Pyridazinone compound and application thereof

Abstract

The invention provides a pyridazinone compound, or a stereoisomer, a tautomer or a pharmaceutically acceptable salt thereof, and also provides a method related to preparation and application of the compound, a pharmaceutical composition containing the compound and a treatment method of related metabolic diseases. The compound disclosed by the invention shows an excellent thyroid hormone receptor excitation effect and has a wide application prospect in the field of metabolic disease treatment.

Description

A pyridazinone compound and its application Technical Field

[0001] The present invention relates to a pyridazinone compound, a preparation method thereof, and application of the compound in disease treating drugs. Background Art

[0002] Thyroid hormones are essential for normal growth and development and for maintaining metabolic homeostasis. Circulating levels of thyroid hormones are tightly regulated through feedback mechanisms within the hypothalamic-pituitary-thyroid (HPT) axis. Abnormal thyroid function, leading to hypothyroidism or hyperthyroidism, clearly demonstrates that thyroid hormones profoundly affect cardiac function, body weight, metabolism, metabolic rate, body temperature, cholesterol, bone, muscle, and behavior.

[0003] The biological activity of thyroid hormones is mediated by thyroid hormone receptors (THRs). Thyroid hormone receptor subtypes can differ in their contribution to specific physiological responses. THRβ plays an important role in regulating thyroid-stimulating hormone and regulating the effects of thyroid hormones in the liver. The development of thyroid analogs that avoid the adverse effects of hyperthyroidism and hypothyroidism while maintaining the beneficial effects of thyroid hormones will open new avenues for treating patients with metabolic diseases such as obesity, hyperlipidemia, hypercholesterolemia, diabetes and other conditions such as hepatic steatosis and non-alcoholic steatohepatitis (NASH), atherosclerosis, cardiovascular disease, hypothyroidism, thyroid cancer, thyroid disease, and related conditions and diseases.

[0004] In recent years, a series of THRβ agonists have been developed. For example, Resmetirom has the effects of reducing liver fat accumulation and inhibiting liver fibrosis. The structural formula of Resmetirom is as follows:

[0005] However, given the obscure structure-activity relationship of pharmaceutical compounds, developing compounds with improved efficacy, selectivity, and pharmacokinetic properties remains a daunting challenge. Following unsuccessful deuteration studies of resmetirom, the present inventors surprisingly discovered that deuteration of certain resmetirom analogs, as well as at certain positions, has greater potential. The resulting compounds exhibit excellent THRβ agonism and selectivity, demonstrating superior reductions in liver fat accumulation and inhibition of liver fibrosis compared to resmetirom.

[0006] Summary of the Invention

[0007] The present invention provides a compound of formula (X), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof,

[0008] Wherein, X is a nitrogen atom or a carbon atom;

[0009] Y, U and T are nitrogen atoms, oxygen atoms, or sulfur atoms;

[0010] Z is each independently -NH-, -O-, -S-, or -CH2-;

[0011] R 1 is selected from hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, alkenyl, alkynyl, cyano, formyl, or alkoxycarbonyl;

[0012] R 2 , R 3 , R 4 , R 5 , R 8 , R 9 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, or alkynyl.

[0013] The present invention provides a compound of formula (I), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof,

[0014] Wherein, X is a nitrogen atom or a carbon atom;

[0015] Z is -NH-, -O-, -S-, or -CH2-;

[0016] R 1 is selected from hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, alkenyl, alkynyl, cyano, formyl, or alkoxycarbonyl;

[0017] R 2 , R 3 , R 4 , R 5 , R 8 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, or alkynyl.

[0018] The present invention provides a compound of formula (II), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof,

[0019] Among them, R2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, or alkynyl.

[0020] Preferably, the present invention provides the following compounds, or stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0021] The present invention provides a pharmaceutical composition comprising a therapeutically effective dose of any one of the compounds of the present invention or its stereoisomers, tautomers or pharmaceutically acceptable salts and a pharmaceutically acceptable carrier.

[0022] The present invention provides the use of a compound of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present invention, in the preparation of a medicament for a thyroid hormone receptor (THR)-related disease. In addition, the present invention provides a method for treating a disease with a thyroid hormone receptor agonist, comprising administering a therapeutically effective amount of a compound of the present invention or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof to a subject in need thereof.

[0023] Preferably, the disease that can be treated by the thyroid hormone receptor agonist is a metabolic disease, more preferably obesity, hyperlipidemia, hypercholesterolemia, diabetes, non-alcoholic steatohepatitis (NASH), atherosclerosis, cardiovascular disease, hypothyroidism or thyroid cancer.

[0024] Detailed Description of the Invention

[0025] All technical and scientific terms used in this specification have the same meanings as commonly understood by those skilled in the art.

[0026] The term "alkyl" refers to a saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms in this article, and the term includes straight and branched chain hydrocarbon groups. Non-limiting examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, etc. Alkyl described herein can be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, acyloxy, oxo, amide, ester, amido, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkenyloxy, alkynyl, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aryl or heteroaryl.

[0027] The term "aryl" herein refers to a 6-10 membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group, a polycyclic (i.e., rings with adjacent pairs of carbon atoms) group having a conjugated π electron system. The aryl group can be covalently attached to the defined chemical structure at any carbon atom that produces a stable structure. The aryl groups described herein may be optionally substituted with one or more of the following substituents: fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl, and cycloalkoxy.

[0028] The term "heterocyclic group" refers to a ring system containing a nitrogen atom or an oxygen atom. The ring system can be "parallel" to aromatic and non-aromatic ring systems, or linked to other ring systems through "spiro carbon atoms", such as the following structure:

[0029]

[0030] etc.

[0031] The term "heteroaryl" as used herein refers to an aromatic group consisting of 5 to 10 atoms and containing at least one heteroatom selected from N, O, or S. The term can have a single ring (non-limiting examples include furan, thiophene, imidazole, pyrazole, pyridine, pyrazine, oxazole, thiazole, etc.) or multiple fused rings (non-limiting examples include benzothiophene, benzofuran, indole, isoindole, etc.), wherein the fused rings may or may not be aromatic groups containing heteroatoms, provided that the point of attachment is through an atom of the aromatic heteroaryl group. The heteroaryl groups described herein may be optionally substituted with one or more of the following substituents: fluoro, chloro, bromo, iodo, cyano, nitro, hydroxy, amino, alkyl, alkoxy, acyl, acyloxy, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl, and cycloalkoxy.

