KR102623218B1 - Phenylacetic acid derivatives and composition for preventing or treating autoimmune diseases comprising the same - Google Patents

Abstract

The present invention provides a phenylacetic acid derivative, a pharmaceutical composition for preventing or treating autoimmune diseases, and a health functional food composition containing the same as an active ingredient. The compound represented by Formula 1 according to the present invention or a pharmaceutically acceptable salt thereof may have an effect of inhibiting LPA1 (Lysophosphatidic acid 1) receptor activation, and is useful as a pharmaceutical composition and health functional food composition for preventing or treating LPA1-related diseases. It can be used effectively. In addition, the pharmaceutical composition and health functional food composition can effectively prevent or treat not only autoimmune diseases but also other immune-related diseases with similar developmental mechanisms.

Description

Phenylacetic acid derivatives and compositions for preventing or treating autoimmune diseases containing the same as active ingredients {PHENYLACETIC ACID DERIVATIVES AND COMPOSITION FOR PREVENTING OR TREATING AUTOIMMUNE DISEASES COMPRISING THE SAME}

The present invention relates to phenylacetic acid derivatives and compositions for preventing or treating autoimmune diseases containing them as active ingredients.

Autoimmune disease is a disease that causes an abnormal immune response by mistaking the body's own substances for antigens, including psoriasis, Sjögren's syndrome, inflammatory bowel disease, rheumatoid arthritis, multiple sclerosis, neuromyelitis optica, Guillain-Barre syndrome, Graves' disease, and Behcet's disease. etc.

The main cause of autoimmune diseases is believed to be the migration of lymphocytes to tissues, triggering abnormalities in the autoimmune system, but the clear pathogenesis of each disease is not yet known.

Recently, with a focus on efficient immunosuppression, research is being conducted to develop treatments for autoimmune diseases, focusing on immunosuppression at the cellular level (suppressing the activation of lymphocytes and cell types responsible for the immune response in diseased lesion tissues) and molecular levels.

Lysophospholipids, represented by lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P), have long been considered phospholipid metabolites of cell membrane components, but are now used as G protein-coupled receptors. It has been reported to be an important extracellular signaling substance that regulates various biological functions through activation of lysophospholipid receptors, which are classified as coupled receptors (GPCRs).

Among them, LPA1 is the first lysophospholipid receptor to be identified (identified in 1996) and is a key factor regulating nervous system development. Its importance in various diseases has been identified through recent studies. Immune response regulation is attracting attention. The signal transduction mechanism of LPA1 and the organic system with in vivo tissues, which have been revealed through various research data, are not only related to autoimmune diseases such as Sjögren's syndrome, stroke, psoriasis, inflammatory bowel disease, diabetic nephropathy and fibrosis, and systemic sclerosis, but also to the neuropsychiatric system. It is becoming important data for the treatment of a variety of diseases, including disability, neuropathic pain, infertility, cardiovascular disease, and cancer. (Reference Park et al, “Inhibition of lysophosphatidic acid receptor ameliorates Sjogren's syndrome in NOD mice” Oncotarget. 2017 , 8(16), 27240-27251, Yung et al, “LPA receptor signaling: pharmacology, physiology, and pathophysiology“ J Lipid Res. 2014, 55, 1192-1214, Choi et al. “LPA receptors: subtypes and biological actions " Annu Rev Pharmacol Toxicol. 2010;50:157-86. Debendra Pattanaik et al, "A Role for Lysophosphatidic Acid and Sphingosine 1-Phosphate in the Pathogenesis of Systemic Sclerosis" Discov Med 10(51):161-167, August 2010 .)

Currently, steroids, Cyclosporine, Rapamycin, Methotrexate, Cyclophosphamide, IV Immunoglobulin, Axathioprine, Infliximab, etc. Although it is treated with a combination of various immunosuppressants, in most cases, persistent inflammation and frequent recurrence occur, and many systemic side effects occur due to treatment.

In particular, rare autoimmune diseases have a small number of patients, but the cost of the treatment applied is very expensive, so the economic burden on patients is very high, and the cost burden is increasing as long-term treatment is required. Although the number of patients with rare autoimmune diseases is increasing in Korea, there is no clear treatment and the cure rate is low, so it continues to be an economic and social problem.

Korean Patent Publication No. 10-2014-0117842 (published on October 8, 2014)

In order to solve the above problems, the present invention provides phenylacetic acid derivatives.

The present invention provides a pharmaceutical composition for preventing or treating autoimmune diseases containing the phenylacetic acid derivative as an active ingredient.

The present invention provides a health functional food composition for improving or preventing autoimmune diseases containing the phenylacetic acid derivative as an active ingredient.

The compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof according to the present invention,

[Formula 1]

In the above formula, R ¹ and R ² may each be the same or different, and are selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, halogen, nitro, or a substituent of the formula 2 below, or R ¹ and R ² are connected to form a 5-ring heterocycle,

[Formula 2]

At this time, R ³ and R ⁴ may be the same or different, and are selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, trifluoromethyl, phenoxy, halogen, or nitro, or ^R3 and R ⁴ can be connected to form a 5-ring heterocycle.

The pharmaceutical composition for preventing or treating LPA1-related diseases according to the present invention contains the above compound or a pharmaceutically acceptable salt thereof as an active ingredient, and may have an effect of inhibiting LPA1 (Lysophosphatidic acid 1) receptor activation.

The health functional food composition for improving or preventing LPA1-related diseases according to the present invention contains the above compound or a pharmaceutically acceptable salt thereof as an active ingredient and may have an effect of inhibiting LPA1 (Lysophosphatidic acid 1) receptor activation.

Since the phenylacetic acid derivative according to the present invention has the effect of inhibiting LPA1 receptor activation, it can be used as a pharmaceutical composition and health functional food composition for preventing or treating LPA1-related diseases such as stroke, psoriasis, Sjögren's syndrome, inflammatory bowel disease, diabetic nephropathy, and fibrosis. It can be useful.

Autoimmune diseases can be effectively prevented and treated through the pharmaceutical composition and health functional food composition for preventing or treating LPA1-related diseases according to the present invention. In addition, it can be applied in a variety of ways to the treatment of not only the above autoimmune diseases, but also other immune-related diseases with similar developmental mechanisms.

Figure 1 shows the results of evaluating the efficacy of inhibiting LPA1 receptor activation.
Figure 2 shows the IC ₅₀ values of the derivatives in Figure 1 that showed an effect of inhibiting LPA1 receptor activation by more than 50%.
Figure 3 shows the results of a middle cerebral artery stenosis/reperfusion (tMCAO) mouse experiment.
Figure 4 shows the results of drug efficacy evaluation for psoriasis using a compound according to the present invention.

Hereinafter, the present invention will be described in detail.

The present invention provides an animal model (in vivo disease model) for autoimmune diseases such as Sjogren's disease, psoriasis, and multiple sclerosis, and abnormal immune response diseases such as diabetic nephropathy and stroke (cerebral ischemia). ), the usefulness of LPA1 as a new target molecule for the treatment of autoimmune diseases was verified through prior research, and the present invention was completed by discovering a compound with LPA1 antagonistic activity.

The present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1, R ¹ and R ² may each be the same or different, and are hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, halogen, nitro, or any one selected from the substituents of Formula 2 below, Alternatively, R ¹ and R ² may be connected to form a 5-ring heterocycle.

[Formula 2]

At this time, R ³ and R ⁴ in the substituents of Formula 2 may each be the same or different, and are selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, trifluoromethyl, phenoxy, halogen, or nitro. Either one may be used, or R ³ and R ⁴ may be connected to form a 5-ring heterocycle. The 5-ring heterocycle formed by connecting R ³ and R ⁴ may be a 1,3-dioxol ring.

