KR20160059158A - Pharmaceutical composition containing 2-Oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivatives for prevention or treatment of metabolic disease - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating metabolic diseases, which comprises a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative inhibiting activity of GPR43 (G protein receptor 43) as an active ingredient. The derivative and an optical isomer or pharmaceutically acceptable salt thereof is an inverse agonist and a surmountable inhibitor and can significantly inhibits human GPR43 (hGPR43), and thus can be useful as an active ingredient for a pharmaceutical composition for preventing and treating GPR43 activity-related inflammatory or metabolic diseases, or for health-aid functional food.

Description

TECHNICAL FIELD The present invention relates to pharmaceutical compositions containing 2-Oxo-1,2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative as an active ingredient, 3,4-tetrahydropyrimidine-5-carboxamide derivatives for prevention or treatment of metabolic disease.

The present invention relates to a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative (2-Oxo-1,2,3,4- tetrahydropyrimidine-5-carboxamide derivatives) as an active ingredient. The present invention also relates to a pharmaceutical composition for preventing or treating metabolic diseases.

There are many types of receptors in humans, but one of the most abundant and therapeutically relevant is the G-protein coupled receptor (GPCR) type. Within the human genome there are hundreds of thousands of genes, of which about 2% or 2,000 genes are presumed to encode GPCRs. Receptors in which endogenous ligands are identified, including GPCRs, are referred to as "known" receptors, and receptors to which endogenous ligands are not identified are referred to as "orphan" receptors. GPCRs represent an important area for the development of pharmaceutical products, with 60% of all prescription drugs developed from approximately 20 of 100 known GPCRs.

GPCRs are receptors that mediate cellular responses in response to various hormones and neurotransmitters, and are known to be involved in various physiological phenomena or pathological phenomena such as inflammation in the cell and are recognized as a major target of the development of new drugs. . The types of GPCRs are very diverse, and ligands that bind by type vary from low molecular weight materials with relatively simple chemical structures to large molecular weight proteins (FIG. 1). In the case of the short chain fatty acid, intestinal microorganisms are not produced by cells in the body but are produced by fermenting dietary fiber (Non-Patent Document 7). Recently, short chain fatty acids and GPCRs with their ligands have been known to be involved in immune responses such as metabolic diseases and inflammation such as obesity and diabetes. The GPCR protein accounts for about 25% of target substances in drugs used for various diseases such as central nervous system diseases, endocrine diseases, heart diseases, inflammation and metabolic disorders. Nearly half of the new drugs being developed are GPCR proteins as target substances And the clinical importance of it is well recognized (Non-Patent Document 8). In the case of GPR43, it is also irrelevant, and the mainly expressed tissues are white adipose tissue, enteroendocrine cells, and neutrophil.

The GPCR is a receptor that interacts with the G protein, a protein that binds to GTP, as the name suggests. The G protein interacting with the receptor consists of G ?, G ?, G ?, of which G? Serves as a secondary messenger. The Gα is again divided according to their functions into _{Gs, Gi, Gq, G 12} /13 , etc. (FIG. 1). Of these, GPR43 it is also known to regulate the Gi and Gq protein that GPR43 activity when activated cAMP (3'-5'-cyclic adenosine monophosphate) concentration in the cell decreases and the increased Ca ^{2 +} concentrations. cAMP is an important secondary messenger essential to the proper biological response of the cells by hormones and other external signals. cAMP is required for cell communication to regulate feedback by the hormone. The cAMP, which is immobilized at various positions inside the cell, is synthesized from ATP by the enzyme adenylate cyclase and decomposed into AMP by phosphodiesterase, an enzyme that decomposes cAMP.

The GTPase activity of these G proteins varies greatly depending on whether the ligand is attached to the receptor or not. G protein not only acts as a signal transducer that connects an external signal to an intracellular effector but also acts as an amplifier that amplifies the transmitted signal. For example, the time for a neurotransmitter such as norepinephrine to interact with receptors on the cell membrane is only a few milliseconds, but the activated Gs is active for tens of seconds, resulting in a tremendous amplification of the signal It is possible. In the case of cholera toxin causing cholera, when the active form of Gs fails to return to the inactive form before signal transduction, the active form of Gs binds to the G protein and does not return to the state before the original signal transmission do. Therefore, the signal continues to be transmitted, and the living body loses its balance. Exogenous regulation of these G proteins has been found outside of these toxins. In particular, in relation to the development of tumors, pituitary adenomas that secrete growth hormone have been shown to have increased Gs and intracellular cAMP levels, and gene mutations have been observed in these tumors, It is strongly suggested that mutations are involved in tumorigenesis. Therefore, studies on GPCRs are currently being used not only as academic research subjects, but also as an important target of new drug development.