[0032] The term "alkenyl" refers to an alkenyl group having 2 to 8 carbon atoms and at least one alkenyl unsaturated site in this article. Non-limiting examples of alkenyl include vinyl, propenyl, allyl, isopropenyl, butenyl, isobutenyl etc. Alkenyl described herein can be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester group, amino, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfydryl, alkyl mercapto, deuterated alkyl mercapto, sulfone, sulfoxide, amino, silicon, phosphono, deuterated alkyl, heterocycloalkyl, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl, ester group.

[0033] The term "alkynyl" is herein intended to refer to an alkyl radical in which two adjacent carbon atoms are connected by a triple bond, wherein the alkyl radical is as defined herein. Alkynyl refers to an unsaturated alkyl radical as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, such as ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, etc. Alkynyl may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amino, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfhydryl, alkylthiol, deuterated alkylthiol, sulfone, sulfoxide, amino, silicon, phosphono, deuterated alkyl, heterocycloalkyl, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl, ester.

[0034] The term "pharmaceutically acceptable carrier" herein refers to a non-toxic solid, semisolid, or liquid filler, diluent, adjuvant, packaging material, or other formulation excipient. The carrier used can be adapted to the respective dosage form and can be formulated into injections, lyophilized powders (for injection), sprays, oral solutions, oral suspensions, tablets, capsules, enteric-coated tablets, pills, powders, granules, sustained-release or delayed-release formulations, and the like using carriers known to those skilled in the art.

[0035] The terms "prevention" and "treatment" herein have the meanings conventionally understood by those skilled in the art, i.e., "prevention" refers to the administration of a pharmaceutical composition or active compound before the onset of a disease or before the appearance of symptoms to prevent, delay and / or alleviate the onset of the corresponding disease or the appearance of symptoms; "treatment" refers to the administration of a pharmaceutical composition or active compound at the time of or after the onset of a disease, or at the time of or after the appearance of symptoms, to eliminate the corresponding condition, alleviate the severity of the corresponding symptoms, or delay the development of the corresponding disease.

[0036] The term "subject" herein preferably refers to a mammal, in particular a human or an experimental animal (such as a mouse, rat or rabbit, etc.).

[0037] The term "effective amount" as used herein refers to the appropriate dosage of a drug or treatment method required to produce the desired effect. A therapeutically effective amount refers to a dosage of a drug capable of treating a disease or symptom, while a prophylactically effective amount refers to a dosage of a drug or treatment method capable of preventing the occurrence of a disease or reducing the risk of a disease. Those skilled in the art can determine the effective amount based on specific factors, such as the individual's age, weight, and severity of the disease.

[0038] Other abbreviations

[0039] The beneficial effects of the present invention include:

[0040] (1) The compounds of the present invention have a good agonist effect on THRβ, and their agonist activity on THRβ is significantly better than that of Resmetirom; on the other hand, the agonist activity of compounds 6, 16, and 17 on THRα is weak, and almost no agonist activity, indicating that compounds 6, 16, and 17 have a selective agonist effect on THRβ and have a significant safety advantage.

[0041] (2) At the same study dose and in the same study animal system, the elimination phase half-life (T 1 / 2 =7.80h) is significantly longer than the elimination phase half-life of Resmetirom (T 1 / 2 =2.98h).

[0042] (3) Compound 6 of the present invention has a more significant liver distribution advantage than Resmetirom and Compound X1.

[0043] (4) Under the same experimental conditions, compound 6 showed better effects in reducing liver fat accumulation and inhibiting liver fibrosis than Resmetirom. Description of the drawings:

[0044] Figure 1: Comparison of chromatograms after incubation of compound 16 and compound 6 in human liver microsomes

[0045] Figure 2: Bar graph of compound concentrations in blood, liver, kidney, and heart of SD rats 1h and 4h after oral gavage

[0046] Figure 3: Dosing regimen for efficacy testing in the B-DIO mouse NASH model

[0047] Figure 4: Body weight measurement results of mice after drug administration in NASH model efficacy test experiment

[0048] Figure 5: On day 26, blood was collected from mice in groups G1-G6 to measure triglyceride levels

[0049] Figure 6: Blood was collected from mice in groups G1-G6 on day 26 to measure transaminase levels

[0050] Figure 7: On day 26, livers of mice in groups G1-G6 were collected to measure total cholesterol and triglyceride levels

[0051] Figure 8: On day 26, livers of mice in groups G1-G6 were collected, embedded and frozen, and stained with Oil Red O to show the degree of fat change in the livers of mice.

[0052] Figure 9: On day 26, the livers of mice in groups G1-G6 were collected, fixed, and stained with Sirius red to show the degree of liver fibrosis in mice. Specific implementation method:

[0053] The present invention is further illustrated by the following examples, but the present invention is not limited thereto. Throughout this application, various examples of the compounds and methods of the present invention are mentioned herein. The present invention is not limited to these examples. The following examples are merely provided to provide methods for practicing the present invention and are not intended to limit the scope of the present invention in any way.

[0054] The compounds provided herein can be prepared by standard synthetic methods known in the art, and this specification provides general methods for preparing the compounds of the present invention. Starting materials can generally be obtained commercially or prepared by methods well known to those skilled in the art.

[0055] The process is as follows:

[0056] Using SM-1 as the starting material, the cyanide group is hydrolyzed to obtain compound (IM-1), esterification reaction is performed to obtain compound (IM-2), reduction reaction is performed to obtain compound (IM-3), and functional group conversion reaction is performed to obtain compound (II).

[0057] The compounds of the present invention and corresponding preparation methods are further explained and listed below by examples and preparations. It should be understood that although typical or preferred reaction conditions are given in the specific examples, those skilled in the art may also use other reaction conditions. Optimum reaction conditions may vary with the specific reaction substrate or solvent used, but the conditions can be determined by conventional optimization by those skilled in the art.

[0058] Intermediate preparation

[0059] first step:

[0060] Add diethyl malonate (40.0 g) and anhydrous ethanol (320 mL) to the reaction flask. Stir at room temperature until dissolved, then add sodium ethoxide (50.6 g). Stir at room temperature for 0.5 h. Add deuterated iodomethane (80.0 g) in batches to the above solution, controlling the temperature between 20 and 30°C. Maintain this temperature for 2 h. Add 200 mL of water and 100 mL of ethyl acetate to the reaction solution, stir and separate. Extract the aqueous layer with 100 mL of ethyl acetate. Combine the ethyl acetate layers, wash with 200 mL of saturated sodium chloride solution, dry over anhydrous sodium sulfate, and evaporate to dryness under reduced pressure to yield 25.4 g of a reddish-brown oil. m / z: 195.20 (M+1).

