C17 nitrogen substituted and methylene substituted oleanane triterpenoid derivative as well as preparation method and application of C17 nitrogen substituted and methylene substituted oleanane triterpenoid derivative

Abstract

The invention discloses a C17 nitrogen substituted and methylene substituted oleanane triterpenoid derivative as well as a preparation method and application of the C17 nitrogen substituted and methylene substituted oleanane triterpenoid derivative. The invention provides an oleanane triterpenoid derivative with nitrogen substituted and methylene substituted at a C17 site. The invention further discloses application of the compound to preparation of an NRF2-Leap1 uncoupling agent. The compound provided by the invention is used for preparing medicines for preventing and / or treating diseases of patients; diseases include small vascular diseases, mitochondrial brain myopathy, autism spectrum disorder, Rett syndrome, Friedel's ataxia, cerebral apoplexy, hemorrhagic cerebral apoplexy, ischemic cerebral apoplexy, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, cognitive impairment of schizophrenia, Parkinson's disease, cognitive impairment of Parkinson's disease, Alzheimer's disease, vascular dementia, and epilepsy. , Huntington's disease, heart failure, myocardial infarction, renal failure, renal ischemia and the like.

Description

Technical Field

[0001] The present invention relates to the field of biology and medical technology, and in particular to oleanane triterpene derivatives substituted with nitrogen and methylene at position C17, and preparation methods and uses thereof. Background Art

[0002] Current studies have shown that Keap-Nrf2-ARE has a protective effect on cells and is one of the important pathways for the body to maintain redox balance and eliminate the damaging effects of toxic and harmful substances. It plays a vital role in anti-stress, anti-apoptosis, anti-inflammatory response and neuroprotection. Reactive oxygen species (ROS) and oxidative stress are associated with the occurrence of many diseases and have therefore become a research hotspot in recent years. Currently, the structures of Nrf2 agonists are mostly derived from the oleanolic acid triterpenoid nucleus, such as 2-cyano-3,12-dioxooleanolic-1,9(11)-diene-28-acid (CDDO), methyl bardoxolone (CDDO-Me; RTA402) and Omaveloxolone (RTA408). However, these molecules only focus on inflammatory treatment through Nrf2 agonism, and have the characteristics of poor inhibitory activity and poor specificity. Therefore, it is particularly important to develop new Nrf2-Leap1 uncouplers that have both anti-inflammatory and antioxidant effects. Summary of the invention

[0003] In order to solve the above technical problems, the present invention provides oleanane triterpene derivatives with nitrogen substitution and methylene substitution at position C17, and preparation methods and uses thereof.

[0004] The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof:

[0005]

[0006] Wherein: R1 is independently selected from: -NH-heteroarene, -NH-heteroarene diyl-R1', -NH-C(=O)-alkane, -NH-C(=O)-substituted alkane, -NH-C(=O)-alkene, -NH-C(=O)-substituted alkene, -NH-C(=O)-alkyne, -NH-C(=O)-substituted alkyne, -NH-C(=O)-arene, -NH-C(=O)-arene diyl-R1', -NH-C(=O)-alkane ... H-C(=O)-heteroarene, -NH-C(=O)-heteroarenediyl-R1', -N(OH)-C(=O)-alkane, -N(OH)-C(=O)-substituted alkane, -N(OH)-C(=O)-alkene, -N(OH)-C(=O)-substituted alkene, -N(OH)-C(=O)-alkyne, -N(OH)-C(=O)-substituted alkyne, -N(OH)-C(=O)-arene, -N (OH)-C(=O)-aromatic diyl-R1', -N(OH)-C(=O)-heteroaromatic, -N(OH)-C(=O)-heteroaromatic diyl-R1', -NH-C(=O)-L-type amino acid-NH-heteroaromatic, -NH-C(=O)-L-type amino acid-NH-heteroaromatic diyl-R1', -CH2-N(OH)-C(=O)-alkane, -CH2-N(OH)-C(=O)-substituted Alkanes, -CH2-N(OH)-C(=O)-olefins, -CH2-N(OH)-C(=O)-substituted olefins, -CH2-N(OH)-C(=O)-alkynes, -CH2-N(OH)-C(=O)-substituted alkynes, -CH2-N(OH)-C(=O)-heteroarene, -CH2-N(OH)-C(=O)-heteroarene diyl-R1'; and R2: hydrogen or methyl; R3: hydrogen or methyl.