More specifically, the compound is 3-(5-(2-(benzo[d][1,3]dioxol-5-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1 ,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(4-methoxyphenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a ]Pyrazin-2-yl)propanoic acid, 3-(5-(2-(4-chlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2- 1) propanoic acid, 3-(5-(2-(4-bromophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid , 3-(5-(2-(2-chlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5 -(2-(3-chlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-( p-Tolyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(3-methoxyphenyl) Acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-([1,1'-biphenyl]- 4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(3-nitrophenyl)acetyl )-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(3-fluorophenyl)acetyl)-4, 5,6,7-Tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(4-fluorophenyl)acetyl)-4,5,6, 7-Tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, and 3-(5-(2-(3,4-dichlorophenyl)acetyl)-4,5,6,7- It may be any one selected from the group consisting of tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid.

The compound may be a compound represented by the following formula (3) or a pharmaceutically acceptable salt thereof.

[Formula 3]

In Formula 3, R ¹ and R ² may each be the same or different, and may be any one selected from hydrogen, (C1~C4)alkyl, (C1~C4)alkoxy, trifluoromethyl, phenoxy, halogen, or nitro. Alternatively, R ¹ and R ² may be connected to form a 5-ring heterocycle.

In the compound represented by Formula 3, R ¹ and R ² may each be the same or different, and any selected from hydrogen, (C1~C4)alkyl, (C1~C2)alkoxy, trifluoromethyl, phenoxy, or fluoro. Alternatively, R ¹ and R ² may be connected to form a 1,3-dioxole ring.

More specifically, the compound represented by Formula 3 is 3-(5-(2-(4'-methoxy-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6 ,7-Tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(3'-methoxy-[1,1'-biphenyl]-4- 1) Acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(4'-phenoxy-[1 ,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2 -(3'-fluoro-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl) Propanic acid, 3-(5-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1, 5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(4'-(tert-butyl)-[1,1'-biphenyl]-4-yl)acetyl)-4, 5,6,7-Tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(4'-(trifluoromethyl)-[1,1' -Biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(4) '-methyl-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, and 3-(5-(2-(4-(benzo[d][1,3]dioxol-5-yl)phenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5- a] pyrazin-2-yl) propanoic acid.

The present invention provides a pharmaceutical composition for preventing or treating LPA1-related diseases, which contains the above compound or a pharmaceutically acceptable salt thereof as an active ingredient and has an effect of inhibiting LPA1 (Lysophosphatidic acid 1) receptor activation. do.

The LPA1-related disease may be any one selected from the group consisting of stroke, psoriasis, Sjögren's syndrome, inflammatory bowel disease, diabetic nephropathy, and fibrosis, but is not limited thereto.

The pharmaceutical composition according to the present invention may be prepared by further including an appropriate pharmaceutically acceptable carrier, excipient, or diluent according to the above formulation. The term “pharmaceutically acceptable” means that the pharmaceutical composition exhibits non-toxic properties to cells or humans exposed to the pharmaceutical composition.

Carriers, excipients or diluents usable in the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Examples include methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil.

The pharmaceutical composition according to the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions according to conventional methods. there is.

When the pharmaceutical composition according to the present invention is formulated in the above form, it can be prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations contain at least one excipient such as starch, calcium carbonate, or sucrose. ) or it can be prepared by mixing lactose, gelatin, etc.

In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. .

Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogeratin, etc. can be used.

In the pharmaceutical composition according to the present invention, the pharmaceutical composition can be administered in a pharmaceutically effective amount. The above “pharmaceutically effective amount” refers to an amount that is sufficient to treat the disease at a reasonable benefit/risk ratio applicable to medical treatment and does not cause side effects, and the effective dose level is determined by the patient's health status, ulcer It can be determined based on factors including type, severity, drug activity, sensitivity to the drug, method of administration, time of administration, route of administration and excretion rate, duration of treatment, drugs combined or used simultaneously, and other factors well known in the medical field. .

The amount of the compound used as an active ingredient of the pharmaceutical composition according to the present invention may vary depending on the patient's age, gender, weight, and disease, but is 0.001 to 100 mg/kg, preferably 0.01 to 10 mg/kg once or several times a day. It can be administered. Additionally, the dosage of the compound according to the present invention may be increased or decreased depending on the route of administration, severity of disease, gender, weight, age, etc. Accordingly, the above dosage does not limit the scope of the present invention in any way.

The pharmaceutical composition can be administered to mammals such as rats, mice, livestock, and humans through various routes. All modes of administration are contemplated, for example, oral, rectal or by intravenous, intramuscular, subcutaneous, endobronchial inhalation, intrathecal or intracerebroventricular injection.

The present invention provides a health functional food composition for improving or preventing LPA1-related diseases, which contains the above compound or a pharmaceutically acceptable salt thereof as an active ingredient and has an effect of inhibiting LPA1 (Lysophosphatidic acid 1) receptor activation. .

The LPA1-related disease may be any one selected from the group consisting of stroke, psoriasis, Sjögren's syndrome, inflammatory bowel disease, diabetic nephropathy, and fibrosis, but is not limited thereto.

The health functional food according to the present invention may be provided in the form of powder, granules, tablets, capsules, syrup or beverage, and the health functional food is used with other foods or food additives in addition to the compound according to the present invention as an active ingredient, It can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on its purpose of use, for example, prevention, health, or therapeutic treatment.

The effective dose of the compound contained in the health functional food according to the present invention can be used in accordance with the effective dose of the pharmaceutical composition, but in case of long-term intake for the purpose of health and hygiene or health control, It may be below this range, and the active ingredient can be used in amounts above the above range because there is no problem in terms of safety.

There is no particular limitation on the type of health functional food according to the present invention, and examples include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, Examples include beverages, tea, drinks, alcoholic beverages, and vitamin complexes.

The compound according to the present invention can be used in the form of a pharmaceutically acceptable salt, and the salt can be used in the form of either a pharmaceutically acceptable basic salt or an acid salt.

Basic salts can be used in the form of any one of organic base salts and inorganic base salts, such as sodium salt, potassium salt, calcium salt, lithium salt, magnesium salt, cesium salt, aminium salt, ammonium salt, and triethyl salt. It may be selected from the group consisting of amidium salts and pyridinium salts, but is not limited thereto.

A useful acid salt is an acid addition salt formed by a free acid. Inorganic acids and organic acids can be used as free acids. Hydrochloric acid, hydrobromic acid, sulfuric acid, sulfurous acid, and phosphoric acid can be used as inorganic acids, and citric acid, acetic acid, maleic acid, fumaric acid, gluconic acid, and methanesulfonic acid can be used as organic acids. , benzenesulfonic acid, camphorsulfonic acid, oxalic acid, malonic acid, glutaric acid, acetic acid, glyconic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid, citric acid, aspartic acid. etc. can be used. Preferably, hydrochloric acid can be used as the inorganic acid, and methanesulfonic acid can be used as the organic acid.

In addition, the compound according to the present invention may include not only pharmaceutically acceptable salts, but also all salts, hydrates, and solvates that can be prepared by conventional methods.

The addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving the compounds represented by Formula 1 to Formula 3 in a water-miscible organic solvent, such as acetone, methanol, ethanol, or acetonitrile. It can be prepared by adding an excess amount of an organic base or an aqueous solution of an inorganic base and then precipitating or crystallizing it. Alternatively, an addition salt can be obtained by evaporating the solvent or excess base from this mixture and drying it, or it can be prepared by suction filtration of the precipitated salt.

