Angiotensin ii type 2 receptor agonist and use thereof

Abstract

An AT2R agonist as represented by the compound of formula I, a pharmaceutical composition containing the compound, and the use thereof in the preparation of a drug for treating AT2R-mediated diseases.

Description

Angiotensin II type 2 receptor agonists and uses thereof

[0001] The present application claims priority to the prior application filed with the China National Intellectual Property Office on September 30, 2024, with the patent application number 202411381786.6, and the title of "Angiotensin II type 2 receptor (AT2R) agonists and uses thereof". The entire contents of the above-mentioned prior application are incorporated herein by reference. TECHNICAL FIELD

[0002] The present application belongs to the field of medicine, and specifically relates to angiotensin II type 2 receptor (AT2R) agonists and uses thereof, as well as the use of such compounds for treating diseases related to the field of pulmonary fibrosis. BACKGROUND

[0003] Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive fibrotic interstitial pneumonia of unknown etiology, with lesions limited to the lungs. It is more common in men than in women. The main manifestations are progressive dyspnea, accompanied by restrictive ventilatory dysfunction and gas exchange dysfunction, leading to hypoxemia and even respiratory failure, with a poor prognosis. The pulmonary histology and high-resolution CT (HRCT) of the chest show usual interstitial pneumonia (UIP). The median survival of IPF patients from diagnosis is 2-5 years, with a 5-year survival rate of 20%-25%. It is known as "cancer without cancer". (Pathogenesis and drug treatment progress of idiopathic pulmonary fibrosis [J]. China Medical Herald, 2021, 18(29): 45-48).

[0004] About 3 million people worldwide have IPF, with an incidence of about 3-13 / 100,000 and a prevalence of about 5-40 / 100,000 in the Asian population. In 2019, the prevalence of IPF in China was 2-29 / 100,000. The 2023 China Idiopathic Pulmonary Fibrosis Drug Industry Market Development Analysis pointed out that from 2015 to 2022, the number of people with IPF in China increased from 217,000 to 264,000. The conservative estimate of the number of people with IPF in our country is about 50-60 million. This disease mainly affects patients over the age of 50, and is more common in middle-aged and elderly male populations. With the progress of aging, improved diagnosis, and increased understanding of the disease, the number of confirmed cases of IPF is on the rise. It is estimated that in countries with inadequate diagnosis, such as China, Brazil, Russia, and India, there may be as many as a million patients who have not been diagnosed. (Data on file. Boehringer Ingelheim. DOF OFEV. RES. IPF / 08 / 11 January 2016. Worldwide Prevalence 2016; Podolanczuk AJ, et al. Eur Respir J. 2023 Apr 20; 61(4):2200957).

[0005] Recent studies have shown that angiotensin II (Ang II) acts on two GPCR (G protein-coupled receptor) subtypes, AT1R (Ang II type 1 receptor) and AT2R (Ang II type 2 receptor), and AT1R and AT2R have opposite physiological functions. AT2R is mainly expressed in type 2 alveolar epithelial cells and enteric glial cells. Under normal physiological conditions, type 2 alveolar epithelial cells are responsible for alveolar function repair.

[0006] Under the pathological condition of pulmonary fibrosis, type 2 alveolar epithelial cells lose their ability to repair and maintain alveolar integrity, scar formation, release of pro-inflammatory and fibrotic mediators, and stimulation of fibroblasts to produce excessive collagen fibers, leading to fibrosis. Agonizing AT2R can counter-regulate the effects of AT1R, exerting vasodilation, anti-fibrosis, anti-proliferation, anti-inflammatory, and anti-hypertensive effects, and offsetting the harmful effects of AT1R. (Catch your breath: The protective role of the angiotensin AT2 receptor for the treatment of idiopathic pulmonary fibrosis. 2023).

[0007] Although the preparation of compounds as Ang II receptor agonists is described in international patent application WO 2002 / 096883. Compound C21 (N-tert-butoxycarbonyl-3-(4-imidazol-1-ylmethylphenyl)-5- isobutylthiophene-2-sulfonamide) described in this document as a selective AT2 receptor agonist has also been indicated to have potential use in the treatment of stroke, spinal cord injury, sickle cell disease, muscular dystrophy, cancer treatment-related cardiotoxicity, peripheral neuropathy and systemic sclerosis (see, for example, international patent applications WO 2004 / 046141, WO 2016 / 092329, WO 2016 / 107879, WO 2016 / 139475, WO 2017 / 221012, WO 2019 / 008393 and US patent application US2012 / 035232), but there are still many problems to be solved in terms of drugability.

[0008] There are only 2 drugs for treating IPF-related diseases on the market in the world at present, namely pirfenidone and nintedanib. Due to the large side effects of the drugs, the drug withdrawal rate is very high. Within one year of use, the drug withdrawal rates of pirfenidone and nintedanib are 48.5% and 50.0%, respectively. In the future, with the progress of aging, the incidence and mortality rates are on the rise. Therefore, it is urgent to develop drugs with good efficacy and high safety to meet the huge clinical needs of IPF. SUMMARY

[0009] In a first aspect, the present application provides a compound of Formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt or prodrug thereof:

[0010] R 1 selected from C 1-6 alkyl;

[0011] Y is selected from CR 7 , R 7 selected from hydrogen, C 1-3 alkyl;

[0012] L is selected from O;

[0013] R 2 selected from C 1-6 alkyl, wherein the C 1-6 alkyl is optionally substituted with one or more hydrogen or C 3-7 cycloalkyl;

[0014] R 3 selected from halogen or C 1-3 alkyl;

[0015] n = 0 or 1;

[0016] R 4 selected from C1-4 alkyl or C 3-7 cycloalkyl, wherein the C 1-4 alkyl and C 3-7 cycloalkyl are each optionally substituted with hydrogen, halogen, hydroxyl, C 3-7 alkyl or C 1-6 cycloalkyl; R 5 and R 6 are each independently selected from hydrogen, halogen, C 1-4 alkyl.

[0017] In some embodiments, R 1 is selected from ethyl, n-propyl, i-propyl, n-butyl or i-butyl.

[0018] In some embodiments, Y is selected from CR 7 , R 7 is preferably selected from hydrogen or methyl.

[0019] In some embodiments, R 3 is selected from fluorine, chlorine or methyl.

[0020] In some embodiments, R 4 is selected from C 1-4 alkyl or C 3-5 cycloalkyl, wherein the C 1-4 alkyl and C 3-5 cycloalkyl can be substituted with halogen, hydroxyl, C 1-6 alkyl or C 3-5 cycloalkyl.

[0021] In some embodiments, the compound of formula I is represented by formula I-1 :

[0022] wherein R 1 is selected from ethyl, n-propyl, i-propyl, n-butyl or i-butyl; R 2 is selected from C 1-6 alkyl, wherein the C 1-6 alkyl can be substituted with C 3-7 cycloalkyl; n = 0 or 1 ; R 3 is selected from fluorine, chlorine or methyl; R 4 is selected from C 1-4 alkyl or C 3-5 cycloalkyl, wherein the C 1-4 alkyl and C 3-5 cycloalkyl can be substituted with halogen, hydroxyl, C 1-6 alkyl or C 3-5 cycloalkyl; R 5 , R 6 is selected from hydrogen; R 7 is selected from hydrogen or methyl.

[0023] In some embodiments, the compound of Formula I is represented by Formula I-2:

[0024] wherein R 2 selected from C 1-6 linear alkyl, said C 1-6 linear alkyl can be substituted with halogen or C 3-5 cycloalkyl; n = 0 or 1 ; R3is selected from methyl, fluoro or chloro; R 4 selected from methyl, ethyl, isopropyl, cyclopropyl, tert-butyl or C 1-4 fluoroalkyl, said methyl, ethyl, isopropyl, cyclopropyl, tert-butyl or C 1-4 fluoroalkyl can be substituted with halogen, hydroxyl, methyl or cyclopropyl; R 7 selected from hydrogen or methyl.

[0025] In some embodiments, the compound of Formula I is represented by Formula I-3:

[0026] wherein R 2 selected from C 1-3 linear alkyl, said C 1-3 linear alkyl can be substituted with one cyclopropyl; n = 0 or 1 ; R3is selected from methyl, fluoro or chloro; R 4 selected from methyl, ethyl, isopropyl, cyclopropyl or tert-butyl, said tert-butyl can be substituted with hydroxyl, said cyclopropyl can be substituted with methyl; R 7 selected from methyl or hydrogen.

[0027] In some embodiments, the compound is selected from any of the following:

[0028] In a second aspect, a pharmaceutical composition comprising a compound, stereoisomer or pharmaceutically acceptable salt thereof according to any of the first aspect, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0029] In a third aspect, use of a compound according to any of the first aspect, solvate, metabolite, co-crystal or prodrug thereof, or the composition according to the second aspect, in the manufacture of a medicament for the treatment or prevention of a disease mediated by AT2R.

[0030] wherein the disease comprises idiopathic pulmonary fibrosis. DETAILED DESCRIPTION

[0031] DETAILED DESCRIPTION

[0032] Unless otherwise indicated, the following terms have the meanings indicated below when used in the specification and claims.

[0033] "Alkyl" refers to saturated aliphatic hydrocarbon groups. Included are straight and branched chain groups having 1 to 20 carbon atoms. Medium size alkyl groups containing 1 to 6 carbon atoms are preferred, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, t-butyl, pentyl, and the like. Alkyl groups can be substituted or unsubstituted.

[0034] "C x-y " is intended to include groups that have from x to y carbons in the chain. For example, the term "C 1-6 Alkyl" refers to saturated aliphatic hydrocarbon groups. Included are straight and branched chain groups having 1 to 20 carbon atoms. Medium size alkyl groups containing 1 to 6 carbon atoms are preferred, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, t-butyl, pentyl, and the like. Alkyl groups can be substituted or unsubstituted.

[0035] "Cycloalkyl" refers to saturated carbocyclic rings. Exemplary cycloalkyl rings include cyclopropyl, cyclohexyl, and norbornane. Cycloalkyl groups can be optionally substituted with one or more substituents such as those described herein.

[0036] "Halo" means fluoro, chloro, bromo, or iodo, preferably fluoro or chloro.

[0037] By "optionally" is meant that the event or circumstance subsequently described can or can not occur, and that the description includes both the occurrence and non-occurrence of the event or circumstance.

[0038] In some embodiments, "substituted with one or more groups" means that one, two, three, or four hydrogen atoms in the designated atom or group are replaced with the same or different group selected from the indicated range of groups. Unless otherwise indicated, the structural formulas described herein include all isomeric forms (e.g., enantiomeric, diastereomeric, and geometric (or conformational) isomers) of the structures: for example, the R, S configuration of asymmetric centers, the (Z), (E) isomers of double bonds, and the (Z), (E) conformational isomers. Thus, individual stereochemical isomers or mixtures thereof, or enantiomeric, diastereomeric, or geometric (or conformational) isomers of the compounds of the present application are within the scope of the present application.

[0039] The use of the terms "stereochemical" and "stereoisomeric" are used in the present application generally refer to the following references: S. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., Stereochemistry of Organic Compounds, John Wiley & Sons, Inc., New York, 1994. The compounds of the present application can contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. All stereoisomeric forms of the compounds of the present application, including but not limited to, diastereomeric, enantiomeric, atropisomeric, and their mixtures, such as racemic mixtures, form part of the present application. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing an optically active compound, the prefixes D, L or R, S are used to denote the absolute configuration of the molecule. The prefixes d and 1 or (+) and (-) are employed to designate the sign of the rotation of plane-polarized light by the compound, with (+) or 1 meaning that the compound is dextrorotary. The (-) or d form is levorotary. The chemical structures of these stereoisomers are identical but their orientations in space are mirror images of each other. A specific stereoisomer can be referred to as an enantiomer if its mirror image is not superimposable. Mixtures of enantiomers are often referred to as enantiomeric mixtures. A 50:50 mixture of enantiomers is referred to as a racemic mixture or racemate, which can result from racemic synthesis or resolution of a single enantiomer. The term "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomeric forms in which the specific orientations of the chiral molecules are random.

[0040] "stereoisomers" refer to compounds which have the same chemical constitution, but differ in the arrangement of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotamers), geometric isomers (cis / trans), atropisomers, and the like.

[0041] The term "tautomer" or "tautomeric forms" refer to structural isomers that exist in equilibrium with each other. For example, prototropic tautomers (i.e., prototropically shiftable tautomers) include tautomers that shift by the movement of a proton, such as keto-enol and imine-enamine isomerizations. Atomic valence (bond order) tautomers include tautomers that shift by the movement of a bond electron.

[0042] "chiral" is a molecule that has the property of not being superimposable with its mirror image; whereas "achiral" refers to a molecule that is superimposable with its mirror image.