[0007] Preferably, the compound is further defined as:

[0008]

[0009] Wherein: R1 is independently selected from: -NH-heteroarene, -NH-heteroarene diyl-R1', -NH-C(=O)-alkane, -NH-C(=O)-substituted alkane, -NH-C(=O)-alkene, -NH-C(=O)-substituted alkene, -NH-C(=O)-alkyne, -NH-C(=O)-substituted alkyne, -NH-C(=O)-arene, -NH-C(=O)-arene diyl-R1', -NH-C(=O)-alkane ... NH-C(=O)-heteroarene, -NH-C(=O)-heteroarene diyl-R1', -N(OH)-C(=O)-alkane, -N(OH)-C(=O)-substituted alkane, -N(OH)-C(=O)-alkene, -N(OH)-C(=O)-substituted alkene, -N(OH)-C(=O)-alkyne, -N(OH)-C(=O)-substituted alkyne, -N(OH)-C(=O)-arene, -N(OH)-C(=O)-aromatic diyl-R1', -N(OH)-C(=O)-heteroaromatic, -N(OH)-C(=O)-heteroaromatic diyl-R1', -NH-C(=O)-L-type amino acid-NH-heteroaromatic, -NH-C(=O)-L-type amino acid-NH-heteroaromatic diyl-R1', -CH2-N(OH)-C(=O)-alkane, -CH2-N(OH)-C(=O) -substituted alkanes, -CH2-N(OH)-C(=O)-olefins, -CH2-N(OH)-C(=O)-substituted olefins, -CH2-N(OH)-C(=O)-alkynes, -CH2-N(OH)-C(=O)-substituted alkynes, -CH2-N(OH)-C(=O)-heteroarene, -CH2-N(OH)-C(=O)-heteroarenediyl-R1'; and R2: methyl; R3: methyl.

[0010] Preferably, R1' in the aromatic hydrocarbon compound is: -Cl, -F, -Br, -OH, isopropyl, straight-chain / branched alkyl (C≤6), straight-chain / branched alkyl (C≤6) substituted with 1 to 5 halogens, -OH, straight-chain / branched alkyl (C≤6) substituted with 1 to 5 -OHs, straight-chain / branched alkenyl (C≤6), straight-chain / branched alkenyl (C≤6) substituted with 1 to 5 halogens, straight-chain / branched alkenyl (C≤6) substituted with 1 to 5 -OHs, straight-chain / branched alkynyl (C≤6), straight-chain / branched alkynyl (C≤6) substituted with 1 to 5 halogens, straight-chain / branched alkynyl (C≤6) substituted with 1 to 5 -OHs, wait.

[0011] Preferably, the aromatic hydrocarbon group is selected from:

[0012] wait.

[0013] Preferably, the substituted alkane, substituted olefin, substituted alkyne, alkane, olefin, alkyne C chain length is ≤ 6, and is linear, branched or cyclic. The substituent is selected from 1 to 5 -SO3H, -OH, -F, -Br, -Cl, -OH, methyl, ethyl, propyl replacement (and / or).

[0014] Preferably, the compound, its pharmaceutically acceptable salt or stereoisomer is as follows:

[0015]

[0016]

[0017] The present invention also provides the use of the above-mentioned compound, its pharmaceutically acceptable salt, and stereoisomer for preparing NRF2-Leap1 uncoupling agent.

[0018] The biological experiments of the present invention show that many of the above compounds of the present invention exhibit single-digit nanomolar human Nrf2 receptor agonist activity, and are significantly better than the marketed Nrf2 agonist Omaveloxolone. At the same time, the compounds have the activities of scavenging DPPH free radicals, inhibiting the production of lipid peroxide MDA, intervening in ferroptosis, etc. to exert antioxidant effects.

[0019] The present invention also provides the use of the above-mentioned compound, its pharmaceutically acceptable salt, and stereoisomers for treating and / or preventing patient diseases. The drugs are used to prevent or treat diseases including cerebral small vessel disease, mitochondrial encephalomyopathy, autism spectrum disorder, Rett syndrome, Friedreich's ataxia, stroke, hemorrhagic stroke, ischemic stroke, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, schizophrenia cognitive impairment, Parkinson's disease, Parkinson's cognitive impairment, Alzheimer's disease, vascular dementia, epilepsy, Huntington's disease, heart failure, myocardial infarction, renal failure, renal ischemia, etc. According to effective test data, the use of the above-mentioned compound, its pharmaceutically acceptable salt, and the drugs prepared by stereoisomers of the present invention, especially for stroke, multiple sclerosis, and amyotrophic lateral sclerosis, has a significant effect.

[0020] Preferably, the compound, its pharmaceutically acceptable salt, stereoisomer is used to prepare a drug for preventing or treating stroke, multiple sclerosis, and amyotrophic lateral sclerosis.