Among many GPCRs, GPR43 (G protein receptor 43) is a receptor having short chain fatty acids (SCFA, carbon length C ₂ -C ₆ ) such as propionic acid and acetic acid as ligands (Non-Patent Document 1). The GPR43 may be used in combination with other cytokines such as polymorphonuclear leukocytes (PMN), purified neutrophils, peripheral blood mononuclear cells (PBMC), purified monocytes, B lymphocytes It is known that expression is observed not only in immunocytes (non-patent document 2) but also in other tissues (for example, intestinal cells or endothelial cells) that have a major influence on the proliferation of inflammatory responses Literature 3). Non-Patent Document 4 discloses that the short chain fatty acids (SCFAs) play a major role in the development of inflammatory conditions such as colitis, arthritis and asthma.

It has been reported that GPR43 knock-out (KO) mice are protected from chronic dextran sodium sulfate (DSS) -induced colitis (Non-Patent Document 5). In the chronic DSS model, GPR43 KO mice are less affected by the reduction of infiltrated polymorphonuclear leukocytes (PMN) against colon inflammation than wild-type (WT) mice And granulocyte-specific myeloperoxidase activity as well as TNFα levels were further reduced in the GPR 43 KO colon than in the WT colon. SCFA has been shown to induce neutrophil migration in vitro and in vivo with neutrophilic granulocytes isolated in vitro and in neutrophil migration in an inflammatory air pouch model. GPR43 KO isolated neutrophilic granulocytes (neutrophilic granulocytes) have lost their ability to migrate to SCFAs, we can expect that GPR43 is closely related to granulocyte granulocytes.

In addition, Non-Patent Document 6 discloses that the GPR43 KO mouse is protected against high fat diet (HFD) -induced obesity and dyslipidemia. More specifically, GPR43 KO mice were shown to have higher lean body mass and lower body fat mass than WT mice. Plasma adiponectin levels were found to increase in GPR43 mice compared to control mice, and liver of GPR43 mice was found to have less triglyceride content. Furthermore, in GPR43 KO mice that performed the GPR43 gene deletion test by the glucose tolerance test, the sensitivity to insulin was increased and the insulin level was lower, while the glucose level was normal. In addition, it can be seen that, compared to WT mice, the decrease in macrophage content in adipose tissue of GPR43 KO mice is less affected by HFD-induced lipid inflammation and HFD-induced hypercholesterolemia.

Patent Document 1 discloses that a compound that acts as an antagonist to GPR43 is used as a therapeutic agent for diseases such as inflammatory condition, infectious diseases, autoimmune diseases, immune cell dysfunction and cardiometabolic diseases proliferative disease, etc. In Patent Document 2 and Patent Document 3, GPR43 agonists are administered to a mouse model of diabetes to confirm anti-obesity and anti-diabetic effects. Thus, metabolic disease treatment And the like.

Accordingly, the present inventors have made efforts to develop an inhibitor of GPR43 that can be used as an important target of development of new drugs. As a result, human GPR43 DNA was introduced into a cell host Glosensor cell capable of detecting cAMP concentration in a cell, Has found a compound that increases the amount of cAMP in a cell line that is continuously expressed and confirmed that the compound is a competitive and surmountable inhibitor for GPR43 and other ligands, thus completing the present invention.

WO 2013/098033 A1 US 2013/0023539 US 7303889

Le Pouls et al., 2003, The Journal of Biological Chemistry, 278, 28, 25481-25489; Brown et al., 2003, The Journal of Biological Chemistry, 278, 13, 13312-11319. The Journal of Biological Chemistry, 278, 28, 25481-25489; Brown et al., 2003, The Journal of Biological Chemistry, 278, 13, 11312-11319; Stoddart et al., 2008, Pharmacological Reviews, 60, 405-417. Dass et al., 2007, Neurogastroenterol Motil., 19: 66-74, Karaki et al., 2006, Cell Tissue Tes., 324/353-360. K.M. Maslowski et al., 2009, Nature, 461, 1282-1287; C. Sina et al., 2009, The Journal of Immunology, 183: 7514-7522 C. Sina et al., 2009, The Journal of Immunology, 183; 7514-7522 Bjursell et al., 2011, Am. J. Physiol. Endocrinol Metab. 300: (11) E211-E220 Sellin et al., 1999, News. Physiol. Sci, 14, 58-64 Carrieria et al., Curr Top Med Chem 2013; 13: 1069-1097