[0043] "Enantiomer" refers to two isomers of a compound that are nonsuperimposable mirror images of one another.

[0044] "Diastereomer" refers to a stereoisomer that has two or more chiral centers and whose molecules are not a mirror image of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties, and reactivity. Mixtures of diastereomers can be separated by high resolution analytical techniques such as electrophoresis and chromatography, for example HPLC.

[0045] In certain embodiments, the compositions of the present disclosure can comprise two or more enantiomers or diastereomers of a compound, wherein a single enantiomer or diastereomer comprises at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 98% by weight, or at least about 99% by weight or more of the total weight of all stereoisomers. Methods of producing substantially pure enantiomers are well known to those skilled in the art.

[0046] In addition, substitution with heavier isotopes, particularly deuterium, i.e., 2 H or D, can afford certain therapeutic advantages. These advantages can result from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements or an improvement in therapeutic index. It is understood that deuterium is regarded as a reasonable substituent in compounds of formula (I), formula (II), formula (III), and formula (IV).

[0047] Unless otherwise stated, the compounds described herein are intended to include both vaults and vaults differing only in the presence of one or more isotopically enriched atoms. For example, compounds having the structure of the present disclosure, except for the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by a carbon enriched in 13 C or 14 C, are within the scope of the present disclosure. The compounds of the present disclosure optionally contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds can be labeled with isotopes such as, for example, deuterium (2H), tritium (3H), iodine 3 H), carbon 125 ( 125 I), or carbon 14 ( 14 C). Compounds 2 H, 11 C, 13 C, 14 C, 15 C, 12 N, 13 N, 15 N, 16 N, 16 O, 17 O, 14 F, 15 F,16 F、 17 F、 18 F、 33 S、 34 S、 35 S、 36 S、 35 Cl、 37 Cl、 79 Br、 81 Br and 125 Isotopic substitutions of all the elements of the compounds of the present application are contemplated. All isotopically labeled compounds of the present application, whether or not radioactive, are within the scope of the present application.

[0048] "Pharmaceutically acceptable salt" means those salts which retain the biological effectiveness and properties of the parent compound. Such salts include:

[0049] (1) salts with acids, by reaction of the free base of the parent compound with an inorganic or organic acid, inorganic acids include hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid and perchloric acid, and organic acids include acetic acid, propionic acid, acrylic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, hydroxybenzoic acid, gamma-hydroxybutyric acid, methoxybenzoic acid, phthalic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene- 1 -sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, mandelic acid, succinic acid or malonic acid, and the like.

[0050] (2) salts of metal ions or of organic bases which replace the acidic protons of the parent compound or coordinate with the parent compound, examples of metal ions include alkali metal ions, alkaline earth metal ions or aluminum ions, examples of organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like.

[0051] "Pharmaceutical composition" means a mixture of one or more of the compounds of the present application or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof with another chemical component, such as a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate administration of the compound to an animal.

[0052] "Solvate" means an association or complex of one or more solvent molecules with one of the compounds of the present application. Solvents which form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid and aminoethanol. The term "hydrate" means a solvate formed by the association or complexing of one or more water molecules with one of the compounds of the present application.

[0053] "Pharmaceutically acceptable carrier" means a carrier, diluent or excipient having no significant inductive or suppressive effect on the biological activity or properties of the compound administered.

[0054] Non-active ingredients in the pharmaceutical composition of matter, such as but not limited to: calcium carbonate, calcium phosphate, various sugars (e.g., lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic or methacrylic polymers, gels, water, polyethylene glycol, propylene glycol, ethylene glycol, castor or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil, etc.

[0055] In addition to the pharmaceutically acceptable carrier, the aforementioned pharmaceutical composition can further include a pharmaceutically common adjuvant, such as: antibacterial agents, antifungal agents, antimicrobial agents, preservatives, toning agents, solubilizers, thickeners, surfactants, complexing agents, proteins, amino acids, fats, sugars, vitamins, minerals, trace elements, sweeteners, pigments, fragrances, or combinations thereof, etc.

[0056] Specific embodiments

[0057] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0058] The structure of the compound is determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). The NMR determination is performed by a (Bruker Avance III 400) nuclear magnetic instrument, and the determination solvent is deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and the internal standard is tetramethylsilane (TMS). The LC-MS determination uses a Shimadzu liquid chromatograph-mass spectrometer (Shimadzu LC-MS 2020 (ESI)). The HPLC determination uses a Shimadzu high-pressure liquid chromatograph (Shimadzu LC-20A). The MPLC (medium pressure preparative chromatography) uses a Gilson GX-281 reverse-phase preparative chromatograph. The thin-layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, and the specification used for thin-layer chromatography separation and purification of the product is 0.4mm-0.5mm. The silica gel chromatographic column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier. The SFC chiral resolution uses a Shimadzu high-pressure liquid chromatograph (Shimadzu LC-30AD SFC).

[0059] Commonly used organic solvents in the examples are expressed using abbreviations well known in the art, such as: DCM for dichloromethane; THF for tetrahydrofuran; Dioxane for 1,4 dioxane; DMF for N,N dimethylformamide; MeOH for methanol; EtOH for ethanol; HOAc for acetic acid, and the like.

[0060] Example 1. N-((5-isobutyl-3-(4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)thiophen-2- yl)sulfonyl)-1-propyloxycyclopropane-1-carboxamide (Compound 1)

[0061] Synthetic Route:

[0062] Step 1: Preparation of N-(tert-butyl)thiophene-2-sulfonamide (1b)

[0063] Tert-butylamine (41.75 g, 570.81 mmol) was slowly added to a stirring solution of 2- thiophenesulfonyl chloride (25.00 g, 136.88 mmol) in dry tetrahydrofuran (250 mL) at 0 °C. The reaction was allowed to warm to room temperature and was left under nitrogen protection for 16 h. The reaction was quenched with water and extracted with ethyl acetate. The combined organic layers were dried, concentrated, and the target compound N-(tert-butyl)thiophene-2-sulfonamide (1b) was obtained. The product was used in the next step without further purification.

[0064] 1 H NMR (400 MHz, CDCl3) δ 7.61 (dd, J = 3.6, 1.2 Hz, 1H), 7.54 (dd, J = 5.2, 1.2 Hz, 1H), 7.04 (dd, J = 5.0, 3.8 Hz, 1H), 4.66 (s, 1H), 1.29 (s, 9H).

[0065] Step 2: Preparation of N-(tert-butyl)-5-isobutylthiophene-2-sulfonamide (1c)

[0066] To a stirred solution of N-(tert-butyl)thiophene-2-sulfonamide (1b) (10.00 g, 45.60 mmol) in dry tetrahydrofuran (50 mL) was added dropwise n-butyllithium (1.6 M in n-hexane) (75 mL, 120.00 mmol) at -78 °C. After the addition was complete, the reaction was allowed to warm to -40 °C and stirred for 2 h under nitrogen. Iodoisobutane (16.45 g, 89.37 mmol) was added, then the reaction was allowed to warm to room temperature and stirred for 16 h under nitrogen. The reaction was quenched by the slow addition of saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The combined organic layers were dried, concentrated and the crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 10 / 1) to give the target compound N-(tert-butyl)-5-isobutylthiophene-2-sulfonamide (1c).

[0067] 1 H NMR (400 MHz, DMSO-d6) δ 7.16 (s, 1H), 6.98 (s, 2H), 6.38 (s, 1H), 2.68 (d, J = 7.0 Hz, 2H), 1.90 - 1.80 (m, 1H), 1.15 (s, 9H), 0.91 (d, J = 6.4 Hz, 6H).

[0068] Step 3: Preparation of (2-(N-(tert-butyl)sulfamoyl)-5-isobutylthiophen-3-yl)boronic acid (1d)

[0069] To a stirred solution of N-(tert-butyl)-5-isobutylthiophene-2-sulfonamide (1c) (3.50 g, 12.71 mmol) in dry tetrahydrofuran (500 mL) was added dropwise n-butyllithium (1.6 M in n-hexane) (23.9 mL, 38.24 mmol) at -78 °C. After the addition was complete, the reaction was stirred for 4 h at -20 °C under nitrogen, and triisopropyl borate (4.78 g, 25.42 mmol) was added. The reaction was then allowed to warm to room temperature and stirred for 16 h under nitrogen. The reaction was quenched by the slow addition of saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The combined organic layers were dried, concentrated and the crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to give the target compound (2-(N-(tert-butyl)sulfamoyl)-5-isobutylthiophen-3-yl)boronic acid (1d).

[0070] 1 H NMR (400 MHz, DMSO-d6) δ 7.16 (s, 1H), 6.98 (s, 2H), 6.38 (s, 1H), 2.68 (d, J = 7.0 Hz, 2H), 1.90 - 1.80 (m, 1H), 1.15 (s, 9H), 0.91 (d, J = 6.4 Hz, 6H).

[0071] Step 4: Preparation of 1-(4-bromobenzyl)-2-methyl-1H-imidazole (1f)

[0072] Sodium hydride (60%, 0.34 g, 8.50 mmol) was added portionwise to a solution of 2-methyl-1H-imidazole (0.66 g, 8.00 mmol) in N,N-dimethylformamide (12 mL) at 0 °C under nitrogen. After stirring for 30 min, 1-bromo-4-(bromomethyl)benzene (2.00 g, 8.00 mmol) was added. The reaction was stirred at room temperature for 1 h under nitrogen. The reaction was quenched by the addition of saturated aqueous ammonium chloride solution. The mixture was extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 100 / 1-10 / 1) to give 1-(4-bromobenzyl)-2-methyl-1H-imidazole (1f).

[0073] LC-MS (ESI) calcd for C 11 H 11 BrN2[M+H] + m / z 251.0; 253.0, found 252.9.

[0074] Step 5: Preparation of N-(tert-butyl)-5-isobutyl-3-(4-((2-methyl-1H-imidazol-1- yl)methyl)phenyl)thiophene-2-sulfonamide (1g):

[0075] Sodium hydride (60%, 0.34 g, 8.50 mmol) was added portionwise to a solution of 2-methyl-1H-imidazole (0.66 g, 8.00 mmol) in N,N-dimethylformamide (12 mL) at 0 °C under nitrogen. After stirring for 30 min, 1-bromo-4-(bromomethyl)benzene (2.00 g, 8.00 mmol) was added. The reaction was stirred at room temperature for 1 h under nitrogen. The reaction was quenched by the addition of saturated aqueous ammonium chloride solution. The mixture was extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 100 / 1-10 / 1) to give 1-(4-bromobenzyl)-2-methyl-1H-imidazole (1f).

[0076] LC-MS (ESI) calcd for C 23 H 31 N3O2S2[M+H] + m / z 446.2, found 446.3.

[0077] Step 6: Preparation of 5-isobutyl-3-(4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)thiophene-2-sulfonamide (1h)

[0078] N-(tert-butyl)-5-isobutyl-3-(4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)thiophene-2-sulfonamide (lg) (1.05 g, 2.52 mmol) was dissolved in trifluoroacetic acid (8 mL) at room temperature under nitrogen protection, anisole (0.12 mL) was added with stirring. The reaction was then heated to 35 °C and stirred for 16 hours before being terminated. The reaction was concentrated, the resulting crude product was purified by reverse phase C18 column chromatography (mobile phase: 0.1% formic acid in water / acetonitrile) to give 5-isobutyl-3-(4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)thiophene-2-sulfonamide (lh).

[0079] LC-MS (ESI) calcd for C 19 H 23 N3O2S2[M+H] + m / z 390.1, found 390.0.

[0080] Step 7: Preparation of N-((5-isobutyl-3-(4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)thiophene-2-yl)sulfonyl)-l-propyloxycyclopropane-l- carboxamide (Compound 1)

[0081] Dissolve 1 -methoxycyclopropane-1 -carboxylic acid (0.07 g, 0.49 mmol) in dichlorosulfoxide (2 mL) at room temperature under nitrogen. Heat the reaction to 80 °C and stir for 2 hours under nitrogen. Concentrate the cooled reaction and dissolve the residue in dichloromethane (5 mL) at room temperature with stirring add N,N-diisopropylethylamine (0.20 g, 1.54 mmol) and 5-isobutyl-3-(4-((2-methyl-1 H-imidazol-1 -yl)methyl)phenyl)thiophene-2-sulfonamide (1h) (0.10 g, 0.26 mmol) sequentially. Terminate the reaction after 30 minutes of stirring at room temperature. Concentrate the reaction and isolate the crude product by high performance liquid preparative chromatography (mobile phase: 0.1% formic acid in water / acetonitrile) to give the target product N-((5-isobutyl-3-(4-((2-methyl-1 H-imidazol-1 -yl)methyl)phenyl)thiophen-2-yl)sulfonyl)-1 - methoxycyclopropane-1 -carboxamide (Compound 2).