Hereinafter, the present invention will be described in detail through examples to aid understanding. However, the following examples only illustrate the content of the present invention and the scope of the present invention is not limited to the following examples. Examples of the present invention are provided to more completely explain the present invention to those skilled in the art.

<Experiment method>

All reagents needed for the experiment were purchased with a purity of 97% or higher from TCI, Sigma Aldrich, and Alfa Aesar. Before performing the experiment, the glassware to be used for the reaction was sufficiently washed with acetone, dried in an oven at 60°C, and the experiment was conducted under a nitrogen stream. Dichloromethane, tetrahydrofuran, methanol, acetone, toluene, diethyl ether, ethyl acetate, and hexane are largely purified. It was purchased from Gold, and the anhydrous solvent was purchased from Sigma Aldrich. The progress of the reaction was confirmed in TLC using a UV lamp and coloring reagents such as AA, CAM, potassium permanganate (KMnO ₄ ), and ninhydrin. H NMR and C NMR spectra were measured using a BRUKER Ascend 600 spectrometer and Varian VnmrS-400, and CDCl3 and DMSO-d6 were used as solvents for analysis. Chemical shift values are expressed in ppm, and coupling constants (J) are expressed in Hz.

<Example 1>

Synthesis of parent tetrahydropyrazolopyrazine

[Scheme 1]

Reaction Scheme 1 is a process for synthesizing tetrahydropyrazolopyrazine, the core mother nucleus according to the present invention. In the same manner as Reaction Scheme 1, propargylamine is used as a starting material to produce tetrahydropyrazole according to the following method. The synthesis of pyrazine (Tetrahydropyrazolopyrazine) was performed.

<Example 1-1> 4-methyl-N-(prop-2-yn-1-yl)benzenesulfonamide [4-methyl-N-(prop-2-yn-1-yl)benzenesulfonamide]

Propargylamine (0.5ml, 7.81mmol) and triethylamine (TEA 1.3ml, 9.37mmol) were added to dichloromethane (7.5ml) at 0°C to create a mixture. P-Toluene Sulfonyl Chloride (1.49 g, 7.81 mmol) dissolved in dichloromethane (22.5 ml) was added dropwise to the mixture, and then stirred at room temperature for 5 hours. After stirring was completed, water was added to terminate the reaction, and the product was extracted with dichloromethane. Water was removed using sodium sulfate, and the filtrate was concentrated under reduced pressure. Afterwards, it was separated and purified by column chromatography to obtain Compound 1 (1.517g, 92.84%).

Compound 1: ^1H NMR (600 MHz, Chloroform-d) δ 7.78 (d, J = 8.3 Hz, 2H), 7.32 (d, J = 8.2 Hz, 2H), 4.71 (s, 1H), 3.83 (dd, J = 6.1, 2.5 Hz, 2H), 2.44 (s, 3H), 2.11 (s, 1H); ¹³ C NMR (151 MHz, Chloroform-d) δ 143.88, 136.49, 129.72, 127.41, 77.95, 73.02, 32.89, 21.58.

<Example 1-2> N-(2-chloroethyl)-4-methyl-N-(prop-2-yn-1-yl)benzenesulfonamide [N-(2-chloroethyl)-4-methyl- N-(prop-2-yn-1-yl)benzenesulfonamide]

Compound 1 (1.517g, 7.25mmol) and potassium carbonate (1.2g, 8.7mmol) were added to acetone (30ml) to create a mixture. Afterwards, 1-bromo-2-chloroethane (1.2ml, 14.5mmol) was added dropwise to the mixture and refluxed for 3 hours. After cooling, the product was extracted with ethyl acetate and water was removed with magnesium sulfate. The filtrate was concentrated under reduced pressure, and then purified through column chromatography to obtain compound 2 (1.29 g, 65.48%).

Compound 2: ¹ H NMR (600 MHz, Chloroform-d) δ 7.74 (d, J = 8.0 Hz, 2H), 7.31 (d, J = 7.9 Hz, 2H), 4.19 (s, 2H), 3.70 (t, J = 7.0 Hz, 2H), 3.50 (t, J = 6.9 Hz, 2H), 2.43 (s, 3H), 2.11 (m, 1H); ¹³ C NMR (151 MHz, Chloroform-d) δ 143.99, 135.48, 129.69, 127.66, 76.64, 74.17, 48.30, 41.78, 38.16, 21.58.

<Example 1-3> Ethyl 5-tosyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate [Ethyl 5-tosyl-4,5,6,7 -tetrahydropyrazolo[1,5-a]pyrazine-2-carboxylate]

Compound 2 (6.05g, 22.26mmol) and diazoacetate (23.7ml, 27.83mmol) were added to toluene (26.3ml) and stirred at 140°C for 12 hours. After stirring, cesium carbonate (10.88g, 33.4mmol) and tetrahydrofuran (10ml) were added to the mixture and the reaction continued for 30 minutes. After completion of the reaction, the mixture was cooled to room temperature and 1 normal hydrochloric acid was added to neutralize the mixture. The product was extracted through ethyl acetate and water removed through magnesium sulfate. The filtrate was concentrated under reduced pressure, and then separated and purified through column chromatography to obtain compound 3 (4.215 g, 54.19%).

Compound 3: ¹ H NMR (600 MHz, DMSO-d ₆ ) δ 7.73 (d, J = 8.3 Hz, 1H), 7.46 (d, J = 8.1 Hz, 1H), 6.59 (s, 1H), 4.33 (s) , 1H), 4.23 (d, J = 7.1 Hz, 1H), 4.18 (t, J = 5.6 Hz, 1H), 3.60 (t, J = 5.6 Hz, 1H), 3.32 (s, 2H), 2.50 (s) , 1H), 2.40 (s, 2H), 1.25 (s, 1H); ¹³ C NMR (151 MHz, Chloroform-d) δ 144.03, 136.09, 129.94, 127.25, 108.44, 61.50, 49.56, 41.45, 21.56, 14.25.

<Example 1-4> (5-tosyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)methanol [(5-tosyl-4,5,6, 7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)methanol]

Compound 3 (1.13 g, 3.23 mmol) and lithium aluminum hydride (159 mg, 4.2 mmol) were added to tetrahydrofuran (10 ml) at 0°C to create a mixture. Afterwards, the mixture was stirred at room temperature for one hour. After completion of the reaction, 1 normal hydrochloric acid was added to neutralize and the product was extracted with dichloromethane. Magnesium sulfate was added to the extracted product to remove water, and the filtrate was concentrated under reduced pressure and separated and purified through column chromatography to obtain compound 4 (940 mg, 94%).

Compound 4: ^1H NMR (600 MHz, Chloroform-d) δ 7.71 (d, J = 8.1 Hz, 2H), 7.36 (d, J = 8.0 Hz, 2H), 6.02 (s, 1H), 4.61 (m, 2H), 4.30 (d, J = 2.1 Hz, 2H), 4.18 (m, 2H), 3.56 (m, 2H), 2.44 (s, 3H); ¹³ C NMR (151 MHz, Chloroform-d) δ 144.52, 134.70, 132.74, 130.05, 127.70, 100.47, 100.45, 58.86, 58.81, 46.82, 43.76, 43.28, 30.95, 21. 59.

<Example 1-5> Ethyl (E)-3-(5-tosyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)acrylate [Ethyl (E )-3-(5-tosyl-4,5,6,7-tetrahydropyrazolo[1,5-a] pyrazin-2-yl)acrylate]

Dimethyl sulfoxide (4.43ml, 61.8mmol) was added to dichloromethane (10ml) to create a mixture. Afterwards, oxalyl chloride (2.651ml, 30.9mmol) dissolved in dichloromethane (15ml) at -60°C was added dropwise to the mixture. After 5 minutes, compound 4 (1.9 g, 6.18 mmol) was added to the mixture, and the reaction continued for 1 hour. After completion of the reaction, the temperature of the mixture was brought close to room temperature and the product was extracted with dichloromethane. Magnesium sulfate was added to the extracted product to remove water, and the filtrate was concentrated under reduced pressure and separated and purified through column chromatography to obtain compound 5 (1.34 g, 60%).

