Pyrazolo oleanolic acid reducing alcohol derivative and preparation method and application thereof

Abstract

The invention relates to pyrazolo oleanolic acid reduced alcohol derivatives as well as a preparation method and application thereof. The pyrazolo oleanolic acid reducing alcohol compound has a structure as shown in a formula I which is described in the specification. The compound disclosed by the invention has exact anti-inflammatory activity and osteoclast differentiation inhibition activity, so that the compound has a prospect of being developed into drugs for treating inflammation and osteoclast activity abnormity related diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a class of pyrazolooleanolic acid reduced alcohol derivatives, and pharmaceutically acceptable salts and hydrates thereof, and preparation methods thereof and their uses in treating inflammation and diseases related to abnormal osteoclast activity. Background Art

[0002] Inflammation refers to the defensive response of the body's tissues to external stimuli. It is a normal and important response mechanism that helps protect the body from infection and damage (Nat Rev Drug Discov. 2016, 15: 551.). However, abnormal or uncontrolled inflammatory responses can lead to the development of acute or chronic inflammatory disorders and conditions. Chronic inflammation can lead to a series of diseases such as rheumatoid arthritis, inflammatory bowel disease, chronic obstructive pulmonary disease, asthma, psoriasis, and idiopathic pulmonary fibrosis. Viral and bacterial infections or other damage (such as toxins, chemicals, etc.) can lead to uncontrolled acute inflammatory responses and damage, such as acute lung injury, acute respiratory distress syndrome, etc. (Nat Rev DisPrimers. 2019, 5: 18.). With the aging of the global population, the incidence of inflammatory-related diseases such as rheumatoid arthritis continues to increase.

[0003] In the acute inflammatory response stage, the body releases nitric oxide (NO), prostaglandins (PGE), platelet-activating factor (PAF), bradykinin and other inflammatory mediators to activate the immune system after tissue damage. After entering the chronic stage, the tissue will release a large number of inflammatory factors such as IL-1, IL-6, TNF-α, IL-8, IL-12, IFN-γ and IL-18, further aggravating the immune response. In severe cases, it can cause septic shock, systemic inflammatory response, multiple organ failure and even death. By inhibiting the production of these inflammatory factors, the pathological symptoms of related diseases can be significantly improved (Front Endocrinol (Lausanne). 2011, 2: 62.).

[0004] Oleanolic acid (OA) is a natural pentacyclic triterpenoid oleanane compound, which is widely distributed in many traditional Chinese medicines, such as olive, Ligustrum lucidum, Forsythia suspensa, Panax notoginseng, Prunella vulgaris, Hedyotis diffusa and Herba scutellariae (Phytochemistry, 2012, 77: 10.), and has pharmacological activities such as liver protection, anti-osteoporosis, anti-diabetes, antioxidant, anti-inflammatory and anti-cancer (Int J Mol Sci, 2017, 18: 643.), and OA has been used as an over-the-counter (OTC) drug in China to treat hepatitis for decades. Due to its inherent pharmacological activity, resource accessibility and cost affordability, OA is considered to be a high-quality starting material for further modification. However, OA itself has defects such as poor water solubility, weak activity and low bioavailability. Therefore, researchers use oleanolic acid as the starting molecule and make appropriate structural modifications to improve its specific biological activity and drugability, which is an effective strategy for the current research and development of oleanolic acid-based active molecules. Summary of the invention

[0005] After long-term and in-depth research, the inventors have provided a class of pyrazolooleanolic acid reduced alcohol derivatives, which can inhibit inflammation and osteoclast activity, and are thus used for diseases related to inflammation and imbalance of osteoclast activity. Based on the above findings, the inventors have completed the present invention.

[0006] The purpose of the present invention is to provide a novel pyrazolooleanolic acid reduced alcohol derivative having anti-inflammatory activity and osteoclast differentiation inhibition activity.

[0007] Another object of the present invention is to provide a method for preparing the above compound.

[0008] Another object of the present invention is to provide a pharmaceutical composition comprising the above compound.

[0009] Another object of the present invention is to provide the use of the above compound or the above pharmaceutical composition in the preparation of a drug having anti-inflammatory activity and osteoclast differentiation inhibition activity.

[0010] The first aspect of the present invention provides a pyrazolooleanolic acid reduced alcohol derivative as shown in the following general formula I, and a pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate thereof:

[0011]

[0012] in,

[0013] R1 is selected from H, -S(=O)2R 11 or -C(=O)R 11 ,

[0014] Among them, R11 are each independently selected from substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-

[0015] C6 cycloalkyl, substituted or unsubstituted C5-C10 aryl, substituted or unsubstituted 5- to 6-membered heteroaryl having 1-3 heteroatoms selected from O, N, S, or -NR 12 R 13 , where R 12 and R 13 Each independently selected from H or C1-C10 alkyl;

[0016] R2 is selected from H, a group derived from an α, β or γ amino acid, -C(=O)R 21 、-C(=O)CH2-R 21 ,;-S(=O)2R 22 , where R 21 are each independently selected from substituted or unsubstituted C3-C6 cycloalkyl, 5- to 6-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S, and

[0017] R 22 For-NR 12 R 13 , where R 12 and R 13 Each independently selected from H or C1-C10 alkyl;

[0018] The above substitution refers to substitution with a substituent selected from halogen, hydroxyl, CN, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C6 cycloalkyl, C1-C10 alkoxy, wherein the C1-C10 alkyl, C3-C6 cycloalkyl or C1-C10 alkoxy may be optionally further substituted with halogen or amino.

[0019] In a specific embodiment, R1 and R2 can be H at the same time.

[0020] In a specific embodiment, when R1 is H, R2 is not H.

[0021] In a specific embodiment, when R1 is not H, R2 is H.

[0022] In a specific embodiment, the compound of the general formula I can be represented by the following formula I-1:

[0023]

[0024] In Formula I-1, R 111is selected from halogen, hydroxyl, CN, C1-C10 alkyl unsubstituted or substituted by halogen, C2-C10 alkenyl unsubstituted or substituted by halogen, C2-C10 alkynyl unsubstituted or substituted by halogen, C3-C6 cycloalkyl unsubstituted or substituted by halogen, C1-C10 alkoxy unsubstituted or substituted by halogen, n is an integer of 1-5, R2 is as defined above, preferably, R2 is H.

[0025] In a specific embodiment, R1 is selected from: H, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted C1-C6 alkylacyl, substituted or unsubstituted C3-C6 cycloalkylacyl, substituted or unsubstituted C5-C10 arylacyl, substituted or unsubstituted 5- to 6-membered heteroarylacyl, substituted or unsubstituted C1-C6 alkylsulfonyl, substituted or unsubstituted C3-C6 cycloalkylsulfonyl, substituted or unsubstituted C5-C10 arylacyl, or substituted or unsubstituted 5- to 6-membered heteroarylsulfonyl, wherein the substituents of the above groups when substituted are as defined above.

[0026] In a specific embodiment, R2 is selected from: H, substituted or unsubstituted α, β or γ amino acids, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted C1-C10 alkyl acyl, substituted or unsubstituted C3-C6 cycloalkyl acyl, substituted or unsubstituted 5- to 6-membered heterocycloalkyl acyl, or substituted or unsubstituted 5- to 6-membered heterocycloalkyl methylene acyl, wherein the substituents of the above groups when substituted are as defined above.

[0027] In a specific embodiment, R2 is a group derived from an α or β or γ amino acid.

[0028] In a specific embodiment, R2 is aminosulfonyl or aminosulfonyl substituted by C-C10 alkyl.

[0029] In a specific embodiment, the compound of formula I can be selected from the following specific compounds:

[0030]

[0031]

[0032]

[0033] In a specific embodiment, the pharmaceutically acceptable salt includes organic acid salts and inorganic acid salts, including but not limited to maleate, succinate, citrate, tartrate, fumarate, acetate, methanesulfonate, hydrochloride, phosphate, nitrate or sulfate.

[0034] The second aspect of the present invention provides a method for preparing the compound according to the first aspect of the present invention, wherein the method can be selected from the following route 1, route 2 and route 3:

[0035] (a) Route 1:

[0036] In the general formula I, when R2 is H, the compound of the general formula I can be prepared by route 1:

[0037]

[0038] In an organic solvent, compound A is reduced to a hydroxymethyl group under the action of a reducing agent in a specific solvent to obtain compound B.

[0039] Preferably, the solvent is an aprotic polar solvent, such as diethyl ether, tetrahydrofuran, dioxane, toluene. The reducing agent is lithium aluminum hydride, borane complex,

[0040] Wherein, in route 1, the substituent R1 in compounds A and B is defined as above.

[0041] (b) Route 2:

[0042] In the general formula I, when R2 is not H, the compound of the general formula I can be prepared by route 2:

[0043]

[0044] In an organic solvent, in the presence of a base, compound B reacts with an electrophilic reagent to generate a target compound P; or

[0045] In an organic solvent, in the presence of a condensing agent and a base, compound B reacts with the corresponding carboxylic acid to generate the target compound P.

[0046] Wherein, in route 2, the substituents R1 and R2 in compounds B and P are defined as above.

[0047] Preferably, the solvent is a non-protonic polar solvent, such as N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, dichloroethane, tetrahydrofuran, and dioxane; the electrophilic reagent is sulfonyl chloride, acyl chloride, and acid anhydride; the base is triethylamine, N,N-diethylisopropylamine, sodium acetate, potassium carbonate, and 4-dimethylaminopyridine; and the condensing agent is common dicyclohexylcarbodiimide (DCC), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBT), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)urea hexafluorophosphate (HATU).

[0048] (c) Route 3:

[0049] In the general formula I, when R1 is not H, the compound of the general formula I can be prepared by route 3:

[0050]

[0051] In an organic solvent, in the presence of a base, compound C reacts with an electrophilic reagent to generate a target compound P; or

[0052] In an organic solvent, in the presence of a condensing agent and a base, compound C reacts with the corresponding carboxylic acid to generate the target compound P.

[0053] Wherein, in route 3, the substituents R1 and R2 in compounds C and P are defined as described above.

[0054] Preferably, the solvent is a non-protonic polar solvent, such as N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, dichloroethane, tetrahydrofuran, and dioxane; the electrophilic reagent is sulfonyl chloride, acyl chloride, and acid anhydride; the base is triethylamine, N,N-diethylisopropylamine, sodium acetate, potassium carbonate, and 4-dimethylaminopyridine; and the condensing agent is common dicyclohexylcarbodiimide (DCC), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBT), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1-yl)urea hexafluorophosphate (HATU).

[0055] The third aspect of the present invention provides a pharmaceutical composition, which contains a therapeutically effective amount of the compound as described in the first aspect of the present invention, its pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate, and optionally contains a pharmaceutically acceptable carrier.

