Application of small molecule compound in treatment of cardiovascular diseases

Abstract

The invention relates to application of a small molecule compound in treatment of cardiovascular diseases. Application of compound as shown in formula I or stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, pharmaceutically acceptable ester or pharmaceutically acceptable solvate thereof in preparation of medicine for treating and / or preventing heart failure with preserved ejection fraction, # imgabs0

Description

Technical Field

[0001] The present invention relates to the field of medicine, and particularly to the application of small molecule compounds in the treatment of cardiovascular diseases. Background Art

[0002] Cardiovascular diseases represent one of the difficult problems in medical challenges. Heart failure (HF) is the main cause of death in patients with cardiovascular diseases and is also a major problem faced clinically. Heart failure includes heart failure with reduced ejection fraction (HFrEF) and heart failure with preserved ejection fraction (HFpEF).

[0003] Currently, HFpEF accounts for about 50% of all heart failure patients, and its prevalence is increasing at an alarming rate. It is also the main cause of the rising mortality rate of cardiovascular diseases. HFpEF is a comprehensive disease involving multiple organ disorders, which is caused by the combined effects of the heart, lungs, kidneys, bones, immune system, inflammation, metabolism and other components, and is often accompanied by symptoms such as obesity, hypertension, myocardial hypertrophy, diabetes or atrial fibrillation. HFpEF is a syndrome with high morbidity and high mortality. According to clinical statistics, the mortality rate due to HF is 35%, and the proportion of deaths caused by HFpEF accounts for 57% of them. However, so far, few drug therapies or medical devices have been proven to be able to change the disease progression and prognosis of HFpEF patients. At present, there is an urgent need in this field to develop a drug and / or treatment method that can effectively treat HFpEF.

[0004] Jun is a transcription factor and one of the members of the AP1 family. It has chromatin-binding activity and transcriptional cis-regulatory region-binding activity, and is involved in regulating processes such as animal organ development, protein phosphorylation and cell proliferation. A Jun inhibitor refers to an inhibitor that can inhibit the expression of the Jun gene, reduce the binding activity of Jun to DNA, reduce the level of the expression product of the Jun gene, or prevent or block the signal transduction of Jun. Currently, studies have shown that Jun inhibitors have therapeutic effects in animal models of endometriosis, breast cancer, sepsis, etc. Summary of the Invention

[0005] The inventors of the present invention have discovered for the first time that inhibiting the high expression of Jun using a Jun inhibitor can prevent and treat HFpEF. Based on this, the inventors further studied and found that the compounds of the present invention have the activity of inhibiting the binding of JUN to DNA, have the activity of inhibiting JUN transcription, can prevent and treat HFpEF, and have broad application prospects.

[0006] Therefore, in the first aspect of the present invention, the present invention provides the use of the compound shown in formula I or its stereoisomers, prodrugs, crystal forms, pharmaceutically acceptable salts, pharmaceutically acceptable esters or pharmaceutically acceptable solvates in the preparation of a drug for treating and / or preventing heart failure with preserved ejection fraction.

[0007] Alternatively, there is provided a compound shown in formula I or its stereoisomers, prodrugs, crystal forms, pharmaceutically acceptable salts, pharmaceutically acceptable esters or pharmaceutically acceptable solvates for treating and / or preventing heart failure with preserved ejection fraction.

[0008] Alternatively, there is provided a method for treating and / or preventing heart failure with preserved ejection fraction, which comprises: administering to a subject in need an effective amount of the compound shown in formula I or its stereoisomers, prodrugs, crystal forms, pharmaceutically acceptable salts, pharmaceutically acceptable esters or pharmaceutically acceptable solvates.

[0009]

[0010] Wherein:

[0011] R is selected from C6-C10 aryl, 6-10-membered heteroaryl, and the C6-C10 aryl and 6-10-membered heteroaryl are each independently optionally substituted by R 1 and / or R 5 ;

[0012] Preferably, R is selected from C6-C10 aryl, 6-10-membered heteroaryl, and the C6-C10 aryl and 6-10-membered heteroaryl are each independently optionally substituted by R 1 or R 1 and R 5 ;

[0013] More preferably, R is selected from C6-C10 aryl, 6-10-membered heteroaryl, and the C6-C10 aryl and 6-10-membered heteroaryl are each independently optionally substituted by R 1 ;

[0014] R 1 is selected from hydrogen, C1-C6 alkyl, -COOH, -C(=O)O-C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl, -NR cR d ;

[0015] R c 、R d are each independently selected from hydrogen, C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl;

[0016] m is selected from 0, 1, 2, 3, 4, 5, 6;

[0017] Preferably, m is selected from 0, 1;

[0018] More preferably, m is 1;

[0019] R 2 is selected from -O-(CH2) n -R 2’ ;

[0020] n is selected from 0, 1, 2, 3, 4, 5, 6;

[0021] Preferably, n is selected from 0, 1;

[0022] R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5- to 6-membered heteroaryl;

[0023] R 3 is selected from hydroxy, -O-C1-C6 alkyl, wherein the C1-C6 alkyl is optionally substituted by a 5- to 6-membered heterocyclic group;

[0024] Preferably, R 3 is selected from hydroxy, -O-C1-C6 alkyl, wherein the C1-C6 alkyl is optionally substituted by a 6-membered heterocyclic group;

[0025] More preferably, R 3 is selected from hydroxy, -O-C1-C6 alkyl, wherein the C1-C6 alkyl is optionally substituted by morpholinyl;

[0026] Most preferably, R 3 is selected from hydroxy, -O-C1-C6 alkyl;

[0027] R 4 is selected from hydroxy, cyano, nitro, carboxyl, mercapto, halogen, C1-C6 alkyl, -NR a R b 、-CONR a R b, C6-C10 aryl, -O-C6-C10 aryl, -C(=O)-C1-C6 alkyl, -CH2-C6-C10 aryl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-C6-C10 aryl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, 5-6 membered heteroaryl;

[0028] R a , R b are each independently selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, said phenyl optionally substituted by C1-C6 alkyl;

[0029] R 5 is selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio;

[0030] and the compound of formula I does not contain

[0031] In some embodiments, R is selected from phenyl, benzimidazolyl, said phenyl, benzimidazolyl, are each independently optionally substituted by R 1 and / or R 5 ;

[0032] Preferably, R is selected from phenyl, benzimidazolyl, said phenyl, benzimidazolyl, are each independently optionally substituted by R 1 or R 1 and R 5 ;

[0033] More preferably, R is selected from phenyl, benzimidazolyl, said phenyl, benzimidazolyl, are each independently optionally substituted by R 1 ;

[0034] Or preferably, R is selected from phenyl, said phenyl, are each independently optionally substituted by R 1 and / or R 5 ;

[0035] Or more preferably, R is selected from phenyl, said phenyl, are each independently optionally substituted by R1 or R 1 and R 5 is replaced;

[0036] or more preferably, R is selected from phenyl, said phenyl, each independently optionally substituted by R 1 is replaced;

[0037] or preferably, R is selected from

[0038] or more preferably, R is selected from

[0039] Preferably, R 1 is selected from hydrogen, -COOH, -C(=O)O-C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl, -NR c R d ;

[0040] More preferably, R 1 is selected from hydrogen, -COOH, -C(=O)O-C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl;

[0041] Preferably, R c , R d each independently is selected from hydrogen, -(CH2) m -O-C1-C6 alkyl;

[0042] More preferably, in R c , R d , any one is hydrogen, and the other is selected from -(CH2) m -O-C1-C6 alkyl;

[0043] Further preferably, R 1 is selected from hydrogen, -COOH, -COOCH3, -COOCH2CH3, -COO(CH2)2CH3, -COO(CH2)3CH3, -COO(CH2)4CH3, -COO(CH2)5CH3,

[0044]

[0045] Even more preferably, R 1 is selected from hydrogen, -COOH, -COOCH3, -COOCH2CH3,

[0046]

[0047] Most preferably, R1 Selected from hydrogen, -COOH, -COOCH2CH3,

[0048] Preferably, R 5 is selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy;

[0049] More preferably, R 5 is selected from C1-C6 alkoxy;

[0050] Most preferably, R 5 is methoxy.

[0051] In some embodiments, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl, pyrazinyl (such as ), furyl (such as );

[0052] Preferably, R 2’ is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyrazinyl (such as ), furyl (such as );

[0053] Alternatively, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl;

[0054] Or preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl;

[0055] Or more preferably, R 2’ is selected from cyclopentyl, cyclohexyl, phenyl;

[0056] Or more preferably, R 2’ is selected from cyclopentyl, phenyl;

[0057] Preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl, -O-CH2-5-6-membered heteroaryl;

[0058] More preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl, -O-CH2-5-6-membered heteroaryl, and the 5-6-membered heteroaryl is selected from pyrazinyl (such as ), furyl (such as );

[0059] Most preferably, R 2 is selected from

[0060]

[0061] Or preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl;

[0062] Or more preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl;

[0063] Or further preferably, R 2 is selected from

[0064] Or most preferably, R 2 is selected from

[0065] In some embodiments, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3,

[0066] Preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3,

[0067] Or preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3;

[0068] Or more preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3;

[0069] Or further preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3;

[0070] Alternatively, most preferably, R 3 is selected from a hydroxyl group, -OCH3, -OCH2CH3.

[0071] In some embodiments, R 4 is selected from a hydroxyl group, a cyano group, a nitro group, a carboxyl group, a mercapto group, a halogen, a C1-C6 alkyl group, -NR a R b , -CONR a R b , a phenyl group, a phenoxy group, -C(=O)-C1-C6 alkyl group, a benzyl group, a C1-C6 alkoxy group, a C2-C6 alkenyl group, a C3-C8 cycloalkyl group, -C(=O)O-C1-C6 alkyl group, -C(=O)O-phenyl group, a C1-C6 alkylthio group, -S(=O)-C1-C6 alkyl group, -S(=O)2-C1-C6 alkyl group, a thienyl group (such as 2-thienyl);

[0072] Preferably, R a , R b are each independently selected from hydrogen, a methyl group,

[0073] Alternatively, preferably, in R a , R b , any one is selected from hydrogen, a C1-C6 alkyl group, and the other is selected from hydrogen, a C1-C6 alkyl group, -C(=O)-C1-C6 alkyl group, -S(=O)2-C1-C6 alkyl group, -S(=O)2-phenyl group, and the phenyl group is optionally substituted by a C1-C6 alkyl group;

[0074] Alternatively, more preferably, in R a , R b , any one is selected from hydrogen, a methyl group, and the other is selected from hydrogen, a methyl group,

[0075] Preferably, R 4 is selected from a hydroxyl group, a cyano group, a nitro group, a carboxyl group, -NH2, -SH, -CONH2, a methyl group, a phenyl group, a benzyl group, a methoxy group, a phenoxy group, an acetyl group, -F, -Cl, -Br, -CH=CH2, a cyclopentyl group, -COOMe, -COOPh, -SMe, -SOMe, -SO2Me, -NMe2, -NHAc, -NHMs, -NHTs, 2-thienyl;

[0076] More preferably, R 4 is a hydroxyl group.

[0077] In some embodiments, the structural formula of the compound is as shown in Formula I-1 or Formula I-1', preferably, the structural formula of the compound is as shown in Formula I-1,

[0078]

[0079] Wherein:

[0080] R 1 is selected from -COOH, -C(=O)O-C1-C6 alkyl;

[0081] Preferably, R 1 is selected from -COOH, -COOCH3, -COOCH2CH3, -COO(CH2)2CH3, -COO(CH2)3CH3, -COO(CH2)4CH3, -COO(CH2)5CH3;

[0082] More preferably, R 1 is selected from -COOH, -COOCH3, -COOCH2CH3;

[0083] Most preferably, R 1 is selected from -COOH, -COOCH2CH3;

[0084] R 2 is selected from -O-(CH2) n -R 2’

[0085] n is selected from 0, 1, 2, 3, 4, 5, 6;

[0086] Preferably, n is selected from 0, 1;

[0087] R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5-6 membered heteroaryl;

[0088] Preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl, pyrazinyl (such as ), furyl (such as );

[0089] More preferably, R 2’ is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,

[0090] phenyl, pyrazinyl (such as ), furyl (such as );

[0091] Or preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl;

[0092] Or more preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl;

[0093] Or most preferably, R 2’ is selected from cyclopentyl, phenyl;

[0094] Preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl, -O-CH2-5-6 membered heteroaryl;

[0095] More preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl, -O-CH2-5-6 membered heteroaryl, and the 5-6 membered heteroaryl is selected from pyrazinyl (such as ), furyl (such as );

[0096] Most preferably, R 2 is selected from

[0097] Or preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl;

[0098] Or more preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl;

[0099] Or most preferably, R 2 is selected from

[0100] R 3 is selected from hydroxyl, -O-C1-C6 alkyl;

[0101] Preferably, R 3 is selected from hydroxyl, -OCH3, -OCH2CH3, -O(CH2)2CH3;

[0102] More preferably, R 3 is hydroxyl;

[0103] R 4 is selected from hydroxyl, cyano, nitro, carboxyl, mercapto, halogen, C1-C6 alkyl, -NR a R b -CONR a R b, C6-C10 aryl, -O-C6-C10 aryl, -C(=O)-C1-C6 alkyl, -CH2-C6-C10 aryl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-C6-C10 aryl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, 5-6 membered heteroaryl;

[0104] Preferably, R 4 is selected from the group consisting of hydroxy, cyano, nitro, carboxyl, mercapto, halogen, C1-C6 alkyl, -NR a R b , -CONR a R b , phenyl, phenoxy, -C(=O)-C1-C6 alkyl, benzyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-phenyl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, thienyl (such as 2-thienyl);

[0105] R a 、R b are each independently selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, said phenyl optionally substituted by C1-C6 alkyl;

[0106] Preferably, R a 、R b are each independently selected from hydrogen, methyl,

[0107] Or preferably, in R a 、R b , any one is selected from hydrogen, C1-C6 alkyl, and the other is selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, said phenyl optionally substituted by C1-C6 alkyl;

[0108] Or more preferably, in R a 、R b , any one is selected from hydrogen, methyl, and the other is selected from hydrogen, methyl,

[0109] More preferably, R 4Selected from hydroxyl, cyano, nitro, carboxyl, -NH2, -SH, -CONH2, methyl, phenyl, benzyl, methoxy, phenoxy, acetyl, -F, -Cl, -Br, -CH=CH2, cyclopentyl, -COOMe, -COOPh, -SMe, -SOMe, -SO2Me, -NMe2, -NHAc, -NHMs, -NHTs, 2-thienyl;

[0110] Most preferably, R 4 is hydroxyl;

[0111] R 5 is selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio;

[0112] Preferably, R 5 is selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy;

[0113] More preferably, R 5 is selected from C1-C6 alkoxy;

[0114] Most preferably, R 5 is methoxy.