It is an object of the present invention to provide a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative which inhibits GPR43 (G protein receptor 43) activity, an optical isomer thereof or a pharmaceutically acceptable A pharmaceutical composition for preventing or treating an inflammatory or metabolic disease containing a salt as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition comprising the above 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient, And to provide a health functional food composition for preventing or ameliorating a metabolic disease.

Still another object of the present invention is to provide a pharmaceutical composition comprising the 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient, (G-protein coupled receptor 43) inhibitor.

In order to achieve the above object, the present invention provides a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative represented by the following formula 1, an optical isomer thereof or a pharmaceutically acceptable There is provided a pharmaceutical composition for preventing or treating an inflammatory or metabolic disease containing a salt as an active ingredient.

 [Chemical Formula 1]

In Formula 1,

And R ₁ is aryl of C _{6 -10,} wherein the aryl is a halogen, at least one member selected from the group consisting of straight or branched chain alkyl, and straight or branched chain alkoxy of C _{1 -5} C _{1 -5} substituents of substituted Can be;

R ₂ is an aryl group of C _{6 -10,} wherein said aryl is a linear or branched C _{1 -5} alkyl, linear or branched C _{1 -5} alkyl, and at least one member selected from the group consisting of -NR ₃ R ₄ The substituent may be substituted;

R ₃ and R ₄ are independently H or C _{1 -5} straight or branched chain alkyl.

The present invention also provides a pharmaceutical composition comprising a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative represented by the following formula 1, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient And a health functional food composition for preventing or ameliorating an inflammatory or metabolic disease contained therein.

[Chemical Formula 1]

In Formula 1,

R ₁ and R ₂ are the same as defined in the above formula (1).

Further, the present invention provides a pharmaceutical composition comprising a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative represented by the following formula 1, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient Lt; RTI ID = 0.0 > (GPR43) < / RTI >

[Chemical Formula 1]

In Formula 1,

R ₁ and R ₂ are the same as defined in the above formula (1).

The 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative according to the present invention, its optical isomer or a pharmaceutically acceptable salt thereof significantly inhibits the activity of human GPR43 Therefore, it can be usefully used as an active ingredient of a pharmaceutical composition for the prevention and treatment of an inflammatory or metabolic disease associated with GPR43 activity, or a health functional food.

FIG. 1 shows intracellular signal transduction pathways of GPCR (G-protein coupled receptor).
Figure 2 shows that the compounds of the present invention are inverse agonists and are specific for GPR43
Figure 3 is a graph showing that the compounds of the present invention are surmountable inhibitors.
Figure 4 is a graph showing that the compounds of the present invention are competitive inhibitors.
Figure 5 is a graph showing that the compounds of the present invention are inhibitors that do not affect GPR41 and ChemR23, but affect only human GPR43 (hGPR43).

Hereinafter, the present invention will be described in detail.

The present invention relates to a pharmaceutical composition comprising a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient A pharmaceutical composition for preventing or treating inflammation or metabolic diseases.

[Chemical Formula 1]

In Formula 1,

And R ₁ is aryl of C _{6 -10,} wherein the aryl is a halogen, at least one member selected from the group consisting of straight or branched chain alkyl, and straight or branched chain alkoxy of C _{1 -5} C _{1 -5} substituents of substituted Can be;

R ₂ is an aryl group of C _{6 -10,} wherein the aryl is a straight-chain or branched alkoxy, and at least one member selected from the group consisting of -NR ₃ R ₄ a linear or branched alkyl, C _1-5 of C _{1 -5} The substituent may be substituted;

R ₃ and R ₄ are independently H or C _{1 -5} straight or branched chain alkyl.

Preferably,

,

,

또는

이고,R ₁ is

,

,

or

ego,

이다.
R ₂ is

to be.

Preferable examples of the compound represented by the formula (1) according to the present invention include the following compounds.

