatment group. Figure 13d shows the results of comparing daily energy intake (kcal / mouse / day), indicating that no significant difference in energy intake was observed depending on whether BI6727 was administered. Figure 13e shows the results of comparing daily feed intake (g / mouse / day), indicating that the administration of BI6727 did not have a significant effect on the food intake itself. Figure 13f shows the ratio of intake to body weight gain, suggesting that the efficiency of body weight gain was suppressed in the HFD+BI6727 treatment group, as the ratio of intake to body weight gain decreased. Figure 13g shows representative images of the external appearance of animals in each experimental group and excised white adipose tissue (eWAT and iWAT), and it is visually confirmed that the size of the adipose tissue decreased in the HFD+BI6727 treatment group. Figure 13h shows the results of a quantitative comparison of the tissue weights of eWAT and iWAT, demonstrating that the weight of both visceral adipose tissues was significantly reduced by BI6727 treatment under HFD conditions.

[0200] Figure 14 shows that lipid metabolism was improved, as serum analysis results confirmed that TG and LDL were significantly reduced and HDL was increased in the drug-treated group of HFD + BI6727 compared to HFD + Vehicle. Figure 14a shows serum alanine aminotransferase (ALT) levels; no significant change in ALT levels was observed following BI6727 treatment, indicating that there was no negative impact on hepatotoxicity indicators. Figure 14b shows serum aspartate aminotransferase (AST) levels, and no statistically significant difference in AST levels was observed between the experimental groups. Figure 14c shows the results of comparing serum total cholesterol (T-Chol) concentrations; although total cholesterol increased in the HFD group compared to the NCD group, no significant difference was observed within the HFD condition following BI6727 treatment. Figure 14d shows serum triglyceride (TG) concentrations, demonstrating that TG levels were significantly reduced in the HFD+BI6727 treatment group compared to the HFD+vehicle group. Figure 14e shows the results of serum high-density lipoprotein (HDL) concentrations, confirming that HDL levels were significantly increased in the BI6727 treatment group. Figure 14f shows the results of comparing serum low-density lipoprotein (LDL) concentrations, indicating that LDL levels were significantly reduced in the HFD+BI6727 treatment group.

[0201] Figure 15 shows that PLK1 expression in visceral adipose tissue was reduced in the drug-treated group of HFD+BI6727 compared to HFD+Vehicle. Figure 15a shows the results of Western blot analysis of the expression of proteins (PPARγ2, C / EBPα, SREBP-1, FAS, SCD1) involved in adipocyte differentiation and lipid synthesis in eWAT tissues of the vehicle-treated group under normal diet (NCD) and high-fat diet (HFD) conditions. Figure 15b is a graph quantifying the Western blot results of Figure 15a, showing that the expression of proteins related to adipocyte differentiation and lipid synthesis significantly increased under HFD conditions compared to NCD. Figure 15c shows the results of Western blot comparison of the expression of PPARγ2, C / EBPα, SREBP-1, FASN, SCD1, and PLK1 in eWAT tissues of the vehicle-treated group and the BI6727-treated group under HFD conditions. Figure 15d is a graph quantifying the results of Figure 15c, showing that the expression of proteins related to adipocyte differentiation and lipid synthesis, including PLK1, is reduced in the HFD+BI6727 treatment group compared to the HFD+vehicle group. Figures 15e to 15g show the H&E staining results for eWAT tissues, representing the NCD+vehicle, HFD+vehicle, and HFD+BI6727 treatment groups, respectively, and it can be confirmed that the size of adipocytes decreased in the HFD+BI6727 treatment group. Figures 15h to 15j are low-magnification images showing the results of immunohistochemical staining (IHC) for CD68 in eWAT tissues, showing that CD68-positive cells increased in the HFD+vehicle group, while their distribution decreased in the BI6727 treatment group. Figures 15k to 15m are high-magnification images of Figures 15h to 15j, showing that crown-like structures (CLS) formed by macrophages accumulating around dying adipocytes under obesity conditions were clearly observed in the HFD+vehicle group, and that CLS formation was reduced in the HFD+BI6727 treatment group.Figure 15n shows the results of a quantitative analysis of the adipocyte area in eWAT tissue, indicating that the adipocyte size in the HFD+BI6727 treatment group is significantly reduced compared to the HFD+vehicle group.