[0115] In some embodiments, the structural formula of the compound is as shown in Formula I-2,

[0116]

[0117] wherein:

[0118] R 1 is selected from hydrogen, -(CH2) m -O-C1-C6 alkyl, -NR c R d ;

[0119] Preferably, R 1 is selected from hydrogen, -(CH2) m -O-C1-C6 alkyl;

[0120] R c , R d are each independently selected from hydrogen, C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl;

[0121] Preferably, R c , R d are each independently selected from hydrogen, -(CH2) m -O-C1-C6 alkyl;

[0122] More preferably, R c , R dwherein any one of them is hydrogen and the other is selected from -(CH2) m -O-C1-C6 alkyl;

[0123] m is selected from 0, 1, 2, 3, 4, 5, 6;

[0124] Preferably, m is selected from 0, 1;

[0125] More preferably, m is 1;

[0126] More preferably, R 1 is selected from hydrogen,

[0127] R 2 is selected from -O-(CH2) n -R 2’ ;

[0128] n is selected from 0, 1, 2, 3, 4, 5, 6;

[0129] Preferably, n is selected from 0, 1;

[0130] R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5- to 6-membered heteroaryl;

[0131] Preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl, pyrazinyl (such as

[0132] ), furyl (such as );

[0133] More preferably, R 2’ is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,

[0134] phenyl, pyrazinyl (such as ), furyl (such as );

[0135] Or preferably, R 2’ is selected from C3-C8 cycloalkyl;

[0136] Or more preferably, R 2’ is selected from cyclopentyl, cyclohexyl;

[0137] Or most preferably, R 2’ is selected from cyclopentyl;

[0138] Preferably, R 2Selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl, -O-CH2-5-6 membered heteroaryl;

[0139] More preferably, R 2 Selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl, -O-CH2-5-6 membered heteroaryl, and the 5-6 membered heteroaryl is selected from pyrazinyl (such as ), furyl (such as );

[0140] Most preferably, R 2 Selected from

[0141] Or preferably, R 2 Selected from -O-C3-C8 cycloalkyl;

[0142] Or more preferably, R 2 Selected from

[0143] Or most preferably, R 2 Selected from

[0144] R 3 Selected from hydroxyl, -O-C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted by a 5-6 membered heterocyclic group;

[0145] Preferably, R 3 Selected from hydroxyl, -O-C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted by a 6 membered heterocyclic group;

[0146] More preferably, R 3 Selected from hydroxyl, -O-C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted by morpholinyl;

[0147] Further preferably, R 3 Selected from hydroxyl, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3,

[0148] Even more preferably, R 3 Selected from -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3,

[0149] Or preferably, R 3Selected from a hydroxyl group, -O-C1-C6 alkyl;

[0150] Or more preferably, R 3 Selected from a hydroxyl group, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3;

[0151] Or even more preferably, R 3 Selected from a hydroxyl group, -OCH3, -OCH2CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3;

[0152] Or preferably, R 3 Selected from -O-C1-C6 alkyl;

[0153] Or more preferably, R 3 Selected from -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3;

[0154] Or most preferably, R 3 Selected from -OCH3, -OCH2CH3;

[0155] R 4 Selected from a hydroxyl group, a cyano group, a nitro group, a carboxyl group, a mercapto group, a halogen, a C1-C6 alkyl, -NR a R b 、-CONR a R b 、a C6-C10 aryl, -O-C6-C10 aryl, -C(=O)-C1-C6 alkyl, -CH2-C6-C10 aryl, a C1-C6 alkoxy group, a C2-C6 alkenyl, a C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-C6-C10 aryl, a C1-C6 alkylthio group, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, a 5-6 membered heteroaryl;

[0156] Preferably, R 4 Selected from a hydroxyl group, a cyano group, a nitro group, a carboxyl group, a mercapto group, a halogen, a C1-C6 alkyl, -NR a R b 、-CONR a R b, phenyl, phenoxy, -C(=O)-C1-C6 alkyl, benzyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-phenyl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, thienyl (such as 2-thienyl);

[0157] R a , R b are each independently selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, where the phenyl is optionally substituted with C1-C6 alkyl;

[0158] Preferably, R a , R b are each independently selected from hydrogen, methyl,

[0159] Or preferably, in R a , R b , any one is selected from hydrogen, C1-C6 alkyl, and the other is selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, where the phenyl is optionally substituted with C1-C6 alkyl;

[0160] Or more preferably, in R a , R b , any one is selected from hydrogen, methyl, and the other is selected from hydrogen, methyl,

[0161] More preferably, R 4 is selected from hydroxy, cyano, nitro, carboxy, -NH2, -SH, -CONH2, methyl, phenyl, benzyl, methoxy, phenoxy, acetyl, -F, -Cl, -Br, -CH=CH2, cyclopentyl, -COOMe, -COOPh, -SMe, -SOMe, -SO2Me, -NMe2, -NHAc, -NHMs, -NHTs, 2-thienyl;

[0162] Most preferably, R 4 is hydroxy;

[0163] And the compound represented by formula I-2 does not contain In some embodiments, the structural formula of the compound is as shown in formula I-3,

[0164]

[0165] Wherein:

[0166] R 1 is selected from hydrogen, C1-C6 alkyl;

[0167] Preferably, R 1 is hydrogen;

[0168] R 2 is selected from -O-C3-C8 cycloalkyl;

[0169] Preferably, R 2 is R 3 is selected from -O-C1-C6 alkyl;

[0170] Preferably, R 3 is -OCH2CH3;

[0171] R 4 is a hydroxyl group.

[0172] In some embodiments, the compound is selected from the following:

[0173]

[0174]

[0175]

[0176]

[0177]

[0178]

[0179] Or,

[0180]

[0181]

[0182]

[0183]

[0184]

[0185] Or,

[0186]

[0187] In a second aspect of the present invention, the present invention provides the use of a pharmaceutical composition in the preparation of a medicament for treating and / or preventing heart failure with preserved ejection fraction, wherein the pharmaceutical composition comprises a compound of formula I or a stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, pharmaceutically acceptable ester or pharmaceutically acceptable solvate thereof,

[0188] Alternatively, there is provided a pharmaceutical composition for treating and / or preventing heart failure with preserved ejection fraction, wherein the pharmaceutical composition comprises a compound of formula I or a stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, pharmaceutically acceptable ester or pharmaceutically acceptable solvate thereof,

[0189] Alternatively, there is provided a method for treating and / or preventing heart failure with preserved ejection fraction, which comprises: administering to a subject in need an effective amount of a pharmaceutical composition, the pharmaceutical composition comprising a compound of formula I or a stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, pharmaceutically acceptable ester or pharmaceutically acceptable solvate thereof,

[0190]

[0191] wherein R, R 2 , R 3 , R 4 are each independently as described in any technical solution of the first aspect;

[0192] Preferably, the structural formula of the compound is as shown in formula I-1, formula I-1', formula I-2 or formula I-3, wherein the compound shown in formula I-1 or formula I-1' is as described in the first aspect, the compound shown in formula I-2 is as described in the first aspect, and the compound shown in formula I-3 is as described in the first aspect;

[0193] More preferably, the structural formula of the compound is as shown in formula I-1, formula I-2 or formula I-3, wherein the compound shown in formula I-1 is as described in the first aspect, the compound shown in formula I-2 is as described in the first aspect, and the compound shown in formula I-3 is as described in the first aspect;

[0194] More preferably, the structural formula of the compound is as shown in formula I-1, wherein the compound shown in formula I-1 is as described in the first aspect;

[0195] More preferably, the structural formula of the compound is as shown in formula I-1', wherein the compound shown in formula I-1' is as described in the first aspect;

[0196] More preferably, the structural formula of the compound is as shown in formula I-2, wherein the compound shown in formula I-2 is as described in the first aspect;

[0197] More preferably, the structural formula of the compound is as shown in Formula I-3, wherein the compound shown in Formula I-3 is as described in the first aspect;

[0198] Most preferably, the compound is as described above. Brief Description of the Drawings

[0199] Figure 1 : Construction of the HFpEF model. A: Schematic diagram of the experimental procedure; B: Detection results of systolic function after 5 weeks of mouse feeding; C: Detection results of diastolic function after 5 weeks of mouse feeding.

[0200] Figure 2 : Relative expression level of myocardial cell Jun after 15 weeks of HFD+L-NAME feeding, indicating high expression of Jun in HFpEF model mice.

[0201] Figure 3 : T-5224 can effectively alleviate the occurrence and development of HFpEF. A: Detection results of systolic function of mice at different time points; B: Detection results of diastolic function of mice at different time points; C: Changes in body weight of mice under different treatments; D: Changes in the expression level of myocardial cell Jun in mice under different treatments.

[0202] Figure 4 : Detection results of the inhibitory effect of the compound on the binding activity of Jun to DNA at different concentrations.

[0203] Figure 5 : Detection results of the inhibitory effect of the compound on the transcriptional activity of cell Jun at different concentrations.

[0204] Figures 6 to 9 : The compound of the present invention can effectively inhibit the occurrence and development of HFpEF.

[0205] Figures 10 to 18 : NMR spectrum of the compound of the present invention. Detailed Description of the Embodiments

[0206] It should be understood that the terms used herein are intended to describe specific embodiments and are not intended to be limiting. In addition, although any methods, devices, and materials similar or equivalent to those described herein can be used to implement or test the present invention, the preferred methods, devices, and materials are described herein.

[0207] In the present invention, unless otherwise explicitly indicated, the description method "each independently selected from" used throughout this article can either mean that among different groups, the specific options expressed between the same or different symbols do not affect each other, or it can mean that within the same group, the specific options expressed between the same or different symbols do not affect each other.

[0208] The substituents of the compounds of the present invention are disclosed according to group types or ranges. In particular, it is noted that the present invention includes each independent secondary combination of each member of these group types and ranges. For example, the term "C1-C6 alkyl" specifically refers to methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl independently disclosed.

[0209] The term "C1-C6 alkyl" refers to an alkyl group having 1 to 6 carbon atoms, preferably "C1-C4 alkyl", more preferably "C1-C3 alkyl", and most preferably "C1-C2 alkyl". Examples of "C1-C6 alkyl" include, but are not limited to, methyl, ethyl, propyl (e.g., n-propyl, isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), etc. Examples of "C1-C4 alkyl" include, but are not limited to, methyl, ethyl, propyl (e.g., n-propyl, isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl), etc. Examples of "C1-C3 alkyl" include methyl, ethyl, propyl (e.g., n-propyl, isopropyl), etc. Examples of "C1-C2 alkyl" include methyl, ethyl.

[0210] The term "C1-C6 alkoxy" refers to any of the above C1-C6 alkyl groups connected to the rest of the molecule through an oxygen atom (-O-), and examples include methoxy, ethoxy, isopropoxy, etc.

[0211] The term "C1-C6 alkylthio" refers to a group obtained by replacing the oxygen atom in any of the above C1-C6 alkoxy groups with a sulfur atom, such as C1-C6 alkylthio, C1-C4 alkylthio, C1-C3 alkylthio, etc.

[0212] The term "C3-C8 cycloalkyl" refers to a saturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 8 carbon atoms, and non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc., preferably "C3-C6 cycloalkyl".

[0213] The term "C2-C6 alkenyl" refers to a straight-chain or branched-chain aliphatic hydrocarbon group having 2 to 6 carbon atoms and containing one or more carbon-carbon double bonds, such as vinyl, allyl, etc.

[0214] The term "aryl" refers to a group of a carbocyclic aromatic system. For example, "C6-C10 aryl" refers to a group of a carbocyclic aromatic system having 6-10 carbon atoms, and non-limiting examples include, but are not limited to, phenyl, naphthyl, etc.

[0215] The term "heteroaryl" refers to aromatic monocyclic groups, bicyclic groups, tricyclic groups or groups with more rings having at least one heteroatom (N, O or S) in at least one ring. The heteroatom-containing ring optionally further has 1, 2 or 3 heteroatoms selected from N, O or S, and for bicyclic or tricyclic or more-ring heteroaryls, each ring in the bicyclic or tricyclic or more rings should form an aromatic system. For example, "5-6 membered heteroaryl" refers to 5- or 6-membered aromatic monocyclic groups, bicyclic groups, tricyclic groups or groups with more rings having at least one heteroatom (N, O or S) in at least one ring. Non-limiting examples thereof are, for example, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, imidazolyl, thiazolyl, isothiazolyl, thiazoxazolyl, pyrrolyl, furyl, oxazolyl, isoxazolyl, pyrazolyl, thienyl. Again, for example, the term "6-10 membered heteroaryl" refers to 6-, 7-, 8-, 9- or 10-membered aromatic monocyclic groups, bicyclic groups, tricyclic groups or groups with more rings having at least one heteroatom (N, O or S) in at least one ring. Non-limiting examples thereof are, for example, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, imidazolyl, phenyl-pyrrolyl, phenyl-furyl, benzofuryl, benzothienyl, benzimidazolyl, indazolyl, quinolinyl, isoquinolinyl, etc.

[0216] The term "heterocyclic group" refers to 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-membered (preferably 3-7 membered or 3-6 membered) saturated or partially unsaturated carbocycles, in which one or more carbon atoms are replaced by heteroatoms such as nitrogen, oxygen and sulfur. Non-limiting examples of heterocyclic groups are, for example, pyran, pyrrolidine, pyrroline, imidazoline, imidazolidine, pyrazolidine, pyrazoline, thiazoline, thiazolidine, dihydrofuran, tetrahydrofuran, 1,3-dioxolane, piperidine, piperazine, morpholine, morpholinyl, tetrahydropyrrolyl, thiomorpholinyl, etc. For example, "6-membered heterocyclic group" refers to a 6-membered saturated or partially unsaturated carbocycle, in which one or more carbon atoms are replaced by heteroatoms such as nitrogen, oxygen and sulfur. Non-limiting examples of 6-membered heterocyclic groups are, for example, pyran, piperidine, piperazine, morpholine, morpholinyl, thiomorpholinyl, etc. Again, for example, "5-membered heterocyclic group" refers to a 5-membered saturated or partially unsaturated carbocycle, in which one or more carbon atoms are replaced by heteroatoms such as nitrogen, oxygen and sulfur. Non-limiting examples of 5-membered heterocyclic groups are, for example, pyrrolidine, pyrroline, imidazoline, imidazolidine, pyrazolidine, pyrazoline, thiazoline, thiazolidine, 1,3-dioxolane, etc.

[0217] The term "heteroatom" refers to N, O or S.

[0218] The term "substituted" means that any one of the hydrogens on the specified atom or group is selectively replaced by the specified group, provided that the normal valence state of the specified atom is not exceeded.

[0219] In the present invention, the term "treatment" generally refers to obtaining the desired pharmacological and / or physiological effects. Such effects can be prophylactic, according to the complete or partial prevention of a disease or its symptoms; and / or therapeutic, according to the partial or complete stabilization or cure of a disease and / or the side effects resulting from the disease. "Treatment" as used herein encompasses any treatment of a disease in a patient, including: (a) preventing the occurrence of a disease or symptoms in a patient who is susceptible to the disease or symptoms but has not been diagnosed with the disease; (b) inhibiting the symptoms of a disease, i.e., preventing its progression; or (c) alleviating the symptoms of a disease, i.e., causing the disease or symptoms to regress.

[0220] In the present invention, "subject" refers to a vertebrate. In certain embodiments, the vertebrate refers to a mammal. Mammals include, but are not limited to, livestock (such as cattle), pets (such as cats, dogs, and horses), primates, mice, and rats. In certain embodiments, the mammal refers to a human.

[0221] In the present invention, "effective amount" refers to the amount that is effective in achieving the desired therapeutic effect at the required dosage and time. The "therapeutically effective amount" of the substance / molecule of the present invention may vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the substance / molecule to elicit the desired response in the individual. The therapeutically effective amount also encompasses the amount in which the therapeutic beneficial effects of the substance / molecule outweigh any toxic or harmful consequences.

[0222] The pharmaceutical compositions of the present invention may contain pharmaceutically acceptable excipients, including but not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycerol, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, beeswax, lanolin, and the like.

[0223] The pharmaceutical compositions described in the present invention can be prepared in various forms according to different administration routes. For example, the pharmaceutical compositions can be administered in any of the following ways: orally, by spray inhalation, rectally, nasally, buccally, vaginally, topically, parenterally such as by subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intracardiac, intrasternal, and intracranial injection or infusion, or by means of an implanted reservoir. Among them, oral or intravenous administration is preferred.

[0224] The compounds described in the present invention may optionally also be used in combination with one or more other active ingredients, and their respective dosages and ratios can be adjusted by those skilled in the art according to the specific disease condition, the specific situation of the patient, and clinical needs, etc.

[0225] As used herein, unless otherwise specified, the term "prodrug" refers to a derivative of a compound of the present invention that can be hydrolyzed, oxidized, or undergo other reactions under biological conditions (in vitro or in vivo) to provide the compound of the present invention. Prodrugs become active compounds only after such reactions under biological conditions, or they have no or only low activity in their non-reactive forms. Prodrugs can generally be prepared using well-known methods, such as those described in Burger's Medicinal Chemistry and Drug Discovery (1995) 172 - 178, 949 - 982 (edited by Manfred E. Wolff, 5th edition).

[0226] For the stereoisomers in the compounds described herein, when specifically designated as (R)- or (S)-isomers by chemical name, they should be understood to be the (R)-isomer or (S)-isomer with the major configuration, respectively. Any asymmetric carbon atom can exist in the (R)-, (S)-, or (R,S)-configuration, preferably in the (R)- or (S)-configuration.

[0227] Tautomerism refers to the phenomenon in which a functional group in certain compounds changes its structure to become another functional group isomer, and these two isomers can rapidly interconvert. This rapid and reversible conversion process enables the two isomers to coexist in a certain proportion under given conditions, forming a dynamic equilibrium state. Taking the following Compound A and Compound B as examples, those skilled in the art can understand that the following Compound A and Compound B are tautomers of each other (the differences in the structures shown in the boxes), and the two isomers can rapidly interconvert to reach a certain equilibrium. Therefore, for those skilled in the art, the following Compound A and Compound B represent the same compound, only with slightly different representation methods. Similarly, for other compounds with similar structures in this application, those skilled in the art can also understand that different tautomers essentially represent the same compound, only with slightly different representation methods.