[0021] Beneficial effects:

[0022] The present invention synthesizes for the first time a compound shown in formula (I) or a pharmaceutically acceptable salt thereof, i.e., a C17 nitrogen-substituted and methylene-substituted oleanolic acid triterpene derivative. At the same time, the present invention provides a method for preparing a compound shown in formula (I) through a specific example. Furthermore, the present invention provides the use of a compound shown in formula (I) or a pharmaceutically acceptable salt thereof for preparing an NRF2-Leap1 uncoupler. The C17 nitrogen-substituted and methylene-substituted oleanolic acid triterpene derivatives of the present invention are a new type of Nrf2-Leap1 uncoupler, which, in addition to maintaining strong agonistic activity on the Nrf2 target, also has the activity of scavenging DPPH free radicals, inhibiting the generation of lipid peroxide MDA, or intervening in ferroptosis to exert an antioxidant effect. Finally, the present invention also provides a pharmaceutical composition, comprising any one of the above compounds or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. The pharmaceutical composition can be prepared into various pharmaceutically acceptable dosage forms, such as tablets, capsules, oral liquids, granules, injections, or various sustained-release preparations. The pharmaceutical composition can be administered orally or parenterally (such as intravenously, subcutaneously or topically). The dosage can be appropriately adjusted according to the patient's age, gender and disease type. The present invention can be used to prevent or treat multiple diseases in the future, including cerebral small vessel disease, mitochondrial encephalomyopathy, autism spectrum disorder, Rett syndrome, Friedreich's ataxia, stroke, hemorrhagic stroke, ischemic stroke, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, schizophrenia cognitive impairment, Parkinson's disease, Parkinson's cognitive impairment, Alzheimer's disease, vascular dementia, epilepsy, Huntington's disease, heart failure, myocardial infarction, renal failure, renal ischemia, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Effects of compound 28 on mNSS scores of rats with ischemic stroke, compared with the sham operation group: ### p<0.001; compared with the model group: *** p<0.001.

[0024] Figure 2 Effects of compound 28 on cerebral infarction area in rats with ischemic stroke, compared with the sham operation group: ### p<0.001; compared with the model group: *** p<0.001.

[0025] Figure 3 Effects of compound 28 on neurological function scores of EAE model mice, compared with the sham operation group: *** p<0.001; compared with the model group: # p<0.05, ## p<0.01, ###p<0.001; compared with Omaveloxolone 15mg / kg group: &&& p<0.001.

[0026] Figure 4 Effects of compound 28 on body weight of EAE model mice, compared with the sham operation group: *** p<0.001; compared with the model group: # p<0.05, ### p<0.001; compared with Omaveloxolone 15mg / kg group: &&& p<0.001.

[0027] Figure 5 .Compound 28 affects the onset time of SOD1 G93A mice, compared with the control group: *** p<0.001; compared with the model group: # p<0.05, ## p<0.01; compared with Omaveloxolone 3mg / kg group: & p<0.05.

[0028] Figure 6 Effects of compound 28 on motor coordination ability of SOD1 G93A mice, compared with the control group: * p<0.05, *** p<0.001; compared with the model group: ### p<0.001; compared with Omaveloxolone 3mg / kg group: & p<0.05.

[0029] Figure 7 Effects of compound 28 on muscle endurance of SOD1 G93A mice, compared with the control group: * p<0.05, *** p<0.001; compared with the model group: ### p<0.001; compared with Omaveloxolone 3mg / kg group: & p<0.05. DETAILED DESCRIPTION

[0030] The present invention is further described below in conjunction with specific embodiments and test examples, but the scope of the present invention is not limited in any form.

[0031] The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS).

[0032] NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using an AVANCE III 600 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD) and deuterated chloroform (CDCl3) as the solvents, and tetramethylsilane (TMS) as the internal standard.

[0033] Liquid chromatography-mass spectrometry (LC-MS) was performed using a Shimadzu LCMS2020 mass spectrometer, and HPLC was performed using a Shimadzu LC20A liquid chromatograph.

[0034] The thin layer chromatography silica gel plate used was Yantai Jiangyou silica gel plate, the specification used for TLC was 0.2mm±0.03mm, and the specification used for thin layer chromatography separation and purification products was 0.4mm-0.5mm.

[0035] Unless otherwise specified in the examples, SFC separation conditions are as follows: column model: DAICEL CHIRALPAK IC (250 mm*30 mm, 10 um); mobile phase: [CO2-i-PrOH / ACN]; B%: 40%, isocratic elution mode.