[0056] The fourth aspect of the present invention provides the use of the compound as described in the first aspect of the present invention, its pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate, or the pharmaceutical composition as described in the third aspect of the present invention in the preparation of a drug for preventing and / or treating diseases caused by inflammation and / or abnormal osteoclast activity.

[0057] In a specific embodiment, the diseases caused by the inflammation and / or abnormal osteoclast activity include but are not limited to rheumatoid arthritis, inflammatory bowel disease, chronic obstructive pulmonary disease, asthma, psoriasis, idiopathic pulmonary fibrosis, osteoarthritis, osteoporosis, periodontitis, tooth loss, Paget's disease of bone, rickets, giant cell tumor of bone, myeloma bone disease, and bone destruction caused by cancer bone metastasis.

[0058] The fifth aspect of the present invention provides a method for inhibiting osteoclast activity, which comprises the following steps: administering to a subject in need thereof a safe and effective amount of a compound as described in the first aspect of the present invention, a pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate thereof, or a pharmaceutical composition as described in the third aspect of the present invention.

[0059] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

[0060] Beneficial Effects

[0061] The present invention develops a class of compounds of general formula I, namely pyrazolooleanolic acid reductol derivatives, which are confirmed by pharmacological experiments to have definite anti-inflammatory activity and activity of inhibiting osteoclast differentiation, and therefore have the prospect of being developed into drugs for treating diseases related to inflammation and abnormal osteoclast activity. DETAILED DESCRIPTION

[0062] the term

[0063] As used herein, unless otherwise specified, the term "substituted" means that one or more hydrogen atoms on a group are replaced by a substituent selected from the group consisting of halogen, hydroxy, CN, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C6 cycloalkyl, C1-C10 alkoxy, and the C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C6 cycloalkyl, C1-C10 alkoxy may be further substituted by halogen.

[0064] Unless otherwise specified, in all compounds of the present invention, each chiral carbon atom may be in R configuration or S configuration, or a mixture of R configuration and S configuration.

[0065] The term "C1-C10 alkyl" refers to a straight or branched alkyl group having 1 to 10 carbon atoms, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.

[0066] The term "C2-C10 alkenyl" refers to a straight or branched alkenyl group having 2 to 10 carbon atoms, such as ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, sec-butenyl, tert-butenyl, or the like.

[0067] The term "C2-C10 alkynyl" refers to a straight or branched alkynyl group having 2 to 10 carbon atoms, such as ethynyl, propynyl, isopropynyl, butynyl, isobutynyl, sec-butynyl, tert-butynyl, or the like.

[0068] The term "C3-C6 cycloalkyl" refers to a cycloalkyl group having 3 to 6 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like.

[0069] The term "C1-C10 alkoxy" refers to an alkoxy group having 1 to 10 carbon atoms, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, or the like.

[0070] The term halogen refers to F, Cl, Br and I.

[0071] The term "5- or 6-membered heterocyclic group" refers to a substituted or unsubstituted 5- or 6-membered heterocyclic group containing oxygen, nitrogen or sulfur.

[0072] The term "5- or 6-membered heteroaromatic ring group" refers to a substituted or unsubstituted unsaturated 5- or 6-membered heteroaromatic ring group containing oxygen, nitrogen or sulfur.

[0073] Hereinafter, the present invention is described in detail through specific embodiments so that those skilled in the art can better understand the present invention, however, these embodiments are not intended to limit the scope of the present invention.

[0074] In all the following examples, the starting compound oleanolic acid and related reagents were purchased from Shaoyuan Technology (Shanghai) Co., Ltd., Beijing Bailingwei Technology Co., Ltd., and Shanghai Titan Technology Co., Ltd. Unless otherwise specified, other starting materials, solvents, and materials were all from Sinopharm Reagent Company. 1 H NMR was recorded by Brucher AM-400 or GEMINI-400 nuclear magnetic resonance instrument, and chemical shift was expressed in δ (ppm). Mass spectra were recorded by Agilent 1200-6110 single quadrupole liquid chromatography-mass spectrometry. 200-300 mesh silica gel for separation was provided by Qingdao Ocean Chemical Plant. The chemical reagents represented by the English abbreviations are as follows:

[0075] DMF: N,N-dimethylformamide

[0076] THF: Tetrahydrofuran

[0077] DCM: dichloromethane

[0078] DMAP: 4-dimethylaminopyridine

[0079] DCC: dicyclohexylcarbodiimide

[0080] Tr: triphenylmethyl

[0081] Preparation and synthesis of compounds

[0082] Synthesis route of compound 1-18

[0083]

[0084] Reaction conditions: (a) Dess-Martin periodinane, CH2Cl2, rt, 1 h; (b) HCOOEt, MeONa, THF, room temperature, overnight; (c) NH2NH2·2HCl, EtOH, reflux, 4 h; (d) BH3, THF, N2, 70°C, reflux, overnight; (e) substituted sulfonyl chloride, Et3N, CH2Cl2, room temperature, overnight; (f) 2-cyclopropylacetic acid, HOBT, EDCI, Et3N, CH2Cl2, room temperature, overnight,

[0085] In this reaction scheme, R is defined as R1 above, except that R1 is H or -C(=O)R 11 options.

[0086] Preparation of intermediate o1

[0087]

[0088] Oleanolic acid (20 g, 43.79 mmol) was dissolved in dichloromethane (250 mL), and Dess-Martin periodinane (24 g, 56.93 mmol) was added in batches. The reaction system was reacted at room temperature for 1.5 hours. After the reaction of oleanolic acid was completed by thin layer chromatography (TLC), a saturated sodium bicarbonate solution (300 mL) mixed with sodium thiosulfate (100 g) was poured into the reaction system, stirred at room temperature for 15 minutes, and dichloromethane (300 mL) was added to extract three times. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to remove the organic solvent to obtain the crude product o1, a yellow powder with a yield of 97%. ESI-MS: 453.6 (M-1).

[0089] Preparation of intermediate o2

[0090]

[0091] Under nitrogen protection, compound o1 (19.5 g, 42.88 mmol) and freshly prepared sodium methoxide (11.6 g, 214.4 mmol) were dissolved in anhydrous tetrahydrofuran. Ethyl formate (17.2 mL, 214.4 mmol) was slowly added to the reaction mixture at 0°C and stirred overnight at room temperature. After TLC monitoring of the reaction was complete, water (200 mL) was poured into the reaction system, 1 M dilute hydrochloric acid was added dropwise to adjust the pH of the system to 3-5, and ethyl acetate (300 mL) was used for extraction three times. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to remove the organic solvent to obtain crude product o2 as a light yellow solid with a yield of 93%.

[0092] Preparation of intermediate o3

[0093]

[0094] Compound o2 (19.2 g, 39.77 mmol) was dissolved in anhydrous ethanol (200 mL), hydrazine dihydrochloride (4.3 g, 41.36 mmol) was added, and the reaction system was heated to 82°C for 4 hours. After the reaction was complete as monitored by TLC, the reaction solution was cooled to room temperature, concentrated in vacuo to remove the organic solvent, and separated and purified by silica gel column chromatography (dichloromethane: methanol = 80: 1 to 30: 1) to obtain compound o3 as a yellow solid with a yield of 72%. 1 H NMR (400MHz, chloroform-d) δ7.19(s,1H),5.36(s,1H),2.93–2.85(m,1H),2.55( d,J=14.8Hz,1H),2.07-1.91(m,4H),1.83–1.71(m,3H),1.69–1.54(m,4H) ),1.52–1.30(m,5H),1.28(s,3H),1.25(s,1H),1.21–1.19(m,1H),1.17( s,6H),1.12(s,1H),0.94(s,3H),0.91(s,3H),0.83(s,3H),0.79(s,3H). 13 C NMR (151 MHz, chloroform-d) δ 183.01, 150.47, 144.04, 132.00, 122.44, 113.02, 53.44, 46.68, 46.32, 46.20, 42.01, 41.44, 39.49, 38.55, 36.28, 34.09, 33.51, 33.27, 32.65, 32.21, 31.38, 30.88, 27.93, 25.91, 24.20, 23.76, 23.50, 23.30, 19.35, 16.78, 15.26. MS (ESI) m / z C 31 H 47 N2O2[M+H] + Calculated value 479.7, measured value 479.5.

[0095] Example 1: Synthesis of Compound 1

[0096]

[0097] Under nitrogen protection, o3 (1.5 g, 3.13 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL), and borane tetrahydrofuran complex (3.07 mL, 31.3 mmol) was slowly added dropwise at 0°C, and the reaction system was refluxed at 70°C for 5 hours. After TLC monitoring of the reaction was complete, the reaction system was cooled to 0°C, and 4.0 M hydrochloric acid methanol solution was added dropwise to the reaction system, and refluxed for 30 minutes. The reaction solution was cooled to room temperature, vacuum concentrated to remove the organic solvent, and separated and purified by silica gel column chromatography (dichloromethane: methanol = 100: 1 to 50: 1) to obtain the target product 1 as a white solid with a yield of 80%. 1H NMR (500 MHz, chloroform-d) δ7.32 (s, 1H), 5.26 (t, J = 3.7 Hz, 1H), 3.58 (d, J = 10.9 Hz, 1H), 3.24 (d, J = 11.0 Hz, 1H), 2.58 (d, J = 14.9 Hz, 1H), 2.06–1.98 (m, 3H), 1.98–1.87 (m, 2H), 1.80–1.70 (m, 3H), 1.6 6–1.58(m,2H),1.58–1.50(m,2H),1.50–1.36(m,2H),1.34(s,3H),1.33–1.28(m,3H),1.24(s, 3H),1.23–1.21(m,1H),1.20(s,3H),1.13–1.03(m,2H),1.01(s,3H),0.90(s,3H),0.89(s,6H). 13 C NMR (125MHz, CDCl3) δ150.41,144.27,122.51,113.11,69.81,53.38,46.65,46.22,42.60,42.08,40.00,38.48,37.14,36.38,3 4.25,33.51,33.32,32.09,31.36,31.18,31.11,25.88,25.71,24.16,23.73,23.66,22.22,19.33,16.63,15.49.HRMS(ESI)m / z C 31 H 49 N2O[M+H] + Calculated value 465.3839, measured value 465.3835.

[0098] Synthesis of Compound 2-17

[0099]

[0100] Compound 1 (66 mg, 0.14 mmol, 1.0 eq), triethylamine (60 μL, 0.42 mmol, 3.0 eq) and the corresponding sulfonyl chloride compound (0.63 mmol, 1.5 eq) were dissolved in dichloromethane solution in turn, and the reaction was stirred at room temperature overnight. After TLC monitoring of the reaction was complete, the reaction mixture was slowly poured into water (100 mL), extracted three times with ethyl acetate (90 mL), and the organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to remove the organic solvent. Silica gel column chromatography was separated and purified (petroleum ether: ethyl acetate = 10:1 to 1:1 gradient elution) to obtain the target product 2-17.