[0228]

[0229] The terms "solvate" or "solvent complex" are used interchangeably and refer to a compound that exists as a combination with a certain solvent molecule. This combination can include a stoichiometric amount of a certain solvent, such as a monohydrate or a dihydrate, or can include any amount of water; for example, methanol or ethanol can form an "alcoholate", which can also be stoichiometric or non-stoichiometric. The term "solvent complex" as used herein refers to the solid form, that is, a compound in a solution of a solvent, although it can be solvated, but it is not a solvent complex as the term is used herein.

[0230] As used herein, the term "pharmaceutically acceptable salt" refers to (i) salts formed by acidic functional groups (such as -COOH) present in the compounds provided by the present invention with appropriate inorganic or organic cations (bases), and includes but is not limited to, alkali metal salts such as sodium salts, potassium salts, lithium salts, etc.; alkaline earth metal salts such as calcium salts, magnesium salts, etc.; other metal salts such as aluminum salts, iron salts, zinc salts, copper salts, nickel salts, cobalt salts, etc.; inorganic base salts such as ammonium salts; organic base salts such as tert-octylamine salts, dibenzylamine salts, morpholine salts, glucosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, N-methylglucosamine salts, guanidine salts, diethylamine salts, triethylamine salts, dicyclohexylamine salts, N,N'-dibenzylethylenediamine salts, chloroprocaine salts, procaine salts, diethanolamine salts, N-benzyl-phenethylamine salts, piperazine salts, tetramethylamine salts, tris(hydroxymethyl)aminomethane salts. And, (ii) salts formed by basic functional groups (such as -NH2) present in the compounds provided by the present invention with appropriate inorganic or organic anions (acids), and includes but is not limited to, hydrohalides such as hydrofluorides, hydrochlorides, hydrobromides, hydroiodides, etc.; inorganic acid salts such as nitrates, perchlorates, sulfates, phosphates, etc.; lower alkanesulfonates such as methanesulfonates, trifluoromethanesulfonates, ethanesulfonates, etc.; arylsulfonates such as benzenesulfonates, p-benzenesulfonates, etc.; organic acid salts such as acetates, malates, fumarates, succinates, citrates, tartrates, oxalates, maleates, etc.; amino acid salts such as glycine salts, trimethylglycine salts, arginine salts, ornithine salts, glutamate salts, aspartate salts, etc.

[0231] As used herein, the term "pharmaceutically acceptable ester" refers to an ester formed by -COOH present in the compounds provided by the present invention with an appropriate alcohol, or an ester formed by -OH present in the compounds provided by the present invention with an appropriate acid (such as a carboxylic acid or an oxygen-containing inorganic acid). Suitable ester groups include but are not limited to, formates, acetates, propionates, butyrates, acrylates, ethyl succinates, stearates or palmitates. The ester can undergo a hydrolysis reaction to form the corresponding acid or alcohol in the presence of an acid or a base.

[0232] As used herein, the term "crystal form" refers to the crystal structure of a substance. When a substance crystallizes, due to the influence of various factors, the bonding mode within or between molecules changes, resulting in different arrangements of molecules or atoms in the lattice space and forming different crystal structures. The compounds of the present invention can exist in one crystal structure or multiple crystal structures, that is, they have "polymorphism". The compounds of the present invention can exist in different crystal forms.

[0233] The present invention will be further explained and illustrated below in conjunction with specific embodiments. Unless otherwise specified, all reagents and raw materials can be obtained commercially, or can be prepared according to the prior art and common general knowledge by conventional technical means in this field, and all instruments are those commonly used by those skilled in the art.

[0234] Example 1

[0235] 1. Materials and Reagents

[0236] In this embodiment, C57BL / 6N wild-type mice were purchased from Vital River Laboratories (Beijing). The sources of the reagents are shown in the following table.

[0237]

[0238] Except as described above, other materials and reagents used in this embodiment are also commercially available products.

[0239] 2. Animal Experiment Guidelines

[0240] In this embodiment, all animal studies were conducted under the guidance of the Experimental Animal Center of the Animal Health and Use Committee of Fuwai Hospital, Chinese National Center for Cardiovascular Diseases. All mice were propagated and raised in the same environment, and the mice were randomly grouped during the experiment. Echocardiogram analysis was performed by independent researchers who were unaware of the research objectives.

[0241] 3. Induction of Heart Failure with Preserved Ejection Fraction Model

[0242] Male C57BL / 6N wild-type mice aged 8 to 10 weeks were divided into three groups, namely the normal group (normal diet and drinking water), the model control group (high-fat diet combined with Nω-nitro-L-arginine methyl ester), and the model treatment group (high-fat diet combined with Nω-nitro-L-arginine methyl ester and treated with T-5224). Among them, the model control group and the model treatment group were modeled in the manner described in the following literature: Gabriele G. Schiattarella et al., Nitrosative stress drives heart failure with preserved ejection fraction, https: / / doi.org / 10.1038 / s41586-019-1100-z. Specifically, a high-fat diet (HFD) (60% kcal from fat (lard)) and Nω-nitro-L-arginine methyl ester (abbreviated as L-NAME, 0.5 g / L in drinking water) were used to induce heart failure with preserved ejection fraction to obtain an HFpEF animal model.

[0243] The systolic function parameter LVEF of the mice was not changed when detected in the fifth week after model induction, while the diastolic function parameter (E / E’) was significantly increased in the fifth week after model induction, indicating that a heart failure model with preserved ejection fraction as described in the aforementioned literature was successfully obtained. At the same time, there was no significant difference in the diastolic function parameter (E / E’) between the model control group and the model treatment group at week 5. Subsequent drug administration was carried out under the condition of the same baseline, such as Figure 1 shown

[0244] The expression of 4.Jun is correlated with HFpEF

[0245] When the model was induced to 15 weeks, cardiomyocytes of normal mice and the HFpEF animal model of mice were extracted and separated by perfusion method, and quantitative RCR detection was carried out. The specific operation is as follows:

[0246] 4.1. Isolation of adult mouse cardiomyocytes:

[0247] To isolate cardiomyocytes from the hearts of adult mice, we used the classical perfusion method to isolate cardiomyocytes. Specifically, 100 μl of sodium heparin (1000 units in 50 ml) was injected into the mice 20 minutes before sacrifice to prevent blood clotting in the heart during the operation and increase the difficulty of digestion. After that, the mice were anesthetized and sacrificed, and the hearts were removed and transferred to calcium-free solution for washing. Then, digestion was carried out using the Langendorff method. The heart was perfused with calcium-free solution using the Langendorff apparatus for 5 minutes, and then digested with digestive enzyme solution (0.7 mg / ml type II collagenase and 0.7 mg / ml bovine serum albumin in calcium-free solution) for about 30 minutes. The heart was continuously touched at about 20 minutes. When the heart became soft and slippery, it indicated that the digestion was basically completed. Then, the tissue from the ventricle was collected, cut into pieces, and gently pipetted to dissociate into single cells. After standing and sedimentation, the supernatant was taken, and the undigested and adhered tissues were removed. The cells were centrifuged at 100 g at 4 °C for 2 minutes to obtain cardiomyocyte precipitate. Most of the supernatant was non-cardiomyocytes. The cardiomyocytes were resuspended in calcium-free solution containing 10% FBS for subsequent experiments. The non-cardiomyocytes could be reselected with medium or PBS for subsequent experiments. If purer cardiomyocytes and non-cardiomyocytes were to be obtained, the cell suspension could be centrifuged (100 g, 2 minutes at room temperature) three times to separate cardiomyocytes from non-cardiomyocytes. The cardiomyocytes were collected for further experiments.

[0248] 4.2. Quantitative PCR detection:

[0249] Total RNA was extracted from cells using the GeneJet RNA Purification Kit (Thermo Scientific, K0732), and 0.1 μg of total RNA was reverse-transcribed to generate cDNA using the iScript™ cDNA Synthesis Kit (Bio-Rad, 1708890). qPCR was performed using the iTaq Universal SYBR Green supermix (1725121, Bio-Rad) on an ABI Vii7 real-time system (Life Technologies, Q6), with β-Actin used for normalization of quantitative analysis. As Figure 2 shown, a significantly higher expression of Jun was observed in the mouse HFpEF animal model compared with normal mice. This indicates that there is a correlation between the expression level of Jun and HFpEF in mice, and Jun is highly expressed in HFpEF.

[0250] 5. Administration method of T-5224

[0251] After obtaining the animal model, starting from the fifth week of inducing the HFpEF model, the model treatment group was treated with T-5224, and the model control group was treated with a drug-free solvent as a control. Mice with normal diet and water were used as negative controls throughout the induction process. When the mice were raised to 5 weeks old, drug administration was carried out. For the treatment group, T-5224 was administered at 250 mg / kg according to the body weight of the mice. Specifically, it was administered every other day. Each time, 0.8 mg of T-5224 was dissolved in 200 μL of 1% PVP solution. Administration started from the fifth week and ended at the thirteenth week (a total of 15 administrations), with a total dose of 250 mg / kg; the control group was given an equal volume of 1% PVP solution, and other treatment methods were the same.

[0252]

[0253] 6. Conventional ultrasound detection

[0254] After being fed under different conditions for five weeks, all mice began to undergo routine ultrasound examinations, which were performed every two weeks until the end of the 15-week examination. Specifically, transthoracic echocardiography was performed using a VisualSonics Vevo 2100 system equipped with an MS400 transducer (Visual Sonics). Left ventricular ejection fraction (LVEF) and other systolic function indices were obtained from short-axis M-mode scans at the mid-ventricular level, as indicated by the presence of the papillary muscles, in conscious, gently restrained mice. Apical four-chamber views were obtained in anesthetized mice for diastolic function measurements using pulsed-wave and tissue Doppler imaging at the mitral valve level. Anesthesia was induced with 2.5% isoflurane and confirmed by the lack of response to firm pressure on one of the hind paws. During echocardiography acquisition (under temperature-controlled conditions), isoflurane was reduced to 1.0 - 1.5% and adjusted to maintain the heart rate within the range of 500 beats per minute. The parameters collected included: heart rate, left ventricular end-diastolic diameter, left ventricular end-systolic diameter, end-diastolic interventricular septal wall thickness, left ventricular end-diastolic posterior wall, left ventricular fractional shortening, LVEF, peak Doppler blood flow velocity in early diastole across the mitral valve, peak Doppler blood flow velocity in late diastole across the mitral valve, isovolumic relaxation time, early diastolic and early filling deceleration times, and tissue Doppler peak of myocardial relaxation velocity at the mitral annulus. At the end of the procedure, all mice recovered from anesthesia without any abnormalities. All parameters were measured at least 3 times and the mean values were given. The ultrasound examination included systolic function and diastolic function examinations.

[0255] 7. Experimental Results and Conclusions

[0256] First, the systolic and diastolic functions of the mice were examined at 5 weeks. There were no significant changes in systolic function, but the diastolic function parameter E / E' increased significantly, demonstrating diastolic dysfunction and indicating that the model described in the aforementioned literature was successfully obtained. At the same time, taking the 5th week as the starting point for drug administration, there were no significant differences in cardiac diastolic function between the model control group and the model treatment group before drug administration ( Figure 1 A - C), and drug administration treatment was carried out on this basis.

[0257] Second, the expression of Jun was shown to be upregulated when compared between the normal group and the model control group, indicating the correlation between the expression of Jun and HFpEF. In the HFpEF mouse model, Jun was highly expressed ( Figure 2 ). Based on this correlation, it can be judged that Jun inhibitors can be used for the prevention and treatment of HFpEF.

[0258] Furthermore, T-5224 was used to verify the effect of Jun inhibitor on the prevention and treatment of HFpEF. When it was determined that the model was successfully constructed and the baselines of the model control group and the model treatment group were consistent, the model treatment group was treated with T-5224. The results of cardiac function detection showed that the occurrence and development of HFpEF in the model treatment group (after administration of T-5224) were well suppressed. Specifically, after treatment with T-5224, the diastolic function of the mice treated with high-fat diet combined with L-NAME (HFD+0.5g / L L-NAME) was significantly improved and could continue until the fifteenth week. However, in the model control group of mice without treatment with T-5224, continuous deterioration of diastolic function was observed ( Figure 3 A-B); meanwhile, in the model treatment group, the expression of Jun was down-regulated compared with the model control group ( Figure 3 D), and the obesity of the mice was improved ( Figure 3 C). This indicates that T-5224, as a Jun inhibitor, can play a role in the prevention and treatment of HFpEF in the mouse HFpEF model.

[0259] Example 2

[0260] Based on the experimental results of Example 1, the inventors further explored the efficacy of other compounds on HFpEF using T5524 as the positive control.

[0261] 1. Synthesis of Compounds

[0262] 1.1 Synthesis of Compound Target 1 (abbreviated as T1)

[0263]

[0264] 1) Synthesis of Compound 2

[0265]

[0266] Under nitrogen protection, a solution of compound 1 (10.0 g, 34.9 mmol, 1.00 eq), compound 1A (2.85 g, 38.4 mmol, 3.55 mL, 1.10 eq) and triphenylphosphine (11.0 g, 41.9 mmol, 1.20 eq) in tetrahydrofuran (100 mL) was cooled to 0 °C, and diisopropyl azodicarboxylate (8.48 g, 41.9 mmol, 8.14 mL, 1.20 eq) was added dropwise. The mixture was stirred at 25 °C for 10 hours. LCMS (EC20183-4-P1A1) monitored the complete reaction of compound 1, and there was a single main peak with the target molecular weight (R t = 0.36 min, MS cal.: 340.13, MS observed: [M+H]+ When (= 341.0), water (300 mL) was added to the reaction system at 25 °C to quench the reaction, and then it was extracted with ethyl acetate (200 mL * 3). All the organic phases were combined, washed with brine (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was purified by reverse-phase high-performance liquid chromatography (neutral conditions) to obtain the light yellow solid compound 2 (4.68 g, 13.3 mmol, 38.1% yield, 97.1% purity).

[0267] LCMS: EC20183-4-P1A1, R t = 0.36 min, MS cal.: 340.13, MS observed: [M+H] +

[0268] = 341.0.

[0269] LCMS: EC20183-4-P1A3, R t = 0.39 min, MS cal.: 340.13, MS observed: [M+H] +

[0270] = 341.1.

[0271] HPLC: EC20183-4-P1A4, R t = 2.34 min, 97.1% purity.

[0272] 1 1H NMR: EC20183-4-P1B, 400 MHz, DMSO-d6

[0273] δ: 11.84 (s, 1H), 7.63 (s, 1H), 7.57 (d, J=8.4 Hz, 1H), 7.44 (d, J=9.6 Hz, 1H), 7.19 (d, J=8.4 Hz, 1H), 6.54 - 6.53 (m, 2H), 3.83 (d, J=8.8 Hz, 2H), 3.07 (t, J=6.8 Hz, 2H), 2.84 (t, J=8.0 Hz, 2H), 2.07 - 1.98 (m, 1H), 0.97 (d, J=6.4 Hz, 6H).

[0274] 2) Synthesis of Compound 3

[0275]

[0276] Compound 2 (4.68 g, 13.3 mmol, 1.00 eq) was dissolved in methanol (20.0 mL), and then a solution of sodium methoxide (1.73 g, 32.0 mmol, 2.40 eq) in methanol (30.0 mL) was added. The mixture was stirred at 0 °C for 1 hour. LCMS (EC20183-6-P1A) monitored the complete reaction of Compound 2, and there was a single main peak with the target molecular weight (R t = 0.53 min, MScal.: 372.16, MS observed: [M+H] + = 373.2). The reaction solution was poured into 1N hydrochloric acid (50.0 mL) solution and diluted with water (200 mL). It was extracted with ethyl acetate (100 mL * 3), and all the organic phases were combined, washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain light yellow solid Compound 3 (4.97 g, 13.0 mmol, 97.4% yield, 97.5% purity).

[0277] LCMS: EC20183-6-P1A, R t = 0.53 min, MS cal.: 372.16, MS observed: [M+H] +

[0278] = 373.2.

[0279] LCMS: EC20183-6-P1A1, R t = 0.53 min, MS cal.: 372.16, MS observed: [M+H] +

[0280] = 373.2.

[0281] HPLC: EC20183-6-P1A2, R t = 3.60 min, 97.5% purity.

[0282] 3) Synthesis of Compound 4 - Notebook Page: EC20183-7

[0283]

[0284] A solution of compound 3 (2.48 g, 6.49 mmol, 1.00 eq), compound 3A (1.86 g, 8.12 mmol, 1.25 eq) and potassium carbonate (1.79 g, 12.9 mmol, 2.00 eq) in N,N - dimethylformamide (15.0 mL) was stirred at 50 °C for 2 h. When the reaction of compound 3 was monitored by LCMS (EC20183 - 7 - P1A1) and the only main peak with the target molecular weight appeared (R t = 0.47 min, MS cal.: 520.21, MS observed: [M + H] + = 521.3), the reaction solution was poured into 1 N HCl (50.0 mL) solution at 0 °C, then diluted with water (150 mL), and extracted with ethyl acetate (100 mL × 3). All the organic phases were combined, washed with water (200 mL × 2) and brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain light yellow solid compound 4 (3.47 g, crude product).