[0082] LC-MS (ESI) calcd for C 26 H 33 N3O4S2[M+H] + m / z 516.2, found 516.3.

[0083] 1 H NMR (400 MHz, DMSO-d6) δ 7.74 (d, J = 8.2 Hz, 2H), 7.65 (d, J = 1.8 Hz, 1H), 7.50 (s, 1H), 7.28 (d, J = 8.3 Hz, 2H), 6.79 (s, 1H), 5.35 (s, 2H), 3.26 (t, J = 6.7 Hz, 2H), 2.62 (d, J = 6.9 Hz, 2H), 2.57 (s, 3H), 1.89 - 1.81 (m, 1H), 1.38 - 1.30 (m, 2H), 0.94 (d, J = 6.6 Hz, 6H), 0.79 - 0.73 (m, 5H), 0.57 (s, 2H).

[0084] Example 2. N-((5-isobutyl-3-(4-((2-methyl-1 H-imidazol-1 -yl)methyl)phenyl)thiophen-2- yl)sulfonyl)-1 -methoxycyclopropane-1 -carboxamide (Compound 2)

[0085] Synthetic Route:

[0086] Step 1 : Preparation of N-((5-isobutyl-3-(4-((2-methyl-1 H-imidazol-1 -yl)methyl)phenyl)thiophen- 2-yl)sulfonyl)-1 -methoxycyclopropane-1 -carboxamide (Compound 2)

[0087] Using 1 -methoxy cyclopropanecarboxylic acid as reactant, the synthetic protocol of Step 7 of Reference Example 1 was referred to give the target compound N-((5-isobutyl-3-(4-((2-methyl-1 H-imidazol-1 -yl)methyl)phenyl)thiophen-2-yl)sulfonyl)-1 - methoxycyclopropane-1 -carboxamide (Compound 2).

[0088] LC-MS (ESI) calcd for C 24 H 29 N3O4S2[M+H] + m / z 488.2, found 488.3.

[0089] 1 H NMR (400 MHz, DMSO-d6) δ 7.73 (d, J = 8.1 Hz, 2H), 7.66 (d, J = 1.7 Hz, 1H), 7.51 (s, 1H), 7.30 (d, J = 8.2 Hz, 2H), 6.79 (s, 1H), 5.35 (s, 2H), 3.00 (s, 3H), 2.63 (d, J = 7.0 Hz, 2H), 2.58 (s, 3H), 1.88 - 1.81 (m, 1H), 0.93 (d, J = 6.6 Hz, 6H), 0.75 - 0.72 (m, 2H), 0.59 (s, 2H).

[0090] Example 3. N-((5-isobutyl-3-(3-methyl-4-((2-methyl-1 H-imidazol-1 - yl)methyl)phenyl)thiophen-2-yl)sulfonyl)-1 -propoxycyclopropane-1 -carboxamide (Compound 3)

[0091] Synthetic route:

[0092] Step 1: Preparation of (4-bromo-2-methylphenyl)methanol (3b)

[0093] To a stirred solution of 4-bromo-2-methylbenzoic acid (11.00 g, 51.16 mmol) in dry tetrahydrofuran (50 mL) was added borane tetrahydrofuran solution (1 M) (103 mL, 103.00 mmol) slowly under nitrogen atmosphere at 0 °C. The reaction was terminated after stirring at room temperature under nitrogen atmosphere for 16 hours. The reaction was quenched by adding water and extracted with ethyl acetate. The combined organic phase was washed with saturated aqueous ammonium chloride solution and saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the crude product was isolated by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 10 / 1) to give (4-bromo-2-methylphenyl)methanol (3b).

[0094] LC-MS (ESI) calcd for C8H9BrO [M-OH] + m / z 183.0; 185.0, found 182.8.

[0095] 1 H NMR (400 MHz, CDC13) δ 7.30 - 7.26 (m, 2H), 7.16 (d, J = 8.8 Hz, 1H), 4.54 (s, 2H), 2.25 (s, 3H).

[0096] Step 2: Preparation of 4-bromo-1-(chloromethyl)-2-methylbenzene (3c)

[0097] To a stirred solution of (4-bromo-2-methylphenyl)methanol (3b) (9.00 g, 45.00 mmol) in dichloromethane (90 mL) was added triethylamine (10.50 mL, 77.50 mmol) at 0 °C, followed by the slow addition of methylsulfonyl chloride (6.30 mL, 77.50 mmol). The reaction was stirred at room temperature for 16 h before being quenched. Water was added to the reaction and extracted with dichloromethane. The combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 10 / 1) to give 4-bromo-1-(chloromethyl)-2-methylbenzene (3c).

[0098] 1 H NMR (400 MHz, CDC13) δ 7.34 (d, J = 1.5 Hz, 1H), 7.30 (dd, J = 8.1, 2.0 Hz, 1H), 7.16 (d, J = 8.1 Hz, 1H), 4.53 (s, 2H), 2.38 (s, 3H).

[0099] Step 3: Preparation of 1-(4-bromo-2-methylbenzyl)-2-methyl-1H-imidazole (3d)

[0100] To a stirred solution of 2-methyl-lH-imidazole (3.76 g, 45.85 mmol) in N,N- dimethylformamide (50 mL) was added 60% sodium hydride (1.74 g, 43.5 mmol) portionwise at 0 °C under nitrogen. The reaction was stirred at 0 °C for 30 min. 4-Bromo-l- (chloromethyl)-2-methylbenzene (3c) (5.00 g, 22.94 mmol) was added. The reaction was stirred at room temperature for 16 h. The reaction was quenched with water and extracted with ethyl acetate. The combined organic phase was washed with saturated aqueous ammonium chloride and brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 5 / 1) to give l-(4-bromo-2-methylbenzyl)-2-methyl-lH-imidazole (3d).

[0101] LC-MS (ESI) calcd for C 12 H 13 BrN2[M+H] + m / z 265.0; 267.0, found 266.7.

[0102] Step 4: Preparation of N-(tert-butyl)-5-isobutyl-3-(3-methyl-4-((2-methyl-lH- imidazol-l-yl)methyl)phenyl)thiophene-2-sulfonamide (3e)

[0103] To a stirred solution of l-(4-bromo-2-methylbenzyl)-2-methyl-lH-imidazole (3d) (1.40 g, 5.30 mmol) in 1,4-dioxane (10 mL) and water (2 mL) was added (2-(N-(tert- butyl)sulfamoyl)-5-isobutylthiophen-3-yl)boronic acid (Id) (2.00 g, 6.26 mmol), potassium carbonate (2.20 g, 15.94 mmol) and [l,l'-bis(diphenylphosphino) ferrocene] palladium dichloride (0.39 g, 0.53 mmol) sequentially at room temperature under nitrogen. The reaction was stirred at 100 °C for 16 h under nitrogen. The cooled reaction was filtered and the filtrate was concentrated. The crude product was purified by column chromatography on silica gel (eluent: dichloromethane / methanol = 10 / 1) to give N-(tert-butyl)-5-isobutyl-3-(3-methyl-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)thiophene-2-sulfonamide (3e).

[0104] LC-MS (ESI) calcd for C 24 H 33 N3O2S2[M+H] + m / z 460.2, found 460.2.

[0105] 1 H NMR (400 MHz, CDC13) δ 7.47 (s, 1H), 7.38 (d, J = 7.9 Hz, 1H), 6.97 (s, 1H), 6.79-6.72 (m, 3H), 5.04 (s, 2H), 2.68 (d, J = 7.1 Hz, 2H), 2.36 (s, 3H), 2.34 (s, 3H), 1.96-1.88 (m, 1H), 1.03 (s, 9H), 0.97 (d, J = 6.6 Hz, 6H).

[0106] Step 5: Preparation of 5-isobutyl-3-(3-methyl-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)thiophene-2-sulfonamide (3f)

[0107] To a stirred solution of N-(tert-butyl)-5-isobutyl-3-(3-methyl-4-((2-methyl-lH- imidazol-l-yl)methyl)phenyl)thiophene-2-sulfonamide (3e) (2.20 g, 4.67 mmol) in trifluoroacetic acid (20 mL) was added anisole (2.2 mL) at room temperature. The reaction was then heated to 30 °C and stirred for 16 h before it was terminated. The reaction was concentrated and the resulting crude product was purified by reverse phase C18 column chromatography (eluent: 0.1% formic acid in water / acetonitrile = 7 / 3) to give 5-isobutyl-3-(3-methyl-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)thiophene-2-sulfonamide (3f).

[0108] 1 H NMR (400 MHz, CDC13) δ 7.45 (s, 1H), 7.36 (d, J = 7.9 Hz, 1H), 6.93 (d, J = 1.2 Hz, 1H), 6.77-6.74 (m, 2H), 6.71 (d, J = 8.0 Hz, 1H), 5.03 (s, 2H), 2.68 (d, J = 7.2 Hz, 2H), 2.34 (s, 6H), 1.97-1.88 (m, 1H), 0.98 (d, J = 6.8 Hz, 6H).

[0109] Step 6: Preparation of N-((5-isobutyl-3-(3-methyl-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)thiophene-2-yl)sulfonyl)-l-propyloxycyclopropane-l-carboxamide (Compound 3)

[0110] Dissolve 1-ethoxy cyclopropane-1 -carboxylic acid (0.08 g, 0.52 mmol) in sulfurous anhydride (1 mL) at room temperature, heat to 80 °C and stir for 2 hours. Concentrate the reaction under reduced pressure to give a yellow oil. To the resulting oil, add dry dichloromethane (1 mL) and add N,N-diisopropylethylamine (0.13 g, 1.04 mmol) followed by 5-isobutyl-3-(3-methyl-4-((2-methyl-1 H-imidazol-1 -yl)methyl)phenyl)thiophene-2-sulfonamide (3f) (0.07 g, 0.17 mmol) at room temperature with stirring. The resulting mixture is stirred at room temperature for 2 hours and then quenched. Concentrate the reaction and separate the resulting residue by high performance liquid chromatography, preparative (mobile phase: 0.1% formic acid in water / acetonitrile) to give N-((5-isobutyl-3-(3-methyl-4-((2-methyl-1 H-imidazol-1 -yl)methyl)phenyl)thiophen-2-yl)sulfonyl)-1 -ethoxycyclopropane-1 -carboxamide (Compound 4).

[0111] LC-MS (ESI) calcd for C 27 H 35 N3O4S2[M+H] + m / z 530.2, found 530.1.

[0112] 1 H NMR (400 MHz, DMSO-d6) δ 7.57 (d, J = 8.1 Hz, 1H), 7.54 (s, 1H), 7.51 (d, J = 1.9 Hz, 1H), 7.41 (d, J = 1.9 Hz, 1H), 6.79 (s, 1H), 6.74 (d, J = 8.0 Hz, 1H), 5.37 (s, 2H), 3.28 (t, J = 6.7 Hz, 2H), 2.63 (d, J = 7.0 Hz, 2H), 2.53 (s, 3H), 2.31 (s, 3H), 1.91 - 1.76 (m, 1H), 1.41 - 1.29 (m, 2H), 0.94 (d, J = 6.6 Hz, 6H), 0.84 - 0.79 (m, 2H), 0.76 (t, J = 7.4 Hz, 3H), 0.66 (d, J = 2.9 Hz, 2H).

[0113] Example 4. 1 -Ethoxy-N-((5-isobutyl-3-(3-methyl-4-((2-methyl-1 H-imidazol-1 - yl)methyl)phenyl)thiophen-2-yl)sulfonyl)cyclopropane-1 -carboxamide (Compound 4)

[0114] Synthetic Route:

[0115] Step 1: Preparation of l-ethoxy-N-((5-isobutyl-3-(3-methyl-4-((2-methyl-lH- imidazol-l-yl)methyl)phenyl)thiophen-2-yl)sulfonyl)cyclopropane-l-carboxamide (Compound 4)

[0116] Using l-ethoxycyclopropane carboxylic acid as the reactant, the synthetic scheme of Example 3, Step 6 was referenced to obtain the target compound, l-ethoxy-N-((5-isobutyl-3-(3-methyl-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)thiophen-2-yl)sulfonyl)cyclopropane-l-carboxamide (Compound 4).

[0117] LC-MS (ESI) calcd for C 26 H 33 N3O4S2[M+H] + m / z 516.2, found 516.3.

[0118] 1 H NMR (400 MHz, DMSO-d6) δ 7.58 - 7.55 (m, 2H), 7.36 (s, 1H), 7.32 (s, 1H), 6.77 (s, 1H), 6.68 (d, J = 8.5 Hz, 1H), 5.32 (s, 2H), 3.35 (q, J = 7.0 Hz, 2H), 2.61 (d, J = 7.0 Hz, 2H), 2.47 (s, 3H), 2.30 (s, 3H), 1.86 - 1.81 (m, 1H), 0.97 - 0.92 (m, 9H), 0.79 (dd, J = 7.1, 3.9 Hz, 2H), 0.62 (dd, J = 6.8, 3.7 Hz, 2H).