Compound 5: ¹ H NMR (600 MHz, Chloroform-d) δ 7.71 (d, J = 8.0 Hz, 2H), 7.57 (d, J = 16.1 Hz, 1H), 7.36 (d, J = 7.9 Hz, 2H) , 7.27 (s, 1H), 6.35 (d, J = 16.1 Hz, 1H), 6.25 (s, 1H), 4.33 (s, 2H), 4.23 (t, J = 6.6 Hz, 4H), 3.59 (t, J = 5.4 Hz, 2H), 2.44 (s, 3H), 1.31 (t, J = 7.1 Hz, 3H); ¹³ C NMR (151 MHz, Chloroform-d) δ 166.80, 148.18, 144.62, 136.06, 135.13, 132.71, 130.09, 127.70, 119.60, 101.14, 60.49, 47.23, 43.66, 43.17, 21.59, 14.28.

<Example 1-6> Ethyl 3-(5-tosyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoate [Ethyl 3-(5- tosyl-4,5,6,7-tetrahydropyrazolo[1,5-a] pyrazin-2-yl)propanoate]

Compound 5 (1.3 g, 3.46 mmol) and 10% Pd/C (0.1 g) were added to methanol (20 ml) to create a mixture, and stirred for 2 hours at room temperature under a hydrogen stream. After completion of the reaction, the mixture was filtered through Celite. The filtrate was concentrated under reduced pressure and purified through column chromatography to obtain compound 6 (1 g, 76%).

Compound 6: ¹ H NMR (600 MHz, Chloroform-d) δ 7.70 (m, 2H), 7.35 (m, 2H), 5.84 (d, J = 1.0 Hz, 1H), 4.27 (s, 2H), 4.14 ( m, 2H), 3.54 (m, 2H), 2.89 (t, J = 7.6 Hz, 2H), 2.61 (dd, J = 8.0, 7.2 Hz, 2H), 2.44 (s, 3H), 1.24 (t, J) = 7.1 Hz, 3H); ¹³ C NMR (151 MHz, Chloroform-d) δ 172.89, 151.73, 134.20, 132.76, 129.99, 127.71, 100.67, 60.41, 46.63, 43.83, 43.28, 33.96, 23.51, 21. 58, 14.23.

<Example 1-7> Methyl 3-(4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoate [Methyl 3-(4,5,6, 7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoate]

Compound 6 (388 mg, 1.03 mmol) and magnesium powder (247 mg, 10.3 mmol) were added to methanol (6 ml) to create a mixture. Afterwards, the mixture was allowed to react at 0°C for 1 hour. After completion of the reaction, the mixture is filtered through Celite. The filtrate was concentrated under reduced pressure and separated and purified through column chromatography to obtain compound 7 (283 mg, 73%), tetrahydropyrazolopyrazine, the core parent of the compound according to the present invention.

Compound 7: ^1H NMR (600 MHz, Chloroform-d) δ 5.78 (s, 1H), 4.05 (s, 2H), 4.01 (s, 2H), 3.69 (s, 3H), 3.28 (s, 2H), 2.93 (m, 2H), 2.67 (m, 2H); ¹³ C NMR (151 MHz, Chloroform-d) δ 173.53, 150.64, 138.07, 99.55, 51.62, 47.61, 43.62, 43.01, 33.90, 23.59.

<Example 2> Synthesis of parent nucleus and phenylacetic acid derivative

[Scheme 2]

Scheme 2 is a process for synthesizing 13 types of phenylacetic acid derivatives with tetrahydropyrazolopyrazine, the core parent nucleus according to the present invention. In the same manner as Scheme 2, tetrahydropyrazolopyrazine of compound 7 and variously substituted phenyl Compounds 8a to 8m were synthesized by amide coupling with 13 types of acetic acid, and 13 types of compounds 9a to 9m were synthesized by hydrolyzing methyl ester.

<Example 2-1> Amide coupling reaction (8a~8m)

Compound 7 (50 mg, 0.224 mmol), acid (45 mg, 0.269 mmol), and triethylamine (0.13 ml, 0.896 mmol) were added to dichloromethane (0.5 ml) to create a mixture. Hydroxybenzotriazol (37mg, 0.269mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (52mg) were added to the resulting mixture. , 0.269 mmol) was added and stirred at room temperature for 12 hours. After completion of the reaction, the product was extracted with dichloromethane. Magnesium sulfate was added to the extracted product to remove water, and then the filtrate was concentrated under reduced pressure and separated and purified through column chromatography to obtain compound 8 imide (40 mg, 42%).

<Example 2-2> Hydrolysis (9a~9m)

Compound 8a (40 mg, 0.1 mmol) was added to tetrahydrofuran (3 ml) to create a mixture. Afterwards, lithium hydroxide (55 mg, 1.3 mmol) dissolved in water (1 ml) was added and stirred at room temperature for 12 hours. After completion of the reaction, 1 normal hydrochloric acid was added to neutralize and the product was extracted with dichloromethane. Magnesium sulfate was added to the extracted product to remove water, and the filtrate was concentrated under reduced pressure and separated and purified through column chromatography to obtain compound 9a (32 mg, 90%). Compounds 9b-9m were obtained from compounds 8b-8m in the same manner.

[Compounds 9a~9m]

1) Compound 9a: 3-(5-(2-(benzo[d][1,3]dioxol-5-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5- a]pyrazin-2-yl)propanoic acid [3-(5-(2-(benzo[d][1,3]dioxol-5-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1, 5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 6.75 (s, 2H), 6.67 (d, J = 8.1 Hz, 1H), 5.95 (s, 2H), 5.88 (d, J = 36.8 Hz, 1H), 4.78 (s, 1H), 4.64 (s, 1H), 4.09 (d, J = 43.5 Hz, 2H), 3.96 (s, 1H), 3.86 (s, 1H), 3.72 (d, J = 24.5 Hz, 2H) ), 2.92 (s, 2H), 2.70 (s, 2H).

2) Compound 9b: 3-(5-(2-(4-methoxyphenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(4-methoxyphenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.15 (m, 2H), 6.86 (m, 2H), 5.87 (d, J = 46.3 Hz, 1H), 4.70 (d, J = 87.7 Hz, 2H), 4.09 (d, J = 42.6 Hz, 2H), 3.90 (s, 1H), 3.84 (s, 1H), 3.78 (s, 3H), 3.75 (d, J = 25.0 Hz, 2H), 2.91 (t, J = 7.3 Hz, 2H), 2.69 (td, J = 7.3, 2.5 Hz, 2H).

3) Compound 9c: 3-(5-(2-(4-chlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [ 3-(5-(2-(4-chlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.30 (td, J = 8.2, 2.6 Hz, 2H), 7.17 (dd, J = 14.9, 8.1 Hz, 2H), 5.89 (d, J = 40.8 Hz, 1H ), 4.71 (d, J = 83.8 Hz, 2H), 4.10 (dd, J = 39.2, 5.5 Hz, 2H), 4.00 (t, J = 5.5 Hz, 1H), 3.85 (t, J = 5.5 Hz, 1H) ), 3.77 (d, J = 22.9 Hz, 2H), 2.92 (m, 2H), 2.70 (m, 2H).