[0101] Example 2: Synthesis of Compound 2

[0102]

[0103] White solid (yield 95%). 1 H NMR (500MHz, chloroform-d) δ7.89–7.85(m,2H),7.63(d,J=1.3Hz,1H),6.95–6.91(m,2H),5.24(t,J=3.6Hz,1H),3.85(s,3H),3.56 (d,J=11.0Hz,1H),3.23(d,J=11.0Hz,1H),2.60(d,J=15.2Hz,1H),2.04–1.99(m,1H),1.98–1.96(m,1H),1.95–1.85(m,3H) ,1.77–1.73(m,1H),1.72–1.66(m,2H),1.61–1.51(m,4H),1.48–1.33(m,4H),1.32–1.27(m,2H),1.26(s,4H),1.25–1.23(m ,1H),1.22–1.19(m,2H),1.19(s,3H),1.16(s,3H),1.08–1.01(m,2H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.79(s,3H). 13C NMR (125MHz, CDCl3) δ164.45,163.93,144.34,130.07,129.45,128.41,122 .32,118.75,114.31,69.78,55.80,53.28,46.60,45.97,42.57,42.05,39.9 0,37.93,37.10,36.31,34.73,34.21,33.31,32.01,31.40,31.12,31.09,25 .81,25.63,24.65,23.71,23.61,22.17,19.47,16.55,15.43.HRMS(ESI)m / z C 38 H 55 N2O4S[M+H] + Calculated value: 635.3883, measured value: 635.3884.

[0104] Example 3: Synthesis of Compound 3

[0105]

[0106] White solid (yield 92%). 1 H NMR (600MHz, chloroform-d) δ7.63(d,J=1.4Hz,1H),7.49(dt,J=7.8,1.3Hz,1H),7.43(t,J=2.1Hz,1H),7.36(t,J=8.1Hz,1H),7.09(dd,J=8.2,2.1Hz,1H),5 .24(t,J=3.7Hz,1H),3.83(s,3H),3.56(d,J=10.9Hz,1H),3.23(d,J=10. 9Hz,1H),2.61(d,J=15.3Hz,1H),2.03–1.98(m,1H),1.98–1.93(m,2H),1. 93–1.86(m,2H),1.76–1.67(m,3H),1.61–1.51(m,3H),1.47(td,J=12.3, 4.0Hz,1H),1.43–1.39(m,1H),1.38–1.32(m,2H),1.30(d,J=4.1Hz,1H),1 .27(s,3H),1.20(s,4H),1.19–1.17(m,1H),1.16(s,3H),1.10–1.06(m,1H ),1.06–1.02(m,1H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.79(s,3H). 13C NMR (125MHz, CDCl3) δ164.82,159.85,144.36,139.12,130.09,128.72,122.29,120. 88,119.74,119.18,111.83,69.77,55.85,53.26,46.61,45.98,42.57,42.05,39.91 ,37.93,37.11,36.30,34.78,34.22,33.30,32.00,31.42,31.13,31.09,25.80,25.6 4,24.68,23.71,23.61,22.18,19.48,16.55,15.43.HRMS(ESI)m / zC38H55N2O4S[M+H] + Calculated value 635.3877, measured value 635.3872.

[0107] Example 4: Synthesis of Compound 4

[0108]

[0109] White solid (yield 88%). 1 H NMR (600MHz, chloroform-d) δ7.73(s,1H),7.68(d,J=8.3Hz,2H),6.72(d,J=8.3Hz,2H),5.25(t,J=3.7Hz,1H),3.5 8(d,J=10.9Hz,1H),3.24(d,J=11.0Hz,1H),2.64(d,J=15.2Hz,1H),2.04–1.95(m,3H),1.94–1.86(m,2H), 1.77–1.68(m,4H),1.61–1.51(m,4H),1.49–1.39(m,3H),1.39–1.34(m,2H),1.34–1.30(m,2H),1.26(s,4H ),1.17(s,3H),1.11–1.07(m,1H),1.06–1.02(m,1H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.81(s,3H). 13C NMR (125MHz, CDCl3) δ164.56,161.82,144.33,130.36,128.93,127.96,12 2.34,118.44,116.21,69.87,53.32,46.61,46.02,42.59,42.07,39.93,3 7.92,37.12,36.30,34.82,34.21,33.31,32.00,31.30,31.15,31.11,25. 83,25.65,24.60,23.72,23.64,22.18,19.48,16.57,15.49.HRMS(ESI)m / z C 37 H 53 N2O4S[M+H] + Calculated value: 621.3721, measured value: 621.3728.

[0110] Example 5: Synthesis of Compound 5

[0111]

[0112] White solid (yield 93%). 1 H NMR (600MHz, chloroform-d) δ7.99–7.94(m,2H),7.64(d,J=1.4Hz,1H),7.18–7.13(m,2H),5.24(t,J=3.6Hz,1H),3.56(d,J=11.0Hz, 1H),3.23(d,J=11.0Hz,1H),2.61(d,J=15.3Hz,1H),2.03–1.98(m,1H),1.98–1.93(m,2H),1.92–1.86(m,2H),1.77–1.67(m ,4H),1.60–1.51(m,3H),1.49–1.39(m,2H),1.38–1.29(m,3H),1.25(s,3H),1.24–1.22(m,1H),1.18(s,3H),1.16(s,3H),1 .08(ddd,J=13.4,4.7,2.5Hz,1H),1.04(ddd,J=13.8,4.5,2.6Hz,1H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.78(s,3H). 13C NMR (125MHz, CDCl3) δ165.86 (d, J = 256.9Hz), 165.16, 144.38, 133.91 (d, J = 2.9Hz), 130. 67(d,J=9.7Hz),128.61,122.24,119.31,116.48(d,J=22.8Hz),69.75,53.20,46.60,45 .95,42.55,42.04,39.89,37.90,37.09,36.26,34.76,34.20,33.30,31.97,31.38,31.1 1,31.08,25.79,25.62,24.65,23.70,23.59,22.15,19.45,16.53,15.42.HRMS(ESI)m / z C 37 H 52 FN2O3S[M+H] + Calculated value: 623.3683, measured value: 623.3691.

[0113] Example 6: Synthesis of Compound 6

[0114]

[0115] White solid (yield 96%). 1 H NMR (400MHz, chloroform-d) δ7.73 (d, J=7.9Hz, 1H), 7.66–7.60 (m, 2H), 7.47 (td, J=8.1, 5.2Hz, 1H), 7.32–7.27 (m, 1H), 5.24(t,J=4.2Hz,1H),3.56(d,J=10.9Hz,1H),3.23(d,J=11.0Hz,1H),2.62(d,J=15.2Hz,1H),2.05–1.96(m,2H ),1.95–1.84(m,3H),1.78–1.66(m,3H),1.57–1.45(m,3H),1.45–1.36(m,3H),1.35–1.29(m,2H),1.26(s,4H), 1.21(s,1H),1.19(s,3H),1.17(s,3H),1.11–1.04(m,2H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.78(s,3H). 13C NMR (125MHz, CDCl3) δ165.45, 162.20 (d, J = 252.1Hz), 144.38, 139.77 (d, J = 7.3Hz), 130.94 (d, J = 7. 7Hz),128.81,123.52(d,J=3.2Hz),122.26,121.25(d,J=21.2Hz),119.61,115.10(d,J=25.0Hz),6 9.79,53.22,46.61,45.96,42.57,42.06,39.91,37.92,37.11,36.27,34.80,34.21,33.31,31.98, 31.39,31.13,31.10,25.81,25.64,24.67,23.71,23.61,22.17,19.47,16.55,15.43.HRMS(ESI)m / z C 37 H 52 FN2O3S[M+H] + Calculated value: 623.3677, measured value: 623.3683.

[0116] Example 7: Synthesis of Compound 7

[0117]

[0118] White solid (yield 93%). 1 H NMR (500MHz, chloroform-d) δ7.90–7.85(m,2H),7.63(d,J=1.4Hz,1H),7.48–7.43(m,2H),5.24(t,J=3.7Hz,1H),3.55(d,J=11.0 Hz,1H),3.23(d,J=10.9Hz,1H),2.61(d,J=15.2Hz,1H),2.04–1.99(m,1H),1.98–1.93(m,2H),1.92–1.85(m,2H),1.77– 1.67(m,3H),1.59–1.48(m,4H),1.48–1.39(m,3H),1.39–1.35(m,1H),1.35–1.32(m,1H),1.32–1.29(m,1H),1.25(s,3H ),1.23–1.20(m,2H),1.19(s,3H),1.17(s,3H),1.11–1.01(m,2H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.78(s,3H). 13C NMR (125MHz, CDCl3) δ165.29,144.40,140.70,136.42,129.49,129.21,12 8.62,122.28,119.44,69.80,53.24,46.63,45.99,42.59,42.08,39.93,37 .94,37.13,36.30,34.81,34.23,33.32,32.01,31.42,31.14,31.11,25.8 2,25.66,24.70,23.72,23.63,22.20,19.48,16.56,15.47.HRMS(ESI)m / zC 37 H 52 ClN2O3S[M+H] + Calculated value 639.3382, measured value 639.3389.

[0119] Example 8: Synthesis of Compound 8

[0120]

[0121] White solid (yield 94%). 1 H NMR (500MHz, chloroform-d) δ8.07(d,J=8.2Hz,2H),7.75(d,J=8.4Hz,2H),7.65(d,J=1.5Hz,1H),5.24(t,J=3.7Hz,1H) ,3.55(d,J=10.9Hz,1H),3.23(d,J=10.9Hz,1H),2.61(d,J=15.4Hz,1H),2.04–1.99(m,1H),1.98–1.93(m,2H) ,1.93–1.85(m,2H),1.77–1.66(m,3H),1.56–1.51(m,2H),1.50–1.39(m,2H),1.38–1.35(m,1H),1.33(s,1H), 1.26(s,7H),1.19(s,3H),1.16(s,3H),1.12–1.01(m,2H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.78(s,3H). 13C NMR (125MHz, CDCl3) δ165.71,144.42,141.40,135.53,128.77,128.32,126.30 (q,J=3.4Hz),123.15,122.23,119.80,69.78,53.19,46.63,45.98,42.58,42.0 7,39.92,37.92,37.12,36.27,34.83,34.22,33.31,31.98,31.40,31.13,31.10 ,25.81,25.65,24.71,23.72,23.62,22.19,19.46,16.56,15.46.HRMS(ESI)m / z C 38 H 52 F3N2O3S[M+H] + Calculated value 673.3645, measured value 673.3639.

