Retinoid acid receptor agonists, methods of making, intermediates, pharmaceutical compositions, and uses thereof

Abstract

The application discloses a retinoic acid receptor agonist shown in formula (I), a preparation method, an intermediate, a pharmaceutical composition and application. The compound has strong agonist activity of retinoic acid receptor gamma and good hepatocyte stability, and can be used for treating diseases related to the retinoic acid receptor.

Description

TECHNICAL FIELD

[0001] The present application relates to a retinoic acid receptor agonist, its preparation method, intermediates, pharmaceutical compositions and uses. BACKGROUND

[0002] Retinoic acid is a metabolite of vitamin A, which initiates and regulates gene expression by activating nuclear receptors, is a key regulator of cell growth, differentiation proliferation and programmed apoptosis, and is involved in physiological and pathological processes such as embryonic development, tissue differentiation and tumor. The physiological effects of retinoic acid are mediated by two types of receptors, namely retinoic acid receptor (RAR) and retinoid X receptor (RXR). Among them, the retinoic acid receptor RAR belongs to the nuclear receptor superfamily, including α, β, γ three kinds. RARβ is divided into β1, β2, β3, β4, etc. The function of retinoic acid receptor signal is ultimately realized through gene transcription regulation, and a complete retinoic acid signal pathway composition includes ligand retinoic acid, receptor dimerization, retinoic acid response element (RA response element, RARE), and auxiliary regulatory factors, etc.

[0003] During cell development, the distribution of retinoic acid receptor RAR has time and space specificity, and different retinoic acid receptor pathways have different functions on cell development. Generally, RARα is the most widely expressed in cells, and exists in most tissues; RARβ is highly expressed in the central nervous system; and RARγ is only expressed in the skin, epithelium and cartilage tissue. In humans and adult rats, the expression product of RARγ is highly restricted in the skin, and the expression amount in other organs is very low. Studies have shown that RARγ selective agonists have the efficacy of treating diseases such as facial acne, psoriasis, fibrodysplasia ossificans progressiva (FOP), and osteochondroma.

[0004] Trifarotene (trade name Aklief) is a RARγ selective agonist developed by Galderma Research and Development Company, which was approved by the US FDA in 2019 for the treatment of facial acne in teenagers aged 9 and above (Lesley J. Scott, Drugs 2019, 79, 1905-1909; Cosio T, Biomedicines, 2021, 9(3): 237).

[0005] Palovarotene is a small molecule drug developed by Clementia Pharmaceuticals, which is also a RAR-g agonist. In January 2022, the drug was approved for marketing in Canada for the treatment of progressive musculoskeletal fibrosis (Kitoh H, Biomedicines, 2020, 8(9): 325). At the same time, the drug has also submitted marketing applications in China, the United States and other countries.

[0006] SUMMARY

[0007] The present application aims to provide a retinoic acid receptor agonist, a preparation method, an intermediate, a pharmaceutical composition and a use thereof.

[0008] In one aspect, the present application provides a compound represented by formula (I):

[0009]

[0010] or a tautomer, a stereoisomer, an isotopic derivative or a pharmaceutically acceptable salt thereof,

[0011] wherein R1 and R2 are independently H, halogen or C1-C6 alkyl;

[0012] m is 1, 2 or 3;

[0013] n is 1, 2, 3 or 4;

[0014] R3 is H or C1-C6 alkyl.

[0015] In some embodiments, in the definition of R1 and R2, the halogen is independently F, Cl, Br or I, respectively.

[0016] In some embodiments, in the definition of R1, R2 and R3, the C 1-6 alkyl is independently C 1-4 alkyl, for example methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl or t-butyl.

[0017] In some embodiments, R1 and R2 are independently H.

[0018] In some embodiments, R3 is H.

[0019] In some embodiments, the compound represented by formula (I) is:

[0020]

[0021] In another aspect, the present application also provides a method for preparing the compound of formula (I) as described above, comprising the following steps: subjecting a compound of formula (II) to the following reaction in the presence of a base in a solvent to obtain the compound of formula (I);

[0022]

[0023] wherein R3 is H, to a group capable of forming upon hydrolysis (e.g. ), and the remaining groups are as defined above.