[0285] LCMS: EC20183 - 7 - P1A1, R t = 0.47 min, MS cal.: 520.21, MS observed: [M + H] +

[0286] = 521.3.

[0287] LCMS: EC20183 - 7 - P1A2, R t = 0.47 min, MS cal.: 520.21, MS observed: [M + H] +

[0288] = 521.3

[0289] 4) Synthesis of compound T1

[0290]

[0291] To a solution of compound 4 (3.47 g, 6.67 mmol, 1.00 eq) in tetrahydrofuran (7.00 mL) was added a solution of lithium hydroxide monohydrate (1.12 g, 26.6 mmol, 4.00 eq) in water (7.00 mL). The mixture was stirred at 50 °C for 2 h. When the reaction of compound 4 was monitored by LCMS (EC20183 - 8 - P1A), the only main peak with the target molecular weight appeared (R t = 0.39 min, MS cal.: 492.18, MS observed: [M + H] +When (= 493.2), the reaction solution was poured into 1N HCl (50.0 mL), diluted with 150 mL of water, and extracted with ethyl acetate (100 mL * 3). All the organic phases were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. TLC (methylene chloride: methanol = 10:1) monitored 4 spots (R f = 0.25, 0.20, 0.10, 0.00). The residue was purified by column chromatography (silica gel, methylene chloride: methanol = 100:1 to 1:1, R f = 0.20) to obtain the white solid compound T1 (0.90 g, 1.82 mmol, 27.3% yield, 99.6% purity).

[0292] LCMS: EC20183 - 8 - P1A, R t = 0.39 min, MS cal.: 492.18, MS observed: [M + H] +

[0293] = 493.2.

[0294] HRMS: EC20183 - 8 - P1F4.

[0295] HPLC: EC20183 - 8 - P1F, R t = 3.62 min, 99.6% purity.

[0296] 1 H NMR: EC20183 - 8 - P1B, 400 MHz, DMSO - d6

[0297] δ: 12.57 (br s, 2H), 12.00 (s, 1H), 7.98 (d, J = 8.0 Hz, 2H), 7.60 (d, J = 8.0 Hz, 2H), 7.55 - 7.54 (m, 2H), 7.46 (d, J = 9.6 Hz, 1H), 7.17 (br d, J = 8.8 Hz, 1H), 6.53 - 6.53 (m, 2H), 5.34 (s, 2H), 3.82 (d, J = 6.4 Hz, 2H), 2.91 (t, J = 7.2 Hz, 2H), 2.57 (t, J = 7.6 Hz, 2H), 2.07 - 1.97 (m, 1H), 0.97 (d, J = 6.4 Hz, 6H)

[0298] 1.2 Synthesis of Compound Target 2 (abbreviated as T2)

[0299]

[0300] 1) Synthesis of Compound 2

[0301]

[0302] A solution of compound 1 (2.49 g, 6.52 mmol, 1.00 eq), compound 1A (1.98 g, 8.15 mmol, 1.25 eq) and potassium carbonate (1.80 g, 13.0 mmol, 2.00 eq) in N,N-dimethylformamide (15.0 mL) was stirred at 50 °C for 2 h. LCMS (EC20184-4-P1A) monitored the complete reaction of compound 1 and showed a single main peak with the target molecular weight (R t = 0.49 min, MS cal.: 534.23, MS observed: [M+H] + = 535.3). When this occurred, the reaction mixture was poured into 50.0 mL of 1 N hydrochloric acid solution at 0 °C, diluted with 150 mL of water, and extracted with ethyl acetate (100 mL × 3). All the organic phases were combined, washed with water (200 mL × 2) and brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the pale yellow solid compound 2 (3.39 g, crude product).

[0303] LCMS: EC20184-4-P1A, R t = 0.49 min, MS cal.: 534.23, MS observed: [M+H] +

[0304] = 535.3

[0305] LCMS: EC20184-4-P1A1, R t = 0.49 min, MS cal.: 534.23, MS observed: [M+H] +

[0306] = 535.3

[0307] 2) Synthesis of compound T2

[0308]

[0309] Trimethyltin hydroxide (2.58 g, 14.3 mmol, 2.25 eq) was added to a solution of compound 2 (3.39 g, 6.34 mmol, 1.00 eq) in 1,2-dichloroethane (30.0 mL), and the mixture was stirred at 70 °C for 9 h. LCMS (EC20184-5-P1A) monitored the incomplete reaction of compound 2, and the target molecular weight was the main peak (R t = 0.44 min, MS cal.: 520.21, MS observed: [M+H]+ When ( = 521.3), at 25 °C, the pH of the reaction solution was adjusted to 6 with 1N hydrochloric acid solution, then diluted with 150 mL of water, and extracted with ethyl acetate (100 mL * 3). All organic phases were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. TLC (methylene chloride: methanol = 20:1) monitored 4 spots (R f = 0.35, 0.30, 0.25, 0.00). The residue was purified by silica gel column chromatography (methylene chloride: methanol = 100:1 to 1:1, R f = 0.30), and then the crude product was purified by preparative HPLC (TFA conditions) to obtain white solid T2 (0.55 g, 1.06 mmol, 16.6% yield, and 100% purity).

[0310] LCMS: EC20184 - 5 - P1A, R t = 0.44 min, MS cal.: 520.21, MS observed: [M + H] +

[0311] = 521.3.

[0312] HRMS: EC20184 - 5 - P1E4.

[0313] HPLC: EC20184 - 5 - P1E3, R t = 2.85 min, 100% purity.

[0314] 1 H NMR: EC20184 - 5 - P1B, 400 MHz, DMSO - d6

[0315] δ: 12.14 (br s, 1H), 11.99 (s, 1H), 8.00 (d, J = 8.4 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 7.55 - 7.53 (m, 2H), 7.47–7.44 (m, 1H), 7.17 (br d, J = 9.2 Hz, 1H), 6.54 - 6.51 (m, 2H), 5.36 (s, 2H), 4.32 (q, J = 7.2 Hz, 2H), 3.83 (d, J = 6.8 Hz, 2H), 2.91 (t, J = 7.6 Hz, 2H), 2.57 (t, J = 7.2 Hz, 2H), 2.07 - 1.98 (m, 1H), 1.32 (t, J = 6.8 Hz, 3H), 0.98 (d, J = 6.4 Hz, 6H)

[0316] Synthesis of Compound Target 3 (abbreviated as T3)

[0317]

[0318] 1) Synthesis of Compound 2B

[0319]

[0320] Under nitrogen protection, potassium carbonate (311 g, 2.26 mol, 3.00 eq) was added to a solution of compound 2A (125 g, 752 mmol, 1.00 eq) in acetonitrile (1.25 L). Subsequently, methyl iodide (266 g, 1.88 mol, 117 mL, 2.50 eq) was added to the mixture. The mixture was stirred for 12 hours under nitrogen protection and at 25 °C. When TLC plate (petroleum ether: ethyl acetate = 5:1) monitored that compound 2A (Rf = 0.15) had completely reacted and two new spots (Rf = 0.40, 0.60) appeared, the reaction solution was diluted with water (3.00 L) and extracted with ethyl acetate (1.00 L × 3). All organic phases were combined, washed successively with water (1.00 L × 3) and brine (1.00 L × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain brown liquid compound 2B (138 g, 710 mmol, 94.4% yield).

[0321] 1H NMR: EC6536 - 710 - P1A1, 400 MHz, CDCl3

[0322] δ: 7.26 - 7.13 (m, 2H), 6.98 - 6.81 (m, 2H), 3.91 - 3.83 (m, 3H), 3.76 - 3.67 (m, 3H), 3.00 (t, J = 7.6 Hz, 2H), 2.67 (t, J = 7.6 Hz, 2H).

[0323] 2) Synthesis of Compound 2

[0324]

[0325] To a mixed solution of compound 1 (40.0 g, 219 mmol, 1.00 eq) in N,N-dimethylformamide (160 mg, 2.20 mmol, 168 μL, 0.01 eq) and dichloromethane (400 mL), oxalyl chloride (33.4 g, 263 mmol, 23.0 mL, 1.20 eq) was added dropwise. After the mixture was stirred at 25 °C for 2 h, it was concentrated under reduced pressure to obtain a residue. After dissolution with dichloromethane (400 mL), a solution of aluminum chloride AlCl3 (73.1 g, 548 mmol, 30.0 mL, 2.50 eq) and compound 2B (51.1 g, 263 mmol, 1.20 eq) in dichloromethane (50.0 mL) was added at -30 °C. After the mixture was stirred at 0 °C for 1 h, ethyl acetate (61.2 g, 694 mmol, 68.0 mL, 3.16 eq) was added dropwise, and then aluminum chloride (161 g, 1.21 mol, 66.0 mL, 5.50 eq) was added at 0 °C. After the reaction solution was stirred at 40 °C for 12 h (two parallel batches), LCMS (EC6536-741-P1A1) monitored that the reaction of compound 1 was complete, and there was a single main peak with the target molecular weight (Rt = 0.37 min, MS cal.: 316.09, MS observed: [M+H]+ = 317.0). After the reaction solution was cooled to 25 °C, it was poured into ice-cold 6 M hydrochloric acid solution, and the aqueous phase was extracted with ethyl acetate (1.00 L * 3). All the organic phases were combined, washed with brine (1.00 L * 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was purified by column chromatography (silica gel, petroleum ether:ethyl acetate = 100:1 to 0:1), and monitored by TLC plate (petroleum ether:ethyl acetate = 1:1, Rf = 0.30) to obtain light yellow oily compound 2 (121 g, 382 mmol, 87.1% yield).

[0326] LCMS1: EC6536-741-P1A1, Rt = 0.37 min, MS cal.: 316.09, MS observed: [M+H]+ = 317.0.

[0327] LCMS2: EC6536-741-P1A2, Rt = 0.37 min, MS cal.: 316.09, MS observed: [M+H]+ = 317.0.

[0328] 1H NMR: EC6536-741-P1A, 400 MHz, DMSO-d6.

[0329] δ: 12.27 (s, 1H), 10.58 (br s, 1H), 10.37 (br s, 1H), 7.49 - 7.35 (m, 3H), 6.93 (d, J = 8.4 Hz, 1H), 6.45 - 6.35 (m, 2H), 3.59 (s, 3H), 2.90 - 2.83 (m, 2H), 2.61 (t, J = 7.6 Hz, 2H).

[0330] 3) Synthesis of Compound Int A

[0331]

[0332] p - Toluenesulfonic acid monohydrate (3.04 g, 15.9 mmol, 0.05 eq) was added to a toluene (1.00 L) solution of Compound 2 (101 g, 319 mmol, 1.00 eq). The mixture was stirred at 120 °C for 12 h. After monitoring by LCMS (EC6536 - 760 - P1A) that the reaction of Compound 2 was complete and the only main peak with the target molecular weight appeared (Rt = 0.56 min, MS cal.: 284.07, MS observed: [M + H]+ = 285.0), ethyl acetate (1.50 L) and saturated sodium bicarbonate solution (1.50 L) were added. The aqueous phase was extracted with ethyl acetate (1.00 L * 3). All organic phases were combined, washed with brine (1.50 L * 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was triturated with petroleum ether:ethyl acetate = 3:1 (100 mL) at 25 °C for 30 min, filtered, and the filter cake was concentrated under reduced pressure to obtain light brown solid Compound Int A (63.1 g, 220 mmol, 69.1% yield, 99.4% purity).

[0333] LCMS1: EC6536 - 760 - P1A, Rt = 0.56 min, MS cal.: 284.07, MS observed: [M + H]+ = 285.0.

[0334] LCMS2: EC6536 - 760 - P1A1, Rt = 0.56 min, MS cal.: 284.07, MS observed: [M + H]+ = 285.0, 99.4% purity.

[0335] 1H NMR: EC6536 - 760 - P1A, 400 MHz, DMSO - d6

[0336] δ: 12.05 (br s, 1H), 11.13 - 10.42 (m, 1H), 7.72 - 7.47 (m, 2H), 7.40 (br d, J = 8.4 Hz, 1H), 7.18 (br d, J = 8.4 Hz, 1H), 6.50 - 6.28 (m, 2H), 3.07 (br t, J = 7.2 Hz, 2H), 2.84 (br t, J = 7.2 Hz, 2H)

[0337] 4) Synthesis of Compound Int A_3

[0338]

[0339] The tetrahydrofuran solution of compound Int A (25.1 g, 87.9 mmol, 1.00 eq) was cooled to 0 °C, and diisopropyl azodicarboxylate (21.3 g, 105 mmol, 20.4 mL, 1.20 eq), Int-A_4 (7.95 g, 92.3 mmol, 8.38 mL, 1.05 eq) and triphenylphosphine (27.6 g, 105 mmol, 1.20 eq) were added dropwise. Subsequently, the mixture was stirred at 25 °C for 12 hours. After monitoring by LCMS (EC6536 - 768 - P1A) that the reaction of compound Int A was complete and there was a single main peak with the target molecular weight (Rt = 0.51 min, MS cal.: 352.13, MS observed: [M + H]+ = 353.0), the reaction solution was poured into a mixed solution of ethyl acetate (300 mL) and water (300 mL). The aqueous phase was extracted with ethyl acetate (200 mL * 3). All the organic phases were combined, washed with brine (300 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was purified by reverse-phase HPLC (neutral condition) to obtain a pale yellow solid compound Int A_3 (15.2 g, 43.1 mmol, 49.0% yield, 100% purity)

[0340] LCMS1: EC6536 - 768 - P1A, Rt = 0.51 min, MS cal.: 352.13, MS observed: [M + H]+ = 353.0.

[0341] LCMS2: EC6536 - 768 - P1B, Rt = 0.51 min, MS cal.: 352.13, MS observed: [M + H]+ = 353.1, 100% purity.

[0342] 1H NMR: EC6536 - 768 - P1A1, 400 MHz, DMSO - d6

[0343] δ: 11.90 (s, 1H), 7.63 (s, 1H), 7.57 (dd, J = 2.0, 8.4 Hz, 1H), 7.43 (d, J = 8.8 Hz, 1H), 7.19 (d, J = 8.4 Hz, 1H), 6.54 - 6.44 (m, 2H), 4.91 (br t, J = 6.0 Hz, 1H), 3.12 - 3.05 (m, 2H), 2.89 - 2.82 (m, 2H), 2.01 - 1.88 (m, 2H), 1.78 - 1.52 (m, 6H)

[0344] 5) Synthesis of Compound Int D

[0345]

[0346] After cooling a methanol (50.0 mL) solution of Compound Int - A_3 (5.00 g, 14.1 mmol, 1.00 eq) to 0 °C, a methanol (10.0 mL) solution of sodium methoxide (1.84 g, 34.0 mmol, 2.40 eq) was added. The mixture was stirred at 0 °C for 2 h. After monitoring by LCMS (EC6536 - 769 - P1A1) that the reaction of Compound Int - A_3 was complete and the only main peak with the target molecular weight appeared (Rt = 0.49 min, MS cal.: 384.16, MS observed: [M + H]+ = 385.1), the reaction solution was poured into water (200 mL), and the pH value was adjusted to 5 with 1 M hydrochloric acid solution. Subsequently, it was extracted with ethyl acetate (200 mL * 3). All the organic phases were combined, washed with brine (200 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a pale yellow oily Compound Int - D (4.70 g, 12.2 mmol, 86.1% yield, crude product).

[0347] LCMS1: EC6536 - 769 - P1A1, Rt = 0.49 min, MS cal.: 384.16, MS observed: [M + H]+ = 385.1.

[0348] LCMS2: EC6536 - 769 - P1A4, Rt = 0.50 min, MS cal.: 384.16, MS observed: [M + H]+ = 385.0.

[0349] 6) Synthesis of Compound 4

[0350]

[0351] To a solution of compound Int D (2.00 g, 5.20 mmol, 1.00 eq) and compound 1C (1.19 g, 5.20 mmol, 1.00 eq) in N,N-dimethylformamide (20.0 mL) was added potassium carbonate (1.44 g, 10.4 mmol, 2.00 eq). The mixture was stirred at 40 °C for 1 hour. When the reaction of compound Int D was complete as monitored by TLC plate (petroleum ether:ethyl acetate = 2:1) and a new spot appeared, the reaction solution was poured into water (50.0 mL), and the pH value was adjusted to 2 with 1 M hydrochloric acid solution, followed by extraction with ethyl acetate (30.0 mL × 3). All the organic phases were combined, washed with brine (30.0 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a pale yellow oily compound 4 (1.70 g, 3.14 mmol, 60.3% yield).