[0122] Example 9: Synthesis of Compound 9

[0123]

[0124] White solid (yield 91%). 1 H NMR (500MHz, chloroform-d) δ8.07–8.04(m,2H),7.79–7.76(m,2H),7.64(d,J=1.4Hz,1H),5.24(t,J=3.7Hz,1H),3.55(d,J=11.0 Hz,1H),3.23(d,J=10.9Hz,1H),2.62(d,J=15.4Hz,1H),2.04–1.99(m,1H),1.98–1.92(m,2H),1.92–1.85(m,2H),1.77– 1.66(m,3H),1.61–1.51(m,4H),1.50–1.46(m,1H),1.45–1.39(m,2H),1.38–1.35(m,1H),1.35–1.32(m,1H),1.32–1.29 (m,1H),1.24(s,3H),1.18(s,3H),1.16(s,3H),1.11–1.01(m,2H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.78(s,3H). 13C NMR (125MHz, CDCl3) δ166.05,144.45,141.88,132.89,128.79,128.40,122. 19,120.05,117.68,117.12,69.78,53.18,46.63,45.98,42.58,42.08,39.9 2,37.92,37.12,36.25,34.84,34.21,33.31,31.97,31.42,31.12,31.11,25 .81,25.65,24.72,23.72,23.62,22.18,19.46,16.55,15.48.HRMS(ESI)m / zC 38 H 52 N3O3S[M+H] + Calculated value 630.3724, measured value 630.3742.

[0125] Example 10: Synthesis of Compound 10

[0126]

[0127] White solid (yield 90%). 1 H NMR (400MHz, chloroform-d) δ8.02–7.97(m,2H),7.64(d,J=1.3Hz,1H),7.33–7.28(m,2H),5.24(t,J=3.7 Hz,1H),3.56(d,J=10.9Hz,1H),3.23(d,J=10.9Hz,1H),2.61(d,J=15.3Hz,1H),2.04–1.85(m,5H ),1.78–1.66(m,3H),1.62–1.46(m,4H),1.45–1.29(m,5H),1.26(s,3H),1.21(d,J=4.5Hz,1H),1 .19(s,3H),1.16(s,3H),1.12–1.01(m,2H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.78(s,3H). 13C NMR (125MHz, CDCl3) δ165.44,153.09,144.41,136.08,130.06,128.72,12 2.25,120.78,119.56,69.78,53.22,46.63,45.98,42.58,42.07,39.92,37 .93,37.12,36.28,34.80,34.22,33.31,31.99,31.39,31.13,31.10,25.8 1,25.65,24.68,23.71,23.62,22.19,19.47,16.55,15.42.HRMS(ESI)m / zC 38 H 52 F3N2O4S[M+H] + Calculated value: 689.3594, measured value: 689.3611.

[0128] Example 11: Synthesis of Compound 11

[0129]

[0130] White solid (yield 88%). 1 H NMR (400MHz, chloroform-d) δ7.83–7.72(m,2H),7.63(s,1H),7.33–7.28(m,1H),5.24(t,J=3.6Hz,1H),3.5 6(d,J=10.9Hz,1H),3.23(d,J=10.9Hz,1H),2.62(d,J=15.3Hz,1H),2.04–1.84(m,5H),1.79–1.65 (m,3H),1.63–1.39(m,7H),1.39–1.36(m,1H),1.35–1.33(m,1H),1.32–1.29(m,1H),1.26(s,3H), 1.20(s,3H),1.17(s,3H),1.12–1.01(m,2H),0.98(s,3H),0.89(s,3H),0.88(s,3H),0.79(s,3H). 13C NMR (125MHz, CDCl3) δ 165.63, 153.98 (dd, J = 259.4, 12.4Hz), 150.08 (dd, J = 255.4, 13.4Hz), 144.41, 134. 52(d,J=8.6Hz),128.68,125.15(dd,J=7.6,3.9Hz),122.23,119.69,118.31(d,J=18.6Hz),117.86(d,J= 20.3Hz),69.77,53.21,46.62,45.98,42.57,42.06,39.92,37.92,37.11,36.26,34.82,34.21,33.30,31 .98,31.41,31.13,31.09,25.80,25.64,24.70,23.71,23.61,22.18,19.46,16.55,15.46.HRMS(ESI)m / z C 37 H 51 F2N2O3S[M+H] + 641.3583, measured value 641.3592.

[0131] Example 12: Synthesis of Compound 12

[0132]

[0133] White solid (yield 84%). 1 H NMR (500MHz, Methanol-d4) δ7.86(d,J=1.4Hz,1H),5.25(t,J=3.7Hz,1H),3.53(d,J=11.0Hz,1H),3.13(d,J=11.1H z,1H),2.72(s,0.5H),2.69(s,3.5H),2.37(s,3H),2.07–1.96(m,4H),1.89(dd,J=13.4,4.0Hz,1H),1.85–1.75(m, 3H),1.67–1.60(m,2H),1.58–1.50(m,2H),1.49–1.43(m,1H),1.40–1.28(m,5H),1.27(s,3H),1.24–1.22(m,1H),1 .20(s,3H),1.20(s,3H),1.17–1.15(m,1H),1.10–1.07(m,1H),1.04(s,3H),0.89(s,3H),0.88(s,3H),0.84(s,3H). 13C NMR(125MHz,MeOD)δ176.98,166.14,159.04,145.67,129.74,123.37,120. 65,115.65,69.73,54.41,47.87,47.18,43.87,43.05,41.03,38.94,38.16 ,37.03,35.72,35.28,33.75,33.03,32.26,31.86,31.69,26.62,26.31,24 .98,24.58,24.04,23.02,20.49,16.99,15.81,12.92,10.90.HRMS(ESI)m / z C 36 H 54 N3O4S[M+H] + Calculated value 624.3830, measured value 624.3819.

[0134] Example 13: Synthesis of Compound 13

[0135]

[0136] White solid (yield 86%). 1 H NMR (500MHz, chloroform-d) δ7.81(s,1H),7.72(s,1H),7.43(s,1H),5.26(t,J=3.6Hz,1H),3.75(s,3H),3.57(d,J=11.0Hz,1H),3.23(d,J=1 1.0Hz,1H),2.64(d,J=15.1Hz,1H),2.04–1.95(m,3H),1.90(td,J=14.2,4.5Hz,2H),1.78–1.67(m,3H),1.61–1.58(m,1H),1.56–1. 53(m,1H),1.53–1.46(m,2H),1.46–1.39(m,2H),1.37(t,J=3.3Hz,1H),1.36–1.33(m,1H),1.31(d,J=4.0Hz,1H),1.26(s,3H),1.25 –1.23(m,1H),1.19(s,3H),1.18(s,3H),1.12–1.07(m,1H),1.07–1.02(m,1H),0.99(s,3H),0.89(s,3H),0.89(s,3H),0.83(s,3H). 13C NMR (125MHz, CDCl3) δ164.34,144.25,139.47,138.03,129.32,126.26,12 2.42,118.22,69.81,53.39,46.59,45.99,42.59,42.06,39.91,37.94,37 .12,36.36,34.79,34.37,34.23,33.31,32.04,31.43,31.15,31.10,25.8 2,25.66,24.67,23.71,23.62,22.18,19.50,16.57,15.48.HRMS(ESI)m / z C 35 H 53 N4O3S[M+H] + Calculated value 609.3833, measured value 609.3819.

[0137] Example 14: Synthesis of Compound 14

[0138]

[0139] White solid (yield 54%). 1 H NMR (400MHz, DMSO-d6) δ7.63(d,J=1.4Hz,1H),5.26(t,J=3.6Hz,1H),3.57(d,J=11.0Hz,1H),3.41(q,J =7.4Hz,2H),3.24(d,J=11.0Hz,1H),2.65(d,J=15.3Hz,1H),2.06–1.99(m,3H),1.98–1.86(m,3H),1.80 –1.70(m,3H),1.63–1.50(m,5H),1.49–1.44(m,1H),1.44–1.40(m,1H),1.40–1.37(m,1H),1.34(s,4H), 1.26(s,3H),1.22–1.18(m,6H),1.13–1.06(m,2H),1.01(s,3H),0.90(s,3H),0.89(s,3H),0.87(s,3H). 13C NMR (125MHz, CDCl3) δ164.26,144.39,128.84,122.34,117.68,69.82,53.41,48.54,46.64,46.04,42.60,42.10,39.96,38.04,37.14,36. 30,34.80,34.24,33.32,32.06,31.54,31.15,31.12,25.86,25.68,24.67,23.73,23.66,22.22,19.55,16.60,15.48,7.75.HRMS(ESI)m / zC 33 H 53 N2O3S[M+H] + Calculated value 557.3771, measured value 557.3777.

[0140] Example 15: Synthesis of Compound 15

[0141]

[0142] White solid (yield 43%). 1 H NMR (500MHz, chloroform-d) δ7.57(s,1H),5.27(t,J=3.7Hz,1H),3.57(d,J=10.9Hz,1H),3.24(d,J=10.9Hz ,1H),2.65(d,J=15.3Hz,1H),2.08–1.99(m,3H),1.99–1.85(m,3H),1.80–1.70(m,3H),1.68–1.59 (m,3H),1.57–1.48(m,2H),1.48–1.42(m,2H),1.41–1.37(m,3H),1.36(s,3H),1.27(s,3H),1.20( s,5H),1.13–1.10(m,1H),1.09–1.05(m,3H),1.02(s,3H),0.89(s,3H),0.89(s,3H),0.88(s,3H). 13C NMR (125MHz, CDCl3) δ163.92,144.38,128.27,122.34,117.94,69.79,53.42,46.63,46.05,42.60,42.09,39.96,38.02,37.13,36.34,34.80 ,34.23,33.32,32.06,31.46,31.28,31.14,31.11,25.85,25.67,24.6 5,23.72,23.66,22.20,19.54,16.59,15.49,6.63,6.61.HRMS(ESI)m / z C 34 H 53 N2O3S[M+H] + Calculated value 569.3771, measured value 569.3774.

[0143] Example 16: Synthesis of Compound 16

[0144]

[0145] White solid (yield 32%). 1 H NMR (500 MHz, chloroform-d) δ7.63 (s, 1H), 5.27 (s, 1H), 3.62–3.53 (m, 3H), 3.25 (d, J=10.6 Hz, 1H), 2.67 (d, J=15.2 Hz, 1H), 2.55–2.44 (m, 2H), 2.07–2.00 (m, 3H), 2.00–1.86 (m, 3H) ,1.80–1.70(m,3H),1.67–1.51(m,6H),1.51–1.43(m,2H),1.34(s,3H),1.26(s,3H), 1.20(s,4H),1.13–1.04(m,2H),1.02(s,3H),0.90(s,3H),0.89(s,3H),0.87(s,3H). 13 C NMR (125MHz, CDCl3) δ165.18,144.42,128.60,125.10(d,J=276.4Hz),122.22,118.70,69.80,53.27,47.07,46.62,46.00,42.58,42.08,39.94,3 7.99,37.11,36.22,34.82,34.21,33.30,32.00,31.43,31.15,31.09,25 .83,25.66,24.62,23.71,23.64,22.19,19.51,16.59,15.47.MS(ESI)m / z C34 H 51 F3N2NaO3S[M+Na] + Calculated value 647.8, measured value 647.4.