[0119] Example 5. N-((5-isobutyl-3-(3-methyl-4-((2-methyl-lH-imidazol-l-yl)methyl)phenyl)thiophen-2- yl)sulfonyl)-l-methoxycyclopropane-l-carboxamide (Compound 5)

[0120] Synthetic Route:

[0121] Step 1: Preparation of N-((5-isobutyl-3-(3-methyl-4-((2-methyl-lH-imidazol-l-yl)methyl)phenyl)thiophen-2- yl)sulfonyl)-l-methoxycyclopropane-l-carboxamide (Compound 5)

[0122] Using 1-methoxycyclopropane carboxylic acid as the reactant, the synthetic protocol of Reference Example 3, Step 6 was followed to obtain the target compound N-((5-isobutyl-3-(3-methyl-4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)thiophen-2-yl)sulfonyl)-1-methoxycyclopropane-1-carboxamide (Compound 5).

[0123] LC-MS (ESI) calcd for C 25 H 31 N3O4S2[M+H] + m / z 502.2, found 502.3.

[0124] 1 H NMR (400 MHz, CDCl3) δ 7.40-7.36 (m, 2H), 7.16 (s, 1H), 6.87 (d, J = 7.8 Hz, 1H), 6.82 (s, 1H), 6.73 (s, 1H), 5.13 (s, 2H), 3.19 (s, 3H), 2.71 (d, J = 7.0 Hz, 2H), 2.66 (s, 3H), 2.31 (s, 3H), 1.96-1.93 (m, 1H), 1.22 (dd, J = 8.2, 5.0 Hz, 2H), 1.07 (dd, J = 8.2, 5.0 Hz, 2H), 0.99 (d, J = 6.6 Hz, 6H).

[0125] Example 6. N-((3-(3-Fluoro-4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)-1-propoxycyclopropane-1-carboxamide (Compound 6)

[0126] Synthetic Route:

[0127] Step 1: Preparation of 1-(4-bromo-2-fluorobenzyl)-2-methyl-1H-imidazole (6b)

[0128] To a stirred solution of 2-methylimidazole (0.68 g, 8.27 mmol) in N,N- dimethylformamide (10 mL) was added 60% sodium hydride (0.32 g, 7.90 mmol) in portions at 0 °C under nitrogen. The reaction was stirred at room temperature for 30 min under nitrogen, then 4-bromo-1-(bromomethyl)-2-fluorobenzene (6a) (2.00 g, 7.52 mmol) was added. The reaction was stirred at room temperature for 16 h under nitrogen, then quenched with saturated aqueous ammonium chloride solution. The reaction mixture was extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the residue was purified by column chromatography on silica gel (eluent: dichloromethane / methanol = 10 / 1) to give 1-(4-bromo-2-fluorobenzyl)-2-methyl-1H-imidazole (6b).

[0129] LC-MS (ESI) calcd for C 11 H 10 BrFN2[M+H] + m / z 269.0; 271.0, found 269.0.

[0130] 1 H NMR (400 MHz, CDCl3) δ 7.31-7.22 (m, 2H), 6.95 (d, J = 1.3 Hz, 1H), 6.84 (d, J = 1.3 Hz, 1H), 6.73 (t, J = 8.1 Hz, 1H), 5.04 (s, 2H), 2.36 (s, 3H).

[0131] Step 2: Preparation of N-(tert-butyl)-3-(3-fluoro-4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)-5- isobutylthiophene-2-sulfonamide (6c)

[0132] To a mixture of 1-(4-bromo-2-fluorobenzyl)-2-methyl-1H-imidazole (6b) (0.70 g, 2.61 mmol) in 1,4 dioxane (10 mL) and water (2 mL) was added (2-(N-(tert-butyl)sulfamoyl)-5-isobutylthiophen-3-yl)boronic acid (1d) (1.00 g, 3.13 mmol), potassium carbonate (1.10 g, 7.97 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (0.19 g, 0.26 mmol) sequentially at room temperature under nitrogen. The reaction was then heated to 110 °C and stirred for 3 hours under nitrogen before being terminated. To the cooled reaction was added water and extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the residue was separated by column chromatography on silica gel (eluent: dichloromethane / methanol = 10 / 1) to give N-(tert-butyl)-3-(3-fluoro-4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)-5-isobutylthiophene-2-sulfonamide (6c).

[0133] LC-MS (ESI) calcd for C 23 H 30 FN3O2S2[M+H] + m / z 464.2, found 464.5.

[0134] 1 H NMR (400 MHz, CDCl3) δ 7.40 (d, J = 10.8 Hz, 1H), 7.35 (d, J = 7.9 Hz, 1H), 6.96 (t, J = 7.7 Hz, 2H), 6.88 (s, 1H), 6.72 (s, 1H), 5.13 (s, 2H), 2.68 (d, J = 7.1 Hz, 2H), 2.39 (s, 3H), 1.95-1.87 (m, 1H), 1.05 (s, 9H), 0.97 (d, J = 6.6 Hz, 6H).

[0135] Step 3: Preparation of 3-(3-fluoro-4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)-5- isobutylthiophene-2-sulfonamide (6d)

[0136] To a stirred solution of N-(tert-butyl)-3-(3-fluoro-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)-5-isobutylthiophene-2-sulfonamide (6c) (1.20 g, 2.59 mmol) in trifluoroacetic acid (12 mL) was added anisole (1.2 mL) at room temperature. The reaction was then stirred at 30 °C for 16 h before it was quenched. The reaction was concentrated and the residue was purified by reverse phase C18 column chromatography (eluent: 0.1% formic acid in water / acetonitrile = 7 / 3) to give 3-(3-fluoro-4-((2-methyl-lH-imidazol-l-yl)methyl)phenyl)-5- isobutylthiophene-2-sulfonamide (6d).

[0137] LC-MS (ESI) calcd for C 19 H 22 FN3O2S2[M+H] + m / z 408.1, found 408.0.

[0138] Step 4: Preparation of N-((3-(3-fluoro-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)-5-isobutylthiophene-2-yl)sulfonyl)-l-propyloxycyclopropane-l- carboxamide (Compound 6)

[0139] To a stirred solution of 3-(3-fluoro-4-((2-methyl-lH-imidazol-l-yl)methyl)phenyl)-5- isobutylthiophene-2-sulfonamide (6d) (0.12 g, 0.29 mmol) in N,N-dimethylformamide (3 mL) was added l-propyloxycyclopropane-l-carboxylic acid (0.09 g, 0.62 mmol), 1-hydroxybenzotriazole (0.08 g, 0.59 mmol), l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.11 g, 0.59 mmol) and N,N- diisopropylethylamine (0.23 g, 1.77 mmol) at room temperature. The reaction was then stirred at 80 °C for 16 h before it was quenched. The cooled reaction was filtered and the filtrate was separated by high performance liquid preparative chromatography (mobile phase: 0.1% formic acid in water / acetonitrile = 3 / 2) to give the desired N-((3-(3-fluoro-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)-5-isobutylthiophene-2-yl)sulfonyl)-l-propyloxycyclopropane-l- carboxamide (Compound 6).

[0140] LC-MS (ESI) calcd for C 26 H 32 FN3O4S2[M+H] + m / z 534.2, found 534.3.

[0141] LC-MS (ESI) calcd for C1 H NMR (400 MHz, DMSO-d6 + D20) δ 7.80 (d, J = 10.9 Hz, 1H), 7.55 - 7.49 (m, 2H), 7.44 (s, 1H), 7.33 (t, J = 8.1 Hz, 1H), 6.85 (s, 1H), 5.40 (s, 2H), 3.26 (t, J = 6.7 Hz, 2H), 2.63 (d, J = 6.9 Hz, 2H), 2.58 (s, 3H), 1.89 - 1.77 (m, 1H), 1.40 - 1.28 (m, 2H), 0.94 (d, J = 6.6 Hz, 6H), 0.81 - 0.71 (m, 5H), 0.62 - 0.53 (m, 2H).

[0142] Example 7. l-Ethoxy-N-((3-(3-fluoro-4-((2-methyl-lH-imidazol-l-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)cyclopropane-l-carboxamide (Compound 7)

[0143] Synthetic Route:

[0144] Step 1: Preparation of methyl l-ethoxycyclopropane-l-carboxylate (7b)

[0145] Methyl 1-hydroxycyclopropane-1-carboxylate (7a) (10.00 g, 86.20 mmol) was dissolved in dry N,N-dimethylformamide (100 mL) at room temperature under nitrogen protection, sodium hydride (60%, 4.10 g, 102.50 mmol) was added portionwise at 0 °C under stirring. The reaction was stirred at room temperature for 30 minutes, then iodoe thane (16.10 g, 103.40 mmol) was added at 0 °C. The reaction was then allowed to warm up to room temperature slowly and stirred at room temperature for 16 hours under nitrogen protection. The reaction was quenched by the addition of saturated aqueous ammonium chloride solution, the resulting mixture was extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, the resulting crude product was separated by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 50 / 1 ~ 10 / 1) to give methyl 1-ethoxycyclopropane-1-carboxylate (7b) as a crude product, which was used directly in the next step without further purification.

[0146] Step 2: l-Ethoxycyclopropane-l-carboxylic acid (7c):

[0147] Methyl 1-ethoxycyclopropane-1-carboxylate (7b) (2.00 g, 13.90 mmol) was dissolved in methanol / water (5 / 1, 30 mL) at room temperature, and sodium hydroxide (2.80 g, 70.00 mmol) was added with stirring. The reaction solution was then heated to 60°C and stirred for 16 hours, after which it was terminated. Water was added to the cooled reaction solution, which was then extracted with methyl tert-butyl ether. The aqueous phase was adjusted to pH 2 with dilute hydrochloric acid and extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the resulting crude product was separated by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 100 / 1 to 1 / 1) to give 1-ethoxycyclopropane-1-carboxylic acid (7c).

[0148] 1 H NMR (400 MHz, CDC13) δ 3.67 (q, J = 7.1 Hz, 2H), 1.41-1.34 (m, 2H), 1.28-1.22 (m, 2H), 1.21 (t, J = 7.2 Hz, 3H).

[0149] Step 3: Preparation of 1-ethoxy-N-((3-(3-fluoro-4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)cyclopropane-1-carboxamide (7)

[0150] Methyl 1-ethoxycyclopropane-1-carboxylate (7b) (2.00 g, 13.90 mmol) was dissolved in methanol / water (5 / 1, 30 mL) at room temperature, and sodium hydroxide (2.80 g, 70.00 mmol) was added with stirring. The reaction solution was then heated to 60°C and stirred for 16 hours, after which it was terminated. Water was added to the cooled reaction solution, which was then extracted with methyl tert-butyl ether. The aqueous phase was adjusted to pH 2 with dilute hydrochloric acid and extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the resulting crude product was separated by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 100 / 1 to 1 / 1) to give 1-ethoxycyclopropane-1-carboxylic acid (7c).

[0151] LC-MS (ESI) calcd for C 25 H 30 FN3O4S2[M+H] +m / z 520.2, found 520.3.

[0152] 1 H NMR (400 MHz, DMSO-d6) δ 7.81 (d, J = 11.9 Hz, 1H), 7.58 - 7.48 (m, 2H), 7.45 (s, 1H), 7.30 (t, J = 8.1 Hz, 1H), 6.84 (s, 1H), 5.40 (s, 2H), 3.33 (q, J = 6.9 Hz, 2H), 2.62 (d, J = 6.9 Hz, 2H), 2.57 (s, 3H), 1.88 - 1.81 (m, 1H), 0.97 - 0.91 (m, 9H), 0.76 (dd, J = 7.0, 3.9 Hz, 2H), 0.58 (d, J = 3.0 Hz, 2H).

[0153] Example 8. N-((3-(3-fluoro-4-((2-methyl-lH-imidazol-l-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)-l-methoxycyclopropane-l-carboxamide (Compound 8)

[0154] Synthetic Route:

[0155] Step 1: Preparation of N-((3-(3-fluoro-4-((2-methyl-lH-imidazol-l-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)-l-methoxycyclopropane-l-carboxamide (Compound 8)

[0156] Using l-methoxycyclopropanecarboxylic acid as the reactant, referring to the synthetic scheme of Example 6, Step 4, the target compound N-((3-(3-fluoro-4-((2-methyl-lH- imidazol-l-yl)methyl)phenyl)-5-isobutylthiophen-2-yl)sulfonyl)-l-methoxycyclopropane- 1-carboxamide (Compound 8) was obtained.