[0024] In the method for preparing the compound of formula (I) as described above, the reaction conditions and reagents of the reaction can be conventional conditions and reagents for this type of reaction in the art. Preferably, the solvent can be an ether solvent, such as tetrahydrofuran. Preferably, the base can be an inorganic base, such as sodium hydroxide.

[0025] In some embodiments, the method further comprises the following steps: subjecting a compound of formula (III) to the following reaction with a compound of formula (III-1) in the presence of a base in a solvent to obtain the compound of formula (II);

[0026]

[0027] wherein Hal is halogen (e.g. Br), and the remaining groups are as defined above.

[0028] In the method for preparing the compound of formula (II) as described above, the reaction conditions and reagents of the reaction can be conventional conditions and reagents for this type of reaction in the art. Preferably, the solvent can be an amide solvent, such as N,N-dimethylformamide. Preferably, the base can be an inorganic base, such as cesium carbonate.

[0029] In some embodiments, the method further comprises the following steps: subjecting a compound of formula (IV) to the following reaction with a compound of formula (IV-1) in a solvent to obtain the compound of formula (III);

[0030]

[0031] wherein each group is as defined above.

[0032] In the method for preparing the compound of formula (III) as described above, the reaction conditions and reagents of the reaction can be conventional conditions and reagents for this type of reaction in the art. Preferably, the solvent can be a halogenated alkane solvent, such as carbon tetrachloride. Preferably, the reaction can be carried out in the presence of an initiator (e.g. benzoyl peroxide).

[0033] In some embodiments, the preparation method further comprises the following step: subjecting the compound shown as formula (V) to the following reaction with the compound shown as formula (V-1) in the presence of a base in a solvent to obtain the compound shown as formula (IV);

[0034]

[0035] wherein each group is previously defined.

[0036] In the preparation method of the compound shown as formula (IV), the reaction condition and reagent of the reaction can be the conventional condition and reagent of this type of reaction in the art. Preferably, the solvent can be an ether solvent, such as tetrahydrofuran. Preferably, the base can be an inorganic base, such as sodium hydride.

[0037] In another aspect, the present application also provides a compound shown as formula (V), (IV), (III) or (II):

[0038]

[0039] wherein each group is previously defined.

[0040] In another aspect, the present application also provides a pharmaceutical composition comprising (i) the compound shown as formula (I) above, or a tautomer, stereoisomer, isotopic derivative or pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

[0041] In another aspect, the present application also provides the compound shown as formula (I) above, or a tautomer, stereoisomer, isotopic derivative or pharmaceutically acceptable salt thereof for use as a medicament.

[0042] In another aspect, the present application also provides the compound shown as formula (I) above, or a tautomer, stereoisomer, isotopic derivative or pharmaceutically acceptable salt thereof, or the above pharmaceutical composition for use as a retinoic acid receptor (e.g. retinoic acid receptor gamma) agonist.

[0043] The present application also provides the use of the compound shown as formula (I) above, or a tautomer, stereoisomer, isotopic derivative or pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in the preparation of a medicament for treating or preventing a disease associated with a retinoic acid receptor (e.g. retinoic acid receptor gamma).

[0044] In the present application, the disease associated with retinoic acid receptor (e.g., retinoic acid receptor gamma) can be acne (including acne vulgaris, acne rosacea, nodulocystic acne, conglobata acne, and secondary acne caused by sunlight or drug therapy), comedones, psoriasis (psoriases), ichthyosis, keratosis of the skin, fibrodysplasia ossificans progressiva, osteochondroma, pigmentation, or diabetic nephropathy.

[0045] In another aspect, the present application also provides a method for treating a disease associated with retinoic acid receptor (e.g., retinoic acid receptor gamma), which comprises administering to a subject in need of such treatment a compound represented by the above formula (I) (preferably, administering to a subject in need of such treatment a therapeutically effective amount of a compound represented by the above formula (I)), or a tautomer, a stereoisomer, an isotopic derivative, or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition.