[0146] Example 17: Synthesis of Compound 17

[0147]

[0148] White solid (yield 65%). 1 H NMR (400MHz, DMSO-d6) δ7.57 (s, 1H), 5.26 (t, J = 2.7Hz, 1H), 3.57 (d, J = 11.0Hz, 1H), 3.24 (d, J = 10.9Hz, 1H),2.86(s,6H),2.83(d,J=9.7Hz,1H),2.63(d,J=15.1Hz,1H),2.06–1.86(m,5H),1.81–1.70(m,3H),

[0149] 1.66–1.47(m,5H),1.46–1.36(m,2H),1.33(s,3H),1.33–1.30(m,1H),1.25(s,3H),1.23–1.2 1(m,1H),1.20(s,3H),1.13–1.03(m,2H),1.01(s,3H),0.90(s,3H),0.89(s,3H),0.86(s,3H). 13 C NMR (125MHz, CDCl3) δ162.28,144.34,129.08,122.39,116.12,69.82,53.37,46.62,46.05,42.59,42.08,39.95,38.98,38.11,37.13, 36.26,34.64,34.23,33.33,32.07,31.45,31.15,31.11,25.85,25.68,24.55,23.72,23.66,22.19,19.56,16.60,15.48.HRMS(ESI)m / z C 33 H 54 N3O3S[M+H] + Calculated value 572.3880, measured value 572.3882.

[0150] Example 18: Synthesis of Compound 18

[0151]

[0152] Compound 1 (66 mg, 0.14 mmol, 1.0 eq), cyclopropylacetic acid (16 μL, 0.17 mmol, 1.2 eq), HOBT (23 mg, 0.17 mmol, 1.2 eq), EDCI (33 mg, 0.17 mmol, 1.2 eq) and triethylamine (22 μL, 0.16 mmol, 1.1 eq) were added to dichloromethane (10 mL) in sequence and stirred at room temperature overnight. After the reaction was complete as monitored by TLC, the reaction mixture was slowly poured into water (100 mL) and extracted three times with dichloromethane (90 mL). The organic layers were combined, back-extracted once with water (30 mL) and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to remove the organic solvent. The target product 18 was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1 to 1:1 gradient elution) to obtain the target product 18 as a white solid in a yield of 85%. 1 H NMR (600MHz, chloroform-d) δ7.86 (s, 1H), 5.26 (t, J = 3.7Hz, 1H), 3.58 (d, J = 11.0Hz, 1H), 3.24 (d, J = 11.0Hz, 1H), 3.02–2.92 (m, 2H), 2.66 (d, J = 15.2Hz, 1H), 2.04–1.99(m,3H),1.98–1.94(m,1H),1.91(td,J=13.8,12.8,3.5Hz,1H) ,1.78–1.74(m,2H),1.74–1.72(m,1H),1.65–1.59(m,2H),1.59–1.52(m,2 H),1.50(dd,J=12.1,3.0Hz,1H),1.47–1.41(m,2H),1.38(t,J=3.7Hz,1H ),1.36–1.33(m,1H),1.32(s,3H),1.29(d,J=3.1Hz,1H),1.24(s,3H),1.2 2–1.21(m,1H),1.20(s,3H),1.12–1.04(m,2H),1.01(s,3H),0.90(s,3H), 0.89(s,3H),0.87(s,3H),0.59–0.54(m,2H),0.27(dt,J=6.1,4.6Hz,2H). 13C NMR (125MHz, CDCl3) δ172.00,162.61,144.30,124.86,122.45,119.11,6 9.84,53.42,46.63,46.03,42.61,42.10,39.97,39.00,38.08,37.14,36 .58,34.63,34.25,33.32,32.09,31.58,31.17,31.12,25.87,25.70,24. 76,23.73,23.68,22.23,19.59,16.60,15.53,7.05,4.57.HRMS(ESI)m / zC 36 H 55 N2O2[M+H] + Calculated value 547.4258, measured value 547.4271.

[0153] Synthetic route of compound 19-33

[0154]

[0155] Reaction conditions: (a) triphenylmethyl chloride, Et3N, CH2Cl2, room temperature, 3h; (b) corresponding carboxylic acid, DCC, DMAP, CH2Cl2, room temperature, 3h; (c) 4.0M hydrochloric acid dioxane solution, room temperature, 1h; (d) aminosulfonylpyridinium salt, CH2Cl2, N2, room temperature, overnight; (e) iodomethane, cesium carbonate, DMF, room temperature, 1h,

[0156] In the above reaction formula, the definition of R corresponds to the definition of the options of R2 except H in the above text (ie, the remaining groups except C(=O) in the options of R2 except H).

[0157] Preparation of compound o4

[0158]

[0159] Compound 1 (1.0 g, 2.15 mmol) was dissolved in dichloromethane (15 mL), triethylamine (0.9 mL, 6.45 mmol) and triphenylmethane (0.90 g, 3.23 mmol) were added, and the reaction system was reacted at room temperature for 3 hours. After TLC monitoring, water (20 mL) was added, and dichloromethane (60 mL) was extracted three times. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to remove the organic solvent. Compound o4 was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 25: 1 to 10: 1) as a white solid with a yield of 80%.

[0160] Preparation of compound o5

[0161]

[0162] Compound o4 (500 mg, 0.71 mmol), the corresponding carboxylic acid (0.11 mmol), DCC (208 mg, 0.1 mmol) and DMAP (87 mg, 0.71 mmol) were dissolved in dichloromethane (10 mL), and the reaction system was reacted at room temperature for 3 hours. After TLC monitoring, water (20 mL) was added, and dichloromethane (60 mL) was extracted three times. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to remove the organic solvent. Compound o5 was obtained by separation and purification by silica gel column chromatography (petroleum ether: ethyl acetate = 25:1 to 10:1).

[0163] Synthesis of target product 19-31

[0164]

[0165] Compound o5 (100 mg) was dissolved in 4.0 M hydrochloric acid dioxane solution (6 mL), and the reaction system was stirred at room temperature for 1 hour. After TLC monitoring, the reaction was completed, vacuum concentrated, saturated sodium bicarbonate solution (10 mL) was added to adjust the pH to alkaline, and dichloromethane (30 mL) was extracted three times. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and vacuum concentrated to remove the organic solvent. Silica gel column chromatography was separated and purified (dichloromethane: methanol = 80: 1 to 50: 1) to obtain the target product 19-31.

[0166] Example 19: Synthesis of Compound 19

[0167]

[0168] White solid (yield 78%). 1H NMR (500MHz, chloroform-d) δ7.24(s,1H),5.27(t,J=3.7Hz,1H),4.12(d,J=11.0Hz,1H),3.79(d,J=10.9Hz ,1H),3.45(s,2H),2.57(d,J=14.7Hz,1H),2.10–1.91(m,5H),1.78–1.66(m,3H),1.63–1.47(m,4H ),1.45–1.40(m,1H),1.36–1.31(m,1H),1.31(s,3H),1.30–1.28(m,1H),1.27–1.24(m,1H),1.21( s,3H),1.19(s,3H),1.18–1.16(m,1H),1.16–1.03(m,3H),1.01(s,3H),0.89(s,3H),0.88(d,6H). 13 C NMR (126 MHz, chloroform-d) δ 174.54, 143.47, 123.21, 112.61, 71.24, 53.46, 46.37, 46.20, 42.73, 41.96, 39.97, 38.46, 36.54, 36.12, 34.07, 33.48, 33.24, 32.07, 31.50, 31.44, 31.02, 25.91, 25.72, 24.22, 23.67, 22.44, 19.32, 16.58, 15.46. ESI-HRMS (m / z) [M+H] + C 33 H 52 N3O2, calculated value 522.4054; measured value 522.4051.

[0169] Example 20: Synthesis of Compound 20

[0170]

[0171] White solid (yield 80%). 1H NMR(500MHz, chloroform-d)δ7.23(s,1H),5.81–5.73(m,1H),5.27(t,J=3.6Hz,1H),5.14–5.11(m,1H),5.09(s,1H),4.14(d,J=11.0Hz,1H),3. 74(d,J=11.0Hz,1H),3.65(dd,J=7.4,4.8Hz,1H),3.15(d,J=7.2Hz,2H),2.87(dd,J=13.4,4.8Hz,1H),2.70(dd,J=13.4,7.4Hz,1H),2 .56(d,J=14.8Hz,1H),2.09–2.05(m,1H),2.04–1.90(m,4H),1.79–1.67(m,3H),1.62–1.46(m,4H),1.43–1.32(m,2H),1.30(s,3H),1. 29–1.26(m,1H),1.21(s,3H),1.18(s,4H),1.16–1.09(m,2H),1.07–1.02(m,1H),1.00(s,3H),0.89(s,3H),0.88(s,3H),0.87(s,3H). 13 C NMR (126 MHz, chloroform-d) δ 174.24, 143.35, 134.08, 123.27, 117.65, 112.42, 71.65, 54.26, 53.47, 46.27, 46.17, 42.71, 41.91, 39.94, 38.44, 36.54, 36.08, 36.02, 35.27, 34.05, 33.44, 33.22, 32.03, 31.66, 31.40, 30.98, 25.91, 25.69, 24.14, 23.70, 23.64, 22.46, 19.28, 16.54, 15.42. ESI-HRMS (m / z) [M+H] + C 37 H 58 N3O2S, calculated value 608.4244; measured value 608.4240.

[0172] Example 21: Synthesis of Compound 21

[0173]

[0174] White solid (yield 72%). 1H NMR (400MHz, chloroform-d) δ7.24(s,1H),5.28(t,J=3.6Hz,1H),4.16(d,J=11.0Hz,1H),3.78–3.73(m,2H),3.33–3.30( m,2H),3.11(dd,J=13.7,4.8Hz,1H),2.94(dd,J=13.7,7.3Hz,1H),2.57(d,J=14.8Hz,1H),2.26(t,J=2.6Hz,1H) ,2.11–1.91(m,5H),1.80–1.68(m,3H),1.63–1.47(m,4H),1.45–1.35(m,2H),1.31(s,3H),1.28–1.23(m,1H),1 .21(s,3H),1.19(s,4H),1.17–1.12(m,1H),1.11–1.02(m,2H),1.01(s,3H),0.89(s,3H),0.88(d,J=2.8Hz,6H). 13 C NMR (126 MHz, chloroform-d) δ 174.13, 143.39, 123.31, 112.65, 79.76, 71.79, 71.70, 54.42, 53.48, 46.31, 46.21, 42.76, 41.97, 39.99, 38.47, 36.75, 36.55, 36.14, 34.08, 33.50, 33.25, 32.06, 31.70, 31.45, 31.03, 25.94, 25.74, 24.23, 23.74, 23.68, 22.51, 20.04, 19.33, 16.59, 15.46. ESI-HRMS (m / z) [M+H] + C 37 H 56 N3O2S, calculated value 606.4088; measured value 606.4086.