[0157] LC-MS (ESI) calcd for C 24 H 28 FN3O4S2[M+H] + m / z 506.2, found 506.3.

[0158] 1H NMR (400 MHz, DMSO-d6) δ 7.81 (d, J = 12.2 Hz, 1H), 7.50 (dd, J = 8.1, 1.6 Hz, 2H), 7.36 (s, 1H), 7.26 (t, J = 8.1 Hz, 1H), 6.83 (s, 1H), 5.37 (s, 2H), 3.04 (s, 3H), 2.62 (d, J = 7.0 Hz, 2H), 2.53 (s, 3H), 1.91 - 1.79 (m, 1H), 0.93 (d, J = 6.6 Hz, 6H), 0.78 - 0.73 (m, 2H), 0.61 - 0.55 (m, 2H). Example 9. N-((3-(3-chloro-4-((2-methyl-lH-imidazol-l-yl)methyl)phenyl)-5-isobutyl-4- methylthiophen-2-yl)sulfonyl)-l-methoxycyclopropane-l-carboxamide (Compound 9)

[0159] Synthetic Route:

[0160] Procedure:

[0161] Step 1: Preparation of 2-isobutyl-3-methylthiophene (9b)

[0162] Zinc chloride (0.5 M tetrahydrofuran solution, 254.50 mmol, 509 mL) was dissolved in dry tetrahydrofuran (120 mL) at room temperature, and isobutylmagnesium bromide (1 M tetrahydrofuran solution, 226.00 mmol, 226 mL), 2-bromo-3-methyl-thiophene (25.00 g, 141.20 mmol) and bis(triphenylphosphine)palladium (3.60 g, 7.10 mmol) were added successively under nitrogen protection. After addition, nitrogen was replaced, and the temperature was raised to 90 °C and stirred for 1 hour under nitrogen protection before termination. The cooled reaction solution was poured into water, and the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, and the obtained crude product was separated by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 10 / 1) to obtain 2-isobutyl-3-methylthiophene (9b).

[0163] 1 H NMR (400 MHz, CDCl3) δ 7.02 (d, J = 5.1 Hz, 1H), 6.77 (d, J = 5.1 Hz, 1H), 2.59 (d, J = 7.2 Hz, 2H), 2.15 (s, 3H), 1.93 - 1.80 (m, 1H), 0.94 (d, J = 6.6 Hz, 6H).

[0164] Step 2: Preparation of 5-isobutyl-4-methylthiophene-2-sulfonyl chloride (9c)

[0165] Dissolve 2-isobutyl-3-methylthiophene (9b) (15.20 g, 98.70 mmol) in dichloromethane (120 mL) at room temperature, add chlorosulfonic acid (35.60 g, 305.50 mmol) with stirring, and terminate the reaction after stirring at room temperature for 1 hour. Pour the reaction solution into water, extract with ethyl acetate, wash the combined organic phase with saturated brine, dry over anhydrous sodium sulfate, and filter. Concentrate the filtrate, and separate the resulting crude product by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 15 / 1) to obtain 5-isobutyl-4-methylthiophene-2-sulfonyl chloride (9c).

[0166] 1 H NMR (400 MHz, CDC13) δ 7.59 (s, 1H), 2.67 (d, J = 7.2 Hz, 2H), 2.20 (s, 3H), 2.01-1.86 (m, 1H), 0.99 (d, J = 6.6 Hz, 6H).

[0167] Step 3: Preparation of N-(tert-butyl)-5-isobutyl-4-methylthiophene-2-sulfonamide (9d)

[0168] Dissolve 5-isobutyl-4-methylthiophene-2-sulfonyl chloride (9c) (10.80 g, 42.90 mmol) in dichloromethane (100 mL) at room temperature, add N,N-diisopropylethylamine (11.10 g, 85.70 mmol) and tert-butylamine (4.70 g, 64.30 mmol). Warm the reaction solution to 40°C and terminate after stirring for 1 hour. Pour the cooled reaction solution into water, extract with ethyl acetate, wash the combined organic phase with saturated brine, dry over anhydrous sodium sulfate, and filter. Concentrate the filtrate, and separate the resulting crude product by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20 / 1) to obtain N-(tert-butyl)-5-isobutyl-4-methylthiophene-2-sulfonamide (9d).

[0169] 1 H NMR (400 MHz, CDC13) δ 7.29 (s, 1H), 2.60 (d, J = 7.2 Hz, 2H), 2.13 (s, 3H), 1.93-1.84 (m, 1H), 1.28 (s, 9H), 0.95 (d, J = 6.6 Hz, 6H).

[0170] Step 4: Preparation of (2-(N-(tert-butyl)sulfonamido)-5-isobutyl-4-methylthiophen-3-yl)boronic acid (9e)

[0171] N-(tert-butyl)-5-isobutyl-4-methylthiophene-2-sulfonamide (9d) (10.30 g, 35.60 mmol) was dissolved in dry tetrahydrofuran (150 mL) at room temperature, the reaction was cooled to -60 °C and n-butyllithium (2.5 M in n-hexane, 42.8 mL, 107.00 mmol) was added dropwise under nitrogen. After stirring for 0.5 h at 60 °C under nitrogen, triisopropyl borate (12.70 g, 67.70 mmol) was added. The reaction was then stirred for 3 h at -60 °C under nitrogen before being quenched. The reaction was allowed to warm to room temperature, quenched with saturated ammonium chloride and diluted with water before being extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the crude product was isolated by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 10 / 1) to give (2-(N-(tert-butyl)sulfonamido)-5-isobutyl-4-methylthiophen-3-yl)boronic acid (9e).

[0172] 1 H NMR (400 MHz, DMSO-d6) δ 2.61 (d, J = 7.2 Hz, 2H), 2.11 (s, 3H), 1.90 - 1.77 (m, 1H), 1.14 (s, 9H), 0.90 (d, J = 6.6 Hz, 6H).

[0173] Step 5: Preparation of 1-(4-bromo-2-chlorobenzyl)-2-methyl-1H-imidazole (9g)

[0174] 4-bromo-1-(bromomethyl)-2-chlorobenzene (9f) (2.00 g, 7.03 mmol) was dissolved in N,N-dimethylformamide (20 mL) at room temperature, 2-methylimidazole (1.15 g, 14.07 mmol) and potassium carbonate (2.92 g, 21.1 mmol) were added with stirring in an ice-water bath. The reaction was then stirred at room temperature for 16 h before being quenched. The reaction was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the residue was isolated by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 20 / 1 ~ 1 / 1) to give 1-(4-bromo-2-chlorobenzyl)-2-methyl-1H-imidazole (9g).

[0175] LC-MS (ESI) calcd for C 11 H 10 BrClN2[M+H] + m / z 285.0; 287.0, found 287.3.

[0176] Step 6: Preparation of N-(tert-butyl)-3-(3-chloro-4-((2-methyl-1H- imidazol-1-yl)methyl)phenyl)-5-isobutyl-4-methylthiophene-2-sulfonamide (9h)

[0177] To a stirred solution of N-(tert-butyl)-3-(3-chloro-4-((2-methyl-1H- imidazol-1-yl)methyl)phenyl)-5-isobutyl-4-methylthiophene-2-sulfonamide (9h) (0.71 g, 1.44 mmol) in trifluoroacetic acid (10 mL) was added anisole (1 mL) at room temperature. The reaction was stirred at 30 °C for 16 h, then quenched. The reaction was concentrated and the residue was purified by reverse phase C18 column chromatography (mobile phase: acetonitrile / 0.1% formic acid in water = 0 / 1 to 1 / 1) to give 3-(3-chloro-4-((2-methyl-1H- imidazol-1-yl)methyl)phenyl)-5-isobutyl-4-methylthiophene-2-sulfonamide (9i).

[0178] LC-MS (ESI) calcd for C 24 H 32 ClN3O2S2[M+H] + m / z 494.2, found 494.2.

[0179] Step 7: Preparation of 3-(3-chloro-4-((2-methyl-1H-imidazol-1- yl)methyl)phenyl)-5-isobutyl-4-methylthiophene-2-sulfonamide (9i)

[0180] To a stirred solution of N-(tert-butyl)-3-(3-chloro-4-((2-methyl-1H- imidazol-1-yl)methyl)phenyl)-5-isobutyl-4-methylthiophene-2-sulfonamide (9h) (0.71 g, 1.44 mmol) in trifluoroacetic acid (10 mL) was added anisole (1 mL) at room temperature. The reaction was stirred at 30 °C for 16 h, then quenched. The reaction was concentrated and the residue was purified by reverse phase C18 column chromatography (mobile phase: acetonitrile / 0.1% formic acid in water = 0 / 1 to 1 / 1) to give 3-(3-chloro-4-((2-methyl-1H- imidazol-1-yl)methyl)phenyl)-5-isobutyl-4-methylthiophene-2-sulfonamide (9i).

[0181] LC-MS (ESI) calcd for C 20 H24 ClN3O2S2[M+H] + m / z 438.1, found 438.1.

[0182] Step 8: Preparation of N-((3-(3-chloro-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)-5-isobutyl-4-methylthiophen-2-yl)sulfonyl)-l- methoxycyclopropane-l-carboxamide (Compound 9)

[0183] To a solution of 3-(3-chloro-4-((2-methyl-lH-imidazol-l-yl)methyl)phenyl)-5- isobutyl-4-methylthiophene-2-sulfonamide (9i) (0.10 g, 0.23 mmol), l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.09 g, 0.46 mmol), 1- hydroxybenzotriazole (0.06 g, 0.46 mmol), N,N-diisopropylethylamine (0.09 g, 0.69 mmol) and l-methoxycyclopropane-l-carboxylic acid (0.09 g, 0.75 mmol) in N,N- dimethylformamide (2 mL) was heated to 80 °C and stirred for 16 h. The reaction was filtered and the filtrate was separated by high performance liquid preparative chromatography (mobile phase: acetonitrile / 0.1% formic acid in water) to give the target compound N-((3-(3-chloro-4-((2-methyl-lH-imidazol-l- yl)methyl)phenyl)-5-isobutyl-4-methylthiophen-2-yl)sulfonyl)-l- methoxycyclopropane-l-carboxamide (Compound 9).

[0184] LC-MS (ESI) calcd for C 25 H 30 ClN3O4S2[M+H] + m / z 536.1, found 536.2.

[0185] 1 H NMR (400 MHz, DMSO-d6) δ 7.42 (d, J = 1.6 Hz, 2H), 7.33 (s, 1H), 7.27 - 7.16 (m, 1H), 6.99 (d, J = 7.9 Hz, 1H), 5.41 (s, 2H), 3.07 (s, 3H), 2.59 (d, J = 7.0 Hz, 2H), 2.43 (s, 3H), 1.87 - 1.70 (m, 4H), 0.95 (d, J = 6.6 Hz, 6H), 0.80 - 0.71 (m, 2H), 0.70 - 0.57 (m, 2H).

[0186] Example 10. N-((3-(3-fluoro-4-((2-isopropyl-lH-imidazol-l-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)-l-methoxycyclopropane-l-carboxamide (Compound 10)

[0187] Synthetic Route:

[0188] Step 1: Preparation of l-(4-bromo-2-fluorobenzyl)-2-isopropyl-lH-imidazole (10b)

[0189] To a solution of 4-bromo-2-fluorobenzyl bromide (10a) (2.00 g, 7.46 mmol), 2- isopropylimidazole (1.23 g, 11.20 mmol) and potassium carbonate (3.10 g, 22.39 mmol) in N,N-dimethylformamide (12 mL) was added at room temperature. The reaction was stirred at room temperature for 16 hours under nitrogen atmosphere. The reaction was diluted with water and extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the residue was separated by column chromatography on normal phase silica gel (eluent: dichloromethane / methanol = 10 / 1) to give l-(4-bromo-2-fluorobenzyl)-2-isopropyl-lH-imidazole (10b) as yellow oil.

[0190] LC-MS (ESI) calcd for C 13 H 14 BrFN2[M+H] + m / z 297.0; 299.0, found 298.9.

[0191] 1 H NMR (400 MHz, CDC13) δ 7.33 - 7.19 (m, 2H), 7.02 (d, J = 1.2 Hz, 1H), 6.79 (d, J = 1.2 Hz, 1H), 6.67 (t, J = 7.8 Hz, 1H), 5.08 (s, 2H), 2.98 - 2.89 (m, 1H), 1.28 (d, J = 6.8 Hz, 6H).

[0192] The synthesis of Step 2 to Step 4 was referred to the synthetic scheme of Example 6, Step 2 to Step 4, wherein Step 4 used l-methoxycyclopropane carboxylic acid as the reactant, to give the target compound N-((3-(3-fluoro-4-((2-isopropyl-lH-imidazol-l- yl)methyl)phenyl)-5-isobutylthiophen-2-yl)sulfonyl)-l-methoxycyclopropane-l- carboxamide (Compound 10).