[0061] Step 2:

[0062] IM-1 (25.4 g), potassium hydroxide (18.5 g), and 125 mL of water were added to the reaction flask in sequence and heated to 70°C for 1.0 h. The reaction mixture was cooled to room temperature, adjusted to pH 2 with concentrated HCl, and concentrated under reduced pressure. The water was then removed once with 100 mL of methanol. To the concentrate was added 250 mL of methanol and 25 g of anhydrous sodium sulfate, stirred at room temperature for 0.5 h, and filtered. The filtrate was concentrated under reduced pressure to dryness to yield 19.8 g of a pale yellow solid. m / z: 139.10 (M+1), 137.10 (M-1).

[0063] Step 3:

[0064] IM-2 was heated to 185°C for 4 h. After the reaction mixture cooled to room temperature, 80 mL of water and 80 mL of dichloromethane were added, stirred, and the layers separated. The aqueous layer was further extracted with 70 mL of dichloromethane. The combined dichloromethane layers were washed with 100 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, and dried under reduced pressure to obtain 7.9 g of a reddish-brown oil.

[0065] Step 4:

[0066] IM-4 (4.1 g), IM-3 (2.8 g), silver nitrate (2.15 g), and sulfuric acid (12.3 g) were added to 40 mL of water. The temperature was raised to 40°C, followed by the addition of ammonium thiosulfate (21.8 g). After the addition was complete, the temperature was raised to 70°C and the reaction was allowed to react for 1 h. The reaction mixture was cooled to room temperature and filtered. The filter cake was dried to yield 7.35 g of a gray solid. m / z: 211.05 (M+1).

[0067] Compound preparation

[0068] Example 1:

[0069] first step:

[0070] Intermediate 1-1 (3 g) (prepared by the method of reference WO2019144835A1) was dissolved in ethyl acetate (100 mL), and concentrated hydrochloric acid (20 mL) was added with stirring. After the addition was completed, the temperature was raised to 120 ° C and the reaction was carried out for 6 h. After the reaction was completed, the reaction solution was cooled to room temperature, and H2O (200 mL) was added dropwise to the reaction solution. After the addition was completed, the solution was stirred at room temperature for 1 h, filtered, and the filter cake was rinsed with H2O (20 mL × 2). The solid was collected and dried at 45 ° C to obtain 2.0 g of a khaki solid.

[0071] Step 2:

[0072] Intermediate 1-2 (2 g) was dissolved in methanol (100 mL), and SOCl2 (10 mL) was added dropwise in an ice bath. After the addition was complete, the temperature was raised to 100°C and the reaction was carried out for 5 h. After the reaction was completed, the mixture was concentrated under reduced pressure. The residue was slurried with ethyl acetate and filtered. The filter cake was rinsed with ethyl acetate, and the solid was collected and dried at 45°C to obtain 2.3 g of a yellow solid.

[0073] Step 3:

[0074] Intermediate 1-3 was dissolved in tetrahydrofuran (40 mL), and NaBH4 (0.82 g) was added. After the addition was complete, methanol (4 mL) was slowly added dropwise, and a large amount of bubbles were generated. After the addition was complete, the reaction was stirred for 1 h. After the reaction was complete, the reaction was quenched with ice water and the pH was adjusted to about 6 with 1 M HCl. The mixture was extracted with ethyl acetate (20 mL × 2), and the organic phases were combined and washed with saturated NaCl (20 mL). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was passed through a column to obtain 1.6 g of a light yellow solid. 1 H-NMR (400MHz, DMSO-d6) δ: 12.44 (s, 1H), 12.21 (s, 1H), 7.86 (s, 2H), 7.44 (d, 1H, J= 0.8Hz), 5.31~5.27 (t, 1H, J=12.4&6.4Hz), 4.41 (d, 2H, J=5.6Hz), 3.10~3.00 (s, 1H).

[0075] Step 4:

[0076] Compound 7 (100 mg) was dissolved in dichloromethane (20 mL), and diethylaminosulfur trifluoride (DAST) (8 drops) was added dropwise in an ice bath. The mixture was stirred in an ice bath for 30 min. After the reaction was complete, the reaction solution was slowly added dropwise to ice water. The organic phase was separated and temporarily stored. The aqueous phase was extracted with dichloromethane (15 mL). The combined organic phases were washed with saturated NaCl (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was slurried with ethyl acetate to obtain 20 mg of a white solid. m / z: 448.00 (M+H); 1H-NMR (400MHz, DMSO-d6) δ: 12.67 (s, 1H), 12.22 (s, 1H), 7.82 (s, 2H), 7.44 (d, 1H, J=0.8Hz), 5.36 (d, 2H, J=46.8Hz), 3.10~3.00 (s, 1H).

[0077] Example 2:

[0078] first step:

[0079] IM-5 (7.3 g), SM-2-1 (6.2 g), cuprous iodide (3.3 g), potassium carbonate (12.1 g), and 75 mL of dimethyl sulfoxide were added sequentially to a reaction flask. The mixture was heated to 90°C under nitrogen for 5 h. The reaction mixture was cooled and added to water, filtered through celite, and the filter cake was rinsed with methanol. The filtrate and the eluent were combined and extracted with ethyl acetate (500 mL x 2). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and dried under reduced pressure. Flash chromatography was performed to yield 4.8 g of intermediate 2-1 as an off-white solid. m / z: 352.05 (M+I), 350.05 (M-1).

[0080] Step 2:

[0081] Intermediate 2-1 (4.8 g), sodium acetate (5.65 g), and 40 mL of acetic acid were added to a reaction flask in sequence. The temperature was raised to 130°C and the reaction mixture was allowed to react for 7.0 h. The reaction mixture was cooled, compressed to dryness, and 20 mL of sodium acetate was added for dissolution. The mixture was again compressed to dryness and used in the next step without further purification. m / z: 376.10 (M+1), 374.15 (M-1).

[0082] The crude product from the previous step and 50 mL of ethanol were added to a reaction flask, followed by a solution of sodium hydroxide (8.3 g) in water (20 mL). The temperature was raised to 110°C and the reaction mixture was allowed to react for 4 h. After cooling, the reaction mixture was dried under reduced pressure. 50 mL of water and 50 mL of ethyl acetate were added to the concentrate, followed by stirring and separation. The aqueous layer was further extracted with ethyl acetate (50 mL x 2). The combined ethyl acetate layers were washed sequentially with 100 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, and dried under reduced pressure. The concentrate was slurried with a mixture of ethyl acetate and petroleum ether (1:2), filtered, and dried to yield 3.5 g of a yellow-brown solid. m / z: 334.05 (M+1), 332.10 (M-1).