[0175] Example 22: Synthesis of Compound 22

[0176]

[0177] White solid (yield 81%). 1H NMR (600MHz, chloroform-d) δ7.23(s,1H),5.25(t,J=3.6Hz,1H),4.04(d,J=11.0Hz,1H),3.84(d,J=11.0Hz,1H),3.78(dd,J=8.6,5.7Hz ,1H),3.11–3.06(m,1H),2.93–2.88(m,1H),2.56(d,J=14.7Hz,1H),2.18–2.11(m,1H),2.07(dd,J=13.6,4.5Hz,1H),2.04–1.9 0(m,4H),1.88–1.67(m,6H),1.62–1.45(m,4H),1.43–1.39(m,1H),1.35–1.31(m,1H),1.30(s,3H),1.28(s,1H),1.27–1.23(m, 1H),1.21(s,3H),1.18(s,3H),1.17–1.13(m,2H),1.12–1.08(m,1H),1.06–1.02(m,1H),1.00(s,3H),0.88(s,3H),0.87(s,6H). 13 C NMR (151 MHz, chloroform-d) δ 175.60, 143.44, 123.15, 112.39, 71.20, 59.93, 53.46, 47.04, 46.32, 46.18, 42.70, 41.92, 39.93, 38.45, 36.55, 36.18, 34.07, 33.43, 33.24, 32.04, 31.51, 31.40, 31.00, 30.61, 25.89, 25.69, 25.60, 24.15, 23.65, 23.63, 22.43, 19.29, 16.55, 15.44. ESI-HRMS (m / z) [M+H] + C 36 H 56 N3O2, calculated value 562.4367; measured value 562.4371.

[0178] Example 23: Synthesis of Compound 23

[0179]

[0180] White solid (yield 74%). 1H NMR (500MHz, chloroform-d) δ7.51(s,1H),7.23(s,1H),6.82(s,1H),5.27(t,J=3.6Hz,1H),4.13(d,J=10.9Hz,1H),3.81(dd,J=7.9, 4.4Hz,1H),3.74(d,J=11.0Hz,1H),3.11(dd,J=14.8,4.3Hz,1H),2.89(dd,J=14.8,7.9Hz,1H),2.57(d,J=14.8Hz,1H),2.0 9–1.90(m,5H),1.78–1.71(m,2H),1.69–1.54(m,3H),1.53–1.46(m,2H),1.43–1.38(m,1H),1.37–1.31(m,2H),1.30(s,3H) ,1.27–1.23(m,1H),1.20(s,3H),1.18(s,3H),1.16–1.09(m,2H),1.06–1.01(m,1H),0.98(s,3H),0.89(s,3H),0.88(s,6H). 13 C NMR (126 MHz, chloroform-d) δ 174.98, 143.40, 135.09, 123.25, 112.56, 71.67, 54.65, 53.47, 46.30, 46.18, 42.77, 41.95, 39.95, 38.46, 36.56, 36.11, 34.06, 33.52, 33.23, 32.06, 31.64, 31.48, 31.01, 25.91, 25.69, 24.29, 23.69, 22.43, 19.34, 16.56, 15.47. ESI-HRMS (m / z) [M+H] + C 37 H 56 N5O2, calculated value 602.4429; measured value 602.4434.

[0181] Example 24: Synthesis of Compound 24

[0182]

[0183] White solid (yield 77%). 1H NMR (500MHz, chloroform-d) δ7.22(s,1H),5.26(t,J=3.6Hz,1H),4.03(d,J=11.0Hz,1H),3.73(d,J=11.1Hz,1H),2.73(dd,J=12.9,5.1Hz,1H),2.62(dd,J= 12.9,6.7Hz,1H),2.56(d,J=14.8Hz,1H),2.36(dd,J=14.6,7.2Hz,1H),2.30(dd,J=14.6,6.0Hz,1H),2.09(dd,J=13.6,4.6Hz,1H),2.04–1.90(m,5 H),1.79–1.66(m,3H),1.65–1.57(m,3H),1.56–1.47(m,2H),1.45–1.39( m,1H),1.37–1.32(m,1H),1.30(s,3H),1.29–1.21(m,2H),1.21(s,3H),1 .19(s,3H),1.16–1.14(m,1H),1.13–1.08(m,2H),1.07–1.02(m,1H),1.0 0(s,3H),0.89(d,J=2.9Hz,4H),0.89–0.88(m,6H),0.87(d,J=1.7Hz,4H). 13 C NMR (126 MHz, chloroform-d) δ 173.64, 143.57, 123.12, 112.55, 70.88, 53.46, 46.37, 46.21, 45.62, 42.72, 41.95, 41.49, 39.95, 38.46, 37.47, 36.54, 36.09, 35.9 1,34.14,33.47,33.26,32.09,31.65,31.44,31.03,25.91,25.74,25.32,24 .21,23.67,22.99,22.78,22.56,19.32,16.60,15.46.ESI-HRMS(m / z)[M+H] + C 39 H 64 N3O2, calculated value 606.4993; measured value 606.4995.

[0184] Example 25: Synthesis of Compound 25

[0185]

[0186] White solid (yield 80%). 1H NMR (600MHz, chloroform-d) δ7.24(s,1H),5.28(t,J=3.6Hz,1H),4.98–4.94(m,1H),4.19(d,J=11.0Hz,1H),3.86(d,J=11.0Hz,1 H),2.64–2.52(m,4H),2.35–2.30(m,1H),2.08(dd,J=13.6,4.6Hz,1H),2.05–1.99(m,2H),1.98–1.92(m,2H),1.79–1.73 (m,2H),1.71–1.65(m,1H),1.63–1.55(m,2H),1.54–1.46(m,2H),1.44–1.40(m,1H),1.36–1.31(m,2H),1.31(s,3H),1. 28–1.24(m,2H),1.21(s,3H),1.19(s,3H),1.17–1.10(m,2H),1.10–1.06(m,1H),1.01(s,3H),0.90(s,3H),0.89(s,6H). 13 C NMR (126 MHz, chloroform-d) δ 176.09, 170.10, 143.23, 123.42, 112.73, 76.12, 72.29, 53.42, 46.26, 46.17, 42.68, 41.95, 39.97, 38.45, 36.52, 36.24, 34.01, 33.50, 33.23, 32.04, 31.53, 31.44, 31.00, 29.84, 26.94, 26.20, 25.94, 25.67, 24.24, 23.68, 23.62, 22.44, 19.31, 16.60, 15.47. ESI-HRMS (m / z) [M+H] + C 36 H 53 N2O4, calculated value 577.4000; measured value 577.4006.

[0187] Example 26: Synthesis of Compound 26

[0188]

[0189] White solid (yield 85%). 1H NMR (400MHz, chloroform-d) δ7.24(s,1H),5.27(t,J=3.6Hz,1H),4.11(d,J=10.9Hz,1H),3.78(d,J=11.0Hz,1H),3.17–3.10(m,2H),3. 03–2.95(m,2H),2.67(p,J=6.1Hz,1H),2.57(d,J=14.8Hz,1H),2.41(d,J=6.5Hz,2H),2.38(d,J=6.6Hz,2H),2.09–2.05(m,1H) ,2.04–1.93(m,4H),1.79–1.69(m,2H),1.67–1.58(m,2H),1.57–1.46(m,2H),1.44–1.38(m,1H),1.35–1.32(m,1H),1.30(s,3 H),1.29–1.23(m,2H),1.21(s,3H),1.19(s,3H),1.15–1.12(m,1H),1.11–1.03(m,2H),0.99(s,3H),0.90(s,3H),0.88(s,6H). 13 C NMR (126 MHz, chloroform-d) δ 172.57, 143.23, 123.39, 112.53, 71.66, 53.43, 49.94, 46.22, 46.15, 42.76, 41.94, 39.93, 39.33, 38.44, 36.52, 36.16, 34.00, 33.47, 33.21, 32.03, 31.70, 31.41, 31.00, 26.51, 26.49, 25.92, 25.67, 24.19, 23.64, 22.56, 19.28, 16.59, 15.45. ESI-HRMS (m / z) [M+H] + C 37 H 57 N2O4S, calculated value 625.4034; measured value 625.4034.

[0190] Example 27: Synthesis of Compound 27

[0191]

[0192] White solid (yield 62%). 1H NMR (500MHz, chloroform-d) δ7.19(s,1H),5.27(t,J=3.6Hz,1H),4.13(d,J=11.0Hz,1H),3.71(d,J=11.0Hz,1H),3.47–3.37(m,2H) ,3.09–3.01(m,2H),2.64–2.58(m,1H),2.56(d,J=14.8Hz,1H),2.24–2.14(m,2H),2.08–1.98(m,5H),1.97–1.91(m,2H),1 .79–1.71(m,2H),1.68–1.58(m,3H),1.57–1.46(m,3H),1.44–1.39(m,1H),1.37–1.34(m,1H),1.31(s,3H),1.29–1.28(m, 1H),1.25(s,2H),1.22(s,2H),1.19(s,3H),1.13–1.08(m,2H),1.07–1.02(m,1H),1.00(s,3H),0.89(s,3H),0.88(s,6H). 13 CNMR (126 MHz, chloroform-d) δ 172.73, 143.41, 123.24, 71.43, 53.41, 46.28, 46.19, 42.94, 42.76, 41.96, 39.95, 38.58, 38.46, 36.51, 36.17, 34.03, 33.48, 33.24, 32.08, 31.75, 31.45, 31.02, 29.83, 25.94, 25.64, 25.09, 24.90, 24.28, 23.68, 22.47, 19.37, 16.59, 15.49. ESI-HRMS (m / z) [M+H] + C 37 H 58 N3O2, calculated value 576.4524; measured value 576.4535.