[0036] In the present invention, if the specific experimental conditions are not indicated, the conventional experimental conditions or the conditions recommended by the manufacturer shall be followed. If the manufacturer of the reagents or instruments is not indicated, conventional products can be obtained by commercial purchase.

[0037] In the present invention, the test results are expressed as average values.

[0038] The detection indexes in the present invention are: human Nrf2 receptor function test (agonist detection); DPPH free radical scavenging ability test; MDA anti-lipid peroxidation ability test.

[0039] ■Example 1

[0040] Compound 1:

[0041]

[0042] Synthesis route:

[0043]

[0044] Step 1: Synthesis of intermediate 3

[0045] The raw material 1' (25g, 244.78mmol) was added to anhydrous methanol (400mL) and dissolved, and then the intermediate 2 (35.57g, 269.25mmol, 30.85mL) and sodium methoxide methanol solution (5.4M, 113.32mL) were added and stirred at 70°C for 12h. After the reaction was completed by LCMS detection, the reaction mixture was added with concentrated hydrochloric acid solution at 0°C to adjust the pH value to less than 7, and then concentrated under reduced pressure to obtain a residue. After purification by column chromatography (silica gel column, petroleum ether / ethyl acetate = 1 / 0 to 7 / 3), intermediate 3 (31g, yield 70.70%, yellow oil) was obtained. LCMS: rt = 0.100min, 169.2[MH] + ; 1 H NMR (400MHz, DMSO-d6) δ = 11.18 (br s, 1H), 4.89-4.76 (m, 1H), 3.56 (s, 2H), 1.31 (d, J = 7.0Hz, 6H).

[0046] Step 2: Synthesis of intermediate 4

[0047] Intermediate 3 (31 g, 182.18 mmol) was added to anhydrous acetonitrile (310 mL) and dissolved, then phosphorus oxychloride (30.73 g, 200.39 mmol, 18.68 mL) was added under nitrogen protection and stirred at 65°C for 12 hours. After the reaction was completed by LCMS, the reaction mixture was added to normal temperature water (300 mL) at room temperature for quenching, and after stirring for 1 hour, it was concentrated and filtered to obtain intermediate 4 (21.4 g, yield 62.28%, yellow solid). 1 H NMR (400MHz, DMSO-d6) δ = 12.20 (br s, 1H), 5.82 (s, 1H), 5.05-4.83 (m, 1H), 1.34 (d, J = 7.0Hz, 6H).

[0048] Step 3: Synthesis of compound 1

[0049] Intermediate 5 (50 mg, 108.07 μmol) and intermediate 4 (30.57 mg, 162.10 μmol) were dissolved in dioxane (1 mL). After nitrogen replacement three times, PEPPSIPd (6.36 mg, 7.56 μmol) and sodium tert-butoxide (31.16 mg, 324.21 μmol) were added. After nitrogen replacement three times, the mixture was stirred at 100 ° C for 12 hours. PEPPSIPd (6.36 mg, 7.56 μmol) and sodium tert-butoxide (31.16 mg, 324.21 μmol) were added again. After nitrogen replacement three times, the mixture was stirred at 100 ° C for 2 hours. After the reaction was completed by LCMS detection, the reaction mixture was filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by a thin layer chromatography plate (dichloromethane / anhydrous methanol = 10 / 1) to obtain a crude product, which was again prepared by chromatography (C 18 Column, 0.1% formic acid solution) and freeze-dried to obtain compound 1 (4.2 mg, white solid, yield <1%). LCMS: rt = 1.502 min, 615.4 [M+H] + ; Purity: 97.48%; 1 H NMR (400MHz, DMSO-d6)δ=9.50(br s,1H),8.66(s,1H),6.26(s,1H),5.80(br s,1H),5.01-4.83(m,1H),4.65(s,1H),3.07(br d,J=3.8Hz,1H),2.08-1.91(m,2H),1.90-1.82(m,2H),1.81-1.64(m,4H),1.6 1-1.53(m,1H),1.49-1.42(m,5H),1.39(s,3H),1.30(d,J=7.0Hz,7H),1.23(br s,3H),1.18(s,3H),1.11-1.04(m,4H),0.96(br d,J=2.4Hz,6H),0.89(s,3H).