[0046] Unless otherwise indicated, the terms used in the present application have the following definitions, and the definitions of terms not mentioned below are as generally understood by those skilled in the art to which the present application pertains.

[0047] The term "tautomer" refers to functional group isomers that result from the rapid movement of an atom in a molecule between two positions. For example, acetone and 1-propen-2-ol can be interconverted by the rapid movement of a hydrogen atom between the oxygen and the alpha-carbon.

[0048] The term "stereoisomer" refers to isomers that have the same order of connectivity of atoms but differ in the orientation of atoms in space. For example, cis-trans isomers (e.g., Z-isomers, E-isomers), optical isomers (e.g., enantiomers, diastereomers), atropisomers, etc. These stereoisomers can be separated, purified, and enriched by asymmetric synthesis methods or chiral separation methods (including but not limited to thin layer chromatography, rotary chromatography, column chromatography, gas chromatography, high pressure liquid chromatography, etc.), and can also be obtained by chiral resolution through bonding (chemical bonding, etc.) or salification (physical bonding, etc.) with other chiral compounds. Optical isomers include enantiomers and diastereomers. All of these isomers and mixtures thereof are included within the scope of the present application.

[0049] The term "isotopic derivative" refers to a compound in which one or more atoms are replaced by one or more atoms having a specific atomic mass or mass number. Examples of isotopes that can be incorporated into the compounds include, but are not limited to, isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, sulfur, and chlorine (e.g., 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 18 F,35 S and 36 Cl). Isotopically-labelled compounds can generally be prepared by substituting one or more of the hydrogen, halogen, etc. moieties present in the compounds described herein with their isotopically-labeled counterparts. Examples of isotopic substitutions include deuterium for hydrogen.

[0050] The term "pharmaceutically acceptable salt" means a salt of a compound prepared from a relatively nontoxic, pharmaceutically acceptable acid or base. Alkali addition salts can be prepared from the neutral forms of the compounds of the disclosure by contacting the neutral forms with a sufficient amount of the pharmaceutically acceptable base in pure solution or in a suitable inert solvent. Acid addition salts can be prepared by contacting the neutral form of the compound of the disclosure with a sufficient amount of the pharmaceutically acceptable acid in pure solution or in a suitable inert solvent. When the compound of the disclosure contains relatively acidic functionalities, base addition salts can be formed by contacting the neutral form of such compounds with a sufficient amount of the pharmaceutically acceptable base in pure solution or in a suitable inert solvent. When the compound of the disclosure contains relatively basic functionalities, acid addition salts can be formed by contacting the neutral form of such compounds with a sufficient amount of the pharmaceutically acceptable acid in pure solution or in a suitable inert solvent. When the compound of the disclosure contains both relatively acidic and relatively basic functionalities, base addition salts or acid addition salts can be formed by contacting the neutral form of such compounds with a sufficient amount of the pharmaceutically acceptable base or acid, respectively, in pure solution or in a suitable inert solvent.

[0051] The term "halogen" denotes fluorine, chlorine, bromine or iodine.

[0052] The term "hydroxy" denotes the -OH group.

[0053] The term "alkyl" refers to a saturated straight or branched chain monovalent hydrocarbon group having a specified number of carbon atoms. C 1-6 Alkyl refers to an alkyl group having from 1 to 6 (e.g., 1, 2, 3, 4, 5, 6) carbon atoms, including C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl, such as methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, s-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like.

[0054] The term "substituted" or "substituent" means that one or more hydrogen atoms are replaced by a designated group. When a substitution position is not specified, substitution can occur at any position, provided that a stable or chemically feasible compound results.

[0055] The term "treatment" refers to therapeutic treatment. With respect to a particular condition, treatment refers to: (1) relieving the disease or condition, or one or more of the biological manifestations thereof, (2) interfering with (a) one or more points in the biological cascade leading to or causing the condition or (b) one or more of the biological manifestations of the condition, (3) ameliorating one or more symptoms, effects or side effects associated with the condition, or one or more symptoms, effects or side effects associated with the condition or treatment thereof, or (4) slowing the development of the condition or one or more of the biological manifestations thereof.