[0352] 1H NMR: EC6536-773-P1A, 400 MHz, DMSO-d6

[0353] δ: 12.01 (s, 1H), 8.05 - 7.87 (m, 2H), 7.69 - 7.49 (m, 4H), 7.43 (br d, J = 8.8 Hz, 1H), 7.21 - 7.08 (m, 1H), 6.72 - 6.40 (m, 2H), 5.42 - 5.26 (m, 2H), 4.90 (br s, 1H), 4.44 - 4.26 (m, 2H), 3.62 - 3.47 (m, 3H), 3.03 - 2.82 (m, 2H), 2.77 - 2.56 (m, 2H), 1.96 - 1.87 (m, 2H), 1.72 (br s, 2H), 1.60 (br s, 2H), 1.37 - 1.29 (m, 3H).

[0354] 7) Synthesis of compound T3

[0355]

[0356] A solution of compound 4 (1.70 g, 3.14 mmol, 1.00 eq) in methanol (20.0 mL) was cooled to 0 °C, and then a solution of sodium hydroxide (627 mg, 15.6 mmol, 5.00 eq) in water (5.00 mL) was added. The mixture was stirred at 25 °C for 12 h. After monitoring by LCMS (EC6536 - 776 - P1A3) that the reaction of compound 4 was complete and there was a single main peak with the target molecular weight (Rt = 0.51 min, MS cal.: 504.18, MS observed: [M + H]+ = 505.3), the reaction solution was poured into water (50.0 mL), and the pH was adjusted to 5 with 1 M hydrochloric acid solution. Then, it was extracted with ethyl acetate (30.0 mL * 3). All the organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue, which was purified by column chromatography (silica gel, dichloromethane:methanol = 1:0 to 10:1), and the crude product was obtained by TLC monitoring (petroleum ether:ethyl acetate = 1:2, Rf = 0.65). At 25 °C, the crude product was triturated with dichloromethane (20.0 mL) for 30 min, filtered, and the filter cake was concentrated under reduced pressure to obtain the white solid compound Target 3 (510 mg, 1.01 mmol, 32.1% yield, 100% purity).

[0357] LCMS: EC6536 - 776 - P1A3, Rt = 0.51 min, MS cal.: 504.18, MS observed: [M + H]+ = 505.3.

[0358] HRMS: EC20403 - 1.

[0359] HPLC: EC20403 - 1 - P1A2, Rt = 3.52 min, 100% purity.

[0360] 1H NMR: EC20403 - 1 - P1A7, 400 MHz, DMSO - d6

[0361] δ: 13.79 - 12.12 (m, 2H), 12.04 (br s, 1H), 7.98 (d, J = 8.4 Hz, 2H), 7.60 (br d, J = 8.0 Hz, 2H), 7.57 - 7.50 (m, 2H), 7.45 (br d, J = 8.8 Hz, 1H), 7.17 (br d, J = 9.2 Hz, 1H), 6.54 - 6.44 (m, 2H), 5.35 (s, 2H), 4.91 (br t, J = 5.2 Hz, 1H), 2.92 (br t, J = 7.2 Hz, 2H), 2.57 (br t, J = 7.2 Hz, 2H), 2.01 - 1.88 (m, 2H), 1.79 - 1.65 (m, 4H), 1.59 (br d, J = 2.4 Hz, 2H)

[0362] Synthesis of Compound Target 4 (abbreviated as T4)

[0363]

[0364] 1) Synthesis of Compound 2B

[0365]

[0366] A solution of Compound 1 (5.00 g, 17.4 mmol, 1.00 eq), benzyl alcohol (2.08 g, 19.2 mmol, 1.99 mL, 1.10 eq) and triphenylphosphine (5.50 g, 20.9 mmol, 1.20 eq) in tetrahydrofuran (50.0 mL) was cooled to 0 °C. Diisopropyl azodicarboxylate (4.24 g, 20.9 mmol, 4.07 mL, 1.20 eq) was added dropwise under nitrogen protection. The mixture was stirred at 25 °C for 10 h. After monitoring by LCMS (EC20185 - 4 - P1A1) that the reaction of Compound 1 was complete and the only main peak with the target molecular weight appeared (Rt = 0.35 min, MS cal.: 374.12, MS observed: [M + H]+ = 375.0), the reaction solution was quenched with water (200 mL) at 25 °C and extracted with ethyl acetate (150 mL * 3). All organic phases were combined, washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was purified by reverse HPLC (neutral condition) to obtain light yellow solid Compound 2A (2.10 g, 4.81 mmol, 27.4% yield, 85.7% purity) and light yellow solid Compound 2B (2.89 g, 4.27 mmol, 24.4% yield, 55.3% purity).

[0367] LCMS: EC20185-4-P1A1, Rt = 0.35 min, MS cal.: 374.12, MS observed: [M+H]+ = 375.0.

[0368] LCMS: EC20185-4-P1C1, Rt = 0.51 min, MS cal.: 374.12, MS observed: [M+H]+ = 375.0.

[0369] LCMS: EC20185-4-P1D1, Rt = 0.50 min, MS cal.: 374.12, MS observed: [M+H]+ = 375.0.

[0370] HPLC: EC20185-4-P1C2, Rt = 4.31 min, 85.7% purity.

[0371] HPLC: EC20185-4-P1C3, Rt = 4.31 min, 55.3% purity.

[0372] 2) Synthesis of Compounds 3B and 3C

[0373]

[0374] After cooling a methanol (15.0 mL) solution of Compound 2 (2.89 g, 4.27 mmol, 1.00 eq) to 0 °C, a methanol (20.0 mL) solution of sodium methoxide (553 mg, 10.2 mmol, 2.40 eq) was added, and the mixture was stirred at 0 °C for 1 hour. When the reaction of Compound 2 (Rf = 0.50) was complete as monitored by TLC plate (petroleum ether: ethyl acetate = 3:1) and two new spots appeared, the reaction solution was poured into 1 M hydrochloric acid (50.0 mL) solution, diluted with water (150 mL), and extracted with ethyl acetate (100 mL * 3). All organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (silica gel, petroleum ether: ethyl acetate = 100:1 to 1:1, Rf = 0.30) to obtain white solid Compound 3B (0.88 g, 2.05 mmol, 48.1% yield, 94.9% purity) and white solid Compound 3C (0.75 g, 1.55 mmol, 36.2% yield, 83.8% purity)

[0375] LCMS: EC20185-6-P1A3, Rt = 0.49 min, MS cal.: 406.14, MS observed: [M+H]+ = 407.1.

[0376] LCMS: EC20185-6-P1A5, Rt = 0.48 min, MS cal.: 406.14, MS observed: [M+H]+ = 407.2.

[0377] HPLC: EC20185-6-P1A4, Rt = 3.49 min, 94.9% purity.

[0378] HPLC: EC20185-6-P1A6, Rt = 3.50 min, 83.8% purity.

[0379] 3) Synthesis of Compound 4

[0380]

[0381] A solution of Compound 3 (1.63 g, 3.60 mmol, 1.00 eq), Compound 3A (1.03 g, 4.50 mmol, 1.25 eq) and potassium carbonate (995 mg, 7.20 mmol, 2.00 eq) in N,N-dimethylformamide (5.00 mL) was heated to 50 °C and stirred for 2 hours. After monitoring by LCMS (EC20185-12-P1A) that the reaction of Compound 3 was complete and the only main peak of the target molecular weight appeared (Rt = 0.43 min, MS cal.: 554.19, MS observed: [M+H]+ = 555.2), the reaction solution was poured into 1 M HCl (20.0 mL) at 0 °C, then diluted with water (50.0 mL), and extracted with ethyl acetate (30.0 mL * 3). All the organic phases were combined, washed successively with water (30.0 mL * 2) and brine (30.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. Four spots (Rf = 0.50, 0.30, 0.25, 0.00) were monitored by TLC plate (petroleum ether:ethyl acetate = 2:1). The residue was purified by column chromatography (silica gel, petroleum ether:ethyl acetate = 100:1 to 1:1, Rf = 0.30). The crude product was purified by reverse HPLC (neutral condition) to obtain white solid Compound 4 (0.68 g, 1.23 mmol, 34.0% yield, 100% purity)

[0382] LCMS: EC20185-12-P1A, Rt = 0.43 min, MS cal.: 554.19, MS observed: [M+H]+ = 555.2.

[0383] LCMS: EC20185-12-P1E, Rt = 0.43 min, MS cal.: 554.19, MS observed: [M+H]+ = 555.2.

[0384] HPLC: EC20185 - 12 - P1E1, Rt = 3.04 min, 100% purity.

[0385] 1H NMR: EC20185 - 12 - P1B, 400 MHz, DMSO - d6

[0386] δ: 11.90 (s, 1H), 8.00 (d, J = 8.0 Hz, 2H), 7.62 (d, J = 8.4 Hz, 2H), 7.56 - 7.54 (m, 2H), 7.47–7.33 (m, 6H), 7.17 (d, J = 8.4 Hz, 1H), 6.63 - 6.58 (m, 2H), 5.35 (s, 2H), 5.19 (s, 2H), 3.86 (s, 3H), 3.56 (s, 3H), 2.94 (t, J = 7.2 Hz, 2H), 2.65 (t, J = 7.6 Hz, 2H).

[0387] 4) Synthesis of Compound T4

[0388]

[0389] To a solution of Compound 4 (0.68 g, 1.23 mmol, 1.00 eq) in tetrahydrofuran (5.00 mL) was added a solution of lithium hydroxide monohydrate (205 mg, 4.90 mmol, 4.00 eq) in water (5.00 mL). The mixture was stirred at 50 °C for 4 hours. After monitoring by LCMS (EC20185 - 15 - P1A4) that the reaction of Compound 4 was complete and there was a single main peak with the target molecular weight (Rt = 0.27 min, MS cal.: 526.16, MS observed: [M + H]+ = 527.2), at 25 °C, the pH of the reaction mixture was adjusted to 1 with 1 M hydrochloric acid solution, and then stirred at this temperature for 1 hour. The reaction mixture was filtered, and the filter cake was collected, dispersed in deionized water, and then lyophilized to obtain white solid Compound Target 4 (0.51 g, 968 μmol, 79.0% yield, 100% purity).

[0390] LCMS: EC20185 - 15 - P1A4, Rt = 0.27 min, MS cal.: 526.16, MS observed: [M + H]+ = 527.2.

[0391] HRMS: EC20185 - 15 - P1E.

[0392] HPLC: EC20185 - 15 - P1C3, Rt = 3.53 min, 100% purity.

[0393] 1H NMR EC20185-15-P1B2, 400 MHz, DMSO-d6

[0394] δ: 12.56 (br s, 2H), 11.95 (s, 1H), 7.98 (d, J = 8.4 Hz, 2H), 7.61 (d, J = 8.0 Hz, 2H), 7.56 - 7.54 (m, 2H), 7.49 - 7.45 (m, 3H), 7.41 (t, J = 6.8 Hz, 2H), 7.36 - 7.33 (m, 1H), 7.17 (d, J = 8.8 Hz, 1H), 6.64 - 6.59 (m, 2H), 5.34 (s, 2H), 5.19 (s, 2H), 2.91 (t, J = 6.8 Hz, 2H), 2.57 (t, J = 7.6 Hz, 2H).

[0395] 1.5 Synthesis of Compound Target 5 (abbreviated as T5)

[0396]

[0397] 1) Synthesis of Compound 2

[0398]

[0399] Into a methanol solution of NH2OH·HCl (31.3 g, 451 mmol, 3.00 eq), a methanol solution of NaOMe (135 g, 752 mmol, 30.0% purity, 5.00 eq) was slowly added dropwise under nitrogen protection and in an ice bath for at least 10 minutes. The reaction solution was warmed to room temperature and reacted for another 10 minutes. The reaction solution was then cooled back to the ice bath, and a methanol solution of 80 mL of Compound 1 (25.0 g, 150 mmol, 1.00 eq) was slowly added dropwise to the reaction system. The mixture was warmed to room temperature and reacted for 1 hour, and then warmed to 70 °C and reacted for another 4 hours. LCMS1 (EC6536 - 775 - P1A1) detection showed that the reaction of Compound 1 was complete, with a yield of 91.7% (Rt = 0.19 min, MS calculated value: 167.0, MS detected value: [M + H]+ = 168.0). The reaction solution was poured into 300 mL of ice water, and the pH was adjusted to 5 with 1 M HCl solution. The mixture was filtered by suction, the filter cake was collected, and washed successively with 500 mL of water, diisopropyl ether, and n - hexane. The filter cake was concentrated under reduced pressure to obtain white solid Compound 2 (21.7 g, 129 mmol, yield 85.7%, purity 99.4%). Detection was performed by LCMS (EC6536 - 775 - P1A2) and 1H NMR (EC6536 - 775 - P1A1).

[0400] LCMS1: EC6536 - 775 - P1A1, R t= 0.19 min, MS cal.: 167.0, MS observed:

[0401] [M+H] + = 168.0.

[0402] LCMS: EC6536-775-P1A2, R t = 0.20 min, MS cal.: 167.0, MS observed: [M+H] +

[0403] = 168.2.

[0404] 1 H NMR: EC6536-775-P1A1, 400 MHz, DMSO-d6

[0405] δ: 14.03 - 10.20 (m, 2H), 10.09 - 8.59 (m, 1H), 7.56 (br d, J = 8.0 Hz, 1H), 6.94 - 6.41 (m, 2H), 2.25 (s, 3H)

[0406] 2) Synthesis of Compound 3

[0407]

[0408] Dissolve Compound 2 (21.7 g, 129 mmol, 1.00 eq) in 400 mL of THF. Under ice bath and nitrogen protection, slowly add dropwise TEA (78.3 g, 774 mmol, 107 mL, 6.00 eq) and SOCl2 (23.0 g, 193 mmol, 14.0 mL, 1.50 eq). The reaction mixture is reacted at room temperature for 30 minutes. LCMS1 (EC20395-3-P1A2) shows that Compound 2 has completely reacted. The yield of Compound 3 is 93.2% (Rt = 0.29 min, MS cal.: 149.0, MS observed: [M+H]+ = 150.0). The reaction system is introduced into 200 mL of ice water, and the pH is adjusted to 1 with 1 M aqueous HCl solution. The precipitated yellow solid is Compound 3 (14.6 g, 97.6 mmol, yield 75.6%, purity 99.7%). The structure is confirmed by LCMS (EC20395-3-P1B1) and 1H NMR (EC20395-3-P1A1).

[0409] LCMS1: EC20395-3-P1A2, R t = 0.29 min, MS cal.: 149.0, MS observed: [M+H] +

[0410] = 150.0.

[0411] LCMS: EC20395-3-P1B1, R t = 0.29 min, MS cal.: 149.0, MS observed: [M+H] +

[0412] = 150.2.

[0413] 1 H NMR: EC20395-3-P1A1, 400 MHz, DMSO-d6

[0414] δ: 7.60 (d, J = 8.0 Hz, 1H), 7.34 (s, 1H), 7.13 (d, J = 8.0 Hz, 1H), 2.43 (s, 3H).

[0415] 3) Synthesis of Compounds 4 and 4A

[0416]

[0417] Dissolve Compound 3 (9.60 g, 64.1 mmol, 1.00 eq) in 100 mL of DCM, under ice bath and nitrogen protection, slowly add dropwise DIEA (12.4 g, 96.2 mmol, 16.7 mL, 1.50 eq) and CH3OCH2Cl (18.4 g, 228 mmol, 17.3 mL, 3.56 eq) in sequence. Stir the reaction system at room temperature for 30 minutes. Detected by LCMS (EC6536-790-P1A2), Compound 3 has completely reacted, and the yield of the target compound is 41.5% (Rt = 0.67 min, MS calculated value: 193.0, MS detected value: [M+H]+ = 194.1). Add 200 mL of ice water to the reaction system, wash the organic phase with 200 mL of water and 200 mL of saturated brine, and dry over anhydrous sodium sulfate. Filter, evaporate to dryness, obtain a solid, and purify it by preparative reverse-phase chromatography (neutral condition) to obtain Compound 4 as a yellow oil (4.14 g, 20.0 mmol, yield 31.2%, purity 93.7%). The structure was confirmed by 1H NMR 1 (EC6536-790-P1A1) and LCMS1 (EC6536-790-P1D1). Obtain Compound 4A as a yellow oil (4.25 g, 21.6 mmol, yield 33.7%, purity 98.3%), and the structure was confirmed by LCMS2 (EC6536-790-P2A1) and 1H NMR 2 (EC6536-790-P1A2).