[0050] ■Example 2

[0051] Compound 2:

[0052]

[0053] Synthesis route:

[0054]

[0055] Step 1: Synthesis of intermediate 2'

[0056] The raw material 1 (3 g, 6.10 mmol) was added to anhydrous toluene (60 mL) and dissolved, and then triethylamine (12.35 g, 122.04 mmol, 16.99 mL) and DPPA (5.04 g, 18.31 mmol, 3.95 mL) were added at 0°C and stirred at 25°C for 2 h. After the reaction was completed by LCMS, the reaction solution was concentrated to obtain a crude product. After purification by column chromatography (silica gel column, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1), intermediate 2' (2.8 g, yield 79.93%, colorless oil) was obtained. LCMS: rt = 0.677 min, 517.4 [M+H] + ; Purity 90.74%.

[0057] Step 2: Synthesis of intermediate 3'

[0058] Intermediate 2' (2.8 g, 5.42 mmol) was added to anhydrous toluene (20 mL) and stirred at 80°C for 2 h. After the reaction was completed by LCMS, the reaction solution was concentrated to obtain intermediate 3' (2.6 g, white solid). LCMS: rt = 0.693 min, 489.3 [M+H] + .

[0059] Step 3: Synthesis of intermediate 4

[0060] Dissolve the intermediate 3' (2.9 g, 5.93 mmol) in anhydrous acetonitrile (30 mL), and add concentrated hydrochloric acid (12 M, 5 mL). The reaction solution was stirred at 25 ° C for 0.5 hours. After the reaction was completed by LCMS detection, the reaction mixture was added with 20% sodium hydroxide solution (50 mL) at 0 ° C to adjust the pH value to greater than 8, and then extracted with ethyl acetate (20 mL * 3). The combined organic layer was washed with saturated brine (20 mL * 2), dried over anhydrous sodium sulfate, and filtered and concentrated under reduced pressure to obtain a residue. Purify by column chromatography (silica gel column, petroleum ether / ethyl acetate = 1 / 1 ~ dichloromethane / anhydrous methanol = 10 / 1). Intermediate 4 (2.2 g, yield 79.32%, white solid) was obtained. LCMS: rt = 0.481, 0.749 min, 485.3 [M + Na] + ;Purity 99.18%; 1H NMR (400MHz, CHLOROFORM-d) δ = 8.06 (s, 1H), 5.98 (s, 1H), 3.60 (d, J = 4.6Hz, 1H), 2.22 (br d,J=13.2Hz,1H),2.15-2.05(m,1H),2.04-1.93(m,1H),1.87-1.78(m,3H),1.78-1.52(m,4H ),1.51-1.42(m,3H),1.37-1.23(m,10H),1.22-1.12(m,4H),1.03-0.96(m,7H),0.90(s,3H).

[0061] Step 4: Synthesis of intermediate 6

[0062] Intermediate 4 (300 mg, 5.93 mmol) and intermediate 5 were dissolved in anhydrous dichloromethane (6 mL), triethylamine (196.84 mg, 1.95 mmol, 270.75 μL) was added, and the reaction was stirred at 25 ° C for 12 hours. After the reaction was completed by LCMS, the reaction solution was filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase column chromatography (C 18 Column, 0.1% formic acid aqueous solution). After spin drying, intermediate 6 (400 mg, yield 97.32%, yellow solid) was obtained. LCMS: rt = 0.631 min, 656.4 [M + Na] + ; Purity 100%.

[0063] Step 5: Synthesis of intermediate 7

[0064] Intermediate 6 (360 mg, 567.95 μmol) was dissolved in anhydrous dichloromethane (4 mL), trifluoroacetic acid (0.4 mL) was added and stirred at 25°C for 3 hours. The reaction solution was concentrated under reduced pressure to obtain Intermediate 7 (360 mg, yellow oil). LCMS: rt = 0.477 min, 534.3 [M+H] + .

[0065] Step 6: Synthesis of Compound 2

[0066] Intermediate 7 (180 mg, 337.24 μmol) and intermediate 8 (95.41 mg, 505.86 μmol) were dissolved in dioxane (2 mL), and diisopropylethylamine (217.93 mg, 1.69 mmol, 293.71 μL) was added and microwaved at 120 ° C for 4 hours. After the reaction was completed by LCMS, the reaction mixture was filtered and concentrated under reduced pressure to obtain a crude product. The crude product was prepared by chromatography (C 18Column, 0.1% formic acid solution) and freeze-dried to obtain compound 2 (18.6 mg, off-white solid, yield 3.84%). LCMS: rt = 1.463 min, 686.4 [M+H] + ; Purity 95.47%; 1 H NMR (400MHz, DMSO-d6) δ = 9.95 (s, 1H), 8.65 (s, 1H), 7.59 (s, 1H), 6.27 (d, J = 7.8Hz, 1H), 6.23 (s, 1H), 4.99-4.89 (m, 1H), 4.42 (d, J = 1.6Hz, 1H), 3.92 (br t,J=7.4Hz,1H),3.10-3.01(m,1H),2.98(br d,J=4.4Hz,1H),2.19-2.10(m,1H),2.07(s,1H),2.01-1.92(m,1H),1.89-1.78(m,3H),1. 74-1.62(m,3H),1.55(brd,J=14.0Hz,1H),1.43-1.35(m,7H),1.32-1.27(m,8H),1.23(br d,J=6.6Hz,3H),1.18(s,4H),1.07-0.98(m,7H),0.94(s,3H),0.86(s,3H).