(1) Synthesis of 4- (4- (dimethylamino) phenyl) -N- (3,5-dimethylphenyl) -6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine- Copy Mid;

(2) Synthesis of 4- (4- (dimethylamino) phenyl) -N- (3,4-dimethylphenyl) -6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine- Copy Mid;

(3) Synthesis of N- (3-chloro-4-methylphenyl) -4- (4- (dimethylamino) phenyl) -6- Carboxamide; And

(4) 4- (4- (Dimethylamino) phenyl) -6-methyl-2-oxo-Nm-tolyl-1,2,3,4-tetrahydropyrimidine-5-carboxamide.

The compound represented by the formula (1) of the present invention can be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid and the like, aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, Derived from organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like. Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, But are not limited to, but are not limited to, but are not limited to, but are not limited to, but are not limited to, halides, halides, halides, halides, halides, halides, But are not limited to, lactose, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene Hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-carboxylate, benzenesulfonate, Sulfonate, naphthalene-2-sulfonate, mandelate and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving a derivative of the formula (1) in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile and the like, Followed by filtration and drying. Alternatively, the solvent and excess acid may be distilled off under reduced pressure, followed by drying and crystallization in an organic solvent.

In addition, the present invention encompasses not only the compound represented by the formula (1) and pharmaceutically acceptable salts thereof, but also solvates, optical isomers and hydrates thereof which can be prepared therefrom.

Further, the pharmaceutical composition is characterized by inhibiting GPR43 (G-protein coupled receptor 43) activity.

Wherein the inflammatory disease is selected from the group consisting of allergic and non-allergic rhinitis, chronic and acute rhinitis, chronic and acute gastritis or enteritis, ulcerative gastritis, acute and chronic nephritis, acute and chronic hepatitis, chronic obstructive pulmonary disease, Atherosclerosis, fungal infections, bacterial infections, fungal infections, burns, wounds due to surgical or dental surgery, prostaglandin E hyperaemia, atherosclerotic arteries Rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, iritis, peritonitis, uveitis, dermatitis, eczema, multiple sclerosis and the like.

In addition, the metabolic diseases include obesity, type 2 diabetes, dyslipidemia, insulin resistance, hepatic steatosis, and non-alcoholic fatty liver.

The present invention also provides a pharmaceutical composition comprising a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative represented by the following formula 1, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient And a health functional food composition for preventing or ameliorating an inflammatory or metabolic disease contained therein.

[Chemical Formula 1]

In Formula 1,

R1 and R2 are the same as defined in the above formula (1).

Further, the present invention provides a pharmaceutical composition comprising a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative represented by the following formula 1, an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient Lt; RTI ID = 0.0 > (GPR43) < / RTI >

[Chemical Formula 1]

In Formula 1,

R1 and R2 are the same as defined in the above formula (1).

At this time, the GPR43 activity inhibitor is a surmountable inhibitor and is an inverse agonist. The surmountable inhibitor is also known as competitive antagonists and is capable of reversibly binding to the same binding site (active site) of the receptor, such as an internal ligand or an agonist, . Once bound, the inhibitor blocks agonist binding and the activity level of the receptor is determined by the relative binding power of the molecule to the position and the relative concentration. The inverse agonist also refers to a substance that binds to the receptor as an agonist but reverses the activity of the receptor.

In a specific embodiment of the present invention, screening for inhibitor screening of GPR43 using high throughput screening (HTS) has been performed to screen compounds inhibiting GPR43 activity, increase the amount of cAMP in the cells , Indicating significant activity (see Example 1).

In addition, the present inventors confirmed that the compound of the present invention is specific for GPR43. As a result, it was confirmed that the compound of the present invention was the inverse effect of GPR43 when the compound was not in the presence of propionic acid, 2A and 2B), confirming that GPR43 activity is also reduced by the compound when propionic acid is present, confirming that the compound of the present invention is specific to GPR43 (see FIGS. 2C and 2D of Experimental Example 1).

Further, it was confirmed that the GPR43 inhibition by the compound of the present invention was overcome by the active agent. As a result, it was confirmed that the compound of the present invention is a surmountable inhibitor that GPR43 is re-activated by the active agent inhibiting GPR43 (See FIG. 3 of Experimental Example 2).

In addition, it was confirmed that the compound of the present invention competitively competes with the active agent of GPR43 (see FIG. 4 of Experimental Example 3).

In addition, it was confirmed that the compound of the present invention is an inhibitor that affects only human GPR43 (hGPR43), although it has no effect on GPR41 and ChemR23 (see FIG. 5 of Experimental Example 4).