[0193] Example 28: Synthesis of Compound 28

[0194]

[0195] White solid (yield 85%). 1H NMR (400MHz, chloroform-d) δ7.24(s,1H),5.27(t,J=3.6Hz,1H),4.12(d,J=11.0Hz,1H),3.79(d,J=11.0Hz,1H),3.39(s,2 H),3.14(d,J=6.0Hz,4H),3.10(d,J=6.2Hz,4H),2.57(d,J=14.7Hz,1H),2.09–2.04(m,1H),2.04–1.93(m,4H),1.7 9–1.67(m,3H),1.66–1.57(m,2H),1.56–1.46(m,2H),1.44–1.38(m,1H),1.36–1.33(m,1H),1.30(s,3H),1.29–1. 22(m,2H),1.21(s,3H),1.18(s,3H),1.16–1.11(m,2H),1.08–1.02(m,1H),1.00(s,3H),0.89(s,3H),0.87(s,6H). 13 C NMR (126 MHz, chloroform-d) δ 170.11, 143.29, 123.31, 112.53, 71.40, 58.40, 53.42, 51.71, 50.79, 46.28, 46.15, 42.70, 41.93, 39.94, 38.44, 36.51, 36.06, 34.02, 33.46, 33.19, 32.03, 31.61, 31.40, 30.99, 25.89, 25.65, 24.17, 23.64, 22.49, 19.29, 16.59, 15.44. ESI-HRMS (m / z) [M+H] + C 37 H 58 N3O4S, calculated value 640.4143; measured value 640.4145.

[0196] Example 29: Synthesis of Compound 29

[0197]

[0198] White solid (yield 82%). 1H NMR (600MHz, chloroform-d) δ7.24(s,1H),5.26(t,J=3.6Hz,1H),4.10(d,J=11.0Hz,1H),3.79(d,J=11.0Hz,1H),3.75(t,J= 4.7Hz,4H),3.26–3.19(m,2H),2.62–2.59(m,4H),2.56(s,1H),2.08(dd,J=13.6,4.6Hz,1H),2.04–1.92(m,4H),1. 78–1.68(m,3H),1.62–1.55(m,2H),1.54–1.45(m,2H),1.35–1.31(m,2H),1.30(s,3H),1.30–1.28(m,1H),1.28–1. 24(m,1H),1.21(s,3H),1.18(s,3H),1.16–1.09(m,3H),1.06–1.02(m,1H),1.00(s,3H),0.89(s,3H),0.88(s,6H). 13 C NMR (126 MHz, chloroform-d) δ 170.41, 143.44, 123.21, 112.63, 70.97, 66.95, 59.73, 53.44, 46.36, 46.19, 42.69, 41.94, 39.96, 38.45, 36.52, 36.07, 34.10, 33.48, 33.25, 32.07, 31.54, 31.43, 31.02, 25.91, 25.69, 24.21, 23.66, 22.52, 19.31, 16.60, 15.46. ESI-HRMS (m / z) [M+H] + C 37 H 58 N3O3, calculated value 592.4473; measured value 592.4481.

[0199] Example 30: Synthesis of Compound 30

[0200]

[0201] White solid (yield 47%). 1H NMR (400MHz, chloroform-d) δ7.22(s,1H),5.26(t,J=3.6Hz,1H),4.10(d,J=11.0Hz,1H),3.79(d,J=11.0Hz,1H),3.24 (d,J=2.3Hz,2H),3.08–3.03(m,4H),2.70–2.64(m,4H),2.56(d,J=14.8Hz,1H),2.07(dd,J=13.7,4.6Hz,1H), 2.04–1.89(m,4H),1.79–1.65(m,3H),1.63–1.46(m,4H),1.44–1.33(m,2H),1.30(s,3H),1.28(s,1H),1.25(s ,1H),1.21(s,3H),1.18(s,3H),1.16–1.11(m,2H),1.10–1.02(m,2H),1.00(s,3H),0.89(s,3H),0.88(s,6H). 13 C NMR (126 MHz, chloroform-d) δ 170.24, 143.34, 123.06, 112.57, 70.87, 59.48, 53.31, 52.68, 46.24, 46.07, 45.03, 42.58, 41.83, 39.84, 38.33, 36.39, 35.96, 33.96, 33.36, 33.12, 31.93, 31.43, 31.32, 30.90, 29.71, 25.79, 25.57, 24.12, 23.55, 22.38, 19.19, 16.48, 15.35. ESI-HRMS (m / z) [M+H] + C 37 H 59 N4O2, calculated value 591.4633; measured value 591.4638.

[0202] Example 31: Synthesis of Compound 31

[0203]

[0204] White solid (yield 84%). 1H NMR (500MHz, chloroform-d) δ7.22(s,1H),5.27(t,J=3.7Hz,1H),4.02(d,J=11.0Hz,1H),3.74(d,J=11.1Hz,1H),2.62(d,J=15.5Hz, 1H),2.57(d,J=14.9Hz,1H),2.30–2.21(m,1H),2.17(s,3H),2.11–2.06(m,1H),2.05–1.98(m,4H),1.97–1.85(m,4H),1.83 –1.67(m,4H),1.62–1.55(m,2H),1.55–1.50(m,1H),1.49–1.46(m,1H),1.45–1.41(m,2H),1.36–1.31(m,2H),1.30(s,3H), 1.26–1.25(m,2H),1.21(s,3H),1.19(s,3H),1.16–1.03(m,3H),1.01(s,3H),0.97–0.93(m,1H),0.89(s,3H),0.88(s,6H). 13 C NMR (126 MHz, chloroform-d) δ 176.03, 143.62, 123.08, 112.76, 70.45, 53.46, 48.23, 46.40, 46.24, 43.84, 42.76, 41.97, 39.97, 38.46, 36.54, 36.21, 34.17, 33.50, 33.31, 32.08, 31.60, 31.46, 31.06, 30.79, 29.92, 29.85, 28.82, 25.94, 25.72, 24.26, 23.69, 22.56, 19.34, 16.60, 15.48. ESI-HRMS (m / z) [M+H] + C 39 H 62 N3O2, calculated value 604.4837; measured value 604.4839.

[0205] Example 32: Preparation of Compound 32

[0206]

[0207] Under nitrogen protection, tert-butyl alcohol (1.0 mL, 11 mmol) was dissolved in anhydrous toluene (17 mL, 163 mmol), and chlorosulfonyl isocyanate (0.87 mL, 10 mmol) was added at 0°C. The reaction mixture was stirred at room temperature for 1 hour, and pyridine (1.8 mL, 22 mmol) was added. The reaction system was reacted at room temperature for 4 hours. After the reaction was completed by TLC monitoring, the Boc-protected aminosulfonyl pyridine salt was filtered.

[0208]

[0209] Under nitrogen protection, o4 (1.0 g, 1.41 mmol) and aminosulfonyl pyridinium salt (0.728 g, 2.82 mmol) were dissolved in anhydrous dichloromethane (10 mL), and the reaction mixture was reacted at room temperature overnight. After TLC monitoring, water (20 mL) was added, and dichloromethane (60 mL) was extracted three times. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to remove the organic solvent. Compound o6 was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15: 1 to 10: 1) as a white solid with a yield of 24%.

[0210]

[0211] Compound o6 (100 mg) was dissolved in 4.0 M hydrochloric acid dioxane solution (6 mL), and the reaction system was reacted at room temperature for 1 hour. After TLC monitoring, the reaction was completed, vacuum concentrated, saturated sodium bicarbonate solution (10 mL) was added to adjust the pH to alkaline, and dichloromethane (30 mL) was extracted three times. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and vacuum concentrated to remove the organic solvent. Silica gel column chromatography was separated and purified (dichloromethane: methanol = 30: 1 to 10: 1) to obtain the white target product 32 (46%). 1 H NMR (500 MHz, chloroform-d) δ7.22 (s, 1H), 5.28 (t, J = 3.7 Hz, 1H), 4.20 (d, J = 9.2 Hz, 1H), 3.76 (d, J = 9.3 Hz, 1H), 2.57 (d, J = 14.8 Hz, 1H), 2.06–2.00 (m, 2H), 1.99–1.92 (m, 2H), 1.80–1.72 (m, 3H),1.64–1.52(m,3H),1.51–1.41(m,3H),1.33–1.31(m,1H),1.30(s,3H),1.28–1.24(m, 3H),1.20(s,3H),1.19(s,3H),1.18–1.05(m,3H),1.02(s,3H),0.90(s,3H),0.88(s,6H). 13C NMR (126 MHz, chloroform-d) δ 143.22, 123.48, 112.70, 77.68, 53.40, 46.24, 46.15, 42.52, 41.93, 40.03, 38.44, 36.50, 36.37, 33.92, 33.50, 33.21, 31.98, 31.46, 31.41, 30.99, 25.94, 25.55, 24.26, 23.70, 23.67, 21.86, 19.30, 16.54, 15.48. ESI-HRMS (m / z) [M+H] + C 31 H 50 N3O3S, calculated value 544.3567; measured value 544.3566.

[0212] Example 33: Synthesis of Compound 33

[0213]

[0214] Compound o6 (200 mg, 0.22 mmol) was dissolved in N,N-dimethylformamide (6 mL), cesium carbonate (143 mg, 0.44 mmol) was added, the reaction mixture was stirred at room temperature for 1 hour, iodomethane (27 μL, 0.44 mmol) was added, and the reaction system was reacted at room temperature for 2 hours. After TLC monitoring, water (10 mL) was added, and ethyl acetate (30 mL) was extracted three times, the organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to remove the organic solvent. Compound o7 was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 15:1 to 10:1) as a white solid with a yield of 98%.

[0215]

[0216] Compound o7 was dissolved in 4.0M hydrochloric acid dioxane solution (6mL), and the reaction system was reacted at room temperature for 1 hour. After TLC monitoring, the reaction was completed, vacuum concentrated, saturated sodium bicarbonate solution (10mL) was added to adjust the pH to alkaline, and dichloromethane (30mL) was extracted three times. The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and vacuum concentrated to remove the organic solvent. Silica gel column chromatography was separated and purified (dichloromethane: methanol = 50: 1 to 20: 1) to obtain the target product 33 as a white solid with a yield of 70%. 1H NMR (400 MHz, chloroform-d) δ7.23 (s, 1H), 5.28 (t, J = 3.6 Hz, 1H), 4.11 (d, J = 9.4 Hz, 1H), 3.67 (d, J = 9.2 Hz, 1H), 2.79 (s, 3H), 2.57 (d, J = 14.8 Hz, 1H), 2.06–1.99 (m, 2H), 1.98–1.91 (m, 2H), 1.80–1.71 (m ,3H),1.64–1.52(m,3H),1.51–1.40(m,3H),1.38–1.32(m,1H),1.30(s,3H),1.29–1.23(m,2H) ,1.21(s,3H),1.19(s,3H),1.18–1.06(m,3H),1.01(s,3H),0.90(s,3H),0.88(d,J=2.4Hz,6H). 13 C NMR (151 MHz, chloroform-d) δ 143.10, 123.32, 76.84, 53.29, 46.11, 46.03, 42.40, 41.79, 39.89, 38.33, 36.38, 36.31, 33.81, 33.08, 31.88, 31.32, 30.86, 29.73, 25.81, 25.41, 24.09, 23.57, 23.54, 21.82, 19.18, 16.37, 15.34. ESI-HRMS (m / z) [M+H] + C 32 H 52 N3O3S, calculated value 558.3724; measured value 558.3730.