4) Compound 9d: 3-(5-(2-(4-bromophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(4-bromophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.45 (m, 2H), 7.11 (dd, J = 14.5, 8.2 Hz, 2H), 5.89 (d, J = 40.2 Hz, 1H), 4.71 (d, J = 82.7 Hz, 2H), 4.09 (m, 2H), 3.93 (dt, J = 100.4, 5.4 Hz, 2H), 3.76 (d, J = 22.3 Hz, 2H), 2.92 (td, J = 7.4, 2.5 Hz) , 2H), 2.69 (q, J = 7.0 Hz, 2H).

5) Compound 9e: 3-(5-(2-(2-chlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [ 3-(5-(2-(2-chlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.40 (ddt, J = 8.3, 5.3, 2.6 Hz, 1H), 7.30 (m, 1H), 7.24 (m, 2H), 5.91 (d, J = 26.5 Hz) , 1H), 4.76 (d, J = 59.0 Hz, 2H), 4.13 (dt, J = 38.0, 5.5 Hz, 2H), 4.05 (t, J = 5.5 Hz, 1H), 3.91 (m, 3H), 2.94 (m, 2H), 2.72 (m, 2H).

6) Compound 9f: 3-(5-(2-(3-chlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [ 3-(5-(2-(3-chlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.26 (m, 4H), 7.13 (m, 1H), 5.90 (d, J = 36.4 Hz, 1H), 4.72 (d, J = 85.5 Hz, 2H), 4.11 (dt, J = 43.6, 5.5 Hz, 2H), 4.02 (t, J = 5.5 Hz, 1H), 3.86 (t, J = 5.5 Hz, 1H), 3.79 (d, J = 22.9 Hz, 2H), 2.93 (t, J = 7.3 Hz, 2H), 2.71 (td, J = 7.2, 4.4 Hz, 2H).

7) Compound 9g: 3-(5-(2-(p-tolyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [3 -(5-(2-(p-tolyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.12 (d, J = 7.1 Hz, 4H), 5.88 (d, J = 45.9 Hz, 1H), 4.70 (d, J = 92.5 Hz, 2H), 4.09 ( dt, J = 47.2, 5.5 Hz, 2H), 3.87 (dt, J = 46.2, 5.4 Hz, 2H), 3.77 (d, J = 25.0 Hz, 2H), 2.91 (t, J = 7.4 Hz, 2H), 2.68 (td, J = 7.3, 2.8 Hz, 2H), 2.32 (s, 3H).

8) Compound 9h: 3-(5-(2-(3-methoxyphenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(3-methoxyphenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.23 (m, 1H), 6.79 (m, 3H), 5.87 (d, J = 52.5 Hz, 1H), 4.70 (d, J = 93.9 Hz, 2H), 4.09 (dt, J = 47.8, 5.4 Hz, 2H), 3.88 (dt, J = 64.5, 5.4 Hz, 2H), 3.78 (m, 5H), 2.91 (t, J = 7.4 Hz, 2H), 2.67 (m , 2H).

9) Compound 9i: 3-(5-(2-([1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a] Pyrazin-2-yl) propanoic acid [3-(5-(2-([1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a] pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.56 (m, 4H), 7.43 (t, J = 7.6 Hz, 2H), 7.32 (m, 3H), 5.89 (d, J = 38.1 Hz, 1H), 4.73 (d, J = 78.0 Hz, 2H), 4.11 (dt, J = 50.8, 5.4 Hz, 2H), 4.00 (t, J = 5.4 Hz, 1H), 3.85 (m, 3H), 2.92 (t, J = 7.5 Hz, 2H), 2.67 (t, J = 7.5 Hz, 2H).

10) Compound 9j: 3-(5-(2-(3-nitrophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [ 3-(5-(2-(3-nitrophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, DMSO-d6) δ 12.12 (s, 1H), 8.13 (m, 2H), 7.70 (m, 1H), 7.60 (q, J = 7.9 Hz, 1H), 5.94 (s, 1H) ), 4.75 (d, J = 100.7 Hz, 2H), 4.12 (t, J = 5.5 Hz, 1H), 4.03 (m, 4H), 3.93 (t, J = 5.5 Hz, 1H), 2.73 (q, J = 7.5 Hz, 2H), 2.52 (d, J = 8.4 Hz, 2H).

11) Compound 9k: 3-(5-(2-(3-fluorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(3-fluorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.31 (m, 1H), 6.99 (m, 3H), 5.90 (d, J = 40.3 Hz, 1H), 4.73 (d, J = 89.6 Hz, 2H), 4.11 (m, 2H), 3.98 (t, J = 5.4 Hz, 1H), 3.86 (t, J = 5.5 Hz, 1H), 3.81 (d, J = 24.7 Hz, 2H), 2.93 (t, J = 7.2 Hz, 2H), 2.72 (td, J = 7.1, 3.0 Hz, 2H).

12) Compound 9l: 3-(5-(2-(4-fluorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(4-fluorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.20 (ddd, J = 15.1, 8.2, 5.3 Hz, 2H), 7.01 (m, 2H), 5.88 (d, J = 38.0 Hz, 1H), 4.71 (d) , J = 76.3 Hz, 2H), 4.08 (m, 2H), 3.97 (s, 1H), 3.85 (d, J = 5.4 Hz, 1H), 3.77 (d, J = 23.4 Hz, 2H), 2.90 (t , J = 7.1 Hz, 2H), 2.68 (s, 2H).

13) Compound 9m: 3-(5-(2-(3,4-dichlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propane Acid [3-(5-(2-(3,4-dichlorophenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Chloroform-d) δ 7.37 (dd, J = 10.4, 8.2 Hz, 1H), 7.32 (dd, J = 15.1, 2.1 Hz, 1H), 7.06 (ddd, J = 12.8, 8.2, 2.1 Hz, 1H), 5.90 (d, J = 26.8 Hz, 1H), 4.71 (d, J = 62.4 Hz, 2H), 4.12 (dt, J = 21.6, 5.5 Hz, 2H), 3.95 (dt, J = 111.3, 5.5 Hz, 2H), 3.73 (d, J = 19.5 Hz, 2H), 2.90 (dt, J = 7.5, 3.7 Hz, 2H), 2.66 (td, J = 7.4, 5.2 Hz, 2H).

<Example 3> Synthesis of biphenylacetic acid derivatives

<Example 3-1> Synthesis of biphenylacetic acid (12a-12i)

To synthesize a biphenylacetic acid derivative, biphenylacetic acid was first synthesized using the method shown in Scheme 3 below.

[Scheme 3]

4-Bromophenylacetic acid (830mg, 3.86mmol), 4-Methylphenylboronic acid (500mg, 3.68mmol), and cesium carbonate (2.4g, 7.36mmol) in methanol (10ml). was added to create a mixture. Afterwards, palladium catalyst (2.5 mg) was added and refluxed for one day. After completion of the reaction, the temperature of the mixture was brought close to room temperature, neutralized using 1 normal hydrochloric acid, and the product was extracted with dichloromethane. Magnesium sulfate was added to the extracted product to remove water, and the filtrate was concentrated under reduced pressure and separated and purified through column chromatography to obtain compound 12h (870 mg, 95%). Compounds 12a to 12i were synthesized using the same method.

[Compounds 12a-12i]

1) Compound 12a: 2-(4'-methoxy-[1,1'-biphenyl]-4-yl)acetic acid [2-(4'-methoxy-[1,1'-biphenyl]-4-yl )acetic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.52 (m, 4H), 7.32 (m, 2H), 6.98 (m, 2H), 3.82 (s, 3H), 3.62 (s, 2H).