[0418] LCMS: EC6536-790-P1A2, R t= 0.67 min, MS cal.: 193.0, MS observed: [M+H] +

[0419] = 194.1.

[0420] LCMS1: EC6536 - 790 - P1D1, R t = 0.66 min, MS cal.: 193.0, MS observed:

[0421] [M+H] + = 194.0.

[0422] LCMS2: EC6536 - 790 - P2A1, R t = 0.56 min, MS cal.: 193.0, MS observed:

[0423] [M+H] + = 194.0.

[0424] 1 H NMR 1: EC6536 - 790 - P1A1, 400 MHz, CDCl3

[0425] δ: 7.54 (d, J = 8.0 Hz, 1H), 7.25 (s, 1H), 7.11 (d, J = 8.0 Hz, 1H), 5.55 (s, 2H), 3.64 (s, 3H), 2.51 (s, 3H)

[0426] 1 H NMR 2: EC6536 - 790 - P1A2, 400 MHz, CDCl3

[0427] δ: 7.72 (br d, J = 8.0 Hz, 1H), 7.10 (br d, J = 7.8 Hz, 1H), 7.04 (br s, 1H), 5.31 (s, 2H), 3.44 (s, 3H), 2.48 (s, 3H)

[0428] 4) Synthesis of Compound 5

[0429]

[0430] Compound 4 (2.07 g, 10.0 mmol, 1.00 eq) was dissolved in 30 mL of acetonitrile. AIBN (164 mg, 1.00 mmol, 0.10 eq) and DBDMH (3.73 g, 13.0 mmol, 1.30 eq) were added. The reaction mixture was reacted at 80 °C for 8 hours. LCMS (EC20395 - 14 - P2C4) detected that the reaction of compound 4 was complete, and the yield of the target compound was 60.5% (Rt = 0.69 min, MS calculated value: 270.9, MS detected value: [M + H]+ = 271.9). The reaction mixture was treated with 50 mL of ice water, and the layers were separated. The organic phase was washed successively with an aqueous solution of Na2S2O3 (50.0 mL), an aqueous solution of NaHCO3 (50.0 mL), and saturated brine (50.0 mL), dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to obtain a yellow oily compound 5 (5.27 g, crude product).

[0431] LCMS: EC20395 - 14 - P2C4, Rt = 0.69 min, MS calculated value: 270.9, MS detected value: [M + H]+ = 271.9.

[0432] LCMS: EC20395 - 14 - P2C4, R t = 0.69 min, MS cal.: 270.9, MS observed: [M + H] +

[0433] = 271.9.

[0434] 5) Synthesis of Compound Int E

[0435]

[0436] Int A_3 (6.00 g, 17.0 mmol, 1.00 eq) was dissolved in 50 mL of ethanol. At room temperature, NaOEt (0.29 M, 146 mL, 2.50 eq) was added. The reaction mixture was reacted at 50 °C for 2 hours. LCMS (EC6536 - 771 - P1A1) detection showed that the reaction of Int A_3 was complete, and the yield was 91.8% (Rt = 0.52 min, MS calculated value: 398.1, MS detected value: [M + H]+ = 399.2). The reaction mixture was poured into 200 mL of hydrochloric acid aqueous solution with pH 5, and extracted with ethyl acetate (200 mL * 3). The combined organic phases were washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound Int E (6.78 g, 16.6 mmol, yield 97.8%, purity 97.9%), which was a yellow oil and was verified by LCMS1 (EC6536 - 771 - P1A3) detection.

[0437] LCMS: EC6536 - 771 - P1A1, Rt = 0.52 min, MS calculated value: 398.1, MS detected value: [M + H]+ = 399.2.

[0438] LCMS1: EC6536 - 771 - P1A3, Rt = 0.53 min, MS calculated value: 398.1, MS detected value: [M + H]+ = 399.2.

[0439] LCMS: EC6536 - 771 - P1A1, R t = 0.52 min, MS cal.: 398.1, MS observed: [M + H] +

[0440] = 399.2.

[0441] LCMS1: EC6536 - 771 - P1A3, R t = 0.53 min, MS cal.: 398.1, MS observed:

[0442] [M + H] + = 399.2.

[0443] 6) Synthesis of Compound 6

[0444]

[0445] Dissolve Compound 5 (5.27 g, 19.3 mmol, 1.43 eq) in 50 mL of DMF, stir at room temperature, add Compound IntE (5.50 g, 13.5 mmol, 1.00 eq) and K2CO3 (3.74 g, 27.0 mmol, 2.00 eq), and raise the reaction solution to 50 °C and stir for 2 hours. LCMS (EC20395 - 15 - P1A3) detection shows that Compound 5 has completely reacted, with a yield of 36.9% (Rt = 0.93 min, MS calculated value: 589.2, MS detected value: [M + H]+ = 590.4). The reaction system is treated with 100 mL of ethyl acetate and 100 mL of water. After phase separation, the organic phase is washed with 100 mL of water and 100 mL of saturated brine, and dried over anhydrous sodium sulfate. Filter, evaporate under reduced pressure to obtain the crude product, and purify it by reverse - phase high - performance liquid chromatography HPLC to obtain yellow oily Compound 6 (3.17 g, 5.06 mmol, yield 37.4%, purity 94.2%). Detection is carried out by LCMS1 (EC20395 - 15 - P1D16) and HPLC (EC20395 - 15 - P1A23).

[0446] LCMS: EC20395 - 15 - P1A3, R t= 0.93 min, MS cal.: 589.2, MS observed: [M+H] +

[0447] = 590.4.

[0448] LCMS1: EC20395-15-P1D16, R t = 2.10 min, MS cal.: 589.2, MS observed:

[0449] [M+H] + = 590.1.

[0450] HPLC: EC20395-15-P1A23, R t = 4.01 min, purity: 94.2%

[0451] 7) Synthesis of Compound T5

[0452]

[0453] Dissolve Compound 6 (2.00 g, 3.20 mmol, 1.00 eq) in a mixed solution of 10.0 mL of dioxane and 10.0 mL of ethanol. Add 6M HCl (14.5 mL, 27.3 eq) at room temperature, and stir the reaction mixture at room temperature for 3 hours. LCMS (EC20395-17-P1A4) detection shows that Compound 6 has completely reacted, with a yield of 92.1% (Rt = 0.66 min, MS calculated value: 545.2, MS detected value: [M+H]+ = 546.4). The reaction mixture is treated with 100 mL of ethyl acetate and 100 mL of water, the liquid phases are separated, the organic phase is washed successively with 100 mL of water and 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness under reduced pressure. The crude product is purified by reverse preparative liquid chromatography to obtain the target compound T5 (750 mg, 1.37 mmol, yield 67.9%, purity 99.4%), a white solid. The structure is confirmed by LCMS1 (EC20395-18-P1A1), HPLC (EC20395-18-P1A2), HRMS (EC20395-18-P1A1), 1H NMR (EC20395-18-P1A3) and 13C NMR (EC20395-18-P1A2).

[0454] LCMS: EC20395-17-P1A4, R t = 0.66 min, MS cal.: 545.2, MS observed: [M+H] +

[0455] = 546.4.

[0456] LCMS1: EC20395-18-P1A1, R t = 0.67 min, MS cal.: 545.2, MS observed:

[0457] [M+H] + = 546.4.

[0458] HRMS: EC20395-18-P1A1

[0459] HPLC: EC20395-18-P1A2, R t = 3.82 min, purity: 99.4%

[0460] 1 H NMR: EC20395-18-P1A3, 400 MHz, CDCl3

[0461] 13 C NMR: EC20395-18-P1A2

[0462] δ: 12.69 (br s, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.58 (d, J = 2.2 Hz, 1H), 7.57 - 7.53 (m, 2H), 7.51 (d, J = 9.0 Hz, 1H), 7.39 (d, J = 8.4 Hz, 1H), 6.96 (d, J = 8.4 Hz, 1H), 6.49 (d, J = 2.4 Hz, 1H), 6.38 (dd, J = 2.4, 8.9 Hz, 1H), 5.35 (s, 2H), 4.83 (tt, J = 2.8, 5.8 Hz, 1H), 4.15 (q, J = 7.2 Hz, 2H), 3.11 (t, J = 7.8 Hz, 2H), 2.70 (t, J = 7.6 Hz, 2H), 2.00 - 1.76 (m, 6H), 1.71 - 1.60 (m, 2H), 1.24 (t, J = 7.2 Hz, 3H)

[0463] Synthesis of Compound Target 6 (abbreviated as T6)

[0464]

[0465] 1) Synthesis of Compound 7

[0466]

[0467] Compound 4A (2.00 g, 10.3 mmol, 1.00 eq) was dissolved in 20 mL of acetonitrile and stirred at room temperature. AIBN (169 mg, 1.04 mmol, 0.10 eq) and DBDMH (3.85 g, 13.4 mmol, 1.30 eq) were added, and the reaction mixture was reacted at 80 °C for 8 h. LCMS (EC20419-5-P2A4) detection showed that the reaction of 4A was complete, and the yield was 32.6% (Rt = 0.58 min, MS cal.: 270.9, MS observed: [M+H]+ = 271.9). The reaction mixture was treated with 50 mL of ice water, layered, and the organic phase was washed with an aqueous solution of Na2S2O3 (50.0 mL), an aqueous solution of NaHCO3 (50.0 mL), and saturated brine (50.0 mL), and dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated under reduced pressure to obtain 5.74 g of the crude product of compound 7, which was a yellow oil.

[0468] LCMS: EC20419-5-P2A4, Rt = 0.58 min, MS calculated value: 270.9, MS detected value: [M+H]+ = 271.9.

[0469] 2) Synthesis of compound T6

[0470]

[0471] Compound 7 (5.74 g, 21.1 mmol, 1.35 eq) was dissolved in 50 mL of DMF and stirred at room temperature. Int D (6.41 g, 15.6 mmol, 1.00 eq) and K2CO3 (4.31 g, 31.2 mmol, 2.00 eq) were added, and the reaction mixture was heated to 50 °C and reacted for 1 h. LCMS (EC20419-6-P1A) showed that the reaction of compound 7 was complete, and the yield was 17.5% (Rt = 0.87 min, MS cal.: 575.2, MS observed: [M+H]+ = 576.2). The reaction mixture was diluted with 50 mL of water and extracted with ethyl acetate (50.0 mL * 3). The combined organic phases were washed with water (50.0 mL * 5), washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to obtain a solid, which was purified by reverse preparative liquid chromatography to obtain the target compound T6 (683 mg, 1.18 mmol, yield 7.56%, purity 99.5%), a yellow solid. The structure was confirmed by HPLC (EC20419-6-P1D2), 1H NMR (EC20419-6-P1A1), 12C NMR (EC20419-6-P1A2), and HRMS (EC20419-6-P1A1) detections.

[0472] LCMS: EC20419-6-P1A, R t= 0.87 min, MS cal.: 575.2, MS observed: [M+H] + = 576.2.

[0474] HRMS: EC20419-6-P1A1

[0475] HPLC: EC20419-6-P1D2, R t = 4.19 min, purity: 99.5%

[0476] 13 C NMR 1: EC20419-6-P1A2

[0477] 1 H NMR 1: EC20419-6-P1A1, 400 MHz, CDCl3

[0478] δ: 12.68 (s, 1H), 7.90 (d, J = 8.0 Hz, 1H), 7.57 (d, J = 2.0 Hz, 1H), 7.56 - 7.53 (m, 1H), 7.51 (d, J = 8.8 Hz, 1H), 7.40 (s, 1H), 7.35 (d, J = 8.0 Hz, 1H), 6.94 (d, J = 8.4 Hz, 1H), 6.49 (d, J = 2.4 Hz, 1H), 6.38 (dd, J = 2.4, 8.8 Hz, 1H), 5.36 (s, 2H), 5.31 (s, 2H), 4.83 (td, J = 2.8, 5.6 Hz, 1H), 3.69 (s, 3H), 3.47 (s, 3H), 3.10 (t, J = 7.6 Hz, 2H), 2.71 (t, J = 7.6 Hz, 2H), 2.01 - 1.76 (m, 6H), 1.71 - 1.64 (m, 2H)

[0479] Synthesis of Compound Target 7 (abbreviated as T7)

[0480]

[0481] Synthesis of Compound T7

[0482]

[0483] At 0 °C, Compound 2 (1.78 g, 12.0 mmol, 1.10 eq), Compound 1 (4.45 g, 10.9 mmol,

[0484] 1.00 eq) and triphenylphosphine (3.44 g, 13.1 mmol, 1.20 eq) were dissolved in tetrahydrofuran (40.0 mL). Diisopropyl azodicarboxylate (2.65 g, 13.1 mmol, 2.54 mL, 1.20 eq) was added dropwise to the reaction solution, and the mixture was stirred at 25 °C for 10 hours. LCMS (EC19915 - 16 - P1A) monitored that compound 1 (Rt = 0.400 min) was not completely reacted. When the main peak with the target molecular weight appeared (Rt = 0.32 min, MS cal.: 528.23, MS observed: [M + H]+ = 529.3), the reaction solution was concentrated under reduced pressure to obtain a residue. TLC plate (dichloromethane:methanol = 15:1) detected 5 spots (Rf = 0.50, 0.30, 0.20, 0.15, 0.00). The residue was purified by column chromatography (silica gel, dichloromethane:methanol = 100:1 to 1:1, Rf = 0.20), and then the crude product was purified by preparative - HPLC (trifluoroacetic acid condition) to obtain white solid compound Target 7 (0.65 g, 1.23 mmol, yield 11.2%, purity 100%).

[0485] LCMS: EC19915 - 16 - P1A, R t = 0.32 min, MS cal.: 528.23, MS observed: [M + H] +

[0486] = 529.3.

[0487] HRMS: EC21255 - 1 - P1A2.

[0488] HPLC: EC21255 - 1 - P1A1, R t = 3.03 min, 100% purity.

[0489] 1 1H NMR: EC21255 - 1 - P1B, 400 MHz, DMSO - d6

[0490] δ: 11.98 (s, 1H), 9.32 (s, 1H), 7.93 (s, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.64 (dd, J1 =

[0491] 1.2 Hz, J2 = 8.4 Hz, 1H), 7.57 - 7.53 (m, 2H), 7.42 (d, J = 8.8 Hz, 1H), 7.23 (d, J = 8.8 Hz, 1H), 6.50 - 6.45 (m, 2H), 5.43 (s, 2H), 4.90 (t, J = 5.6 Hz, 1H), 4.00 (q, J = 7.2 Hz, 2H), 2.92 (t, J = 7.6 Hz, 2H), 2.62 (t, J = 7.2 Hz, 2H), 1.98 - 1.90 (m, 2H), 1.73–1.59 (m, 6H), 1.09 (t, J = 7.2 Hz, 3H).

[0492] 1.8 Synthesis of Compound Target 8 (abbreviated as T8)

[0493]

[0494] Compound C230915014-C (1 g) was added to anhydrous methanol (5 ml), and 10 μl of concentrated sulfuric acid was added dropwise. The reaction was carried out at 60 °C for 3 h. TLC (Plate1: PE:EA = 1:1, 254 nm) showed a small amount of raw material remaining. Saturated sodium bicarbonate aqueous solution was added to the system to adjust the pH to 8 - 9, and it was extracted 3 times with EA (20 ml * 3), washed with 20 ml of saturated brine, the organic phase was collected, and concentrated to obtain 0.35 g of T8. The NMR spectrum of the product is as Figure 10 shown.

[0495] 1.9 Synthesis of Compound Target 9 (abbreviated as T9)

[0496]

[0497] Compound C230915014-C (0.7 g) was added to isopropanol (30 ml), and 0.29 ml (4 eq) of concentrated sulfuric acid was added dropwise. The reaction was carried out at 85 °C for 2 h. TLC (Plate1: PE:EA = 1:1, 254 nm) showed the formation of the target product. It was concentrated, extracted 3 times with 40 ml of EA and 30 ml of water, washed with 20 ml of saturated brine, the organic phase was loaded onto a column, and separated by column chromatography with PE:EA = 50:1 to 1:1. The target fraction was concentrated to collect the organic phase, and concentrated to obtain 0.52 g of T9. The NMR spectrum of the product is as Figure 11 shown.