Thus, by confirming that the GPR43 activity inhibitor of the present invention is capable of significantly inhibiting human GPR43 (hGPR43) activity as an inverse agonist and a surmountable inhibitor, And can be usefully used as an active ingredient of a pharmaceutical composition for the prevention and treatment of metabolic diseases.

The compound of formula (I) according to the present invention may be administered orally or parenterally in a variety of formulations at the time of clinical administration. In the case of formulation, the compound of the present invention may be used as a filler, an extender, a binder, a wetting agent, a disintegrant, Diluents or excipients.

Solid form preparations for oral administration include tablets, patients, powders, granules, capsules, troches and the like, which may contain one or more excipients such as starch, calcium carbonate, Sucrose, lactose, gelatin or the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions or syrups. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like are included in addition to commonly used simple diluents such as water and liquid paraffin. .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used.

The effective dose of the compound of the present invention on the human body may vary depending on the age, weight, sex, dosage form, health condition and disease severity of the patient, and is generally about 0.001-100 mg / kg / 0.0 > mg / kg / day. ≪ / RTI > It is generally from 0.07 to 7000 mg / day, preferably from 0.7 to 2500 / day, based on adult patients weighing 70, and may be administered once to several times per day It may be administered in divided doses.

Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation Examples.

However, the following Examples, Experimental Examples and Preparation Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples, Experimental Examples and Production Examples.

< Example  1> High-speed bulk screening ( high throughput screening ( HTS )) For the screening of inhibitors of GPR43

<1-1> GPR43 Establishment of stable animal cell line

A human GPR43-Myc gene was transfected into a Glosensor cell line to establish an animal cell line stably expressing GPR43. Specifically, GPR43 gene (SEQ ID NO: 1) was amplified from cDNA extracted from a human cell line using a PCR method, and then inserted into a glargensensor cell line (Promega) and hGPR43-Myc gene (Stratagene) into a pIRES-neo3 vector The gluosensor cell line was transfected with pIRES-neo3-hGPR43-Myc using Fugene6 (Promega) reagent. Single colony cells were separated and cultured in DMEM (Welgene) supplemented with 10% fetal bovine serum (FBS, Invitrogen) and 200 μg / ml hygromycin (Goldbio) and 500 μg / ml G418 (Goldbio) And subcultured in one medium under the conditions of 5% CO 2 and 37 ° C. One cell line with relatively high activity among the established cell lines was selected and used in the subsequent experiments.

<1-2> Glow sensor / hGPR43  Using cell lines GPR43  Screening inhibitor

In order to screen for GPR43-inhibiting compounds, the test substance library was treated with Glosensor cell line which consistently expresses human GPR43 prepared in Example 1, and the amount of cAMP in the cells was measured and cAMP The substances that increase the amount were selected.

Specifically, the gluosensor / hGPR43 cell line of Example <1-1> was dispensed into a 96-well plate at 100,000 cells / well and after 16-20 hours, the cells were cultured in a CO ₂ independent medium (CO ₂ independent media, Invitrogen) was replaced with a solution of 2% cAMP reagent (Promega) containing luciferin. After incubation at 20 ° C for two hours, the luminescence was measured with a luminometer. The test substance library and 1 mM of propionic acid were treated. After 5 minutes, 1 μM of Forskolin, which directly activates the adenyl cyclase, was treated After incubation at 20 ° C for 15 minutes, the luminescence was measured with a luminometer instrument capable of measuring luminescence. The emission value obtained later was divided by the value measured before the material treatment to calculate the final value.

As a result, as shown in Tables 1 and 2, the following compounds that inhibit GPR43 activity from the test substance library and increase the amount of cAMP were screened (Table 1), and these compounds inhibited GPR43 activity and cAMP And exhibit significant activity, it has been confirmed that the compound of the present invention can be used for prevention and treatment of inflammation or metabolic diseases (Table 1 and Table 2).

In Table 2 below ** indicates that 10 μM ≤ IC ₅₀ ≤ 20 μM, *** represents 100 nM ≤ IC ₅₀ ? 10? M.

compound Chemical structural formula compound Chemical structural formula One

3

2

4

compound Degree of activity Compound 1 ** Compound 2 *** Compound 3 ** Compound 4 **

< Experimental Example  1> GPR43 Identification of the specificity of the compound against

In order to confirm that the compound of Example 1 is specific to GPR43, the following experiment was conducted.