[0056] The term "therapeutically effective amount" refers to the amount of a compound that, when administered to a patient, is sufficient to effect treatment or prevention of the diseases or conditions described herein. The "therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted by those skilled in the art as needed. Amounts in ranges beyond those recited above can also be used depending on the dosage form and the severity of the disease.

[0057] The pharmaceutical composition can be prepared into various types of administration unit dosage forms according to the purpose of treatment.

[0058] The compound of the present application can be clinically administered in a conventional administration manner.

[0059] The term "subject" refers to any animal to which a compound or composition is to be or has been administered, preferably a mammal, and most preferably a human. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, and the like, with humans being most preferred.

[0060] On the basis of common general knowledge in the art, the above-mentioned preferred conditions can be combined in any manner, i.e. to obtain each preferred embodiment of the present application.

[0061] The reagents and materials used in the present application are commercially available.

[0062] The positive progress effect of the present application is that the compound of the present application has strong agonist activity, selectivity and good hepatocyte stability of retinoic acid receptor γ, and can be used for treating diseases related to retinoic acid receptor (e.g. retinoic acid receptor γ). DETAILED DESCRIPTION

[0063] The present application is further illustrated by the following examples, but the present application is not limited to the scope of the examples described. The experimental methods in the following examples, unless otherwise specified, are selected according to conventional methods and conditions, or according to the instructions of the goods.

[0064] Example 1

[0065]

[0066] Step 1 : Synthesis of Intermediate 2

[0067] Compound 1 (4 g, 18.58 mmol) was dissolved in 40 mL of tetrahydrofuran, and the system was stirred at 25 °C. Ethynylmagnesium chloride (0.5 M, 83.20 mL) was added dropwise. After the addition was completed, the system was continuously stirred at 25 °C for 1 h. Then it was heated to 95 °C and refluxed for 1 h. TLC monitoring showed that the raw material was completely reacted and the main product was generated. The reaction system was quenched with excess saturated ammonium chloride solution and extracted with ethyl acetate (20 mL*3). The organic phase was washed successively with water (20 mL), saturated sodium bicarbonate solution (20 mL*2), and saturated brine (10 mL*2), dried over anhydrous sodium sulfate, and filtered. The filtrate was evaporated under reduced pressure to obtain an oil. Silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether, gradient 0%~1%) gave Intermediate 2 as a white oil (2 g, yield: 55.32%).

[0068] 1 H NMR (400 MHz, CD3Cl) δ ppm 2.34-2.40 (m, 2H), 0.61 (s, 4H), 0.16-0.17 (m, 12H)

[0069] Step 2: Synthesis of Intermediate 3

[0070] Cobalt diiodide (80.43 mg, 257.17 μmol) was suspended in 20 mL of acetonitrile. At 25 °C, Intermediate 2 (1 g, 5.14 mmol) and Compound 2a (877.66 mg, 6.17 mmol) were added dropwise to the above system in 10 mL of acetonitrile. After the addition was completed, zinc powder (33.63 mg, 514.35 μmol) was added to the above solution at once. The reaction system was protected by nitrogen and stirred at 25 °C for 1 min. Then the temperature was lowered to 0 °C and stirring was continued for 5 min. Elemental iodine (65.27 mg, 257.17 μmol) was added to the above system. The reaction system was slowly raised to 25 °C and stirred for 5 h. TLC monitoring showed that the raw material was completely reacted and the main product was generated. The system was diluted with 10 mL of 2M hydrochloric acid and extracted with ethyl acetate (10 mL*2). The organic phase was dried over anhydrous sodium sulfate and filtered after being combined. The filtrate was evaporated under reduced pressure to obtain an oily residue. Silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether, gradient 0%~5%) gave Intermediate 3 as a white solid (350 mg, yield: 25.94%).