[0083] Step 3:

[0084] Intermediate 2-2 (3.5 g) was dissolved in 150 mL of water. 177 mL of concentrated hydrochloric acid was added, and the temperature was lowered to 0-10°C. A solution of sodium nitrite (2.2 g) in water (15 mL) was added while maintaining the temperature at 0-10°C. After complete addition, the mixture was stirred in an ice-water bath for 1 hour. This was designated Solution A. SM-2-2 (1.98 g) was added to 75 mL of water, followed by 177 mL of pyridine. This was designated Solution B. Solution A was slowly added to Solution B at a temperature of 0-10°C. After completion of the addition, the temperature was maintained at 0-10°C and the reaction was allowed to proceed for 1.5 hours. The reaction mixture was filtered, and the filter cake was washed with water and n-hexane and dried to yield 4.7 g of an orange-red solid. m / z: 501.20 (M+1), 499.15 (M-1).

[0085] Step 4:

[0086] Intermediate 2-3 (4.7 g), sodium acetate (7.85 g), and 145 mL of acetic acid were added to a reaction flask and heated to 130°C for 3 h. The reaction mixture was cooled, dried under reduced pressure, and 100 mL of water was added. The mixture was extracted with ethyl acetate (75 mL x 3). The combined ethyl acetate layers were washed with 200 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, and dried under reduced pressure. The concentrate was slurried with a mixture of ethyl acetate and n-hexane (1:2), filtered, and dried to yield 3.5 g of a light brick-red solid. m / z: 455.10 (M+1), 453.10 (M-1).

[0087] Step 5:

[0088] Intermediate 2-4 (3.5 g) and 53 mL of acetic acid were added to a reaction flask. Concentrated hydrochloric acid (24 mL) was then added and the temperature was raised to 120°C for 2.5 h. The reaction solution was cooled to room temperature and poured into 110 mL of water. The mixture was stirred for 0.5 h and filtered. The filter cake was washed with water and dried to yield 3.2 g of a light brick-red solid. m / z: 474.05 (M+1), 472.05 (M-1).

[0089] Step 6:

[0090] Intermediate 2-5 (3.2 g) and 60 mL of methanol were added to a reaction flask. Thionyl chloride (820 mg) was slowly added with stirring, and the temperature was raised to 100°C for 4 h. The reaction solution was cooled to room temperature and dried under reduced pressure. The concentrate was slurried with a mixture of ethyl acetate and petroleum ether (1:2), filtered, and dried to obtain 3.0 g of compound 8 as a light brick-red solid. m / z: 488.10 (M+1), 486.10 (M-1).

[0091] Step 7:

[0092] Compound 8 (3.0 g) was dissolved in a mixture of 150 mL of tetrahydrofuran and 15 mL of methanol. Sodium borohydride (1.87 g) was slowly added with stirring and allowed to react at room temperature for 0.5 h. The reaction mixture was quenched with 90 mL of water, adjusted to pH 6 with 1 M hydrochloric acid, and concentrated to approximately half its volume. The mixture was then extracted with ethyl acetate (90 mL x 3). The combined ethyl acetate layers were washed sequentially with 200 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, and dried under reduced pressure. The concentrate was slurried with a mixture of ethyl acetate and n-hexane (1:1), filtered, and dried to yield 2.6 g of compound 12 as an off-white solid. m / z: 460.10 (M+1), 458.05 (M-1).

[0093] Step 8:

[0094] Compound 12 (2.4 g) and 200 mL of dichloromethane were added to a reaction flask. Diethylaminosulfur trifluoride (DAST, 2.5 g) was slowly added at 0-5°C. After the addition was complete, the mixture was allowed to react at room temperature for 0.5 h. The reaction mixture was quenched by adding 200 mL of ice water, stirred, and separated. The aqueous phase was extracted with dichloromethane (100 mL x 2). The combined dichloromethane layers were washed sequentially with 300 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and dried under reduced pressure. The concentrate was purified by flash chromatography to yield 395 mg of a white solid. m / z: 462.10 (M+1), 460.10 (M-1); 1 H NMR (600MHz, DMSO) δ12.00 (s, 1H), 7.81 (s, 2H), 5.30 (d, J = 46.8Hz, 2H), 3.25 (s, 1H), 2.32 (d, J = 10.5Hz, 3H); 13 C NMR (150MHz, DMSO) δ159.74, 156.59, 150.93, 148.41, 147.82, 145.36, 141.17 , 141.07, 138.45, 131.15, 128.42, 126.91, 79.55, 78.44, 27.99, 18.40, 12.64.

[0095] Example 3:

[0096] Compound 12 (60 mg) and tetrahydrofuran were added to a reaction flask and stirred at room temperature until dissolved. Phosphorus tribromide (71 mg) was added and the temperature was raised to 50°C for 0.5 h. After cooling, the reaction solution was purified by column chromatography to yield 48 mg of a white solid. m / z: 524.05 (M+1), 522.05 (M-1).

[0097] Example 4:

[0098] Compound 10 (290 mg), azetidine (40 mg), and potassium carbonate (116 mg) were weighed, toluene (15 mL) and 1,4-dioxane (5 mL) were added, and the mixture was heated to 50°C for 2 h. The reaction mixture was filtered, and the filtrate was concentrated to dryness and purified by column chromatography to yield 25 mg of a white solid. m / z: 500.10 (M+H), 499.05 (MH); 1 H-NMR (600MHz, DMSO-d6) δ: 12.20 (s, 1H), 7.80 (s, 2H), 3.57 (s, 2H), 3.37 ( t, 4H, J=21.0, 10.2Hz), 3.09~3.00 (s, 1H), 2.34 (s, 3H), 2.06~1.99 (m, 2H).

[0099] Example 5:

[0100] first step:

[0101] 3,6-Dichloro-4-isopropyl-5-methylpyridazine (1.0 g), SM-2-1 (873 mg), cuprous iodide (466 mg), potassium carbonate (1.69 g), and 10 mL of dimethyl sulfoxide were added to a reaction flask in that order. Under nitrogen, the temperature was raised to 90°C and the reaction was allowed to proceed for 16 hours. The reaction mixture was cooled and added to water, followed by extraction with ethyl acetate (50 ml x 2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 400 mg of a yellow oil. m / z: 348.05 (M+1).

[0102] Step 2:

[0103] Intermediate 5-1 (800 mg), sodium acetate (951 mg), and 8 mL of acetic acid were added to the reaction flask in sequence. The temperature was raised to 130°C and the reaction mixture was reacted for 16 h. The reaction mixture was cooled, dried, and used directly in the next step without purification. m / z: 370.05 (M+1), 368.05 (M-1).