Specifically, Compound 1 and Compound 3 among the compounds screened from Example 1 were purchased from ChemBridge, and Compound 2 and Compound 4 were synthesized in-house.

The propionic acid and acetic acid which are the original ligands of GPR43 and the compound 1 of the present invention and CATPB which is a substance synthesized in the Euroscreen are treated to give GPR43 with propionic acid, acetic acid, compound 1 of the present invention or The binding sites of the CATPB inhibitor were the same or the binding sites were different. The specificity of these substances for GPR43 was measured by the change in luminescence caused by luciferase when the concentration of Compound 1 was changed.

First, cell lines expressing human GPR43 and glensor (glensor) genes and cell lines expressing only glossensor were cultured in DMEM medium for about 15 hours at 37 ° C. On the other hand, Luciferin, a substrate of luciferase, was mixed in a new medium to a concentration of 2%. The existing medium in the culture plate of the cells was removed, and the medium was changed to the medium containing the luciferin, followed by incubation at 20 ° C for 2 hours. Compound 1 (50 μM, 16.67 μM, 5.56 μM, 1.85 μM, 0.62 μM, 0.21 μM, 0.07 μM and 0.02 μM) of the present invention and 1 mM of propionic acid were treated in the culture dish. After 5 minutes, 1 μM of forskolin, which directly activates adenyl cyclase, was treated and cultured at 20 ° C for 15 minutes. Luminescence was measured using a luminometer instrument. Phoscholin is mainly used to measure levels of cyclic AMP (cAMP), activates the enzyme adenylyl cyclase and increases intracellular cAMP levels.

As a result, as shown in Fig. 2, only the cells (B, D) expressing GPR43 showed changes in luminescence with respect to Compound 1 and CATPB of the present invention. A and B in Fig. 2 are treated with propionic acid, and C and D are GPR43 inhibitor Compound 1, and CATPB and propionic acid simultaneously treated. Inhibitors that inhibit the activation of GPR43 at basal levels are referred to as antagonists and inhibitors that decrease the activity level of GPR43 that is not activated at the basal level Called inverse agonist. FIG. 2B shows that Compound 1 of the present invention and CATPB decrease the activity of GPR43 even in the absence of propionic acid, confirming that the compound of the present invention is an inverse agonist. Since GPR43 regulates the activity of Gi, when GPR43 is activated, the concentration of cAMP in the cell decreases. As a result, the concentration of cAMP increases as the concentration of this substance increases, (Fig. 2).

< Experimental Example  2> GPR43 &Lt; RTI ID = 0.0 &gt; ( surmountable  Identification as an inhibitor

In order to confirm whether GPR43 inhibition by the compound of the present invention and CATPB is overcome by the active agent, the following experiment was conducted.

First, a cell line expressing human GPR43 and a glow sensor gene and a cell line expressing only glow sensor were cultured in DMEM medium for about 15 hours at 37 ° C. On the other hand, luciferin, a substrate of luciferase, is mixed in a new medium so as to have a concentration of 2%. The existing medium in the culture dish with the cells was removed, and the medium was changed to the medium containing the luciferin, followed by incubation at 20 ° C for 2 hours. The culture plate was treated with Compound 1 (20 μM, 10 μM, and 0 μM) of the present invention and CATPB in advance and cultured at 20 ° C. for 30 minutes so that the substances were preferentially bound to GPR43. Then, GPR43 The activated acetic acid and propionic acid were treated with cells (10 mM, 3.33 mM, 1.11 mM, 0.37 mM, 0.12 mM, 0.04 mM, 0.01 mM, and 0.005 mM) Lt; RTI ID = 0.0 &gt; 20 C &lt; / RTI &gt; for 15 minutes. The luminescence was measured using a luminometer instrument.

As a result, as shown in FIG. 3, A is the compound 1 and acetic acid (C2) of the present invention, B is CATPB and acetic acid (C2), C is the compound 1 of the present invention and propionic acid (C3), D is CATPB and propionic acid C3). &Lt; / RTI &gt; As the concentration of acetic acid and propionic acid increases, the degree of luminescence decreases. Also, it was confirmed that the concentration of acetic acid and propionic acid, which decrease the luminescence, increases as the concentration of the two substances (Compound 1 of the present invention, CATPB) as the GPR43 inhibitor increases (Fig. 3). In addition, it was confirmed that GPR43 was activated in a concentration-dependent manner with respect to acetic acid and propionic acid which were later introduced while GPR43 was previously inhibited (FIG. 3). Thus, the compounds of the present invention exhibit the property of a surmountable inhibitor that GPR43 is inhibited but GPR43 can be activated again by an activator (ligand).