[0202] Figure 16 confirms that the expression of genes related to lipid synthesis in liver tissue was reduced in the drug-treated group of HFD + BI6727 compared to HFD + Vehicle. Additionally, the figure confirms that the mRNA expression of a representative gene for liver fibrosis was also significantly reduced in the drug-treated group of HFD + BI6727 compared to HFD + Vehicle. Figure 16a shows the results of Western blot analysis of the expression of lipid synthesis-related proteins SREBP-1 and FASN in liver tissues of the vehicle-treated and BI6727-treated groups under HFD conditions. Figure 16b is a graph quantifying the Western blot results of Figure 16a, showing that the protein expression of SREBP-1 and FASN in the HFD + BI6727-treated group was significantly reduced compared to the HFD + vehicle group. Figure 16c shows the results of a quantitative analysis of mRNA expression of representative genes (TGF-β, α-SMA, Col1a1) associated with liver fibrosis, indicating that the expression of these fibrosis-related genes is significantly reduced in the HFD+BI6727 treatment group compared to the HFD+vehicle group. Figure 16d shows the results of an analysis of mRNA expression of genes (IL-1β, TNFα) associated with liver inflammation, confirming that the expression of inflammation-related genes tends to decrease in the BI6727 treatment group. Figures 16e to 16g show the results of H&E staining on liver tissue, representing the NCD+vehicle, HF D+vehicle, and HFD+BI6727 treatment groups, respectively, demonstrating that fat droplet accumulation and histological abnormalities observed in the HFD+vehicle group are alleviated in the BI6727 treatment group. Figure 16i shows the results of quantifying the degree of fat accumulation (vacuolated area) in liver tissue based on H&E staining images, indicating that the fat accumulation area in the HFD+BI6727 treatment group is significantly reduced compared to the HFD+vehicle group.

[0203] The present invention will be described in more detail below through examples. These examples are intended solely to explain the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the invention.

[0204]

[0205] Example 1. Cell Culture

[0206] The differentiation process from 3T3-L1 preadipocytes to adipocytes was carried out. Initially, a maintenance medium was prepared by supplementing DMEM high-sugar medium with 10% BCS and 1% P / S, and 8 x 10⁶ cells were placed in a 6-well plate. 4 Cells were seeded. After two days, the maintenance medium was changed, and the cells were cultured for an additional two days. At this time, the condition was set to D0. After removing the medium, it was replaced with differentiation medium (a medium containing 10% FBS, 1% P / S, 0.52 mM IBMX, 1 μM Dexamethasone, and 1 μg / ml insulin in DMEM), and cultured for two days. Subsequently, the medium was removed and replaced with adipocyte maintenance medium (a medium containing 10% FBS, 1% P / S, and 1 μg / ml insulin in DMEM), and the cells were continuously cultured for 8 days while replacing the medium with fresh adipocyte maintenance medium every two days.

[0207] For the culture of hMGECs, a maintenance medium using keratinocyte serum-free media was prepared, and 2 x 10⁶ were placed in a 6-well plate. 5 Cells were seeded. The next day, the differentiation medium was replaced with a medium containing 10% FBS, 10 ng / ml EGF, and 1% P / S in DMEM F / 12, and cultured for 8 days while replacing it with a new differentiation medium every two days.

[0208] For drug treatment, BI6727 and BI2536 were used at a concentration of 100 nM each. During the differentiation process from 3T3-L1 preadipocytes to adipocytes, the drugs were administered along with the differentiation medium, and the drugs were added fresh whenever the medium was changed. During hMGEC culture, the drugs were also added fresh along with the exchange of the differentiation medium to ensure continuous treatment. In this manner, the cellular response was observed while maintaining the continuous effect of the drugs.

[0209]

[0210] Example 2. Oil Red O Staining (Oil Red O Staining, based on a 6-well plate)

[0211] After differentiating 3T3-L1 preadipocytes to D10, the medium in the plate was removed and washed once with deionized water (DW). After removing the DW, 2 ml of 4% formaldehyde solution (Paraformaldehyde, PFA) was added and fixed for 20 minutes. After removing the 4% PFA, the cells were washed three times again with deionized water. After removing all the DW, 2 ml of 100% propylene glycol was added and reacted for 2 minutes. After removing the 100% propylene glycol, 2 ml of Oil Red O solution heated at 60°C for 10 minutes was added and reacted at 37°C for 2 hours. After removing the Oil Red O solution, 2 ml of 85% propylene glycol was added and reacted again for 2 minutes. The 85% propylene glycol was removed, and the cells were washed twice with deionized water. Finally, images of the cells were taken using a microscope while the cells were in the presence of deionized water.