[0193] LC-MS (ESI) calcd for C 26 H32 FN3O4S2[M+H] + m / z 534.2, found 534.2.

[0194] 1 H NMR (400 MHz, DMSO-d6) δ 7.81 (d, J = 12.1 Hz, 1H), 7.64 - 7.44 (m, 3H), 7.28 (t, J = 8.1 Hz, 1H), 6.86 (s, 1H), 5.49 (s, 2H), 3.54 - 3.47 (m, 1H), 3.05 (s, 3H), 2.63 (d, J = 7.0 Hz, 2H), 1.88 - 1.82 (m, 1H), 1.26 (d, J = 6.9 Hz, 6H), 0.93 (d, J = 6.6 Hz, 6H), 0.81 (dd, J = 7.2, 4.0 Hz, 2H), 0.65 (s, 2H).

[0195] Example 11. N-((3-(3-fluoro-4-((2-(2-hydroxypropan-2-yl)-lH-imidazol-l- yl)methyl)phenyl)-5-isobutylthiophen-2-yl)sulfonyl)-l-methoxycycloprop-l- carboxamide (Compound 11)

[0196] Synthetic Route:

[0197] Step 1: Preparation of methyl l-(4-bromo-2-fluorobenzyl)-lH-imidazole-2-carboxylate (lib)

[0198] To a stirred solution of 4-bromo-l-(bromomethyl)-2-fluorobenzene (lla) (2.00 g, 7.46 mmol) in N,N-dimethylformamide (20 mL) was added successively imidazole-2-carboxylic acid methyl ester (1.40 g, 11.19 mmol) and potassium carbonate (3.10 g, 22.39 mmol) at room temperature. The reaction was terminated after stirring at room temperature for 16 hours. The reaction was diluted with water and extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the residue was separated by column chromatography on silica gel (eluent: dichloromethane / methanol = 10 / 1) to give methyl l-(4-bromo-2-fluorobenzyl)-lH-imidazole-2-carboxylate (lib).

[0199] 1 H NMR (400 MHz, CDCl3) δ 7.31 - 7.22 (m, 2H), 7.19 (s, 1H), 7.15 (s, 1H), 7.04 (t, J = 8.0 Hz, 1H), 5.65 (s, 2H), 3.93 (s, 3H).

[0200] Step 2: Preparation of 2-(l-(4-bromo-2-fluorobenzyl)-lH-imidazol-2-yl)propan-2-ol (lie)

[0201] To a stirred solution of methyl l-(4-bromo-2-fluorobenzyl)-lH-imidazole-2-carboxylate (lib) (2.20 g, 7.03 mmol) in dry tetrahydrofuran (15 mL) was added methyl magnesium bromide (3 M, 5.2 mL, 15.60 mmol) slowly at -70 °C under nitrogen protection. The reaction was then allowed to warm up to room temperature slowly and was terminated after stirring at room temperature for 2 hours under nitrogen protection. The reaction was quenched by the addition of saturated aqueous ammonium chloride solution and was extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the residue of the crude product was separated by column chromatography on silica gel (eluent: dichloromethane / methanol = 10 / 1) to give 2-(l-(4-bromo-2-fluorobenzyl)-lH-imidazol-2-yl)propan-2-ol (lie).

[0202] 1 H NMR (400 MHz, DMSO-d6) δ 7.59 (dd, J = 9.8, 1.9 Hz, 1H), 7.40 (dd, J = 8.2, 1.8 Hz, 1H), 6.98 (d, J = 0.8 Hz, 1H), 6.86 - 6.77 (m, 2H), 5.54 (s, 2H), 5.39 (s, 1H), 1.46 (s, 6H).

[0203] Step 3 - Step 5 refer to the synthetic scheme of Step 2 - Step 4 of Example 10 to give the target compound N-((3-(3-fluoro-4-((2-(2-hydroxypropan-2-yl)-lH-imidazol-l-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)-l-methoxycycloprop-l-earboxamide (Compound 11).

[0204] LC-MS (ESI) calcd for C 26 H 32 FN3O5S2[M+H] + m / z 550.2, found 550.2.

[0205] 1H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.84 (d, J = 12.3 Hz, 1H), 7.42 (d, J = 8.1 Hz, 1H), 6.93 (s, 1H), 6.86 (t, J = 8.2 Hz, 1H), 6.82 (s, 1H), 6.78 (s, 1H), 5.58 (s, 2H), 5.43 (s, 1H), 3.08 (s, 3H), 2.60 (d, J = 6.9 Hz, 2H), 1.87 - 1.79 (m, 1H), 1.49 (s, 6H), 0.93 (d, J = 6.6 Hz, 6H), 0.80 (d, J = 3.0 Hz, 2H), 0.60 (d, J = 3.1 Hz, 2H).

[0206] Example 12. N-((3-(4-((2-tert-Butyl-lH-imidazol-l-yl)methyl)-3-fluorophenyl)-5- isobutylthiophen-2-yl)sulfonyl)-l-methoxycyclopropane-l-carboxamide (Compound 12)

[0207] Synthetic Route:

[0208] Step 1 - Step 4 refer to the synthetic scheme of Example 10, Step 1 - Step 4, wherein Step 1 uses 2-tert-butylimidazole as the reactant, to yield the target compound N-((3-(4-((2-tert-butyl-lH-imidazol-l-yl)methyl)-3-fluorophenyl)-5-isobutylthiophen-2- yl)sulfonyl)-l-methoxycyclopropane-l-carboxamide (Compound 12).

[0209] LC-MS (ESI) calcd for C 27 H 34 FN3O4S2[M + H] + m / z 548.2, found 548.3.

[0210] 1 H NMR (400 MHz, DMSO-d6) δ 13.30 (br s, 1H), 7.81 (d, J = 11.1 Hz, 1H), 7.50 (d, J = 8.2 Hz, 1H), 7.44 (s, 1H), 7.32 (s, 1H), 6.98 (t, J = 8.0 Hz, 1H), 6.88 (s, 1H), 5.59 (s, 2H), 3.08 (s, 3H), 2.64 (d, J = 7.2 Hz, 2H), 1.92 - 1.77 (m, 1H), 1.46 (s, 9H), 0.94 (d, J = 6.6 Hz, 6H), 0.88 - 0.80 (m, 2H), 0.79 - 0.58 (m, 2H).

[0211] Example 13. N-((3-(3-fluoro-4-((2-(l-methylcyclopropyl)-lH-imidazol-l- yl)methyl)phenyl)-5-isobutylthiophen-2-yl)sulfonyl)-l-methoxycyclopropane-l- carboxamide (Compound 13)

[0212] Synthetic Route:

[0213] Step 1: Preparation of l-methylcyclopropane-l-carbaldehyde (13b)

[0214] To a stirred solution of oxalyl chloride (5.60 g, 44.12 mmol) in dry dichloromethane (30 mL) was added dropwise at -78 °C under nitrogen atmosphere dimethylsulfoxide (6.90 g, 88.24 mmol). After stirring at -78 °C for 30 min under nitrogen atmosphere, (l-methylcyclopropyl)methanol (13a) (1.90 g, 22.06 mmol) and triethylamine (17.8 g, 176.5 mmol) were added. After stirring for 30 min, the temperature was raised to 0 °C and stirred for 30 min before quenching. The reaction mixture was diluted with water and extracted with dichloromethane. The combined organic phase was washed successively with dilute hydrochloric acid and saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give crude l-methylcyclopropane-l-carbaldehyde (13b). The product was used directly in the next step without further purification.

[0215] Step 2: Preparation of 2-(l-methylcyclopropyl)-lH-imidazole (13c)

[0216] To a stirred solution of l-methylcyclopropane-l-carbaldehyde (13b) (0.7 g, 8.32 mmol) and aqueous ammonia (1.8 mL) in methanol (7 mL) was added glyoxal (1.30 g, 9.15 mmol) at 0 °C. The reaction mixture was then stirred at room temperature for 16 h before quenching. The reaction mixture was concentrated and the residue was purified by reverse phase C18 column chromatography (mobile phase: acetonitrile / 0.1% formic acid in water = 1 / 20) to give 2-(l-methylcyclopropyl)-lH-imidazole (13c).

[0217] 1 H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 2H), 1.41 (s, 3H), 0.99 (q, J = 3.8 Hz, 2H), 0.70 (q, J = 3.9 Hz, 2H).

[0218] Step 3 - Step 6 refer to the synthetic scheme of Example 10 Step 1 - Step 4, wherein Step 3 uses 2-(1-methylcyclopropyl)-1H-imidazole (13c) as the reactant, to give N-((3-(3-fluoro-4-((2-(1-methylcyclopropyl)-1H-imidazol-1-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)-1-methoxycyclopropane-1-carboxamide (Compound 13).

[0219] LC-MS (ESI) calcd for C 27 H 32 FN3O4S2[M+H] + m / z 546.2, found 546.4.

[0220] 1 H NMR (400 MHz, DMSO-d6) δ 8.13 (s, HCOOH), 7.86 (d, J = 12.4 Hz, 1H), 7.50 (d, J = 7.6 Hz, 1H), 7.18 (s, 1H), 7.06 (s, 2H), 6.84 (s, 1H), 5.41 (s, 2H), 3.08 (s, 3H), 2.61 (d, J = 7.0 Hz, 2H), 1.89 - 1.78 (m, 1H), 1.26 (s, 3H), 0.97 (s, 2H), 0.94 (d, J = 6.6 Hz, 6H), 0.80 (s, 4H), 0.62 (s, 2H).

[0221] Example 14. 1-(2-cyclopropylethoxy)-N-((3-(3-fluoro-4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)cyclopropane-1-carboxamide (Compound 14)

[0222] Synthetic Route:

[0223] Step 1: Preparation of methyl 1-(2-cyclopropylethoxy)cyclopropane-1-carboxylate (14b)

[0224] Methyl 1-hydroxycyclopropane-1-carboxylate (0.25 g, 2.20 mmol) was dissolved in dry N,N-dimethylformamide (10 mL) at room temperature, 60% sodium hydride (0.10 g, 2.60 mmol) was added portionwise with stirring at 0 °C and stirred for 30 minutes. (2-Iodoethyl)cyclopropane (14a) (0.51 g, 2.60 mmol) was added. The reaction was terminated after stirring at room temperature for 16 hours. The reaction was diluted with water slowly and extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the resulting crude product was separated by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 19 / 1-1 / 1) to give methyl 1-(2-cyclopropylethoxy)cyclopropane-1-carboxylate (14b). The product was used in the next step without further purification.

[0225] Step 2: Preparation of 1-(2-cyclopropylethoxy)cyclopropane-1-carboxylic acid (14c)

[0226] Methyl 1-(2-cyclopropylethoxy)cyclopropane-1-carboxylate (14b) (0.41 g, 2.23 mmol) was dissolved in methanol / water (5 / 1, 12 mL) at room temperature, sodium hydroxide (0.45 g, 11.15 mmol) was added. The reaction was then heated to 60 °C and stirred for 3 hours before termination. The cooled reaction was diluted with water and extracted with petroleum ether. The aqueous phase was adjusted to pH about 2-3 with dilute hydrochloric acid. The resulting mixture was extracted with ethyl acetate. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give 1-(2-cyclopropylethoxy)cyclopropane-1-carboxylic acid (14c). The product was used in the next step without further purification.

[0227] LC-MS (ESI) calcd for C9H 14 O3[M-H] - m / z 169.1, found 169.2.

[0228] Step 3: Preparation of 1-(2-cyclopropylethoxy)-N-((3-(3-fluoro-4-((2-methyl-1H- imidazol-1-yl)methyl)phenyl)-5-isobutylthiophen-2-yl)sulfonyl)cyclopropane-1- carboxamide (Compound 14)

[0229] To a microwave tube containing N,N-dimethylformamide (1 mL) were added 1-(2- cyclopropylethoxy)cyclopropane-1-carboxylic acid (14c) (0.13 g, 0.74 mmol), 3-(3- fluoro-4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)-5-isobutylthiophene-2- sulfonamide (6d) (0.15 g, 0.37 mmol), 1-hydroxybenzotriazole (0.10 g, 0.74 mmol), 1- (3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.14 g, 0.74 mmol), and N,N- diisopropylethylamine (0.29 g, 2.22 mmol) at room temperature. After the microwave tube was sealed, it was heated to 80 °C and stirring was terminated after 16 h. The cooled reaction was filtered, and the filtrate was separated by preparative high-performance liquid chromatography (mobile phase: 0.1% aqueous ammonia / acetonitrile) to give the title compound, 1-(2-cyclopropylethoxy)-N-((3-(3-fluoro-4-((2-methyl-1H- imidazol-1-yl)methyl)phenyl)-5-isobutylthiophene-2-yl)sulfonyl)cyclopropane-1-carboxamide (Compound 14).