< Experimental Example  3> GPR43 Identification of the competitive properties of the compounds against the active agents of

In order to measure the degree of luminescence of GPR43 by simultaneously treating the compound of the present invention and CATPB with acetic acid (C2) and propionic acid (C3), the following experiment was conducted.

Specifically, the same procedure was performed as in Experiment 2, except that Compound 1 of the present invention, which is an inhibitor of GPR43, and CATPB were treated simultaneously with acetic acid (C2) or propionic acid (C3) to measure the degree of luminescence .

As a result, as shown in FIG. 4, the pattern of the graph showed a tendency similar to that of FIG. Propionic acid and acetic acid showed a concentration (EC ₅₀ ) of 50% activation of GPR43 of 244 μM and 349 μM, respectively. When the inhibitor concentrations of GPR43 were increased to 10 μM and 20 μM, respectively, the EC ₅₀ value gradually increased. The graph pattern of Figure 3 shows that Compound 1 and CATPB of the present invention are both competitive and acetic acid for propionic acid and acetic acid since the relationship of two different ligands to one receptor, for example, GPR43, And as a surmountable inhibitor. Therefore, it was confirmed that the compound of the present invention exhibited competitive properties with respect to binding of propionic acid and acetic acid (FIG. 4).

< Experimental Example  4> GPR43 , GPR41  And ChemR23 &Lt; / RTI &gt; CATPB Specificity comparison Confirm

In order to confirm that the compound of the present invention is specific to GPR43 as an inhibitor in comparison with CATBP, the following experiment was conducted.

Specifically, as in GPR43, GPR41 or ChemR23, known as GPCR, which regulates the activity of Gi protein, was used to carry out experiments using the same method used in Experimental Example 1 above.

As a result, the compound 1 and CATPB all of the invention as shown in Figure 5, but there was no noticeable effect on GPR41 and ChemR23, only IC _50, each human GPR43 (hGPR43) And the values were 12.3 μM and 130 nM, respectively. It was confirmed that neither Compound 1 nor CATPB of the present invention, which are the above inhibitors, affected mouse GPR43 (mGPR43). Therefore, it was confirmed that the compound of the present invention and CATPB are inhibitors that affect only human GPR43 (FIG. 5).

< Manufacturing example  1> Preparation of pharmaceutical preparations

1. Manufacturing of powder

2 g of the compound of formula 1 according to the invention

Lactose 1 g

The above components were mixed and packed in airtight bags to prepare powders.

2. Preparation of tablets

100 mg of the compound of formula 1 according to the present invention

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. Preparation of capsules

100 mg of the compound of formula 1 according to the present invention

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

4. Manufacture of granules

The compound of formula 1 according to the invention in an amount of 150 mg

Soybean extract 50 mg

Glucose 200 mg

Starch 600 mg

After mixing the above components, 100 μl of 30% ethanol was added and the mixture was dried at 60 ° C to form granules, which were filled in a capsule.

<110> Korea Research Institute of Bioscience and Biotechnology <120> Novel compounds modulating GPR43 receptor and use thereof <130> 14P-06-056 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 993 <212> DNA <213> Homo sapiens <400> 1 atgctgccgg actggaagag ctccttgatc ctcatggctt acatcatcat cttcctcact 60 ggcctccctg ccaacctcct ggccctgcgg gcctttgtgg ggcggatccg ccagccccag 120 cctgcacctg tgcacatcct cctgctgagc ctgacgctgg ccgacctcct cctgctgctg 180 ctgctgccct tcaagatcat cgaggctgcg tcgaacttcc gctggtacct gcccaaggtc 240 gtctgcgccc tcacgagttt tggcttctac agcagcatct actgcagcac gtggctcctg 300 gcgggcatca gcatcgagcg ctacctggga gtggctttcc ccgtgcagta caagctctcc 360 cgccggcctc tgtatggagt gattgcagct ctggtggcct gggttatgtc ctttggtcac 420 tgcaccatcg tgatcatcgt tcaatacttg aacacgactg agcaggtcag aagtggcaat 480 gaaattacct gctacgagaa cttcaccgat aaccagttgg acgtggtgct gcccgtgcgg 540 ctggagctgt gcctggtgct cttcttcatc cccatggcag tcaccatctt ctgctactgg 600 cgttttgtgt ggatcatgct ctcccagccc cttgtggggg cccagaggcg gcgccgagcc 660 gtggggctgg ctgtggtgac gctgctcaat ttcctggtgt gcttcggacc ttacaacgtg 720 tcccacctgg tggggtatca ccagagaaaa agcccctggt ggcggtcaat agccgtggtg 780 ttcagttcac tcaacgccag tctggacccc ctgctcttct atttctcttc ttcagtggtg 840 cgcagggcat ttgggagagg gctgcaggtg ctgcggaatc agggctcctc cctgttggga 900 cgcagaggca aagacacagc agaggggaca aatgaggaca ggggtgtggg tcaaggagaa 960 gggatgccaa gttcggactt cactacagag tag 993