[0071] 1H NMR (400 MHz, CD3Cl) δ ppm 10.28 (s, 1 H), 7.87 (s, 1 H), 7.37 (s, 1 H), 2.67 (s, 3 H), 1.04 (s, 4 H), 0.27 (s, 6 H), 0.26 (s, 6 H)

[0072] Step 3: Synthesis of intermediate 4

[0073] Intermediate 3 (180 mg, 685.73 μmol) was dissolved in 4 mL of tetrahydrofuran, and sodium hydride (219.41 mg, 5.49 mmol) (60% dispersion in mineral oil) and compound 3a (206 mg, 720.01 μmol) were added successively at 0 °C. The reaction system was slowly raised to 25 °C with stirring, and stirring was continued for 5 hours. LCMS monitoring showed that the reaction was complete. The system was quenched with 4 mL of saturated ammonium chloride solution, and extracted with ethyl acetate (3 mL*3). The organic phase was washed with saturated brine (2 mL*2) after being combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure to give a residue. Further purification was performed by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether, gradient 0%~10%) to give intermediate 4 as a white solid (170 mg, yield: 62.81%).

[0074] 1 H NMR (400 MHz, CD3Cl) δ ppm 7.98 (br d, J=8.00 Hz, 2 H), 7.63 (s, 1 H), 7.54 (br d, J=8.13 Hz, 2 H), 7.40 (br d, J=16.13 Hz, 1 H), 7.27 (s, 1 H), 6.98 (br d, J=16.13 Hz, 1 H), 3.88 (s, 3 H), 2.39 (s, 3 H), 0.97 (s, 4 H), 0.21 (s, 6 H), 0.18 (s, 6 H)

[0075] Step 4: Synthesis of intermediate 5

[0076] Intermediate 4 (60 mg, 152.03 μmol) was dissolved in 2 mL of carbon tetrachloride. To the system was added N-bromosuccinimide (32.47 mg, 182.44 μmol) and benzoyl peroxide (3.68 mg, 15.20 μmol). The reaction was stirred at 80 °C for 5 hours. TLC monitoring showed that the starting material was consumed completely and the main product was generated. The system was cooled to 25 °C and the reaction was poured into 10% sodium bisulfite solution (5 mL). The organic phase was separated and the aqueous phase was extracted with ethyl acetate (2 mL*2). The organic phases were combined and washed with saturated brine (2 mL*2). After drying over anhydrous sodium sulfate, the mixture was filtered and the filtrate was evaporated under reduced pressure to give a residue, which was purified by column chromatography on silica gel (mobile phase: ethyl acetate / petroleum ether, gradient 0% to 10%) to give intermediate 5 as a white solid (43 mg, yield: 59.72%).

[0077] 1 H NMR (400 MHz, CD3Cl) δ ppm 8.08 (br d, J = 8.00 Hz, 2H), 7.72 (s, 1H), 7.64-7.58 (m, 3H), 7.44 (s, 1H), 7.12 (br d, J = 16.00 Hz, 1H), 4.63 (s, 2H), 3.75 (s, 3H), 1.01 (s, 4H), 0.28 (s, 6H), 0.24 (s, 6H)

[0078] Step 5: Synthesis of intermediate 6

[0079] Intermediate 5 (30 mg, 63.35 μmol) was dissolved in 1 mL of N,N- dimethylformamide. To the system was added cesium carbonate (30.96 mg, 95.03 μmol) and pyrazole (8.63 mg, 126.7 μmol). The reaction was stirred at 90 °C for 2 hours. LCMS monitoring showed that the starting material was consumed completely. The reaction was filtered and the filtrate was concentrated under reduced pressure to give a residue, which was purified by column chromatography on silica gel (mobile phase: ethyl acetate / petroleum ether, gradient 0% to 20%) to give intermediate 6 (18 mg, yield: 61.65%).