[0067] ■Example 3

[0068] Compound 3:

[0069]

[0070] Synthesis route:

[0071]

[0072] ■Example 4

[0073] Compound 4:

[0074]

[0075] Synthesis route:

[0076]

[0077] Step 1: Synthesis of intermediate 2

[0078] Intermediate 1 (150 mg, 1.18 mmol, 1 eq) was added to anhydrous toluene (1.5 mL) to dissolve, and then thionyl chloride (702.04 mg, 5.90 mmol, 428.59 μL, 5 eq) was added and stirred at 70°C for 1 h. After the reaction was completed by LCMS, the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product (150 mg) was directly used for the next reaction.

[0079] Step 2: Synthesis of compound 4

[0080] Intermediate 3 (198.68 mg, 429.42 μmol, 0.5 eq) was added to anhydrous dichloromethane (2 mL), followed by diisopropylethylamine (333.00 mg, 2.58 mmol, 448.79 μL, 3 eq), the reaction solution was cooled to 0 ° C and intermediate 2 (125 mg, 858.85 μmol, 1 eq) was slowly added. The reaction solution was stirred at 25 ° C for 1 h. After LCMS detection, the reaction solution was concentrated to obtain a crude product. The crude product was purified by chromatographic preparation (C18 column, 0.1% formic acid solution) and freeze-dried to obtain compound 4 (73.4 mg, off-white solid, yield 14.85%). LCMS: rt = 1.649 min, 594.3 [M + Na] +, purity: 99.367%. 1 H NMR (400MHz, DMSO-d6) δ = 8.96 (s, 1H), 8.66 (s, 1H), 7.34 (s, 1H), 6.23 (s, 1H), 3.23-3.17 (m, 1 H),2.97-2.87(m,1H),2.60(s,3H),2.14-2.05(m,1H),2.05-1.90(m,3H),1.90-1.71(m,4H), 1.71-1.58(m,2H),1.55-1.44(m,2H),1.44-1.41(m,3H),1.36-1.33(m,3H),1.33-1.26(m,2H ),1.23-1.18(m,1H),1.18-1.15(m,3H),1.06(s,3H),1.00(s,3H),0.96(s,3H),0.88(s,3H).

[0081] ■Example 5

[0082] Compound 5:

[0083]

[0084] Synthesis route:

[0085]

[0086] ■Example 6

[0087] Compound 6:

[0088]

[0089] Synthesis route:

[0090]

[0091] Step 1: Synthesis of Intermediate 1

[0092] To a toluene solution (40.0 mL) of material 218600-44-3 (2.00 g, 4.07 mmol, 1.00 eq) was added DPPA (3.36 g, 12.2 mmol, 2.63 mL, 3.00 eq) and Et3N (8.24 g, 81.4 mmol, 11.3 mL, 20.0 eq). The mixture was stirred dropwise at 0°C for 6 h. LC-MS (EB11687-32-P1A2) showed that about 84.0% of the product was detected. The mixture was concentrated to give a crude product. The residue was purified by column chromatography (SiO2, 0-10% EtOAc in CH2C l2 Solution), TLC (CH2Cl2: EtOH = 10: 1, Rf = 0.5) purification to give Intermediate 1 (1.40 g, 2.71 mmol, yield 67.0%) as a white solid. LCMS: (EB11687-32-P1A1_LCMS_SH), RT = 1.236 min, MS (ESI) m / z = 517 [M+1] +.

[0093] Step 2: Synthesis of Intermediate 2

[0094] To a toluene solution (104 mL) of intermediate 1 (1.40 g, 2.71 mmol, 1.00 eq), the mixture was stirred at 110 ° C for 2 h. LC-MS (EB11687-34-P1 C1) showed that about 97.0% of the expected compound was detected. The mixture was concentrated to obtain a crude product. The crude compound 2 (1.20 g, 2.46 mmol, yield 90.6%) was a white solid and was used directly in the next step without further purification. LCMS: (EB11687-34-P1C1_LCMS_SH), RT = 1.231 min, MS (ESI) m / z = 489 [M+1] + .