Claims (10)

A pharmaceutical composition comprising an effective amount of a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof, Pharmaceutical composition for preventing or treating disease:
[Chemical Formula 1]

(In the formula 1,
And R ₁ is aryl of C _{6 -10,} wherein the aryl is a halogen, at least one member selected from the group consisting of straight or branched chain alkyl, and straight or branched chain alkoxy of C _{1 -5} C _{1 -5} substituents of substituted Can be;
R ₂ is an aryl group of C _{6 -10,} wherein the aryl is a straight-chain or branched alkoxy, and at least one member selected from the group consisting of -NR ₃ R ₄ a linear or branched alkyl, C _1-5 of C _{1 -5} The substituent may be substituted;
R ₃ and R ₄ are independently H or C _{1 -5} straight or branched chain alkyl.
The method according to claim 1,
R ₁ is

,

,

or

ego,
R ₂ is

Or a pharmaceutically acceptable salt or solvate thereof.
The method according to claim 1,
A pharmaceutical composition for preventing or treating an inflammatory or metabolic disease, wherein the compound represented by the formula (1) is selected from the following group of compounds:
(1) Synthesis of 4- (4- (dimethylamino) phenyl) -N- (3,5-dimethylphenyl) -6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine- Copy Mid;
(2) Synthesis of 4- (4- (dimethylamino) phenyl) -N- (3,4-dimethylphenyl) -6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine- Copy Mid;
(3) Synthesis of N- (3-chloro-4-methylphenyl) -4- (4- (dimethylamino) phenyl) -6- Carboxamide; And
(4) 4- (4- (Dimethylamino) phenyl) -6-methyl-2-oxo-Nm-tolyl-1,2,3,4-tetrahydropyrimidine-5-carboxamide.
The method according to claim 1,
Wherein said pharmaceutical composition inhibits GPR43 (G-protein coupled receptor 43) activity.
The method according to claim 1,
Wherein the metabolic disease is any one selected from the group consisting of obesity, type 2 diabetes, dyslipidemia, insulin resistance, hepatic steatosis and non-alcoholic fatty liver. A pharmaceutical composition for preventing or treating diseases.
The method according to claim 1,
The inflammatory disease is selected from the group consisting of allergic and non-allergic rhinitis, chronic and acute rhinitis, chronic and acute gastritis or enteritis, ulcerative gastritis, acute and chronic nephritis, acute and chronic hepatitis, chronic obstructive pulmonary disease, , Inflammatory pain, migraine headache, back pain, fibromyalgia, fascia disease, viral infection, bacterial infection, fungal infection, burn, wound due to surgical or dental surgery, prostaglandin E hyperaemia, atherosclerosis, Inflammatory or metabolic disease characterized in that it is selected from the group consisting of gout, degenerative arthritis, rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, iritis, peritonitis, uveitis, dermatitis, eczema and multiple sclerosis &Lt; / RTI &gt;
A pharmaceutical composition comprising an effective amount of a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof, A health functional food composition for preventing or ameliorating a disease:
[Chemical Formula 1]

(In the formula 1,
R ₁ And R &lt; ₂ &gt; are as defined in formula (1) of claim 1).
A pharmaceutical composition comprising a 2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide derivative represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient: GPR43 -protein coupled receptor 43) Inhibitors of activity:
[Chemical Formula 1]

(In the formula 1,
R ₁ and R ₂ are the same as defined in the formula (1).
9. The method of claim 8,
Wherein the GPR43 activity inhibitor is a surmountable inhibitor.
9. The method of claim 8,
Wherein said GPR43 activity inhibitor is an inverse agonist.

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