[0080] LCMS (ESI): m / z C 26 H 33 N2O2Si2 + .[M+H] + Calculated = 461.21, Found = 461.1. 1H NMR (400 MHz, CD3Cl) δ ppm 8.02 (d, J = 8.3 Hz, 2 H), 7.73 (s, 1 H), 7.57 (d, J = 1.5 Hz, 1 H), 7.52 (d, J = 8.5 Hz, 2 H), 7.38 (d, J = 16.1 Hz, 1 H), 7.28 (br s, 1 H), 7.25 (s, 1 H), 6.99 (d, J = 16.1 Hz, 1 H), 6.26 (t, J = 2.1 Hz, 1 H), 5.48 (s, 2 H), 3.93 (s, 3 H), 1.03 (s, 4 H), 0.28 (s, 6 H), 0.22 (s, 6 H)

[0081] Step 6: Synthesis of compound 7

[0082] Intermediate 6 (20 mg, 43.41 μmol) was dissolved in 2 mL of tetrahydrofuran. To the solution was added dropwise 3 mL of sodium hydroxide solution (6.95 mg, 173.64 μmol). The reaction system was stirred at 40 °C for 4 hours. LCMS monitoring showed that the reaction was complete. The reaction system was adjusted to pH 6.0 with 1 M aqueous hydrochloric acid solution, and then extracted with ethyl acetate (1 mL*3). The organic phase was combined and washed with saturated brine (1 mL*2). After drying over anhydrous sodium sulfate, filtration, and evaporation of the filtrate under reduced pressure, a residue was obtained. Silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether, gradient 0%~20%) gave compound 7 as a white solid (14.2 mg, yield: 73.20%).

[0083] LCMS (ESI): m / z C 25 H 31 N2O2Si2 + .[M+H] + Calculated = 447.19, Found = 447.1. 1 H NMR (400 MHz, CD3Cl) δ ppm 8.04 (d, J = 8.31 Hz, 2 H), 7.75 (s, 1 H), 7.62 (d, J = 1.59 Hz, 1 H), 7.55 (d, J = 8.31 Hz, 2 H), 7.43 (d, J = 16.02 Hz, 1 H), 7.30-7.34 (m, 2 H), 7.00 (d, J = 16.02 Hz, 1 H), 6.29 (t, J = 2.02 Hz, 1 H), 5.53 (s, 2 H), 1.06 (s, 4 H), 0.28 (s, 6 H), 0.23 (s, 6 H)

[0084] Biological test example 1: RARγ agonist activity

[0085] The HEK-293 cell line (ATCC # CRL-1573) was used to test the compounds for RARγ agonist activity. The experimental materials are shown in Table 1.

[0086] Table 1

[0087] Experimental Material Name Supplier Catalogue Number MEM Gibco 11090081 Fetal Bovine Serum Biological Industries 04-001-1A Trypsin-EDTA Gibco 25200072 FuGENE HD Transfection Reagent Promega E231A Dual Luciferase Reporter Assay System Promega E2940 96-Well Plates Costar 3903

[0088] Experimental Procedure:

[0089] 1. Day 1: Cell culture. Culture dishes were trypsinized and cells were seeded at the appropriate concentration in 10 mL of media. Cell culture conditions: 37 degrees C, 5% CO2, humidified environment.

[0090] 2. Day 3: Transfection with FuGENE transfection reagent. Transfection mix was prepared according to the manufacturer's instructions, and the test tube was mixed well by vortexing. The transfection mix was allowed to incubate at room temperature for 15 minutes. Cells were trypsinized, and the cell density was determined. The cells were diluted to the appropriate volume and density. The appropriate amount of transfection reagent mix was added, and the cell suspension was added to the 96-well plate in a volume of 100 μL per well. The 96-well plate was incubated for 24 hours under the conditions described in Step 1.

[0091] 3. Day 4: Addition of test compound. The compound was prepared as a 210x stock solution in DMSO, and a 9x dilution was made to a 21x solution. Five microliters of the 21x compound solution was added to each well, and the plate was incubated for 24 hours under the conditions described in Step 1.