[0212]

[0213] Example 3. Cell viability and Adipored assay (Cell viability & Adipored assay, based on a 24-well plate)

[0214] The Presto Blue reagent was equilibrated at room temperature before use. The Presto Blue reagent was mixed with the existing culture medium at a ratio of 9:1 and added to the cell culture medium. The plates were then covered with foil and incubated in a cell culture incubator at 37°C and 5% CO2 for 1 hour. After incubation, fluorescence was measured (Excitation: 560 nm, Emission: 590 nm).

[0215] The medium and Presto Blue mixture were removed from the plate after cell viability measurement was completed. The cells were washed once with DPBS. After removing the DPBS, DPBS and Adipored were mixed at a ratio of 40:1 and 500 μl was added to each well. After adding the mixture, the plate was covered with foil and incubated at room temperature for 10 minutes. After the reaction, fluorescence was measured (Excitation: 485 nm, Emission: 572 nm). The Adipored / Cell viability ratio was calculated using the measured fluorescence values, and the degree of TG (Triglyceride) accumulation was confirmed.

[0216]

[0217] Example 4. RNA Prep and qPCR (based on a 12-well plate)

[0218] The medium was removed from the culture plate in which the cells were growing, and 1 ml of DPBS was added to wash the cells, after which the DPBS was removed. 1 ml of Trizol was added to the cells and homogenized using a vortex for 15 seconds. 200 μl of chloroform was added and vortexed for 30 seconds. Subsequently, the mixture was placed on ice for 2 minutes, and centrifuged at 12,000 rpm for 15 minutes at 4°C to separate the supernatant and subnatant. 400 μl of the supernatant was taken, added along with 400 μl of isopropanol, and inverted to mix. 800 μl of the mixed solution was placed into a Qiagen kit column. It was centrifuged at 12,000 rpm for 1 minute at room temperature, the flow-through was discarded, and 700 μl of RW1 was added to the column. It was centrifuged again at 12,000 rpm for 1 minute at room temperature. The flow-through was discarded, and 500 μl of RPE was added to the column. Subsequently, centrifugation was performed at 12,000 rpm for 1 minute at room temperature. The above two processes were repeated once more.

[0219] The column was transferred to a new collection tube and dried by centrifuging at 12,000 rpm for 1 minute at room temperature. The column was transferred to a new 1.5 ml tube, 30 μl of RNase-free water was added to the center of the column, and it was left for 1 minute. RNA was extracted by centrifuging at 12,000 rpm for 1 minute at room temperature. The concentration of the extracted RNA was measured using a Nanodrop, and the extracted RNA was stored on ice. 20 μl of cDNA was synthesized using 1,000 ng of RNA, and a Takara EcoDry (Cat#639543) was used. The synthesized cDNA was diluted to a concentration of 5 ng / μl and stored at -20℃. 10 ng of cDNA was used for qPCR.

[0220] 10 ng of cDNA, SYBR, and primers were added, and qPCR was performed using a viia7 instrument.

[0221]

[0222] Example 5. Western blot (based on a 6-well plate)

[0223] In the preparation of the cell lysate, 100 μl of a mixture of Lysis buffer and 1 mM PMSF protease inhibitor was added to the cells, vortexed for 15 seconds, and then placed on ice for 5 minutes. This process was repeated a total of three times. Afterward, the supernatant obtained by centrifugation at 12,000 rpm for 15 minutes at 4°C was used as the cell lysate.

[0224] For protein quantification, BCA reagents A and B were mixed in a ratio of 50:1. The cell lysate was diluted fivefold to prepare the sample for quantification. 10 μl each of the sample to be quantified and standard BSA solutions (0.125, 0.25, 0.5, 1 mg / ml) were added to each well of a 96-well plate, followed by the addition of 200 μl of BCA solution. After reacting this mixture at 37°C for 30 minutes, the protein concentration was determined by reading the absorbance at 562 nm. The protein loading amount was set to 10 μg, and after adding 5XSB and lysis buffer, the mixture was boiled at 100°C for 10 minutes.