[0217] Biological activity test experiment

[0218] 1. ELISA method was used to determine the effect of compounds on inhibiting the secretion of inflammatory cytokines by RAW 264.7 cells induced by lipopolysaccharide (LPS).

[0219] Experimental materials: RAW264.7 cells were purchased from the American Type Culture Collection (ATTC, Manassas, VA, USA), and LPS was purchased from Sigma-Aldrich (St. Louis, MO, USA).

[0220] Experimental principle:

[0221] Succinate dehydrogenase in the mitochondria of living cells can reduce methylthiazolyl blue (MTT) to water-insoluble blue-purple crystalline formazan, while dead cells do not have this function. Dissolving formazan with DMSO and measuring its absorbance at a wavelength of 570nm can indirectly reflect the number of living cells. Within a certain range of cell numbers, the amount of MTT crystals formed is proportional to the number of cells. It can be used for cytotoxicity analysis.

[0222] LPS, a component of the cell wall of Gram-negative bacteria, is an important biological inflammatory factor that can induce the massive release of inflammatory factors such as IL-1β, IL-6, and TNF-α by activating NF-κB. Therefore, the LPS-induced RAW264.7 cell classic inflammation model has also been widely used in the evaluation of the anti-inflammatory activity of small molecule compounds.

[0223] RAW 264.7 cells were collected at 1×10 5 Cells / well were pre-seeded in 96-well plates in triplicate and allowed to adhere for 12 hours. Then 10 μg / mL lipopolysaccharide was added, and 6 concentrations were set: 200.000, 66.667, 22.222, 7.407, 2.469, 0.823 μM. The cells were cultured in a 37°C, 5% CO2 incubator for 48 hours with corresponding cell controls and blank controls. The supernatant was collected and the IL-1β content was determined by ELISA, and the IC 50 The results are shown in Table 1 below. Selection index SI = CC 50 / IC 50 , to determine whether the sample has biologically significant activity, and SI greater than 2 is considered to have biological effects. All index inspection results were processed by Excel 2019 and Graphpad Prism 8 software. Dose-response curves were drawn based on the experimental results, and CC was calculated. 50 The 50% Cytotoxic concentration is the lethal concentration for 50% of cells, and the IC 50 The value (50% inhibitory concentration) refers to the concentration of the compound that has a 50% inhibitory effect.

[0224] Table 1

[0225]

[0226] “-” means not tested, “ / ” means no meaning.

[0227] From the results in Table 1, it can be seen that some compounds in this series, such as compounds 1, 15, 24, 27, 31 and 32, have significantly enhanced inhibitory activity against IL-1β factor compared with oleanolic acid and the marketed drug diacerein (IL-1β inhibitor), especially compound 1, which shows that this series of compounds has good anti-inflammatory activity and the compound selection index is significantly improved.

[0228] 2. TRAP staining to detect the effect of compounds on the osteoclast differentiation of bone marrow-derived macrophages (BMM)

[0229] Bone marrow cells were obtained by flushing the femur and tibia of 6-8 week old female BALB / c mice with RPMI 1640 medium and cultured in BMM cell culture medium overnight. Non-adherent cells were collected and cultured in BMM cell culture medium for proliferation. After 4 days of culture, fresh BMM cell culture medium was replaced for another 3 days. Adherent cells of BMM cells were washed 3 times with PBS and then sterilized with EDTA-PBS (2mM) at 4°C for 15 minutes. BMM cells were centrifuged and cultured in DMEM medium for subsequent experiments. BMM cells were cultured at 8×10 4 Cells / well were seeded in triplicate in a 96-well plate and allowed to adhere overnight. Then 30 ng / mL M-CSF, 50 ng / mL RANKL, and 6 concentrations were added: 200.000, 66.667, 22.222, 7.407, 2.469, and 0.823 μM. Corresponding cell controls and blank controls were also set up and cultured in a 37°C, 5% CO2 incubator for 11 days. The culture medium was changed every 3 days. At the end of the culture, the cells were stained with TRAP, and the number of positively stained cells was counted under a microscope to calculate the IC 50 The results are shown in Table 2 below. Selection index SI = CC 50 / IC 50 , to determine whether the sample has biologically significant activity, and SI greater than 2 is considered to have biological effects. All index inspection results were processed by Excel 2019 and Graphpad Prism 8 software. Dose-response curves were drawn based on the experimental results, and CC was calculated. 50 The 50% Cytotoxic concentration is the lethal concentration for 50% of cells, and the IC 50 The value (50% inhibitory concentration) refers to the concentration of the compound that has a 50% inhibitory effect. The activity of some compounds has not been tested.

[0230] Table 2

[0231]

[0232] From the results in Table 2, it can be seen that some compounds in this series, such as compounds 1, 27, 29, 31 and 33, have greatly improved osteoclast differentiation inhibition activity compared with oleanolic acid and the marketed drug diacerein, and the selectivity index has also been significantly improved, showing potential for the treatment of bone-related diseases and good safety.

Claims

1. A pyrazolooleanolic acid reduced alcohol derivative as shown in the following general formula I, or a pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate thereof: in, R1 is selected from H, -S(=O)2R 11 or -C(=O)R 11 , Among them, R 11 Each is independently selected from substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C5-C10 aryl, substituted or unsubstituted having 1-3 selected from O, N, S heteroatom 5 to 6 membered heteroaryl, or -NR 12 R 13 , where R 12 and R 13 Each independently selected from H or C1-C10 alkyl; R2 is selected from H, a group derived from an α, β or γ amino acid, -C(=O)R 21 、-C(=O)CH2-R 21 ,;-S(=O)2R 22 , where R 21 are each independently selected from substituted or unsubstituted C3-C6 cycloalkyl, 5- to 6-membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S, and R 22 For-NR 12 R 13 , where R 12 and R 13 Each independently selected from H or C1-C10 alkyl; The above substitution refers to substitution with a substituent selected from halogen, hydroxyl, CN, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C6 cycloalkyl, C1-C10 alkoxy, wherein the C1-C10 alkyl, C3-C6 cycloalkyl or C1-C10 alkoxy may be optionally further substituted with halogen or amino.

2. The pyrazolooleanolic acid reduced alcohol derivative according to claim 1, or a pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate thereof, wherein: R1 and R2 are both H.

3. The pyrazolooleanolic acid reduced alcohol derivative according to claim 1, or a pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate thereof, in, When R1 is H, R2 is not H, or When R1 is not H, R2 is H.

4. The pyrazolooleanolic acid reduced alcohol compound according to claim 1, or a pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate thereof, wherein: The compound of the general formula I is represented by the following formula I-1: In the above formula I-1, R 111 is selected from halogen, hydroxy, CN, C1-C10 alkyl unsubstituted or substituted by halogen, C2-C10 alkenyl unsubstituted or substituted by halogen, C2-C10 alkynyl unsubstituted or substituted by halogen, C3-C6 cycloalkyl unsubstituted or substituted by halogen, C1-C10 alkoxy unsubstituted or substituted by halogen, n is an integer from 1 to 5, and R2 is as defined in claim 1.

5. The pyrazolooleanolic acid reduced alcohol derivative according to claim 1, or a pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate thereof, wherein: The compound of general formula I is selected from the following specific compounds:

6. The pyrazolooleanolic acid reduced alcohol derivative according to claim 1, or a pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate thereof, wherein: The pharmaceutically acceptable salts include maleate, succinate, citrate, tartrate, fumarate, acetate, methanesulfonate, hydrochloride, phosphate, nitrate or sulfate.

7. A method for preparing the compound according to any one of claims 1 to 6, wherein the method is selected from the following route 1, route 2 and route 3: (a) Route 1: In the general formula I, when R2 is H, the compound of the general formula I is prepared by route 1: In a solvent, compound A is reduced to a hydroxymethyl group under the action of a reducing agent to obtain compound B. in, In the first route, the solvent is selected from diethyl ether, tetrahydrofuran, dioxane, and toluene; the reducing agent is selected from lithium aluminum hydride and borane complex. Wherein, the substituent R1 in compounds A and B is defined as described in claim 1; (b) Route 2: In the general formula I, when R2 is not H, the compound of the general formula I is prepared by route 2: In a solvent, in the presence of a base, compound B reacts with an electrophilic reagent to generate a target compound P; or In a solvent, in the presence of a condensing agent and a base, compound B reacts with the corresponding carboxylic acid to generate the target compound P. Wherein, in route 2, the substituents R1 and R2 in compounds B and P are defined as in claim 1; (c) Route 3: In the general formula I, when R1 is not H, the compound of the general formula I is prepared by route 3: In an organic solvent, in the presence of a base, compound C reacts with an electrophilic reagent to generate a target compound P; or In an organic solvent, in the presence of a condensing agent and a base, compound C reacts with the corresponding carboxylic acid to generate the target compound P. Wherein, in route 3, the substituents R1 and R2 in compounds C and P are defined as described in claim 1, Wherein, in routes 2 and 3, the solvent is an aprotic polar solvent selected from N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane, dichloroethane, tetrahydrofuran, and dioxane; The electrophilic reagent is selected from sulfonyl chloride, acyl chloride, and acid anhydride; The base is selected from triethylamine, N,N-diethylisopropylamine, sodium acetate, potassium carbonate, 4-dimethylaminopyridine; and The condensing agent is selected from dicyclohexylcarbodiimide (DCC), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), 1-hydroxybenzotriazole (HOBT), and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (HATU).

8. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 6, a pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate thereof, and optionally a pharmaceutically acceptable carrier.

9. Use of the compound according to any one of claims 1 to 6, its pharmaceutically acceptable salt, geometric isomer, stereoisomer, solvate or hydrate, or the pharmaceutical composition according to claim 8 in the preparation of a medicament for preventing and / or treating diseases caused by inflammation and / or abnormal osteoclast activity.

10. The use according to claim 9, wherein Diseases caused by inflammation and / or abnormal osteoclast activity include rheumatoid arthritis, inflammatory bowel disease, chronic obstructive pulmonary disease, asthma, psoriasis, idiopathic pulmonary fibrosis, osteoarthritis, osteoporosis, periodontitis, tooth loss, Paget's disease of bone, rickets, giant cell tumor of bone, myeloma bone disease and bone destruction caused by cancer bone metastasis.

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