[0104] The product from the previous step and 20 mL of ethanol were added to a reaction flask, followed by a solution of sodium hydroxide (9.28 g) in water (10 mL). The temperature was raised to 110°C and the reaction mixture was allowed to react for 3 h. The reaction mixture was cooled, reduced to semi-dryness, slurried with water, and filtered. The filter cake was washed with ethyl acetate and dried to yield 600 mg of a yellow-brown solid. m / z: 328.00 (M+1), 326.05 (M-1).

[0105] Step 3:

[0106] Dissolve intermediate 5-2 (500 mg) in 60 mL of water, add 30 mL of hydrochloric acid, then cool to 0-10°C and add a solution of sodium nitrite (379 mg) in water (3 mL). Stir in an ice-water bath for 1 hour. This is referred to as solution A. Add SM-2-2 (342 mg) to 15 mL of water, followed by 30 mL of pyridine. This is referred to as solution B. Slowly add solution A to solution B while maintaining the temperature at 0-10°C. After the addition is complete, continue stirring for 1.5 hours. Filter the reaction mixture, wash the filter cake with water, and dry to yield 650 mg of an orange solid. m / z: 495.10 (M+1), 493.10 (M-1).

[0107] Step 4:

[0108] Intermediate 5-3 (650 mg), sodium acetate (1.08 g), and 33 mL of acetic acid were added to a reaction flask and heated to 130°C for 3 h. The reaction solution was cooled and dried, and 25 mL of water was added. The mixture was extracted with ethyl acetate (25 mL x 2). The combined organic phases were washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to yield 190 mg of a white solid. m / z: 449.00 (M+1), 447.00 (M-1); 1 H-NMR (600MHz, DMSO-d6) δ12.02 (s, 1H), 7.78 (s, 2H), 3.32N3.24 (m, 1H), 2.34 (s, 3H), 1.31 (d, J = 6.6Hz, 6H).

[0109] Step 5:

[0110] Intermediate 5-4 (350 mg) and 20 mL of acetic acid were added to a reaction flask. Concentrated hydrochloric acid (10 mL) was added and the temperature was raised to 120°C for 2.5 h. The reaction solution was cooled to room temperature and poured into 50 mL of water. After stirring for 0.5 h, it was filtered. The filter cake was washed with water and dried to obtain 200 mg of a light brick-red solid.

[0111] Step 6:

[0112] Intermediate 5-5 (200 mg) and 20 mL of methanol were added to a reaction flask, and thionyl chloride (300 mg) was slowly added with stirring. The temperature was raised to 100°C and the reaction was allowed to react for 4 h. The reaction solution was cooled to room temperature and dried under reduced pressure. The concentrate was slurried with a mixture of ethyl acetate and petroleum ether, filtered, and dried to obtain 100 mg of a light brick-red solid.

[0113] Step 7:

[0114] Compound 5-6 (100 mg) was dissolved in a mixture of 15 mL of tetrahydrofuran and 2 mL of methanol. Sodium borohydride (200 mg) was slowly added with stirring and allowed to react at room temperature for 0.5 h. The reaction solution was quenched with 30 mL of water, adjusted to pH 6 with hydrochloric acid (1 M), concentrated to approximately half the remaining volume, and extracted with ethyl acetate (20 mL × 3). The ethyl acetate layers were combined and washed sequentially with 20 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and dried under reduced pressure. The concentrate was slurried with a mixture of ethyl acetate and n-hexane, filtered, and dried to obtain 80 mg of an off-white solid compound.

[0115] Step 8:

[0116] Compound 5-7 (80 mg) and 20 mL of dichloromethane were added to a reaction flask. Diethylaminosulfur trifluoride (DAST, 100 mg) was slowly added at 0-5°C. After the addition was complete, the mixture was allowed to react at room temperature for 0.5 h. The reaction mixture was quenched by adding 20 mL of ice water, stirred, and separated. The aqueous phase was extracted with dichloromethane (20 mL x 2). The combined dichloromethane layers were washed sequentially with 30 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, and dried under reduced pressure. The concentrate was purified by column chromatography to yield 40 mg of a white solid. m / z: 457.36 (M+1); 1 H NMR (600MHz, DMSO) δ12.69 (s, 1H), 12.00 (s, 1H), 7.81 (s, 2H), 5.30 (d, J=46.8Hz, 2H), 3.30 (d, J=7.5Hz, 1H), 2.33 (s, 3H), 1.30 (d, J=6.7Hz, 6H).

[0117] Using a similar preparation scheme, the following compounds can be prepared;

[0118] m / z: 457.16 (M+1); 1 H NMR (600MHz, DMSO) δ 12.68 (s, 1H), 12.14 (s, 1H), 7.81 (s, 2H), 5.30 (d, J = 46.8Hz, 2H), 3.38 (d, J = 7.7Hz, 1H), 2.16 (s, 3H), 1.42 (d, J = 6.8Hz, 6H).

[0119] Example 6:

[0120] first step:

[0121] Intermediate 6-4 (prepared with reference to Example 2 in WO2021104288A1) (3 g) was dissolved in acetic acid (AcOH, 100 mL). Concentrated hydrochloric acid (20 mL) was added with stirring. After addition, the temperature was raised to 120°C and the reaction was allowed to react for 6 h. After completion of the reaction, the reaction solution was cooled to room temperature and H2O (200 mL) was added dropwise. After addition, the solution was stirred at room temperature for 1 h and filtered. The filter cake was rinsed with H2O (20 mL x 2). The solid was collected and dried at 45°C to obtain 2.0 g of a khaki solid. m / z: 454.00 (M+H), 452.00 (MH).

[0122] Step 2:

[0123] Intermediate 6-5 (2 g) was dissolved in methanol (CH3OH, 100 mL) and thionyl chloride (SOCl2, 10 mL) was added dropwise in an ice bath. After addition, the mixture was heated to 100°C and reacted for 5 h. After completion of the reaction, the mixture was concentrated under reduced pressure. The residue was slurried with petroleum ether:ethyl acetate (1:1) for 1 h and filtered. The filter cake was rinsed with petroleum ether and the solid was collected and dried at 45°C to obtain 2.3 g of a yellow solid. m / z: 468.00 (M+H), 466.00 (MH); 1 H-NMR (400MHz, DMSO-d6) δ: 12.77 (s, 1H), 12.21 (s, 1H), 7.81 (s, 2H), 7.45 ( d, 1H, J=1.2Hz), 3.84 (s, 3H), 3.10~3.00 (m, 1H), 1.21 (s, 3H), 1.19 (s, 3H).