[0095] Step 3: Synthesis of Intermediate 3

[0096] To a MeCN solution (7.70 mL) of intermediate 2 (1.20 g, 2.46 mmol, 1.00 eq) was added dropwise HCl (12 M, 13.00 mL, 58.6 eq), stirred at 25 ° C for 20 min, EtOAc (11.0 mL) was added and the mixture was cooled to 0 ° C, NaOH (0.67 mL, 10.0% aq) and saturated NaHCO3 (4.60 mL) were added, and stirred for 5 min. LC-MS (EB11687-45-P1A1) showed that about 96.6% of the desired compound was detected. The organic phase was separated, washed with brine, and then dried and concentrated over NaSO4. The crude intermediate 3 (1.10 g, 2.38 mmol, yield 93.3%) was a yellow solid. It was used directly in the next step without further purification. LCMS: (EB11687-45-P1A1_LCMS_SH)RT=1.730min, MS(ESI)m / z=463[M+1] + .

[0097] Step 4: Synthesis of compound 6

[0098] To a DCM solution (2 mL) of intermediate 3 (200 mg, 432 μmol, 1.00 eq) and TME (131 mg, 1.30 mmol, 181 μL, 3.00 eq) was slowly added material 3a (91.2 mg, 648 μmol, 75.3 μL, 1.50 eq). The reaction mixture was stirred at 0-25 °C for 1 h. LCMS (EB11739-8-P1A1) showed that intermediate 3 had been consumed and the desired MS peak was detected. The mixture was concentrated at 25 °C to give a residue. The residue was purified by prep-HPLC: Xtimate C18 150*40mm*10μm; mobile phase: [water (NH4HCO3)-ACN]; gradient: 52%-92% B / 25 to obtain compound 6 (68.0 mg, 119μmol, yield 27.5%, purity 99.2%) as a white solid (LCMS: EB11739-8-P1B2y, HNMR: EB11739-8-P1D1, HPLC: EB11739-8-P1 C2). .LCMS: EB11739-8-P1A1, RT = 1.681min, MS (ESI) m / z = 567.4 [M+1] + .EB11739-8-P1B2y,RT=1.361min,MS(ESI)m / z=567.2[M+1] + .HPLC: EB11739-8-P1C2, RT=4.788min, purity 99.2%. 1H NMR:EB11739-8-P1D1,400MHz,CDCl3-d1δ8.03(s,1H)7.70-7.76(m,2H)7.47-7.53(m,1H)7.39-7.4 5(m,2H)5.98(s,1H)5.85(s,1H)3.22(d,J=4.64Hz,1H)2.78(d,J=13.00Hz,1H)2.44(d,J=14.00Hz,1 H)2.19(dt,J=13.74,3.28Hz,1H)1.96-2.05(m,2H)1.71-1.94(m,7H)1.51-1.64(m,3H)1.42(s,3H)1 .29-1.39(m,3H)1.25(s,3H)1.20(d,J=13.14Hz,1H)1.16(s,3H)1.05(d,J=6.74Hz,6H)0.91(s,3H). .

[0099] ■Example 7

[0100] Compound 7:

[0101]

[0102] Synthesis route:

[0103]

[0104] Step 1: Synthesis of Intermediate 1

[0105] To a toluene solution (40.0 mL) of material 218600-44-3 (2.00 g, 4.07 mmol, 1.00 eq) was added DPPA (3.36 g, 12.2 mmol, 2.63 mL, 3.00 eq) and Et3N (8.24 g, 81.4 mmol, 11.3 mL, 20.0 eq). The mixture was stirred dropwise at 0°C for 6 h. LC-MS (EB11687-32-P1A2) showed that about 84.0% of the product was detected. The mixture was concentrated to give a crude product. The residue was purified by column chromatography (SiO2, 0-10% EtOAc in CH2C l2 Solution), TLC (CH2Cl2: EtOH = 10: 1, Rf = 0.5) purification to give Intermediate 1 (1.40 g, 2.71 mmol, yield 67.0%) as a white solid. LCMS: (EB11687-32-P1A1_LCMS_SH), RT = 1.236 min, MS (ESI) m / z = 517 [M+1] +.