[0225] For SDS-PAGE, the separation solution was placed in the gel cassette, 1 mL of isopropanol was added, and the gel was allowed to solidify. The isopropanol was removed and the sample washed with distilled water (DW). Then, the stacking gel was placed in the cassette, and 15 combs were inserted to solidify the gel. Once the gel solidified, the combs were removed, the gel plate was mounted in the tank, filled with running buffer, and the markers and samples were loaded. Electrophoresis was performed at 90 V for stacking and 120 V for separation.

[0226] During the transfer process, after all the samples in the gel had been transferred, the gel was separated from the cassette, and transfer was performed using an iblot3 instrument. Once transfer was complete, the cell membrane was separated, placed in Ponceau S solution to stain the protein locations, and then washed.

[0227] Blocking was performed at room temperature for 1 hour using 5% BSA (in TBST) or 5% Skim milk (in TBST). Afterward, the samples were washed with TBST for 5 seconds, and this process was repeated two more times.

[0228] To attach the primary antibody, the primary antibody was diluted in 5% BSA or 5% Skim milk (in TBST) and incubated overnight at 4°C. Afterward, the cells were washed three times with TBST for 10 minutes each. To attach the secondary antibody, the secondary antibody was diluted in 5% BSA or 5% Skim milk (in TBST) and incubated at room temperature for 1 hour. Subsequently, the cells were washed three times with TBST for 10 minutes each. For the detection process, the required amount of ECL solution was mixed in a 1:1 ratio and reacted with the washed cell membranes. The membranes were then placed on an OHP film, and Western blot detection was performed using an Image Quant 800 instrument.

[0229]

[0230] Example 6. Animal experiment

[0231] Eight-week-old male C57BL / 6J mice were used, and drug administration was carried out for 8 weeks with a 60% high-fat diet (HFD) at a low concentration of BI6727 (0.3 mg / kg / day) and 0 mg / kg / day (Vehicle) every 5 days. Body weight was measured once a week.

[0232]

[0233] Foregoing, specific parts of the present invention have been described in detail. It is evident to those skilled in the art that such specific descriptions are merely preferred embodiments and do not limit the scope of the invention. Accordingly, the actual scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A pharmaceutical composition for preventing or treating obesity, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, which inhibits the formation of fat cells. Chemical formula 1 In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, and R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

2. In Paragraph 1, In the above chemical formula 1 R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, and R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group, or a piperidine derivative, and A pharmaceutical composition in which R3 is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocycle.

3. In Paragraph 2, The above R3 is a pharmaceutical composition further comprising cyclopropane.

4. In Paragraph 1, A pharmaceutical composition in which the above compound is one or more of BI 6727 and BI 2536.

5. In Paragraph 1, The above compound is a pharmaceutical composition having a concentration of 0.1 mg / kg / day or more and 0.5 mg / kg / day or less.

6. In Paragraph 1, The above compound is a pharmaceutical composition that downregulates one or more selected from the group consisting of PPARγ, SREBP-1, FASN PI3K, p-AKT(Ser473) / AKT, and ACC genes.

7. In Paragraph 1, A pharmaceutical composition in which the above compound is administered orally, or by intravenous injection, intramuscular injection, intra-articular injection, intra-synovial injection, intraretinal injection, intrahepatic injection, intralesional injection, or intracranial injection.

8. A pharmaceutical composition for the prevention or treatment of eye disorders, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient and inhibiting the formation of fat cells. Chemical formula 1 In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocyclic group, and R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

9. In Paragraph 8, In the above chemical formula 1 R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, and R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group, or a piperidine derivative, and A pharmaceutical composition in which R3 is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocycle.

10. In Paragraph 9, The above R3 is a pharmaceutical composition further comprising cyclopropane.

11. In Paragraph 8, A pharmaceutical composition in which the above compound is one or more of BI 6727 and BI 2536.

12. In Paragraph 8, The above compound is a pharmaceutical composition having a concentration of 0.1 mg / kg / day or more and 0.5 mg / kg / day or less.

13. In Paragraph 8, The above compound is a pharmaceutical composition that downregulates one or more selected from the group consisting of PPARγ, SREBP-1, FAS N PI3K, p-AKT(Ser473) / AKT, and ACC genes.

14. In Paragraph 8, The above composition is a pharmaceutical composition that inhibits the production of fat cells in myosem epithelial cells.