[0124] Step 3:

[0125] Intermediate 6-6 was dissolved in tetrahydrofuran (THF, 40 mL), and sodium borohydride (NaBH4, 0.82 g) was added. After the addition was complete, methanol (4 mL) was slowly added dropwise, generating a large amount of bubbles. The reaction was stirred for 1 h. After completion, the reaction was quenched with ice water and the pH was adjusted to approximately 6 with 1 M HCl. The mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined and washed with saturated NaCl (20 mL). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (dichloromethane:methanol = 15:1 to 10:1) to obtain 1.6 g of a light yellow solid. m / z: 440.00 (M+H), 438.05 (MH); 1H-NMR (400MHz, DMSO-d6) δ: 12.44 (s, 1H), 12.21 (s, 1H), 7.86 (s, 2H), 7.44 (d, 1H, J = 0.8Hz), 5.31 ~5.27 (t, 1H, J=12.4&6.4Hz), 4.41 (d, 2H, J=5.6Hz), 3.10~3.00 (m, 1H), 1.21 (s, 3H), 1.19 (s, 3H).

[0126] Step 4:

[0127] Intermediate 6-7 (100 mg) was dissolved in dichloromethane (20 mL), and diethylaminosulfur trifluoride (DAST, 8 drops) was added dropwise in an ice bath. The mixture was stirred in an ice bath for 30 min. After the reaction was complete, the reaction solution was slowly added dropwise to ice water. The organic phase was separated and temporarily stored. The aqueous phase was extracted with dichloromethane (15 mL). The combined organic phases were washed with saturated NaCl (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was slurried with EA to obtain 20 mg of compound X1 as a white solid. m / z: 442.00 (M+H), 440.05 (MH); 1 H-NMR (400MHz, DMSO-d6) δ: 12.67 (s, 1H), 12.22 (s, 1H), 7.82 (s, 2H), 7.44 (d, 1H) , J=0.8Hz), 5.36 (d, 2H, J=46.8Hz), 3.10~3.00 (m, 1H), 1.21 (s, 3H), 1.19 (s, 3H).

[0128] Biological testing

[0129] 1. Compound THRβ and THRα Binding Assay

[0130] Test Principle: Utilizing HTRF, the effects of test compounds on THRα and THRβ are assessed. The MAb Anti-GST-Eu cryptate antibody binds to the GST-tagged THRα-LBD (THRβ-LBD), while Streptavidin-XL665 binds to the biotin-tagged SRC3-2 coactivator peptide. Agonist binding to the THRα-LBD (THRβ-LBD) causes conformational changes in the THRα-LBD (THRβ-LBD), thereby increasing recruitment of the SRC3-2 coactivator peptide. Simultaneously, the resulting decrease in the distance between the XL665-labeled SRC3-2 and the Eu-anti-GST antibody increases the THRFRET signal. Compounds' effects on THRα-LBD (THRβ-LBD) activity at varying concentrations are used to assess their agonistic potential.

[0131] Test method: THRβ test process:

[0132] (1) Prepare 1× reaction solution.

[0133] (2) Compound preparation: The compound test concentration starts at 20 μM, and is diluted 3-fold to 10 concentrations, with replicates for each concentration. The test concentration starts at 20 μM, and is diluted 3-fold to 10 concentrations, with replicates for each concentration. Dilute the solution to a 100-fold final concentration in a 384-well Source plate, and then transfer 200 nL to a 384-well reaction plate using an Echo550. Transfer 200 nL of 100% DMSO and the test sample to the Min and Max wells, respectively.

[0134] (3) Prepare 2× protein solution using 1× reaction solution.

[0135] (4) Prepare 4×SRC3-2 solution using 1× reaction solution.

[0136] (5) Prepare a 4× GST-Eu cryptate and XL665 mixed solution using 1× reaction solution.

[0137] (6) Add 10 μL of 2× protein solution to each well of the reaction plate, centrifuge at 1000 rpm for 1 min, and incubate at room temperature for 30 min.

[0138] (7) Add 5 μL of 4×SRC3-2 solution to each well of the reaction plate and centrifuge at 1000 rpm for 1 min.

[0139] (8) Add 5 μL of 4×GST-Eu cryptate and XL665 mixed solution to each well of the reaction plate, centrifuge at 1000 rpm for 1 min, and incubate at 25°C.

[0140] (9) Fluorescence signals were read using EnVision.

[0141] The THRα test process is the same as above.

[0142] Test results:

[0143] The compounds of the present invention have activity against THRβ and THRα

[0144] The above data show that compounds 1, 6, 16, and 17 of the present invention have good agonist effects on THRβ, and their agonist activity on THRβ is significantly better than that of Resmetirom; on the other hand, compounds 6, 16, and 17 have weak agonist activity on THRα, with almost no agonist activity, indicating that compounds 6, 16, and 17 have selective agonist effects on THRβ and have excellent safety.

[0145] 2. Comparison of compound metabolites using human liver microsome incubation method

[0146] The purpose of this study was to evaluate the content of the main metabolites of compounds incubated in human liver microsomes.

[0147] Human liver microsome incubation:

[0148] Sample Preparation: The sample was precipitated with acetonitrile (ACN) containing 0.1% formic acid at a ratio of 1:2 and further centrifuged. The supernatant was dried using a stream of nitrogen. The residue was reconstituted in 300 L of 10% ACN and 90% H2O containing 0.1% formic acid. 10 L was injected into the LC-MS / MS system for analysis.

[0149] Sample chromatography analysis

[0150] Gradient elution table

[0151] Sample test results: See Figure 1 for the specific test spectrum, and the data are as follows.

[0152] The percentage of the main oxidative metabolites of the compounds in human liver microsome incubation study (%)

[0153] The above research data show that at the same research dose and the same human liver microsome incubation system, the content of the main metabolite of compound 16 (monooxidation product: 9.7%) was significantly higher than that of the main metabolite of compound 6 (monooxidation product 3%), indicating that the replacement of the hydrogen atom in the methyl group of compound 6 by a deuterium atom significantly increased the metabolic stability.

[0154] 3. Pharmacokinetic study of compounds in Sprague Dawley rats Elimination phase half-life (T 1 / 2 Features

[0155] The purpose of this study was to evaluate the elimination half-life characteristics of the test compound after a single intravenous bolus administration.

[0156] Administration route: Group 1: Resmetirom intravenous push; Group 2: Compound 6 intravenous push;

[0157] Dosage: Group 1: 5 mg / kg; Group 2: 5 mg / kg;

[0158] Dosing frequency: single dose;

[0159] Dosage method: Use a suitable disposable sterile syringe and intravenous infusion needle to accurately draw out the test sample of the required concentration and inject it into the tail vein. The administration should be completed in about 30 seconds.