[0092] 4. Day 5: Plate reading. The Dual-Luciferase Reporter Assay System was allowed to equilibrate at room temperature for 30 minutes, and 100 μL of Luciferase Reagent II was added to each well of the 96-well plate. The plate was read for firefly luciferase intensity in the appropriate manner. Stop & Glo reagent was then added, and the plate was read for Renilla luciferase intensity.

[0093] 5. Data analysis. Signal = firefly luciferase intensity / Renilla luciferase intensity, fold activation of agonist = compound signal / baseline signal, and the baseline signal was obtained from DMSO without compound. The data were analyzed using GraphPad Prism to obtain the pEC50 values, which were converted to EC50 values.

[0094] The results of the experiments for some of the compounds of the application are shown in Table 2.

[0095] Table 2

[0096]

[0097] As can be seen from the data in Table 2, the compound 7 prepared in Example 1 has a greater agonist activity for RARγ than Palovarotene.

[0098] Biological Test Example 2: Hepatocyte stability

[0099] The in vitro metabolic stability of the compounds was tested using human hepatocytes.

[0100] The main experimental materials are shown in Table 3 below:

[0101] Table 3

[0102]

[0103] Experimental procedure:

[0104] 1. 10 micromolar of the test compound and human hepatocytes (1.0 x 10 6 cells per milliliter) were added to WEM medium, total volume 200 microliters, incubated at 37 degrees Celsius for 240 minutes.

[0105] 2. Centrifugation, supernatant was detected by LC-MS / MS.

[0106] 3. Chromatographic conditions:

[0107]

[0108] Mass spectrometric conditions:

[0109]

[0110]

[0111] The experimental results of some of the compounds of the present application are shown in Table 4:

[0112] Table 4

[0113]

[0114] As can be seen from the data in Table 4, the compound 7 prepared in Example 1 has better stability in hepatocytes compared to Palovarotene.

[0115] Biological Test Example 3: Activation of retinoic acid receptors (RARa and, RARβ)

[0116] The agonistic activity of the compounds on RARa and RARβ was tested using the HEK-293 cell line.

[0117] Experimental procedure:

[0118] 1. Cell culture

[0119] HEK293 cells were cultured in DMEM + 10% FBS medium at 37 degrees Celsius in a humidified carbon dioxide incubator with a carbon dioxide concentration of 5%.

[0120] 2. Plasmid transfection and cell plating

[0121] 1) Digest HEK293 cells with trypsin, adjust the cell suspension to 200,000 cells per ml with culture medium (100 μl / well for 96-well plate).

[0122] 2) Prepare the transfection reagent mix according to Table 5, mix well and let stand at room temperature for 20 minutes.

[0123] Table 5

[0124]

[0125]

[0126] 3) Add the prepared transfection mix to 10 ml of the cell suspension with adjusted cell density, mix well by inverting the tube, and plate into 96-well plate with a volume of 100 μl / well.

[0127] 4) Incubate the 96-well plate in a 37°C, 5% CO2 humidified incubator for 24 hours.

[0128] 3. Compound treatment

[0129] 1) Dilute the test compound and positive control compound with DMSO to a final concentration of 210 times.

[0130] 2) Transfer the compound solution from 1) above to the second well of a 96-well plate, respectively, and add 30 μl of 100% DMSO to the other wells. Take 15 μl of the compound solution from the second well and add to the third well, and continue to dilute by 3 times to get 10 concentrations.

[0131] 3) Add 5 μl of the DMSO solution from 2) above to 45 μl of culture medium, mix well, to get the dilution solution of 10 concentrations.

[0132] 4) Add 5 μl of the compound dilution solution from 3) above to each well of the detection plate (with 100 μl of cells).

[0133] 5) Put the detection plate back into the CO2 incubator for 24 hours.

[0134] 4. Luciferase detection

[0135] Use the Promega Dual-Glo Luciferase Assay System kit to detect the detection plate, and use Enspire to read the Firefly luciferase fluorescence signal and the Renilla Luciferase fluorescence signal, respectively.