2) Compound 12b: 2-(3'-methoxy-[1,1'-biphenyl]-4-yl)acetic acid [2-(3'-methoxy-[1,1'-biphenyl]-4-yl )acetic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.54 (d, J = 8.1 Hz, 2H), 7.33 (dd, J = 17.4, 8.2 Hz, 3H), 7.16 (d, J = 7.0 Hz, 1H), 7.12 (m, 1H), 6.88 (ddd, J = 8.2, 2.5, 1.0 Hz, 1H), 3.82 (s, 3H), 3.63 (s, 2H).

3) Compound 12c: 2-(4'-phenoxy-[1,1'-biphenyl]-4-yl)acetic acid [2-(4'-phenoxy-[1,1'-biphenyl]-4-yl )acetic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.60 (m, 2H), 7.55 (m, 2H), 7.36 (m, 4H), 7.12 (m, 1H), 7.03 (m, 4H), 3.64 (s) , 2H).

4) Compound 12d: 2-(3'-fluoro-[1,1'-biphenyl]-4-yl)acetic acid [2-(3'-fluoro-[1,1'-biphenyl]-4-yl )acetic acid]

¹ H NMR (600 MHz, Methanol-d4) δ 7.57 (m, 2H), 7.42 (m, 2H), 7.35 (m, 4H), 3.65 (s, 2H).

5) Compound 12e: 2-(4'-fluoro-[1,1'-biphenyl]-4-yl)acetic acid [2-(4'-fluoro-[1,1'-biphenyl]-4-yl )acetic acid]

¹ H NMR (600 MHz, Methanol-d4) δ 7.61 (m, 2H), 7.54 (m, 2H), 7.36 (m, 2H), 7.15 (m, 2H), 3.64 (s, 2H).

6) Compound 12f: 2-(4'-(tert-butyl)-[1,1'-biphenyl]-4-yl)acetic acid [2-(4'-(tert-butyl)-[1,1' -biphenyl]-4-yl)acetic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.54 (m, 4H), 7.45 (d, J = 8.6 Hz, 2H), 7.33 (m, 2H), 3.63 (s, 2H), 1.34 (s, 9H) ).

7) Compound 12g: 2-(4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)acetic acid [2-(4'-(trifluoromethyl)-[1,1'- biphenyl]-4-yl)acetic acid]

¹ H NMR (600 MHz, Methanol-d4) δ 7.80 (m, 2H), 7.72 (m, 2H), 7.64 (m, 2H), 7.41 (m, 2H), 3.67 (s, 2H).

8) Compound 12h: 2-(4'-(methyl)-[1,1'-biphenyl]-4-yl)acetic acid [2-(4'-methyl-[1,1'-biphenyl]-4- yl)acetic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.54 (m, 2H), 7.48 (m, 2H), 7.33 (m, 2H), 7.23 (m, 2H), 3.63 (s, 2H), 2.36 (s) , 3H).

9) Compound 12i: 2-(4-(benzo[d][1,3]dioxol-5-yl)phenyl)acetic acid [2-(4-(benzo[d][1,3]dioxol-5- yl)phenyl)acetic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.49 (m, 2H), 7.32 (m, 2H), 7.08 (m, 2H), 6.87 (m, 1H), 5.97 (s, 2H), 3.62 (s) , 2H).

<Example 3-2> Synthesis of biphenylacetic acid derivatives

[Scheme 4]

In the same manner as Scheme 4, compounds 13a to 13i were synthesized through an amide coupling reaction between compound 7 synthesized in Example 1 and compounds 12a to 12i, and compounds 14a to 14i were synthesized by hydrolyzing the methyl ester. . The amide coupling reaction was the same as Example 2-1, and the hydrolysis was the same as Example 2-2.

[Compounds 14a-14i]

1) Compound 14a: 3-(5-(2-(4'-methoxy-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[ 1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(4'-methoxy-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6 ,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.53 (m, 4H), 7.33 (dd, J = 8.2, 2.9 Hz, 2H), 6.98 (dd, J = 8.6, 3.9 Hz, 2H), 5.96 (m , 1H), 4.79 (s, 2H), 4.03 (m, 3H), 3.91 (m, 3H), 3.82 (d, J = 2.0 Hz, 3H), 2.83 (dt, J = 15.7, 7.5 Hz, 2H) , 2.58 (dt, J = 20.2, 7.7 Hz, 2H).

2) Compound 14b: 3-(5-(2-(3'-methoxy-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[ 1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(3'-methoxy-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6 ,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.57 (m, 2H), 7.32 (m, 3H), 7.14 (m, 2H), 6.90 (m, 1H), 5.99 (s, 1H), 4.79 (d) , J = 10.2 Hz, 2H), 3.97 (m, 6H), 3.83 (d, J = 1.8 Hz, 3H), 2.83 (dt, J = 14.2, 7.6 Hz, 2H), 2.58 (dt, J = 17.3, 7.6 Hz, 2H).

3) Compound 14c: 3-(5-(2-(4'-phenoxy-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[ 1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(4'-phenoxy-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6 ,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.56 (m, 4H), 7.34 (m, 4H), 7.11 (tt, J = 7.4, 1.1 Hz, 1H), 7.02 (m, 4H), 5.95 (d) , J = 44.6 Hz, 1H), 4.78 (d, J = 11.1 Hz, 2H), 3.95 (m, 6H), 2.83 (dt, J = 14.2, 7.6 Hz, 2H), 2.58 (dt, J = 17.4, 7.6 Hz, 2H).

4) Compound 14d: 3-(5-(2-(3'-fluoro-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[ 1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(3'-fluoro-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6 ,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

¹ H NMR (600 MHz, Methanol-d4) δ 7.60 (m, 3H), 7.38 (m, 10H), 7.08 (m, 3H), 5.96 (m, 1H), 4.80 (m, 2H), 4.04 (m , 3H), 3.93 (m, 3H), 2.84 (dt, J = 12.9, 7.6 Hz, 2H), 2.58 (dt, J = 15.8, 7.6 Hz, 2H).

5) Compound 14e: 3-(5-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[ 1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6 ,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

¹ H NMR (600 MHz, Methanol-d4) δ 7.34 (m, 4H), 7.15 (m, 4H), 5.96 (d, J = 45.3 Hz, 1H), 4.80 (d, J = 13.8 Hz, 2H), 4.04 (m, 3H), 3.93 (m, 3H), 2.83 (dt, J = 14.5, 7.6 Hz, 2H), 2.58 (dt, J = 18.0, 7.6 Hz, 2H).

6) Compound 14f: 3-(5-(2-(4'-(tert-butyl)-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydro pyrazolo[1,5-a]pyrazin-2-yl)propanoic acid[3-(5-(2-(4'-(tert-butyl)-[1,1'-biphenyl]-4-yl)acetyl )-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.58 (m, 1H), 7.52 (m, 3H), 7.46 (m, 2H), 7.31 (dd, J = 30.2, 8.0 Hz, 2H), 5.95 (d) , J = 47.1 Hz, 1H), 4.78 (d, J = 8.9 Hz, 2H), 3.96 (m, 6H), 2.83 (dt, J = 15.0, 7.6 Hz, 2H), 2.58 (dt, J = 19.2, 7.6 Hz, 2H), 1.34 (s, 9H).

7) Compound 14g: 3-(5-(2-(4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetra Hydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid[3-(5-(2-(4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)acetyl) -4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.80 (d, J = 8.0 Hz, 3H), 7.72 (dd, J = 8.7, 3.5 Hz, 3H), 7.64 (m, 3H), 7.41 (m, 2H) ), 5.96 (m, 1H), 4.78 (m, 2H), 4.04 (m, 3H), 3.94 (m, 3H), 2.83 (dt, J = 13.5, 7.5 Hz, 2H), 2.58 (m, 2H) .