[0160] Sample Collection: Approximately 0.3 mL of whole blood was collected from the jugular vein of each group of animals at each time point. Blood samples were collected from animals (intravenous group) before drug administration, and at 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, and 24 h after drug administration.

[0161] Sample Processing: Before blood collection, place a centrifuge tube containing 10 μL of EDTA-K2 and 1.2 mL of 50% acetonitrile-DMSO (so that the volume ratio of whole blood to 50% acetonitrile-DMSO is approximately 1:4) in an ice box filled with crushed ice. Add the collected blood to the centrifuge tube and vortex for at least 5 minutes. Then, temporarily store and transport the sample in an ice box filled with crushed ice. Centrifuge at 12,000 g for 10 minutes at 4°C, completing the centrifugation within 1 hour of blood collection. Transfer the supernatant of the whole blood to a newly labeled tube and store at or below -60°C for analysis.

[0162] Pharmacokinetic study of the elimination phase half-life of the compound in rats (T 1 / 2 )

[0163] The above research data show that at the same research dose and in the same research animal system, the elimination phase half-life (T 1 / 2 =7.80h) is significantly longer than the elimination phase half-life of Resmetirom (T 1 / 2 =2.98h).

[0164] 4. Study on the distribution of compounds in major organs of Sprague Dawley rats

[0165] The purpose of this study was to evaluate the distribution characteristics of the test compound in major organs after a single oral bolus administration.

[0166] Administration route: Group 1: Resmetirom administered by oral gavage; Group 2: Compound X1 administered by oral gavage; Group 3: Compound 6 administered by oral gavage;

[0167] Dosage: Group 1: 5 mg / kg; Group 2: 5 mg / kg; Group 3: 5 mg / kg;

[0168] Dosing frequency: Single dose

[0169] Blood and organ collection:

[0170] At each blood collection point, 0.25-0.3 ml of systemic blood (jugular vein) was collected in an EDTA-K2 anticoagulant tube, gently inverted to mix, and then placed on crushed ice for storage and transportation; collection plan: PO blood collection point: 1 hour and 4 hours after administration; animals were euthanized, and tissues and organs were collected, and the liver (0.3-0.5 g was collected from a fixed position in the liver lobe), kidney, and heart were weighed, placed in a 5 ml centrifuge tube, quickly frozen with dry ice, and stored at -20°C.

[0171] Sample preparation:

[0172] Solvent (methanol:acetonitrile:water at 25:25:50) was added and the slurry was ground and homogenized. This process was performed quickly using a tissue grinder.

[0173] For liver, add solvent to 3 ml, add one large steel ball and one small steel ball, and grind at 50 Hz for 60 seconds. Vortex to mix, and centrifuge at 8000 rpm for 5 minutes. Remove 0.3 ml of the supernatant, add solvent to 3 ml, vortex to mix, and centrifuge at 8000 rpm for 5 minutes. Remove 0.1 ml of the supernatant, add 0.2 ml of acetonitrile, vortex for 30 seconds, and centrifuge at 10000 rpm for 5 minutes. Remove the supernatant and analyze for drug content.

[0174] For kidneys, divide the tubes into two centrifuge tubes and add solvent to each tube to 3 ml. Add one large steel ball and one small steel ball, and grind at 50 Hz for 60 seconds. Observe the grinding effect and perform multiple grindings. After grinding, vortex to mix thoroughly and centrifuge at 8000 rpm for 5 minutes. Take 0.3 ml of the supernatant from each tube, add 3 ml of solvent to the same tube, vortex to mix thoroughly, and centrifuge at 8000 rpm for 5 minutes. Take 0.1 ml of the supernatant and add 0.2 ml of acetonitrile. Vortex for 30 seconds, centrifuge at 10,000 rpm for 5 minutes, and remove the supernatant for drug content analysis.

[0175] For the heart, add solvent to 3ml, grind with one large steel ball and one small steel ball at 50Hz for 30 seconds, remove the heart and mince with scissors. Grind at 50Hz for 60 seconds, observing the grinding effect. Repeat the grinding process several times. After grinding, vortex to mix thoroughly and centrifuge at 8000rpm for 5 minutes. Collect 0.1ml of the supernatant, add 0.2ml of acetonitrile, vortex for 30 seconds, centrifuge at 10000rpm for 5 minutes, and remove the supernatant for drug content analysis.

[0176] For plasma, 0.1 ml of plasma was taken, 0.2 ml of acetonitrile was added, vortexed for 30 seconds, centrifuged at 10,000 rpm for 5 minutes, and the supernatant was collected and sent for analysis of drug content.

[0177] Detection method:

[0178] Chromatographic column: C18 Ultimate UHPLC XB-C18 (2.1×50mm) 377# with a guard column of the same model

[0179] Mobile phase: A: 0.1% formic acid solution; B: acetonitrile

[0180] Gradient elution

[0181] Flow rate: 0.2 mL / min

[0182] Column temperature: 40°C

[0183] Injection volume: 10 μL

[0184] Results and Analysis:

[0185] According to the test data, the summary chart is shown in Figure 2.

[0186] Test conclusion:

[0187] The above research data show that the compound 6 of the present invention has a more significant advantage in liver distribution compared with Resmetirom and compound X1.

[0188] 5. Efficacy testing of compound B-DIO in NASH mouse model

[0189] Purpose of the test:

[0190] The purpose of this experiment was to evaluate the pharmacodynamics of the compounds of the present invention using the NASH model of B-DIO mice.

[0191] Test method:

[0192] Five C57BL / 6 mice fed a normal diet were assigned to the G1 experimental group. Twenty-five B-DIO mice were randomly assigned to five experimental groups based on body weight, with five mice in each group. The dosing schedule is shown in Figure 3 . The experimental groupings are shown in the following table:

[0193] Grouping was performed on Day 1. Beginning on Day 0, CCL4 was intraperitoneally injected into the G2-G6 groups to establish the model. The concentration of the modeling reagent was 0.2 μl / g. Modeling was performed twice weekly, starting on Day 0. Body weights were measured twice weekly after grouping. At the end of the experiment (Day 26), mice were fasted for 4 hours. Non-anticoagulated blood was collected from the G1-G6 groups and centrifuged for serum analysis. Livers were collected and weighed from the G1-G6 groups. 20 mg of livers were used for TC and TG determination, and the remainder was fixed in formalin for pathological observation.