[0136] 5. Data processing

[0137] 1) The final measurement is the normalized value "F / R" obtained by dividing the Firefly luciferase fluorescent signal by the Renilla Luciferase fluorescent signal.

[0138] 2) Copy and paste the data into Excel, and get the activation rate by equation.

[0139] Activation rate (%) = (Final measurement - Min mean value) / (Max mean value - Min mean value) x 100%

[0140] 3) Calculate EC50 by GraphPrism5.0 software. Import data into Graphpad Prism and use Log (agonist) vs. response - variable slope for curve fitting,

[0141] Fitting equation: Y = Bottom + (Top - Bottom) / (1 + (EC50 / X) ^ HillSlope

[0142] Y is the agonist rate, and X is the compound concentration.

[0143] The experimental results of some compounds of the present application are shown in Table 6.

[0144] Table 6

[0145]

[0146] From the data in Table 6, it can be seen that the agonist activity of compound 7 prepared in Example 1 on RARa is weaker than Palovarotene. The agonist activity on RARβ is similar to Palovarotene.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, ， wherein R1and R2are independently H, halogen or C1-C6alkyl; m is 1, 2 or 3; n is 1, 2, 3 or 4; R3is H.

2. The compound of formula (I) as claimed in claim 1, characterized in that, the halogen in R1and R2is independently F, Cl, Br or I, respectively; and / or, the C1-C6alkyl in R1and R2is independently C1-C4alkyl, respectively.

3. The compound of formula (I) as claimed in claim 1, characterized in that, the C1-C6alkyl in R1and R2is independently methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl or t-butyl, respectively.

4. The compound of formula (I) as claimed in claim 1, characterized in that, the compound represented by formula (I) is: 。 5. A process for the preparation of a compound of formula (I) as claimed in any one of claims 1 to 4, characterized in that, comprising the following steps: subjecting a compound represented by formula (II) to the following reaction in the presence of a base in a solvent to obtain a compound represented by formula (I); ， wherein R3is H, is a group which can form upon hydrolysis, R1, R2, m and n are as defined in any one of claims 1 to 4.

6. The process for the preparation of a compound of formula (I) according to claim 5, characterized in that, To .

7. Process for the preparation of a compound of formula (I) according to claim 5 or 6, characterized in that, comprising one or more of the following reactions 1-3: reaction 1: comprising the following steps: subjecting a compound represented by formula (III) to the following reaction with a compound represented by formula (III-1) in the presence of a base in a solvent to obtain a compound represented by formula (II); ， reaction 2: comprising the following steps: subjecting a compound represented by formula (IV) to the following reaction with a compound represented by formula (IV-1) in a solvent to obtain a compound represented by formula (III); ， reaction 3: comprising the following steps: subjecting a compound represented by formula (V) to the following reaction with a compound represented by formula (V-1) in the presence of a base in a solvent to obtain a compound represented by formula (IV); ， wherein Hal is halogen, R1, R2, m and n are as defined in any one of claims 1 to 4, as defined in claim 5 or 6.

8. The process for the preparation of a compound of formula (I) according to claim 7, characterized in that, Hal is Br.

9. A compound represented by formula (IV), (III) or (II): 、 or , wherein R1, R2, n and m are as defined in any one of claims 1 to 4, R1, R2, n and m are as defined in any one of claims 1 to 4, 10. A pharmaceutical composition comprising (i) a compound represented by formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4; and (ii) a pharmaceutically acceptable carrier.

11. Use of a compound represented by formula (I) or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, or a pharmaceutical composition according to claim 10, in the manufacture of a medicament for the treatment or prevention of a disease associated with retinoic acid receptor. wherein the disease associated with retinoic acid receptor is a disease associated with retinoic acid receptor gamma.

12. The use according to claim 11, wherein the compound is ###00006### 11 the disease associated with retinoic acid receptor is acne, comedones, psoriasis, ichthyosis, keratosis of the skin, fibrodysplasia ossificans progressiva, osteochondroma, pigmentation or diabetic nephropathy.

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