8) Compound 14h: 3-(5-(2-(4'-methyl-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1 ,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(4'-methyl-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6, 7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.55 (m, 3H), 7.48 (dd, J = 11.1, 7.9 Hz, 3H), 7.34 (dd, J = 8.3, 2.1 Hz, 2H), 7.29 (d) , J = 7.9 Hz, 1H), 7.23 (m, 3H), 5.95 (m, 1H), 4.80 (d, J = 9.3 Hz, 2H), 4.07 (d, J = 5.1 Hz, 2H), 4.01 (m , 1H), 3.92 (m, 3H), 2.83 (dt, J = 15.3, 7.6 Hz, 2H), 2.58 (dt, J = 20.0, 7.6 Hz, 2H), 2.36 (s, 3H), 1.29 (d, J = 3.3 Hz, 2H).

9) Compound 14i: 3-(5-(2-(4-(benzo[d][1,3]dioxol-5-yl)phenyl)acetyl)-4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazin-2-yl)propanoic acid [3-(5-(2-(4-(benzo[d][1,3]dioxol-5-yl)phenyl)acetyl)-4, 5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid]

^1H NMR (600 MHz, Methanol-d4) δ 7.48 (m, 2H), 7.28 (dd, J = 30.8, 7.9 Hz, 2H), 7.06 (m, 2H), 6.86 (dd, J = 8.0, 4.3 Hz) , 1H), 5.97 (m, 3H), 4.77 (d, J = 7.0 Hz, 2H), 3.96 (m, 6H), 2.82 (dt, J = 15.8, 7.6 Hz, 2H), 2.57 (dt, J = 21.0, 7.6 Hz, 2H).

<Experimental Example 1> Evaluation of efficacy of inhibiting LPA1 receptor activation

The following experiment was performed to evaluate the degree of inhibitory effect of the compound according to the present invention on LPA1 receptor activation.

RH7777 cells overexpressing the human LPA1 receptor were transfected with STAT3-luciferase, and the activities of the compounds were measured through luciferase activation assay. To activate the receptor, the cells were treated with 2 μM LPA, and compounds according to the present invention were administered to the cells 30 minutes before the LPA treatment. Analysis of luciferase activation was performed 24 hours after the LPA treatment. At this time, the concentration of the reference substance and compound was 100 nM. (However, when AM095 was used as a reference material, 10 μM was used as the treatment concentration.)

Figure 1 shows the results of evaluating the efficacy of inhibiting LPA1 receptor activation.

Referring to Figure 1, when the LPA1 receptor is in an inactive state (vehicle treated group (veh)), activity 1 is set, and compounds that inhibit the activity by about 50% or more based on the activation value by LPA treatment can be identified. Five types of compounds were selected as effective compounds: Compound 9a, Compound 9d, Compound 9g, Compound 9h, and Compound 9i.

Figure 2 shows the IC ₅₀ values of the derivatives in Figure 1 that showed an effect of inhibiting LPA1 receptor activation by more than 50%.

Referring to FIG. 2, the five compounds selected as effective compounds in the experiment for evaluating the efficacy of inhibiting LPA1 receptor activation were treated with cells at different concentrations, and the IC ₅₀ values for LPA1 activation of these compounds were obtained. The IC ₅₀ value of each compound derived from these results was expressed as a relative percentage (%) with the IC ₅₀ value of AM152, a reference material, as 100%, and the results are shown in Table 1.

division
compound LPA1 inhibitory activity (IC ₅₀ , nM)
note
IC ₅₀ (nM) Compared to AM152
Equivalent activity concentration
Experiment set 1 AM152 0.1001 100% reference material 9a 0.1334 85.5% test substance 9d 2.1 4.70% test substance

experiment set 2 AM152 0.32 100% reference material 9g 0.47 68.2% test substance 9h 0.62 51.9% test substance 9i 0.12 263% test substance

<Experimental Example 2> Evaluation of drug efficacy for cerebral ischemia

Compound 9a and compound 9d according to the present invention were administered to experimental mice (7-8 weeks old) for MCAO (middle cerebral artery occlusion), a model of transient focal cerebral ischemia, and middle cerebral artery stenosis/reperfusion (tMCAO) to evaluate the symptoms and drug efficacy. did.

For tMCAO, male experimental mice (7 weeks old, Orient Co., Ltd) were subjected to middle cerebral artery occlusion (MCAO) for 90 minutes, followed by reperfusion, and each compound was orally administered to the mice.

Specifically, MCAO was caused by blocking the right middle cerebral artery blood flow by inserting a silicone-coated single fiber (9 mm long 5-0 nylon single fiber) from the branching area of the cerebral artery of the mouse, and the single fiber was placed 90 minutes after the occurrence of occlusion. It was later removed to allow blood to flow again, thereby inducing transient focal cerebral ischemia (tMCAO). For drug administration, mice that developed tMCAO immediately after reperfusion were randomly divided into three groups and administered vehicle (10% Tween80, p.o.), compound 9a (5 mg/kg, p.o.), or compound 9d (5 mg/kg, p.o.), respectively. It was administered, and 22 hours after reperfusion, the degree of damage was expressed numerically.

First, the neurological aftereffects of the mice after the experiment were quantified based on the modified neurological severity score scale (mNSS). The total score for neurological sequelae was 18 points, of which each was subdivided into motor function (total 6 points), sensory function (total 2 points), balance sense function (total 6 points), and reflex function (total 4 points). Aftereffect scores were assigned to each item and then summed. The modified neurological severity score scale is based on previous literature (Chen et al., 2001. Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats. Stroke; a journal of cerebral circulation 32:2682-2688.) Reported analysis methods were used.

In addition, brains were removed from mice that completed neurological sequelae tests, stained with 2,3,5-triphenyltetrazolium (TTC), and cerebral infarct volume was measured and analyzed using ImageJ (NIH, Bethesda, MD, USA). did.

Figure 3 shows the results of a middle cerebral artery stenosis/reperfusion (tMCAO) mouse experiment.

Referring to Figure 3, A and B show the volume of the infarcted area in the brain in photographs and numbers, respectively, and C shows the neurological sequelae in numbers. Through A and B, it can be seen that the cerebral infarction volume was statistically significantly reduced by the compounds according to the present invention, and when the compound according to the present invention was administered through C, neurological symptoms were reduced compared to the control group treated with vehicle. It can be seen that aftereffects have decreased.

Therefore, it was confirmed that Compound 9a and Compound 9d according to the present invention can effectively treat cerebral ischemia by effectively reducing brain damage occurring in cerebral ischemia, and thus can be usefully used as a pharmaceutical composition containing them as an active ingredient.

<Experimental Example 3> Evaluation of drug efficacy for psoriasis

Compound 9i according to the present invention was administered to experimental mice (7-8 weeks old) in which psoriasis was induced by applying imiquimod to the skin, and the symptoms and efficacy were evaluated.

Specifically, the imiquimod-induced psoriasis experiment uses Aldara cream containing 5% imiquimod. Psoriasis is induced by applying 62.5 mg of cream to the back and right ear of mice anesthetized with isoflurane every day for 7 days (for normal control groups, petroleum jelly is applied instead of Aldara cream). From the 1st day of application of the cream, compound 9i (10 mg/kg in 1% DMSO in 10% Tween 80, p.o.) was administered to experimental mice once a day for 6 days, and 1% dimethyl sulfoxide (compound 9i) dissolved in A 10% Tween 80 solution (10 mL/kg, p.o.) containing dimethyl sulfoxide (DMSO) was administered to mice equally as a negative control.