[0230] LC-MS (ESI) calcd for C 28 H 34 FN3O4S2[M+H] + m / z 560.2, found 560.3.

[0231] 1 H NMR (400 MHz, Methanol-d4) δ 7.54 (dd, J = 11.4, 1.4 Hz, 1H), 7.48 (dd, J = 7.9, 1.5 Hz, 1H), 7.40 (s, 1H), 7.32 (dd, J = 17.3, 9.4 Hz, 2H), 6.80 (s, 1H), 5.39 (s, 2H), 3.48 (t, J = 6.8 Hz, 2H), 2.70 (d, J = 7.0 Hz, 2H), 2.62 (s, 3H), 2.24 - 2.15 (m, 1H), 2.05 - 2.01 (m, 2H), 1.97 - 1.88 (m, 1H), 0.98 (d, J = 6.6 Hz, 6H), 0.95 (t, J = 3.5 Hz, 2H), 0.90 (t, J = 6.8 Hz, 2H), 0.83 (t, J = 5.9 Hz, 2H), 0.42 - 0.34 (m, 2H).

[0232] Example 15. 1-(Cyclopropylmethoxy)-N-((3-(3-fluoro-4-((2-methyl-1H-imidazol-1- yl)methyl)phenyl)-5-isobutylthiophene-2-yl)sulfonyl)cyclopropane-1-carboxamide (Compound 15)

[0233] Synthesis route:

[0234] Step 1: Preparation of methyl 1-(cyclopropylmethoxy)cyclopropane-1-carboxylate (15b)

[0235] To a stirred solution of methyl 1-hydroxycyclopropane-1-carboxylate (15a) (2.00 g, 17.22 mmol) in N,N-dimethylformamide (20 mL) was added slowly sodium hydride (60%, 1.38 g, 34.45 mmol) at 0 °C, followed by the addition of bromomethylcyclopropane (4.65 g, 34.45 mmol). The resulting mixture was stirred at 0 °C for 20 min, then at room temperature for 16 h before being quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the residue was separated by normal silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 10 / 1) to give methyl 1-(cyclopropylmethoxy)cyclopropane-1-carboxylate (15b).

[0236] 1 H NMR (400 MHz, CDC13) δ 3.73 (s, 3H), 3.46-3.34 (m, 2H), 1.31-1.26 (m, 2H), 1.19-1.12 (m, 2H), 1.09-1.01 (m, 1H), 0.57-0.49 (m, 2H), 0.29-0.18 (m, 2H).

[0237] Step 2: Preparation of 1-(cyclopropylmethoxy)cyclopropane-1-carboxylic acid (15c)

[0238] Methyl 1-(cyclopropylmethoxy)cyclopropane-1-carboxylate (15b) (0.70 g, 4.11 mmol) was dissolved in methanol / water (13 mL / 5 mL) at room temperature and sodium hydroxide (1.64 g, 41.13 mmol) was added with stirring. The reaction was then heated to 60 °C and stirred for 3 h before being quenched with water and extracted with ethyl acetate. Dilute hydrochloric acid (3 M) was added dropwise to the resulting aqueous phase until the pH was about 3 and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the residue was separated by silica gel column chromatography (eluent: petroleum ether / ethyl acetate) to give the product 1-(cyclopropylmethoxy)cyclopropane-1-carboxylic acid (15c).

[0239] 1H NMR (400 MHz, CDC13) δ 3.44 (d, J = 7.1 Hz, 2H), 1.40 - 1.33 (m, 2H), 1.26 - 1.20 (m, 2H), 1.12 - 0.98 (m, 1H), 0.59 - 0.47 (m, 2H), 0.27 - 0.17 (m, 2H).

[0240] Step 3 refers to the synthetic scheme of Example 14, Step 3, wherein 1-(cyclopropylmethoxy)cyclopropane-1 -carboxylic acid (15c) is used as the reactant, to give the target compound 1-(cyclopropylmethoxy)-N-((3-(3-fluoro-4-((2-methyl-1H-imidazol-1-yl)methyl)phenyl)-5- isobutylthiophen-2-yl)sulfonyl)cyclopropane-1 -carboxamide (Compound 15).

[0241] LC-MS (ESI) calcd for C 27 H 32 FN3O4S2[M+H] + m / z 546.2, found 546.1.

[0242] 1 H NMR (400 MHz, DMSO-d6) δ 7.81 (d, J = 11.8 Hz, 1H), 7.57 - 7.47 (m, 2H), 7.41 (s, 1H), 7.31 - 7.21 (m, 1H), 6.83 (s, 1H), 5.39 (s, 2H), 3.16 (d, J = 6.9 Hz, 2H), 2.62 (d, J = 7.0 Hz, 2H), 2.55 (s, 3H), 1.91 - 1.78 (m, 1H), 0.93 (d, J = 6.6 Hz, 6H), 0.87 - 0.79 (m, 1H), 0.78 - 0.72 (m, 2H), 0.58 (d, J = 2.9 Hz, 2H), 0.39 - 0.29 (m, 2H), 0.08 - 0.02 (m, 2H).

[0243] Example 16. N-((3-(4-((2-cyclopropyl-1H-imidazol-1-yl)methyl)-3-fluorophenyl)-5- isobutylthiophen-2-yl)sulfonyl)-1 -propoxy cyclopropane-1 -carboxamide (Compound 16)

[0244] Synthetic Route:

[0245] Step 1 - Step 4 refer to the synthetic scheme of Example 10 Step 1 - Step 4, wherein Step 1 uses 1 -(4-bromo-2-fluorobenzyl)-2-cyclopropyl- 1H-imidazole (16b) as the reactant, and Step 4 uses 1 -propyloxycyclopropane- 1 -carboxylic acid as the reactant, to give the target compound N-((3-(4-((2-cyclopropyl- 1H-imidazol- 1 -yl)methyl)-3 -fluorophenyl)-5 -isobutylthiophen-2-yl)sulfonyl)- 1 -propyloxycyclopropane- 1 -carboxamide (Compound 16).

[0246] LC-MS (ESI) calcd for C 28 H 34 FN3O4S2[M+H] + m / z 560.2, found 560.3.

[0247] 1 H NMR (400 MHz, DMSO-d6) δ 7.80 (d, J = 12.1 Hz, 1H), 7.50 (d, J = 7.1 Hz, 2H), 7.35 (s, 1H), 7.30 - 7.21 (m, 1H), 6.85 (s, 1H), 5.49 (s, 2H), 3.27 (d, J = 6.7 Hz, 2H), 2.63 (d, J = 6.9 Hz, 2H), 2.26 (s, 1H), 1.91 - 1.77 (m, 1H), 1.40 - 1.27 (m, 2H), 1.10 (d, J = 8.0 Hz, 2H), 1.02 (s, 2H), 0.94 (d, J = 6.6 Hz, 6H), 0.82 (t, J = 6.5 Hz, 2H), 0.76 (t, J = 7.4 Hz, 3H), 0.62 (s, 2H).

[0248] Example 17. N-((3-(4-((2-(tert-Butyl)-lH-imidazol-l-yl)methyl)-3-fluorophenyl)-5- isobutylthiophen-2-yl)sulfonyl)-l-ethoxycyclopropane-l-carboxamide (Compound 17)

[0249] Synthetic Route:

[0250] Step 1: Preparation of N-((3-(4-((2-(tert-Butyl)-lH-imidazol-l-yl)methyl)-3-fluorophenyl)-5- isobutylthiophen-2-yl)sulfonyl)-l-ethoxycyclopropane-l-carboxamide (Compound 17)

[0251] N-((3-(4-((2-(tert-butyl)-lH-imidazol-l-yl)methyl)-3-fluorophenyl)-5- isobutylthiophene-2-yl)sulfonyl)-l-ethoxycyclopropane-l-carboxamide (Compound 17) was obtained according to the synthetic protocol of Reference Example 12, Step 4.

[0252] LC-MS (ESI) calcd for C 28 H 36 FN3O4S2[M+H] + m / z 562.2, found 562.3.

[0253] 1 H NMR (400 MHz, DMSO-d6) δ 7.85 (d, J = 11.5 Hz, 1H), 7.50 (d, J = 7.9 Hz, 1H), 7.33 - 7.12 (m, 2H), 6.97 - 6.81 (m, 2H), 5.54 (s, 2H), 3.40 - 3.36 (m, 2H), 2.62 (d, J = 7.0 Hz, 2H), 1.93 - 1.78 (m, 1H), 1.42 (s, 9H), 1.00 - 0.90 (m, 9H), 0.87 - 0.79 (m, 2H), 0.71 - 0.57 (m, 2H).

[0254] In order to demonstrate the beneficial effects of the present application, the following biological test examples are provided.

[0255] Biological Test

[0256] Test Example 1: AT2R binding assay

[0257] The purpose of this assay is to assess the ability of compounds to bind AT2R in vitro.

[0258] Prepare assay buffer 1X TLB (Cisbio, Cat# LABMED), 4X Buloxibutid (CAS: 477775-14-7) and test compound stock, and 4X Tag-lite angiotensin receptor red agonist (Cisbio, Cat# L0007RED). Thaw 1 vial of AT2R labeled cells in 37℃ water bath for about 1-2 min until all ice is melted, quickly transfer the thawed cells to 5 mL 1X TLB, mix gently and centrifuge at 200g for 5 min. Discard the supernatant, resuspend the cells with 1 mL 1X TLB, mix well, add 1.7 mL 1X TLB, mix well and keep at room temperature. Add 10 μL cells to a 384-well plate (Greiner, Cat# 784075), centrifuge at 200g for 3 s. Add 5 μL 4X compound stock to the 384-well plate. Add 5 μL 4X Tag-lite angiotensin receptor red agonist to all test wells. Incubate the reaction plate at room temperature for 1 h, centrifuge at 200g for 60 s at room temperature, and collect data using Envision HTRF detector.

[0259] Inhibition rate calculation:

[0260] %inhibition = 100 - (Signal cmpd -Signal Ave_PC ) / (Signal Ave_VC -Signal Ave_PC ) x 100.

[0261] Compound IC 50 is calculated using GraphPad nonlinear fit formula:

[0262] Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope)).

[0263] X: compound concentration log value; Y: % inhibition.

[0264] The AT2R binding results of some examples are shown in Table 1.

[0265] Table 1: Compound AT2R binding

[0266] The above results show that the compounds of the present application have strong AT2R binding activity.

[0267] Test Example 2: CYP enzyme inhibition study of some compounds

[0268] Inhibitors, probe substrates and reaction time for each subzyme are shown in Table 2. Commercially available mixed human liver microsomes were used, with a liver microsomal concentration of 0.1 mg / mL for dextromethorphan, midazolam; and 0.2 mg / mL for phenacetin and testosterone. Probe substrates, specific inhibitors and test compounds were diluted to the corresponding concentrations in working solutions, and a mixture solution containing liver microsomes, MgCl2, substrate, specific inhibitor, test compound was prepared, 50 μL of 4 mM NADP solution was added to initiate the reaction, and the reaction time is shown in the table. After incubation, 600 μL of ice-cold ACN solution containing IS was added to terminate the reaction, and after centrifugation, LC-MS / MS was used for detection.

[0269] Table 2 Inhibitors, probe substrates and reaction time for each subzyme

[0270] At different concentration points of the test compound or positive inhibitor, the percentage of residual activity was obtained from the ratio of the amount of characteristic metabolite of the probe substrate generated to the amount generated in the absence of the test compound or positive inhibitor. If a significant decrease in the amount of metabolite generated was found at the highest concentration setting point, the half-inhibitory concentration (IC 50 ) was calculated using the log(inhibitor) vs. response - Variable slope formula of GraphPad Prism software:

[0271] Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC 50 -X)*Hillslope))

[0272] X: Log(concentration of the test compound or positive inhibitor);

[0273] Y: percentage of residual activity;

[0274] Top and Bottom: refer to the theoretical maximum and minimum percentage of residual activity, respectively;

[0275] Hillslope: slope coefficient or slope.

[0276] If there is no significant decrease in the amount of metabolite generated at the highest concentration setting point (percentage of residual activity of the enzyme > 50%), the half-inhibitory concentration (IC 50 ) cannot be accurately calculated, and the IC 50 is reported to be greater than the highest concentration tested.

[0277] The rat CYP enzyme inhibition data of some examples are shown in Table 3:

[0278] CYP450 enzyme inhibition results of some of the compounds in Table 3 Note: CYP3A4(T) is the CYP3A4 enzyme inhibition experiment when the probe substrate is Testosterone; CYP3A4(M) is the CYP3A4 enzyme inhibition experiment when the probe substrate is Midazolam.