[0498] 1.10 Synthesis of Compound Target 10 (abbreviated as T10)

[0499]

[0500]

[0501] Compound C230915014-SM1E (8 g) was added to DMF (80 ml), potassium carbonate (15.6 g, 4 eq) and bromocyclohexane (36.9 g, 8 eq) were added. The reaction was carried out at 70 °C for 12 h. LCMS showed the formation of the target product. It was extracted with 240 ml of ethyl acetate and 300 ml of H2O. The organic phase was washed with 200 ml of saturated sodium chloride aqueous solution. The organic phase was concentrated. The concentrate was filtered through silica gel (300 - 400 mesh) with ethyl acetate:n-heptane = 1:4 (500 mL). 500 ml of n-heptane was further added to the filtrate, and the filtrate was filtered through 300 - 400 mesh silica gel to obtain 1.95 g of product fraction TM-10A.

[0502] Compound TM-10A (1.7 g) was added to THF (30 ml), wet Pd(OH)2 (0.5 g, 0.75 eq) and H2 (under 15 psi) were added. The reaction was carried out at 25 °C for 2 h. HPLC showed the formation of the target product. The reaction solution was concentrated to obtain 1.6 g of TM-10B.

[0503] Compound TM-10B (0.8 g) was added to methanol (10 ml), sodium methoxide (0.36 g, 3 eq) was added. The reaction was carried out at 25 °C for 3 h. LCMS showed the formation of the target product. The reaction solution was concentrated to remove MeOH. 40 ml of EA and 40 ml of water were added to the system for extraction and liquid separation. The organic phase was collected. 30 ml of EA and 20 ml of ammonium chloride aqueous solution were added to the aqueous phase for extraction and liquid separation. The organic phases were combined, washed with 60 ml of saturated brine, dried over anhydrous sodium sulfate, to obtain 0.9 g of TM-10C.

[0504] Compound TM-10C (0.9 g) was added to acetone (20 ml), potassium carbonate (0.3 g, 1 eq) and C230915014-SM4 1.06 g (1 eq) were added. The reaction was carried out at 55 °C for 16 h. LCMS showed the formation of the target product. The reaction solution was filtered and concentrated. 40 ml of EA and 30 ml of water were added to the reaction solution for extraction and liquid separation. The organic phase was collected. The organic phase was washed with 30 ml of saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography on TLC (Plate1: PE:EA = 3:1, 254 nm), PE:EA = 50:1 - 5:1. The fraction was concentrated to obtain 1.0 g, and the fraction was freeze-dried to obtain 1 g of TM-10D.

[0505] Compound TM-10D (1 g) was added to DCM (20 ml), and TFA (10 ml) was added. The reaction was carried out at 25 °C for 16 h. LCMS showed the formation of the target product. 40 ml of water was added to the reaction solution, and the mixture was extracted with 40 ml of DCM. The organic phase was collected, washed with 40 ml of saturated brine, separated, and the organic phase was collected. The product was purified by column chromatography TLC (Plate1 PE:EA = 1:1, 254 nm), PE:EA = 25:1 - 2:1, and the fraction was concentrated to obtain 0.1 g of T10. The NMR spectrum of the product is as Figure 12 shown.

[0506] 1.11 Synthesis of Compound Target 11 (abbreviated as T11)

[0507]

[0508] Compound TM-10B (1.6 g) was added to ethanol (20 ml), sodium ethoxide (0.6 g, 2 eq) was added, and H2 (under 15 psi) was added. The reaction was carried out at 25 °C for 12 h. LCMS showed the formation of the target product. The reaction solution was concentrated to remove EtOH. 40 ml of EA and 40 ml of water were added to the system, and the mixture was extracted and separated. The organic phase was collected. 30 ml of EA and 20 ml of ammonium chloride aqueous solution were added to the aqueous phase, and the mixture was extracted and separated. The organic phases were combined, washed with 60 ml of saturated brine, and dried over anhydrous sodium sulfate to obtain 1.8 g of TM-11A.

[0509] Compound TM-11A (1.8 g) was added to acetone (30 ml), potassium carbonate (0.6 g, 1.1 eq) and C230915014-SM4 2.15 g (1.05 eq) were added. The reaction was carried out at 55 °C for 2 h. LCMS showed the formation of the target product. The reaction solution was filtered and concentrated. 40 ml of EA and 30 ml of water were added to the reaction solution, and the mixture was extracted and separated. The organic phase was collected, washed with 30 ml of saturated brine, and dried over anhydrous sodium sulfate. The organic phase was purified by column chromatography TLC (Plate1:PE:EA = 3:1, 254 nm), PE:EA = 10:1 - 5:1 (10% DCM), and the fraction was concentrated to obtain 2 g of TM-11B.

[0510] Compound TM-11B (0.6 g) was added to D (10 ml), HCl / Dioxane 0.37 ml (2 M) was added, and the reaction was carried out at 25 °C for 12 h. LCMS showed the formation of the target product. The reaction solution was concentrated, extracted with 40 ml of ethyl acetate and 30 ml of H2O. The organic phase was washed with 20 ml of saturated sodium chloride aqueous solution. The organic phase was concentrated, and slurried with 10 mL of ethanol to obtain 330 mg of T11. The NMR spectrum of the product is as Figure 13 shown.

[0511] 1.12 Synthesis of Compound Target 12 (abbreviated as T12)

[0512]

[0513] Add compound C230915014 - SM2 (1 g) into anhydrous methanol (5 ml), dropwise add 0.03 ml (0.2 eq) of concentrated sulfuric acid, react at 60 °C for 3 h. LCMS shows the formation of the target product. Add saturated sodium bicarbonate solution to the system to adjust the pH to 8 - 9, extract with EA (20 ml * 3). Wash the organic phase with saturated brine, dry the organic phase with anhydrous sodium sulfate, and concentrate to obtain 0.75 g of TM - 12A.

[0514] Add compound TM - 12A (1 g) into acetone (20 ml), add potassium carbonate (0.4 g, 1.1 eq), tert - butyl 4 - (bromomethyl)benzoate 0.77 g (1.1 eq). React at 55 °C for 10 h. LCMS shows the formation of the target product. Filter and concentrate the reaction solution. Add 20 mL of water to the reaction solution, extract with 20 mL of EA. Concentrate the organic phase and mix the sample, purify by column chromatography TLC (Pate1: PE: EA = 5:1, 254 nm), PE: EA = 50:1 - 20:1, and concentrate the fractions to obtain 1.2 g of TM - 12B.

[0515] Add compound TM - 12B (1.2 g) into DCM (20 ml), add TFA (6 ml, 39 eq), react at 25 °C for 3 h. LCMS shows the formation of the target product. Add 40 ml of water to the reaction solution, extract with 20 ml of DCM, collect the organic phase. Wash the organic phase with 30 ml of water, separate the layers, collect the organic phase, and concentrate to obtain 0.8 g of T12. The NMR spectrum of the product is as Figure 14 shown.

[0516] 1.13 Synthesis of Compound Target 13 (abbreviated as T13)

[0517]

[0518]

[0519] Add compound TM - 13 (1.3 g) into methanol (20 ml), add sulfuric acid (0.37 g, 1.5 eq), react at 80 °C for 0.5 h. TLC (Plate1: PE: EA = 1:1, 254 nm) shows the formation of a new spot. Concentrate the reaction solution, extract with 40 ml of ethyl acetate and 30 ml of H2O. Wash the organic phase with 20 ml of saturated sodium chloride aqueous solution, concentrate the organic phase, and slurry with ethyl acetate: n - heptane = 1:5 to obtain 1.0 g of T13. The NMR spectrum of the product is as Figure 15 shown.

[0520] 1.14 Synthesis of Compound Target 14 (abbreviated as T14)

[0521]

[0522] Add compound C230915014-C (2 g) to 2-bromoethanol (30 ml), dropwise add 0.5 ml (2.5 eq) of concentrated sulfuric acid, react at 85 °C for 2 h. LCMS shows the formation of the target product. Concentrate the reaction mixture, extract with 40 ml of EA and 30 ml of water three times, wash with 20 ml of saturated brine, concentrate the organic phase and triturate with 50 ml of CAN to obtain 1.6 g of TM-14A.

[0523] Add compound TM-14A (1.4 g) to DMF (14 ml), add potassium carbonate (0.25 g, 0.8 eq) and morpholine 0.2 ml (1 eq). React at 5 °C for 0.5 h. LCMS shows the formation of the target product. Add 15 ml of H2O to the reaction mixture, filter, and send the filtrate for preparative separation. The fraction is lyophilized to obtain 0.28 g of T14. The NMR spectrum of the product is as Figure 16 shown.

[0524] 1.15 Synthesis of Compound Target 15 (abbreviated as T15)

[0525]

[0526] Add compound 2-methoxy-4-methylbenzoic acid (4 g) to ethanol (20 ml), dropwise add 3.5 ml (2 eq) of SOCl2, react at 25 °C for 14 h. TLC (Plate1: PE:EA = 3:1, 254 nm) shows the formation of the target product. Slowly add 12 mL of water to the reaction mixture, and adjust the pH of the reaction mixture to 7-8 with saturated aqueous NaHCO3. Extract three times with 40 mL * 3 EA, and combine the organic phases. Wash the organic phase twice with 20 mL * 2 saturated NaCl aqueous solution, dry over anhydrous Na2SO4 and filter. The filtrate is rotary evaporated under reduced pressure at 45 °C to obtain 4 g of TM-15B.

[0527] Add compound TM-15B (0.6 g) to chloroform (9 ml), add NBS 0.55 g (1 eq) and benzoyl peroxide 0.04 g (0.05 eq), react at 35 °C for 1 h. HPLC shows the formation of the target product. Add 20 mL of saturated aqueous NaHCO3 to the reaction mixture, and collect the organic phase. Extract the aqueous phase twice with 15 mL * 2 DCM, and combine the organic phases. Wash the organic phase three times with 20 mL * 3 water, dry the organic phase over 10 g of anhydrous Na2SO4, filter and collect the filtrate, and concentrate to obtain 0.7 g of TM-15C.

[0528] Compound TM-15C (0.44 g) was added to acetone (7 ml), potassium carbonate (0.22 g, 1 eq) and C230915014-A 0.7 g (1 eq) were added. The reaction was carried out at 65 °C for 14 h. LCMS showed the formation of the target product. 21 mL of water was added to the reaction solution, and the aqueous phase was extracted three times with 10 mL * 3 EA. The organic phases were combined. The organic phase was washed twice with 20 mL * 2 water, dried over 10 g of anhydrous Na2SO4 and filtered. The filtrate was concentrated to obtain 1 g of TM-15A.

[0529] Compound TM-15A (1 g) was added to DCM (10 ml), TFA (2.4 ml, 20 eq) was added. The reaction was carried out at 25 °C for 2 h. TLC (Plate1: PE: EA = 3:1, 254 nm) showed that the reaction was complete. 30 ml of water was added to the reaction solution, and it was extracted with 20 ml of DCM. The organic phase was collected, washed with 30 ml of water, separated, and the organic phase was collected. After concentration, 6 mL of n-heptane: ethyl acetate was added and slurried at 15 °C for 20 min. Filtration gave 0.53 g of T15. The NMR spectrum of the product is as Figure 17 shown.

[0530] 1.16 Synthesis of compound Target 16 (abbreviated as T16)

[0531]

[0532] Compound C230915014-C (0.7 g) was added to n-propanol (30 ml), 0.29 ml (4 eq) of concentrated sulfuric acid was added dropwise. The reaction was carried out at 85 °C for 2 h. TLC (Plate1: PE: EA = 1:1, 254 nm) showed the formation of the target product. It was concentrated, extracted three times with 40 ml of EA and 30 ml of water, washed with 20 ml of saturated brine. The organic phase was loaded onto a column and eluted with PE: EA = 50:1 to 1:1 to separate the product. The target fractions were concentrated to collect the organic phase, and concentration gave 0.6 g of T16. The NMR spectrum of the product is as Figure 18 shown.

[0533] 2. Inhibition of DNA-binding activity assay

[0534] 2.1. Test method

[0535] c-jun transcription factor DNA-binding activity assay

[0536] The binding ability of DNA and transcription factors was tested using TransAM kits (Active Motif 46096), and the transcription factor was c-Jun / AP-1. The drugs with final concentrations of 500 μM, 250 μM, 125 μM, 62.5 μM, and 31.25 μM and 1 μL of cell extract containing the transcription factor were sequentially added to a 96-well plate pre-coated with double-stranded DNA oligomers, incubated for 1 h, incubated with an antibody against the c-JUN transcription factor, the absorbance at 450 nm was measured, and the absorbance at 655 nm was used as the background. The binding activity of the dsDNA sequence and the transcription factor was detected. The lower the binding activity, the stronger the inhibitory activity of the compound.

[0537] 2.2 Test results

[0538] The results are as follows in the table and Figure 4 shown below.

[0539] DNA&JUN binding activity

[0540]

[0541] It can be seen from the above results that except for T7, the other 6 compounds have the activity of inhibiting the binding of JUN to DNA. Among them, T3, T4, and T5 are better or equivalent to the positive control T-5224. It can be seen that the compounds of the present invention (especially the compounds of formula I-1, formula I-1', and formula I-2 series) have the activity of inhibiting the binding of JUN to DNA.

[0542] 3. Test for inhibitory transcriptional activity

[0543] 3.1 Test method

[0544] The dual-luciferase reporter system was used to detect the inhibitory DNA binding activity

[0545] 293T cells were plated at a ratio of 1:3 and incubated for 24 h, then changed to antibiotic-free medium Opti-MEM (Gibco, 11058021), and transiently transfected with the luciferase reporter plasmid pGL4.44 [luc2P / AP1 RE / Hygro] (Promega E411) for 24 h. The cells were incubated in 1% PS / 10% FBS / DMEM (Gibco, 11965092) containing the drug for 1 h, and the final concentrations of the drug were 50 μM, 25 μM, 12.5 μM, and 6.25 μM in sequence, and then stimulated with PMA (final concentration 10 ng / ml) (Sigma, 79346-5MG) and cultured for 34 h, and the lysate was measured using the dual-luciferase reporter gene detection system (Promega). Both the drug and PMA were DMSO stock solutions and were added to the culture medium after dilution with the medium.

[0546] 3.2 Test Results

[0547] The results are shown in the following table and Figure 5 as follows.

[0548]

[0549]

[0550] From the above results, it can be seen that T3 and T5 further have the activity of inhibiting JUN transcription. Obviously, the compounds of the present invention (especially the compounds of formula I-1, formula I-1' and formula I-2 series) have the activity of inhibiting JUN transcription.

[0551] 4. Efficacy Test on Animal Models

[0552] The materials and reagents, animal experiment guidelines, and induction of the heart failure model with preserved ejection fraction were the same as in Example 1.

[0553] The administration method is as follows:

[0554] After successful induction of the animal model, the model treatment group was treated with the compounds of the present invention (such as compounds T3, T5, T8-T16) and T-5524, and the model control group was treated with a drug-free solvent for control. Mice with normal diet and water were used as negative controls throughout the induction process. When the mice were raised to 5-8 weeks old and the model was successfully constructed by ultrasonic detection, drug administration was started. For the treatment group, the compounds T3, T5, T8-T16 and T-5524 were administered at a single dose of 12 mg / kg according to the body weight of the mice. Specifically, the administration was carried out every other day. Each time, 0.54 mg of the compound was dissolved in 100 μL of 0.5% PVP solution (specifically, the actual drug preparation process was prepared together according to the number of mice. For example, for 14 mice, 8 mg of the drug was dissolved in 1.5 ml of the solution, and each mouse was administered 100 μl). Drug administration was started from the successful induction of the model. The control group was given an equal volume of 0.5% PVP solution, and other treatment methods were the same.

[0555] The experimental results are as Figures 6 to 9 shown (where the 0 week on the abscissa represents the day when drug administration started after successful modeling, that is, the starting point of drug administration). From Figure 6It can be clearly seen that at 3 weeks after drug administration, the positive control T-5524 has not shown the effect of inhibiting the disease development. However, the compounds T3 and T5 of the present invention have significantly inhibited the disease development at 3 weeks after drug administration, showing obvious advantages. Thus, compared with the positive control T-5524, the compounds T3 and T5 of the present invention can inhibit the disease development earlier, with obvious advantages. Similarly, it can be seen that the compounds T8 to T16 of the present invention also have obvious therapeutic effects on HFpEF and show quick effects. It can be seen that the compounds of the present invention (especially the compounds of the series of formula I-1, formula I-1' and formula I-2) have obvious therapeutic effects on HFpEF and show quick effects.

[0556] In summary, the inventors of the present application have found that the compounds of the present invention have the activity of inhibiting the binding of JUN to DNA, have the activity of inhibiting JUN transcription, can produce therapeutic effects on HFpEF, have obvious advantages in the treatment of HFpEF, and have broad application prospects.