Symptoms of disease in mice were measured every day starting from the day after the first compound administration. Disease symptoms were measured by PASI (Psoriasis area and severity index) parameters. PASI parameters include ear thickness, skin erythema, and skin scaling.

Figure 4 shows the results of drug efficacy evaluation for psoriasis using a compound according to the present invention.

Referring to Figure 4, the severity of skin erythema and skin scaling was evaluated by giving a score from 0 to 4 (0=asymptomatic, 1=mild, 2=moderate, 3=severe, 4=very severe).

A is a photo of the skin condition on the final day of the experiment when vehicle (10% Tween80 containing 1% DMSO, p.o.) or compound 9i (10 mg/kg, p.o.) was administered to a vaseline-applied mouse and an experimental mouse with psoriasis, respectively, and B and C are photos of the skin condition on the final day of the experiment. The skin erythema and skin scaling symptom scores are shown, respectively, and D shows the thickness measurement results of the right ear to which Aldara cream was applied.

Looking at the changes in disease severity in B and C, it can be seen that skin erythema and skin scaling symptoms began to significantly decrease after 4 days of compound 9i administration, and the drug efficacy was maintained until the end of the experiment, and the change in ear thickness in D Looking at , it can be seen that when compound 9i is administered, the thickness of the ear thickened by the disease is reduced on the 5th and 6th days of administration.

Through this, it was confirmed that compound 9i according to the present invention can prevent or treat psoriasis by effectively reducing the severity of psoriasis, and can be usefully used as a pharmaceutical composition containing it as an active ingredient.

<Experimental Example 4> Toxicity evaluation

Compound 9a, Compound 9d, Compound 9h, Compound 9g, and Compound 9i were each suspended in 0.5% methylcellulose solution and administered once orally at doses of 0.5 g/kg, 1 g/kg, and 2 g/kg to male Balb/c mice. The survival rate and body weight of mice were examined for 7 days after administration.

After this administration, the animals were observed for death, clinical symptoms, and weight changes, and hematological and blood biochemical tests were performed. An autopsy was performed to visually observe abnormalities in the abdominal and thoracic organs.

As a result, there were no notable clinical symptoms or dead animals in any of the animals, and no toxic changes were observed in body weight changes, blood tests, blood biochemistry tests, and autopsy findings.

As a result of the above results, the compounds of the present invention showed no toxic changes in rats up to 2 g/kg, and therefore, they were judged to be safe substances with an oral lethal dose (LD50) of 2 g/kg or more.

Below, a formulation example of a composition containing compound 9a according to the present invention is described, but the present invention is not intended to be limited, but merely explained in detail.

<Prescription Example 1> Prescription example of pharmaceutical composition

<Prescription Example 1-1> Preparation of powder

A powder was prepared by mixing 20 mg of compound 9a, 100 mg of lactose, and 10 mg of talc and filling it in an airtight bubble.

<Prescription Example 1-2> Preparation of tablets

Tablets were prepared by mixing 20 mg of compound 9a, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate and compressing them according to a conventional tablet manufacturing method.

<Prescription Example 1-3> Preparation of capsules

A capsule was prepared by mixing 10 mg of compound 9a, 100 mg of corn starch, 100 mg of lactose, and 2 mg of magnesium stearate, mixing the ingredients according to a typical capsule preparation method, and filling a gelatin capsule.

<Prescription Example 1-4> Preparation of injections

After mixing 10 mg of compound 9a, an appropriate amount of sterilized distilled water for injection, and an appropriate amount of a pH adjuster, the mixture was prepared with the above ingredient content per ampoule (2 ml) according to a typical injection preparation method.

<Prescription Example 1-5> Preparation of ointment

Prepare an ointment by mixing 10 mg of compound 9a, 250 mg of PEG-4000, 650 mg of PEG-400, 10 mg of white petrolatum, 1.44 mg of methyl parahydroxybenzoate, 0.18 mg of propyl paraoxybenzoate, and the remaining amount of purified water, and then prepare an ointment according to a conventional ointment preparation method. did.

<Prescription Example 2> Health functional food

<Prescription Example 2-1> Manufacturing of health food

Compound 9a 1 mg, appropriate amount of vitamin mixture (vitamin A acetate 70 μg, vitamin E 1.0 mg, vitamin B 1 0.13 mg, vitamin B 2 0.15 mg, vitamin B 6 0.5 mg, vitamin B 12 0.2 μg, vitamin C 10 mg, biotin 10 ㎍, nicotinic acid amide 1.7 ㎎, folic acid 50 ㎍, calcium pantothenate 0.5 ㎎) and appropriate amount of mineral mixture (ferrous sulfate 1.75 ㎎, zinc oxide 0.82 ㎎, magnesium carbonate 25.3 ㎎, potassium phosphate monobasic 15 ㎎, calcium phosphate dibasic) 55 mg, potassium citrate 90 mg, calcium carbonate 100 mg, magnesium chloride 24.8 mg) were mixed to prepare granules, and health food was prepared according to a conventional method.

<Prescription Example 2-2> Manufacturing of health drinks

Add 1 mg of compound 9a, 1000 mg of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, 1 g of taurine, and purified water to make a total of 900 ml. Mix the above ingredients according to a normal health drink manufacturing method, then add about 1 g. After stirring and heating at 85°C for an hour, the resulting solution was filtered, placed in a sterilized 2 L container, sealed, sterilized, and refrigerated.

As the specific parts of the present invention have been described in detail above, it is clear to those skilled in the art that these specific techniques are merely preferred embodiments and do not limit the scope of the present invention. do. That is, the practical scope of the present invention is defined by the appended claims and their equivalents.

Claims (10)

delete delete delete A compound represented by the following formula (3) or a pharmaceutically acceptable salt thereof:
[Formula 3]

In the above formula, R ¹ and R ² may each be the same or different and are selected from hydrogen, (C1~C4)alkyl, (C1~C2)alkoxy, trifluoromethyl, phenoxy, or halogen, or R ¹ and R ² are linked to form a 1,3-dioxol ring (except for compounds where both R ¹ and R ² are hydrogen). delete In claim 4,
The compound is 3-(5-(2-(4'-methoxy-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1, 5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(3'-methoxy-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6 ,7-Tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(4'-phenoxy-[1,1'-biphenyl]-4- I) Acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(3'-fluoro-[1 ,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2 -(4'-fluoro-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl) Proanoic acid, 3-(5-(2-(4'-(tert-butyl)-[1,1'-biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo [1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(4'-(trifluoromethyl)-[1,1'-biphenyl]-4-yl)acetyl )-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, 3-(5-(2-(4'-methyl-[1,1'- Biphenyl]-4-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid, and 3-(5-(2-(4) -(benzo[d][1,3]dioxol-5-yl)phenyl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)propanoic acid A compound or a pharmaceutically acceptable salt thereof, characterized in that it is selected from the group consisting of. 3-(5-(2-(benzo[d][1,3]dioxol-5-yl)acetyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazine-2 -1) Contains propanoic acid or a compound represented by the following formula (3) or a pharmaceutically acceptable salt thereof as an active ingredient, and is characterized by having an effect of inhibiting LPA1 (Lysophosphatidic acid 1) receptor activation, for stroke, psoriasis, and Sjögren's Pharmaceutical composition for preventing or treating LPA1-related diseases selected from the group consisting of syndrome, inflammatory bowel disease, diabetic nephropathy and fibrosis:
[Formula 3]

In the above formula, R ¹ and R ² may each be the same or different and are selected from hydrogen, (C1~C4)alkyl, (C1~C2)alkoxy, trifluoromethyl, phenoxy, or halogen, or R ¹ and R ² are connected to form a 1,3-dioxole ring. delete delete delete