[0194] Results and Analysis:

[0195] During the study, no animals experienced abnormal mortality or obvious clinical symptoms. On Day 26, the average body weight of the G2 model group was significantly higher than that of the G1 vehicle control group. Compared to the G2 model group, the body weights of the remaining treatment groups remained unchanged (see Figure 4).

[0196] On Day 26, compared with the G1 vehicle control, the values ​​of ALT, AST, and TG in the G2 model control group were significantly increased. Compared with the G2 model group, the above indicators of mice in the G3-G6 drug-treated groups were significantly reduced, among which the reduction range of ALT was more than 80%, the reduction range of AST was more than 75%, and the reduction range of TG was more than 45%. Specific values ​​are shown in the table below; see Figures 5 and 6.

[0197] Day 26 Effects of the test product on blood lipids and liver injury indicators in NASH mice induced by HFD combined with carbon tetrachloride Note: a: mean ± standard error; b: ALT, AST, TG of G1 vehicle control group, G3-G6 drug-treated groups and G2 model group were statistically analyzed (One-way ANOVA), *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

[0198] On Day 26, TC values ​​in the G2 model control group were significantly increased compared to the G1 vehicle control. These values ​​were significantly decreased in the G3-G6 treatment groups by 9.3%, 24.9%, 25.5%, and 27.7%, respectively, compared to the G2 model group. TG values ​​in the G2 model control group remained unchanged compared to the G1 vehicle control. These values ​​were significantly decreased in the G4-G6 treatment groups by 30.4%, 35.9%, and 35.9%, respectively, compared to the G2 model group. See the table below and Figure 7 for specific values.

[0199] Day 26 Effects of the test product on liver TC and TG in NASH mice induced by HFD combined with carbon tetrachloride Note: a: mean ± standard error; b: TC and TG contents in the liver of G1 vehicle control group, G3-G6 drug-treated groups and G2 model group were statistically analyzed (One-way ANOVA), *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[0200] On Day 26, the livers of animals in groups G1-G6 were histopathologically examined with Oil Red O. While the G1 vehicle control group showed no significant staining, the area ratio of lipid droplets in the livers of mice in the G2 model group increased significantly. Compared to the G2 model group, the area ratio of lipid droplets in the livers of animals treated with test compound 6 (0.3 mg / kg) (G4), compound 6 (1 mg / kg) (G5), and compound 6 (2 mg / kg) (G6) all decreased significantly. The area ratio of lipid droplets in the livers of animals treated with resmetirom (3 mg / kg) (G3) decreased slightly, but no significant difference was observed. This suggests that under the conditions of this experiment, test compound 6 had a significant improvement effect on hepatic steatosis at doses of 0.3 mg / kg, 1 mg / kg, and 2 mg / kg, while resmetirom had a slight improvement effect on hepatic steatosis at a dose of 3 mg / kg, as shown in Figure 8.

[0201] On Day 26, the livers of animals in groups G1-G6 were histopathologically examined with Sirius Red. The liver tissue in the G1 vehicle control group was normal, with no fibrosis observed. Microscopically, fibrosis in the portal tracts of the livers of the G2 model group was commonly observed, accompanied by significant fibrous bridging. Compared with the G2 model group, the test products Resmetirom (3 mg / kg) (G3), Compound 6 (0.3 mg / kg) (G4), Compound 6 (1 mg / kg) (G5), and Compound 6 (2 mg / kg) (G6) all had a moderately positive effect on liver fibrosis, as shown in Figure 9.

[0202] Test conclusion:

[0203] In this study, a NASH model was successfully established in C57 mice using a high-fat diet combined with CCL4 injections, and efficacy testing of two test products was successfully completed. Both test products, Resmetirom (G3) and Compound 6 (G4-G6), significantly improved the elevated transaminases, blood lipids, and hepatic lipid abnormalities associated with modeling, as demonstrated by significantly reducing blood ALT, AST, and TG levels, as well as liver TC and TG. Furthermore, Resmetirom (G3) and Compound 6 (G5-G6) significantly reduced hepatic steatosis; Resmetirom (G3) and Compound 6 (G4-G6) significantly improved liver fibrosis; and Compound 6 (G4-G6) significantly reduced the proportion of liver lipid droplet area. Under the same experimental conditions, Compound 6 (G4-G6) demonstrated superior efficacy in reducing liver fat accumulation and inhibiting liver fibrosis compared to Resmetirom (G3).

Claims

1. A compound of formula (X), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, in, X is N or C; Y, U and T are each independently N, O or S; -Z- is -NH-, -O-, -S-, or -CH2-; R 1 is selected from hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, alkenyl, alkynyl, cyano, formyl, or alkoxycarbonyl; R 2 , R 3 , R 4 , R 5 , R 8 , R 9 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, or alkynyl.

2. A compound of formula (I), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, in, X is N or C; -Z- is -NH-, -O-, -S-, or -CH2-; R 1 is selected from hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, alkenyl, alkynyl, cyano, formyl, or alkoxycarbonyl; R 2 , R 3 , R 4 , R 5 , R 8 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, or alkynyl.

3. A compound of formula (II), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, in, R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, or alkynyl.

4. A compound of any one of the following formulae 1 to 12 and 16 to 18, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, 5. A pharmaceutical composition comprising the compound according to any one of claims 1 to 4 or its stereoisomer, tautomer or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

6. Use of the compound according to any one of claims 1 to 4 or its stereoisomer, tautomer or pharmaceutically acceptable salt or the pharmaceutical composition according to claim 5 in the preparation of a thyroid hormone receptor (THR) agonist.

7. The use according to claim 6, wherein The disease that can be treated by thyroid hormone receptor agonists is a metabolic disease, preferably obesity, hyperlipidemia, hypercholesterolemia, diabetes, non-alcoholic steatohepatitis (NASH), atherosclerosis, cardiovascular disease, hypothyroidism or thyroid cancer.

8. A method for treating a disease with a thyroid hormone receptor agonist, comprising administering to an individual in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 4 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof.

9. The method according to claim 8, wherein: The disease is a metabolic disease, preferably obesity, hyperlipidemia, hypercholesterolemia, diabetes, non-alcoholic steatohepatitis (NASH), atherosclerosis, cardiovascular disease, hypothyroidism or thyroid cancer.

10. A method for preparing the compound according to any one of claims 1 to 4, comprising the following reaction scheme: Using SM-1 as the starting material, the cyanide group is hydrolyzed to obtain compound (IM-1), esterification reaction is performed to obtain compound (IM-2), reduction reaction is performed to obtain compound (IM-3), and functional group conversion reaction is performed to obtain compound (II), wherein R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 Definition as in claim 3.

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