Claims

1. Use of a compound of formula I or a stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, pharmaceutically acceptable ester or pharmaceutically acceptable solvate thereof in the preparation of a medicament for the treatment and / or prevention of heart failure with preserved ejection fraction, wherein: R is selected from C6-C10 aryl, 6-10 membered heteroaryl, and each of the C6-C10 aryl and 6-10 membered heteroaryl is independently optionally substituted by R 1 and / or R 5 ; Preferably, R is selected from C6-C10 aryl and 6-10 membered heteroaryl, and each of the C6-C10 aryl and 6-10 membered heteroaryl is independently optionally substituted by R 1 or R 1 and R 5 substituted; More preferably, R is selected from C6-C10 aryl, 6- to 10-membered heteroaryl, and each of the C6-C10 aryl and 6- to 10-membered heteroaryl is independently optionally substituted by R 1 ; R 1 selected from hydrogen, C1-C6 alkyl, -COOH, -C(=O)O-C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl, -NR c R d ; R c 、R d each independently selected from hydrogen, C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl; m is selected from 0, 1, 2, 3, 4, 5, 6; Preferably, m is selected from 0, 1; More preferably, m is 1; R 2 selected from -O-(CH2) n -R 2’ ; n is selected from 0, 1, 2, 3, 4, 5, 6; Preferably, n is selected from 0, 1; R 2’ selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5-6 membered heteroaryl; R 3 selected from a hydroxyl group, -O-C1-C6 alkyl, wherein the C1-C6 alkyl is optionally substituted with a 5-6 membered heterocyclic group; Preferably, R 3 is selected from hydroxy, -O-C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted by a 6-membered heterocyclic group; More preferably, R 3 is selected from a hydroxyl group, -O-C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted by a morpholinyl group; Most preferably, R 3 is selected from hydroxy, -O-C1-C6 alkyl; R 4 selected from hydroxy, cyano, nitro, carboxyl, mercapto, halogen, C1-C6 alkyl, -NR a R b 、-CONR a R b , -O-C6-C10 aryl, -C(=O)-C1-C6 alkyl, -CH2-C6-C10 aryl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-C6-C10 aryl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, 5-6 membered heteroaryl; R a 、R b are each independently selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, wherein the phenyl is optionally substituted by C1-C6 alkyl; R 5 selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio; and the compound represented by Formula I does not contain 2. The use according to claim 1, wherein, R is selected from phenyl, benzimidazolyl, said phenyl, benzimidazolyl, each independently optionally substituted by R 1 and / or R 5 ; Preferably, R is selected from phenyl, benzimidazolyl, the phenyl, benzimidazolyl, each independently optionally substituted by R 1 or R 1 and R 5 substituted; More preferably, R is selected from phenyl, benzimidazolyl, said phenyl, benzimidazolyl, each independently optionally substituted by R 1 ; Alternatively, preferably, R is selected from phenyl, said phenyl, each independently optionally substituted by R 1 and / or R 5 ; Alternatively, more preferably, R is selected from phenyl, said phenyl, each independently optionally substituted by R 1 or R 1 and R 5 substituted; Alternatively, more preferably, R is selected from phenyl, said phenyl, each independently optionally substituted by R 1 ; Alternatively, preferably, R is selected from Alternatively, more preferably, R is selected from Preferably, R 1 is selected from hydrogen, -COOH, -C(=O)O-C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl, -NR c R d ; More preferably, R 1 is selected from hydrogen, -COOH, -C(=O)O-C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl; Preferably, R c , R d are each independently selected from hydrogen, -(CH2) m -O-C1-C6 alkyl; More preferably, R c , R d , any one of them is hydrogen, and the other is selected from -(CH2) m -O-C1-C6 alkyl; More preferably, R 1 is selected from hydrogen, -COOH, -COOCH3, -COOCH2CH3, -COO(CH2)2CH3, -COO(CH2)3CH3, -COO(CH2)4CH3, -COO(CH2)5CH3, More preferably, R 1 is selected from hydrogen, -COOH, -COOCH3, -COOCH2CH3, Most preferably, R 1 is selected from hydrogen, -COOH, -COOCH2CH3, Preferably, R 5 is selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy; More preferably, R 5 is selected from C1-C6 alkoxy groups; Most preferably, R 5 is a methoxy group.

3. Use according to any one of claims 1-2, wherein, R 2’ selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl, pyrazinyl (such as ), furyl (such as ); Preferably, R 2’ is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyrazinyl (such as ), furyl (such as ); Alternatively, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl; Alternatively, preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl; Alternatively, more preferably, R 2’ is selected from cyclopentyl, cyclohexyl, phenyl; Alternatively, more preferably, R 2’ is selected from cyclopentyl, phenyl; Preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl, -O-CH2-5-6 membered heteroaryl; More preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl, -O-CH2-5-6 membered heteroaryl, and the 5-6 membered heteroaryl is selected from pyrazinyl (such as ), furyl (such as ); Most preferably, R 2 is selected from Alternatively, preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl; Or more preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl; Alternatively, more preferably, R 2 is selected from Or most preferably, R 2 is selected from 4. Use according to any one of claims 1-3, wherein R 3 Selected from hydroxy, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3, Preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, Alternatively, preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3; Or more preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3; Alternatively, more preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3; Alternatively, or most preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3.

5. Use according to any one of claims 1 - 4, wherein, R 4 selected from hydroxyl, cyano, nitro, carboxyl, mercapto, halogen, C1-C6 alkyl, -NR a R b 、-CONR a R b 、phenyl, phenoxy, -C(=O)-C1-C6 alkyl, benzyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-phenyl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, thienyl (such as 2-thienyl); Preferably, R a , R b are each independently selected from hydrogen, methyl, Alternatively, preferably, R a , R b is independently selected from hydrogen, C1-C6 alkyl, and the other is selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, where the phenyl is optionally substituted by C1-C6 alkyl; Alternatively, more preferably, R a , R b is selected from the group consisting of hydrogen and methyl, and the other is selected from the group consisting of hydrogen, methyl, Preferably, R 4 is selected from the group consisting of hydroxy, cyano, nitro, carboxyl, -NH2, -SH, -CONH2, methyl, phenyl, benzyl, methoxy, phenoxy, acetyl, -F, -Cl, -Br, -CH=CH2, cyclopentyl, -COOMe, -COOPh, -SMe, -SOMe, -SO2Me, -NMe2, -NHAc, -NHMs, -NHTs, 2-thienyl; More preferably, R 4 is a hydroxyl group.

6. Use according to any one of claims 1-5, wherein The structural formula of the compound is as shown in formula I-1 or formula I-1', preferably, the structural formula of the compound is as shown in formula I-1, wherein: R 1 Selected from -COOH, -C(=O)O-C1-C6 alkyl; Preferably, R 1 is selected from -COOH, -COOCH3, -COOCH2CH3, -COO(CH2)2CH3, -COO(CH2)3CH3, -COO(CH2)4CH3, -COO(CH2)5CH3; More preferably, R 1 is selected from -COOH, -COOCH3, -COOCH2CH3; Most preferably, R 1 is selected from -COOH, -COOCH2CH3; R 2 selected from -O-(CH2) n -R 2’ n is selected from 0, 1, 2, 3, 4, 5, 6; Preferably, n is selected from 0, 1; R 2’ selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5-6 membered heteroaryl; Preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl, pyrazinyl (such as ), furyl (such as ); More preferably, R 2’ is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyrazinyl (such as ), furyl (such as ); Alternatively, preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl; Alternatively, more preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl; Alternatively, most preferably, R 2’ is selected from cyclopentyl, phenyl; Preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl, -O-CH2-5-6 membered heteroaryl; More preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl, -O-CH2-5-6 membered heteroaryl, and the 5-6 membered heteroaryl is selected from pyrazinyl (such as ), furyl (such as ); Most preferably, R 2 is selected from Alternatively, preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl; Or more preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl; Or most preferably, R 2 is selected from R 3 Selected from hydroxy, -O-C1-C6 alkyl; Preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -O(CH2)2CH3; More preferably, R 3 is a hydroxyl group; R 4 selected from hydroxyl, cyano, nitro, carboxyl, mercapto, halogen, C1-C6 alkyl, -NR a R b , -CONR a R b , C6-C10 aryl, -O-C6-C10 aryl, -C(=O)-C1-C6 alkyl, -CH2-C6-C10 aryl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-C6-C10 aryl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, 5-6 membered heteroaryl; Preferably, R 4 is selected from hydroxy, cyano, nitro, carboxyl, mercapto, halogen, C1-C6 alkyl, -NR a R b , -CONR a R b , phenyl, phenoxy, -C(=O)-C1-C6 alkyl, benzyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-phenyl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, thienyl (such as 2-thienyl); R a 、R b are each independently selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, and the phenyl is optionally substituted by C1-C6 alkyl; Preferably, R a and R b are each independently selected from hydrogen, methyl, Alternatively, preferably, R a , R b is selected from the group consisting of hydrogen, C1-C6 alkyl, and the other is selected from the group consisting of hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, and the phenyl is optionally substituted with C1-C6 alkyl; Alternatively, more preferably, R a , R b is selected from the group consisting of hydrogen and methyl, and the other is selected from the group consisting of hydrogen, methyl, More preferably, R 4 is selected from the group consisting of hydroxy, cyano, nitro, carboxyl, -NH2, -SH, -CONH2, methyl, phenyl, benzyl, methoxy, phenoxy, acetyl, -F, -Cl, -Br, -CH=CH2, cyclopentyl, -COOMe, -COOPh, -SMe, -SOMe, -SO2Me, -NMe2, -NHAc, -NHMs, -NHTs, 2-thienyl; Most preferably, R 4 is a hydroxyl group; R 5 selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio; Preferably, R 5 is selected from hydrogen, C1-C6 alkyl, C1-C6 alkoxy; More preferably, R 5 is selected from C1-C6 alkoxy groups; Most preferably, R 5 is a methoxy group.

7. Use according to any one of claims 1-5, wherein The structural formula of the compound is as shown in formula I-2, wherein: R 1 selected from hydrogen, -(CH2) m -O-C1-C6 alkyl, -NR c R d ; Preferably, R 1 is selected from hydrogen, -(CH2) m -O-C1-C6 alkyl; R c 、R d Each independently selected from hydrogen, C1-C6 alkyl, -(CH2) m -O-C1-C6 alkyl; Preferably, R c , R d are each independently selected from hydrogen, -(CH2) m -O-C1-C6 alkyl; More preferably, R c , R d Among them, any one is hydrogen, and the other is selected from -(CH2) m -O-C1-C6 alkyl; m is selected from 0, 1, 2, 3, 4, 5, 6; Preferably, m is selected from 0, 1; More preferably, m is 1; More preferably, R 1 is selected from hydrogen, R 2 Selected from -O-(CH2) n -R 2’ ; n is selected from 0, 1, 2, 3, 4, 5, 6; Preferably, n is selected from 0, 1; R 2’ selected from C1-C6 alkyl, C3-C8 cycloalkyl, C6-C10 aryl, 5- or 6-membered heteroaryl; Preferably, R 2’ is selected from C1-C6 alkyl, C3-C8 cycloalkyl, phenyl, pyrazinyl (such as ), furyl (such as ); More preferably, R 2’ is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, phenyl, pyrazinyl (such as ), furyl (such as ); Alternatively, preferably, R 2’ is selected from C3-C8 cycloalkyl; Alternatively, more preferably, R 2’ is selected from cyclopentyl, cyclohexyl; Or most preferably, R 2’ is selected from cyclopentyl; Preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-C6-C10 aryl, -O-CH2-5-6 membered heteroaryl; More preferably, R 2 is selected from -O-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -O-CH2-phenyl, -O-CH2-5-6 membered heteroaryl, and the 5-6 membered heteroaryl is selected from pyrazinyl (such as ), furyl (such as ); Most preferably, R 2 is selected from Or preferably, R 2 is selected from -O-C3-C8 cycloalkyl; Or more preferably, R 2 is selected from Or most preferably, R 2 is R 3 selected from a hydroxyl group, -O-C1-C6 alkyl, wherein the C1-C6 alkyl is optionally substituted with a 5-6 membered heterocyclic group; Preferably, R 3 is selected from a hydroxyl group, -O-C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted by a 6-membered heterocyclic group; More preferably, R 3 is selected from hydroxy, -O-C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted with a morpholinyl group; More preferably, R 3 is selected from hydroxyl, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3, More preferably, R 3 is selected from -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, Alternatively, preferably, R 3 is selected from hydroxy, -O-C1-C6 alkyl; Or more preferably, R 3 is selected from hydroxy, -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3; Alternatively, more preferably, R 3 is selected from a hydroxyl group, -OCH3, -OCH2CH3, -O(CH2)2CH3, -O(CH2)3CH3, -O(CH2)4CH3, -O(CH2)5CH3; Or preferably, R 3 is selected from -O-C1-C6 alkyl; Alternatively, more preferably, R 3 is selected from -OCH3, -OCH2CH3, -OCH(CH3)CH3, -O(CH2)2CH3; Alternatively, or most preferably, R 3 is selected from -OCH3, -OCH2CH3; R 4 selected from hydroxy, cyano, nitro, carboxyl, mercapto, halogen, C1-C6 alkyl, -NR a R b 、-CONR a R b 、C6-C10 aryl, -O-C6-C10 aryl, -C(=O)-C1-C6 alkyl, -CH2-C6-C10 aryl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-C6-C10 aryl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, 5-6 membered heteroaryl; Preferably, R 4 is selected from hydroxy, cyano, nitro, carboxyl, mercapto, halogen, C1-C6 alkyl, -NR a R b , -CONR a R b , phenyl, phenoxy, -C(=O)-C1-C6 alkyl, benzyl, C1-C6 alkoxy, C2-C6 alkenyl, C3-C8 cycloalkyl, -C(=O)O-C1-C6 alkyl, -C(=O)O-phenyl, C1-C6 alkylthio, -S(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, thienyl (such as 2-thienyl); R a and R b each independently selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, the phenyl optionally substituted by C1-C6 alkyl; Preferably, R a , R b are each independently selected from hydrogen, methyl, Alternatively, preferably, R a , R b is independently selected from hydrogen, C1-C6 alkyl, and the other is selected from hydrogen, C1-C6 alkyl, -C(=O)-C1-C6 alkyl, -S(=O)2-C1-C6 alkyl, -S(=O)2-phenyl, and the phenyl is optionally substituted with C1-C6 alkyl; Alternatively, more preferably, R a , R b Among them, any one is selected from hydrogen and methyl, and the other is selected from hydrogen, methyl, More preferably, R 4 is selected from hydroxy, cyano, nitro, carboxyl, -NH2, -SH, -CONH2, methyl, phenyl, benzyl, methoxy, phenoxy, acetyl, -F, -Cl, -Br, -CH=CH2, cyclopentyl, -COOMe, -COOPh, -SMe, -SOMe, -SO2Me, -NMe2, -NHAc, -NHMs, -NHTs, 2-thienyl; Most preferably, R 4 is a hydroxyl group; and the compound represented by Formula I-2 does not contain 8. The use according to any one of claims 1-5, wherein, The structural formula of the compound is as shown in formula I-3, wherein: R 1 selected from hydrogen, C1-C6 alkyl; Preferably, R 1 is hydrogen; R 2 selected from -O-C3-C8 cycloalkyl; Preferably, R 2 is R 3 selected from -O-C1-C6 alkyl; Preferably, R 3 is -OCH2CH3; R 4 is a hydroxyl group.

9. Use according to any one of claims 1 - 8, wherein, The compound is selected from the following: Or, Or, 10. Use of a pharmaceutical composition in the preparation of a medicament for the treatment and / or prevention of heart failure with preserved ejection fraction, wherein, The pharmaceutical composition comprises a compound of formula I or a stereoisomer, prodrug, crystal form, pharmaceutically acceptable salt, pharmaceutically acceptable ester or pharmaceutically acceptable solvate thereof, Among them, R, R 2 , R 3 , R 4 are each independently defined as in any one of claims 1-5; Preferably, the structural formula of the compound is as shown in formula I-1, formula I-1', formula I-2 or formula I-3, wherein the compound shown in formula I-1 or formula I-1' is defined in claim 6, the compound shown in formula I-2 is defined in claim 7, and the compound shown in formula I-3 is defined in claim 8; More preferably, the structural formula of the compound is as shown in formula I-1, formula I-2 or formula I-3, wherein the compound shown in formula I-1 is defined in claim 6, the compound shown in formula I-2 is defined in claim 7, and the compound shown in formula I-3 is defined in claim 8; Most preferably, the compound is as defined in claim 9.

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