15. In Paragraph 8, A pharmaceutical composition wherein the above ocular disorder is any one selected from the group consisting of dry eye syndrome, blepharitis, conjunctivitis, squamous cell dysplasia and bullous keratoconjunctivitis.

16. In Paragraph 8, A pharmaceutical composition in which the above compound is administered orally, or by intravenous injection, intramuscular injection, intra-articular injection, intra-synovial injection, intraretinal injection, intrahepatic injection, intralesional injection, or intracranial injection.

17. A pharmaceutical composition for the prevention or treatment of liver disease comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, which inhibits the formation of fat cells. Chemical formula 1 In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocyclic group, and R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

18. In Paragraph 17, A pharmaceutical composition in which the above liver disease is any one selected from the group consisting of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and liver cirrhosis.

19. In Paragraph 17, In the above chemical formula 1 R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, and R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group, or a piperidine derivative, and A pharmaceutical composition in which R3 is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocycle.

20. In Paragraph 19, The above R3 is a pharmaceutical composition further comprising cyclopropane.

21. A method for preventing or treating obesity, comprising the step of administering an effective amount of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, to a target individual. Chemical formula 1 In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, and R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

22. In Paragraph 21, In the above chemical formula 1 R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, and R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group, or a piperidine derivative, and R3 is a method for preventing or treating obesity, which is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocyclic group.

23. In Paragraph 22, The above R3 is a method for preventing or treating obesity, further comprising cyclopropane.

24. In Paragraph 21, A method for preventing or treating obesity, wherein the above compound is one or more of BI 6727 and BI 2536.

25. In Paragraph 21, A method for preventing or treating obesity, wherein the above compound has a concentration of 0.1 mg / kg / day or more and 0.5 mg / kg / day or less.

26. A method for preventing or treating ocular disorders, comprising the step of administering an effective amount of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, to a target individual. Chemical formula 1 In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, and R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

27. In Paragraph 26, In the above chemical formula 1 R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, and R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group, or a piperidine derivative, and R3 is a method for preventing or treating eye disorders, which is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocyclic group.

28. In Paragraph 26, A method for preventing or treating an eye disorder, wherein the eye disorder is any one selected from the group consisting of dry eye syndrome, blepharitis, conjunctivitis, squamous dysplasia, and bullous keratoconjunctivitis.

29. A method for preventing or treating liver disease, comprising the step of administering an effective amount of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, to a target individual. Chemical formula 1 In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, and R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

30. In Paragraph 29, A method for preventing or treating liver disease, wherein the above liver disease is any one selected from the group consisting of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and liver cirrhosis.

31. Use of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, for the prevention or treatment of obesity. Chemical formula 1 In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, and R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

32. In Paragraph 31, In the above chemical formula 1 R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, and R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group, or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocyclic group, for the prevention or treatment of obesity.

33. In Paragraph 32, The above R3 is for the prevention or treatment of obesity, further comprising cyclopropane.

34. Use of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, for the prevention or treatment of ocular disorders. Chemical formula 1 In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, and R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

35. In Paragraph 34, In the above chemical formula 1 R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, and R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group, or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocyclic group, for the prevention or treatment of ocular disorders.

36. In Paragraph 34, The above ocular disorder is any one selected from the group consisting of dry eye, blepharitis, conjunctivitis, squamous dysplasia, and bullous keratoconjunctivitis, for the purpose of preventing or treating ocular disorders.

37. Use of a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, for the prevention or treatment of liver disease. Chemical formula 1 In the above chemical formula 1, R1 is any one of hydrogen, a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle, and R2 is any one of a C3-C8 cycloalkyl group, a C3-C8 heterocyclic group, or a piperidine derivative, and R3 is any one of a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 heterocycle.

38. In Paragraph 37, In the above chemical formula 1 R1 is a C2 branched alkyl group or a C5-C6 cycloalkyl group, and R2 is any one of a C5-C6 cycloalkyl group, a C5-C6 heterocyclic group, or a piperidine derivative, and R3 is any one of a methyl group, a C5-C6 cycloalkyl group, or a C5-C6 heterocyclic group, for the prevention or treatment of liver disease.

39. In Paragraph 38, The above R3 is for the prevention or treatment of liver disease, further comprising cyclopropane.

40. In Paragraph 37, The above liver disease is any one selected from the group consisting of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), liver fibrosis, and liver cirrhosis, for the purpose of preventing or treating liver disease.

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