[0279] The results of the study show that the compounds in the application have weaker inhibition on CYP enzymes than the control compounds, indicating a lower possibility of drug-drug interaction in the human body and higher safety.

[0280] Test Example 3: AT2R-CHO cell NO release test

[0281] The purpose of this test is to evaluate the effect of the compound on the amount of NO released from AT2R-CHO cells in vitro.

[0282] AT2R-CHO cells were implanted in a 96-well plate (3603 black bottom wells), F12K + 10% FBS, 5000 cells / 100 uL / well, 37°C, 5% CO2 incubation for 36 hours. The well culture medium was washed once, 50 uL / well, then the DAF-FM probe was added to the cells, the final probe concentration in the well was 5 uM, incubated at 37°C, 5% CO2 for 30 min. Discard the culture medium, add 90 uL of the optimized liquid, then add 10 uL of the diluted compound, incubate at 37°C, 5% CO2 for 15 min, discard the culture medium, wash the cells with PBS once, fix with 50 uL / well (containing DAPI) 4% paraformaldehyde. High content imaging system scanning, data collection.

[0283] Activation rate calculation:

[0284] Relative activation% = (Sample-DMSO) / DMSO*100

[0285] The compound EC is calculated using the GraphPad nonlinear fitting formula 50 :

[0286] Y = Bottom + (Top-Bottom) / (1+10^((LogEC 50 -X)*HillSlope))

[0287] X: compound concentration log value; Y: % Relative activation.

[0288] The in vitro data of some examples are shown in Table 4:

[0289] In vitro AT2R-CHO activation data of some of the compounds of Table 4

[0290] The above results show that the compounds of the present application have strong AT2R agonistic activity in vitro.

[0291] Test Example 4: Efficacy of the compounds on bleomycin-induced pulmonary fibrosis in rats

[0292] The purpose of this test is to evaluate the in vivo efficacy of the compounds on bleomycin-induced pulmonary fibrosis in rats.

[0293] After the animals were purchased and acclimated, they were randomly divided into groups according to body weight. The animals were divided into 4 groups, 12 in each group, namely the model control group, the positive control drug Buloxibutid group (10 mg / kg, bid), the test compound 1 group (10 mg / kg, bid), and the test compound 6 group (10 mg / kg, bid), and 3 animals were selected as the normal control group. After the rats were weighed, they were anesthetized by isoflurane inhalation, and after the animals lost the righting reflex, they were suspended on the upper incisors and fixed on the operating board with the abdomen upwards, ensuring sufficient light to show the vocal cords. The sterile pad surgical forceps gently stretched the tongue to the corner of the mouth on one side, exposing the vocal cords in the direction of the mandible. The delivery device needle loaded with 100 uL of bleomycin was inserted through the vocal cord nodule, and the bleomycin aerosol solution was quickly discharged. Then the delivery device was removed, and the animals were placed back in the cage after they woke up. The control animals were given an equal amount of sterile normal saline. One day after the operation, the animals in each group were given the appropriate solvent or test substance orally by gavage for 12 consecutive days, with a frequency of 2 times / day. After the end of the administration period, the animals were euthanized by deep CO2 anesthesia, the lungs were removed and weighed, and the organ coefficient of the lungs was calculated; the ipsilateral lung was removed, 10% formalin solution was injected through the bronchus, and the bronchus was ligated after the pleura was flattened, and the lung tissue was fixed, and longitudinal sections were taken from the lung apex to the lung base for pathological examination (HE staining, Sirius red staining). The HE-stained sections were observed under a light microscope (x100) for lung tissue pathology and images were collected, and the lung fibrosis score was determined according to the Ashcroft scoring standard, with a higher score indicating a more severe degree of pulmonary fibrosis. The right lung sections after embedding were stained with Sirius red, and the lung tissue pathology was observed under a light microscope (x100) and images were collected. The red collagen area was collected using Image pro plus 6.0 software to obtain the fibrosis ratio. Fibrosis ratio = red collagen area / lung tissue area x 100%.

[0294] The results of the lung coefficient are shown in Table 5; the results of the HE staining score and the lung fibrosis ratio are shown in Table 6.

[0295] Table 5 Effect of test substances on the organ coefficient of the lungs of pulmonary fibrosis model rats after oral gavage administration for 12 consecutive days (Mean ± SD, n = 3 for the normal control group, and n = 12 for the other groups)

[0296] # P<0.05 vs normal control group; ** P<0.01, * P<0.05 vs model control group.

[0297] Table 6. The effect of the test substance on HE staining and Sirius red staining of pulmonary fibrosis model rats after oral gavage administration for 12 consecutive days (Mean ± SD, n = 3 for normal control group, n = 12 for the rest of the groups)

[0298] ### P<0.001 vs normal control group; ** P<0.01, * P<0.05 vs model control group.

[0299] The results of the study showed that the test compound can better reduce the proportion of pulmonary fibrosis in pulmonary fibrosis model rats, and has a therapeutic effect on bleomycin-induced pulmonary fibrosis in SD rats.

[0300] Test Example 5: Efficacy of the compound on bleomycin-induced pulmonary fibrosis in mice

[0301] The purpose of this test is to evaluate the in vivo efficacy of the compound on bleomycin-induced pulmonary fibrosis in mice.

[0302] After the animals were purchased and acclimated, they were randomly divided into groups according to body weight. The animals were divided into 4 groups, 12 in each group, namely the model control group, the positive control drug Buloxibutid group (40 mg / kg, bid), the test compound 1 group (40 mg / kg, bid) and the test compound 6 group (40 mg / kg, bid), and another 3 animals were used as a normal control group. Isoflurane was used to maintain the anesthetized state, and after the tracheal part was found by mouse laryngoscope, a liquid atomizing needle was inserted into the trachea, BLM liquid was quickly pushed in, and after the atomizing needle was withdrawn, the mouse was lifted and gently shaken and the chest was gently massaged to evenly distribute the drug in the lungs. The mouse was placed back in the cage, and the normal control group was given atomized PBS in the trachea, and the other operations were the same. The BLM modeling dose was 2.5 mg / kg. On the 2nd day after modeling, each group was given the solvent or the test substance orally for 14 days, 2 times / day. After the administration period ended, the animals were euthanized with deep CO2 anesthesia, the lungs were removed and weighed, and the organ coefficient of the lungs was calculated. The ipsilateral lung was removed, 0.6 mL of 4% fixing solution was perfused from the trachea, and the lung was fixed in 4% fixing solution. After dehydration and embedding of the fixed lung tissue, the largest surface was used for sectioning, and H&E staining and Sirius red staining were performed. The HE-stained sections were scanned using a slide scanner, and the Ashcroft scoring standard was used for HE fibrosis scoring. The higher the score, the more severe the degree of pulmonary fibrosis. The right lung section after embedding was stained with Sirius red, and the ImageJ software was used to analyze the total lung fibrosis area and the relative expression of fibrosis.

[0303] The lung coefficient results are shown in Table 7; the HE staining score results and the lung fibrosis proportion results are shown in Table 8.

[0304] Table 7. Effect of test substance on lung organ coefficient of pulmonary fibrosis model mice after oral gavage administration for 14 consecutive days (Mean ± SD, n = 3 for the normal control group, and n = 12 for the other groups) #### P < 0.0001 vs the normal control group.

[0305] Table 8. Effect of test substance on HE staining and Sirius red staining of pulmonary fibrosis model mice after oral gavage administration for 14 consecutive days (Mean ± SD, n = 3 for the normal control group, and n = 12 for the other groups) ### P < 0.001 vs the normal control group; *** P < 0.001, ** P < 0.01 vs the model control group.

[0306] The results of the study showed that the test compounds can better reduce the proportion of lung tissue fibrosis in pulmonary fibrosis model mice and have a therapeutic effect on bleomycin-induced pulmonary fibrosis in mice.

[0307] The above detailed description has been given to the purpose of further explaining the object, technical solutions and beneficial effects of the present application. It should be understood that the above is only a specific implementation of the present application and is not used to limit the protection scope of the present application. Any modification, equivalent replacement, improvement, etc. made on the basis of the technical solutions of the present application shall be included in the protection scope of the present application.

Claims

1. A compound of Formula I: ###0001### Formula I or a stereoisomer thereof, or a pharmaceutically acceptable salt or prodrug thereof. R 1 selected from C 1-6 alkyl; Y is selected from CR 7 , R 7 is selected from hydrogen, C 1-3 alkyl; L is selected from O; R 2 selected from C 1-6 alkyl, wherein the C 1-6 alkyl is optionally substituted with one or more hydrogen or C 3-7 cycloalkyl; R 3 selected from halogen or C 1-3 alkyl; n = 0 or 1 ; R 4 selected from C 1-4 alkyl or C 3-7 cycloalkyl, wherein said C 1-4 alkyl and C 3-7 cycloalkyl are each optionally substituted with hydrogen, halogen, hydroxyl, C 3-7 alkyl or C 1-6 alkyl; R 5 and R 6 are each independently selected from hydrogen, halogen, C 1-4 alkyl.

2. The compound of claim 1, or a stereoisomer, or a pharmaceutically acceptable salt or prodrug thereof. R 1 is selected from ethyl, n-propyl, i-propyl, n-butyl or i-butyl.

3. The compound of claim 1, or a stereoisomer, or a pharmaceutically acceptable salt or prodrug thereof. Y is selected from CR 7 , R 7 is selected from hydrogen or methyl.

4. The compound of claim 1, or a stereoisomer, or a pharmaceutically acceptable salt or prodrug thereof, wherein R 3 selected from fluoro, chloro or methyl.

5. The compound of claim 1, or a stereoisomer, or a pharmaceutically acceptable salt or prodrug thereof, wherein R 4 selected from C 1-4 alkyl or C 3-5 cycloalkyl, wherein the C 1-4 alkyl and C 3-5 cycloalkyl can be substituted by halogen, hydroxy, C 1-6 alkyl or C 3-5 cycloalkyl.

6. The compound of any one of claims 1-5, or a stereoisomer, or a pharmaceutically acceptable salt or prodrug thereof, wherein The compound of formula I is represented by formula I-1: R is selected from ethyl, n-propyl, isopropyl, n-butyl or isobutyl; R 1 R is selected from ethyl, n-propyl, isopropyl, n-butyl or isobutyl; R 2 R is selected from C 1-6 alkyl, wherein said C 1-6 alkyl can be substituted by C 3-7 cycloalkyl; n = 0 or 1 ; R 3 R is selected from fluorine, chlorine or methyl; R 4 R is selected from C 1-4 alkyl or C 3-5 cycloalkyl, wherein said C 1-4 alkyl and C 3-5 cycloalkyl can be substituted by halogen, hydroxy, C 1-6 alkyl or C 3-5 cycloalkyl; R 5 , R 6 R is selected from hydrogen; R 7 R is selected from hydrogen or methyl.

7. The compound of any one of claims 1-5, or a stereoisomer, or a pharmaceutically acceptable salt or prodrug thereof, wherein The compound of formula I is represented by formula I-2: wherein R 2 is selected from C 1-6 linear alkyl, said C 1-6 linear alkyl can be substituted by C 3-5 cycloalkyl; n = 0 or 1 ; R3is selected from methyl, fluoro or chloro; R 4 is selected from methyl, ethyl, isopropyl, cyclopropyl, tert-butyl or C 1-4 fluoroalkyl, said methyl, ethyl, isopropyl, cyclopropyl, tert-butyl or C 1-4 fluoroalkyl can be substituted by halogen, hydroxy, methyl or cyclopropyl; R 7 is selected from hydrogen or methyl.

8. The compound of any one of claims 1-5, or a stereoisomer, or a pharmaceutically acceptable salt or prodrug thereof, wherein The compound of formula I is represented by formula I-3: wherein R 2 selected from C 1-3 linear alkyl, said C 1-3 linear alkyl can be substituted by one cyclopropyl; n = 0 or 1 ; R3is selected from methyl, fluoro or chloro; R 4 selected from methyl, ethyl, isopropyl, cyclopropyl or tert-butyl, said tert-butyl can be substituted by hydroxy, said cyclopropyl can be substituted by methyl; R 7 selected from methyl or hydrogen.

9. The compound according to any one of claims 1-8, or a stereoisomer, or a pharmaceutically acceptable salt or prodrug thereof, selected from any of the following compounds:

10. A pharmaceutical composition, wherein, A compound, stereoisomer or pharmaceutically acceptable salt thereof as described in any one of claims 1-9, and one or more pharmaceutically acceptable carriers, diluents or excipients.

11. Use of a compound, solvate, metabolite, co-crystal or prodrug thereof of any one of claims 1-9, or a composition of claim 10, for the manufacture of a medicament for the treatment / prevention of a disease mediated by AT2R.

12. The use according to claim 11, wherein the disease comprises idiopathic pulmonary fibrosis.

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