[0106] Step 2: Synthesis of Intermediate 2

[0107] To a toluene solution (104 mL) of intermediate 1 (1.40 g, 2.71 mmol, 1.00 eq), the mixture was stirred at 110 ° C for 2 h. LC-MS (EB11687-34-P1 C1) showed that about 97.0% of the expected compound was detected. The mixture was concentrated to obtain a crude product. The crude compound 2 (1.20 g, 2.46 mmol, yield 90.6%) was a white solid and was used directly in the next step without further purification. LCMS: (EB11687-34-P1C1_LCMS_SH), RT = 1.231 min, MS (ESI) m / z = 489 [M+1] + .

[0108] Step 3: Synthesis of compound 7

[0109] Pyrazole (453 mg, 6.65 mmol, 5.00 eq) was added to a THF solution (12.0 mL) of intermediate 2 (650 mg, 1.33 mmol, 1.00 eq). The mixture was stirred at 25 ° C for 20 h. LC-MS (EB11687-36-P1A1) showed that the yield was about 88.0%. The crude product was purified by reverse phase HPLC (chromatographic column: Welch Xtimate C1840*200mm 7μm; mobile phase: [water (NH3H2O+NH4HCO3)-ACN]; gradient: 58%-98% B in 25min). Compound 7 (450 mg, 771 μmol, yield 58.0%, purity 95.4%) was obtained as a white solid. LCMS: (EB11687-38-P1A1_LCMS_SH): RT=2.859min, MS(ESI)m / z=557[M+1] + ;RT=3.027min,MS(ESI)m / z=489[M-67] + .HPLC: (EB11687-38-P2C1): RT=3.273min; RT=3.681min. 1HNMR: (EB11415-16-P1A)(CDCl3,400MHz).δppm0.94(s,3H)1.06(s,3H)1.11(s,3H)1.17(s ,3H)1.25-1.28(m,3H)1.31-1.40(m,6H)1.51-1.69(m,6H)1.72-1.83(m,4H)1.87-2.15(m, 4H)2.27(d,J=13.70,9.20Hz,1H)2.94-3.02(m,1H)3.16(d,J=4.80Hz,1H)5.99(s,1H)6.39 (dd,J=2.60,1.63Hz,1H)7.13(s,1H)7.55(d,J=0.88Hz,1H)8.04(s,1H)8.16-8.21(m,1H).

[0110] ■Example 8

[0111] Compound 8:

[0112]

[0113] Synthesis route:

[0114]

[0115] ■Example 9

[0116] Compound 9:

[0117]

[0118] Synthesis route:

[0119]

[0120] ■Example 10

[0121] Compound 10:

[0122]

[0123] Synthesis route:

[0124]

[0125] Step 1: Synthesis of intermediate 3

[0126] Dissolve material 1 (250 mg, 540.35 μmol, 1 eq) and material 2 (136.31 mg, 540.35 μmol, 1 eq) in anhydrous N, N-dimethylformamide (3 mL), add HATU (246.55 mg, 648.42 μmol, 1.2 eq) and diisopropylethylamine (209.50 mg, 1.62 mmol, 282.35 μL, 3 eq), protect with nitrogen, and then react at 25 ° C for 12 hours. After the reaction of the raw materials is completed by LCMS monitoring, the reaction solution is poured into water (10 mL) and extracted with ethyl acetate (10 mL*3), the organic phase is washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to mix. The crude product was purified by normal phase column (petroleum ether:ethyl acetate=1:0-1:1) to obtain intermediate 3 (369.4 mg, 710.27 μmol, yield 98.09%) as a yellow solid.

[0127] Step 2: Synthesis of intermediate 4

[0128] Dissolve intermediate 3 (445 mg, 638.53 μmol, 1 eq) in anhydrous dichloromethane (5 mL) and add trifluoroacetic acid (1.54 g, 13.46 mmol, 1 mL, 21.08 eq) to react at room temperature for 1 hour. After the reaction of the raw material was completed by LCMS, the reaction solution was directly concentrated to obtain intermediate 4 (380 mg, 636.73 μmol, yield 99.72%) as a yellow solid, which was directly used in the next step. LCMS: rt = 0.530 min, 597.4 [M + H] + .

[0129] Step 3: Synthesis of compound 10

[0130] After the intermediate 4 (380 mg, 636.73 μmol, 1 eq) was dissolved in anhydrous acetonitrile (5 mL), ethyl acetoacetate (82.86 mg, 636.73 μmol, 80.61 μL, 1 eq) was added, and then the temperature was raised to 60 ° C for 1 hour. After the reaction of the raw material was completed, the reaction solution was concentrated to remove part of the solvent and then lyophilized by reverse phase preparation to obtain compound 10 (147.8 mg, 215.17 μmol, yield 33.79%) as a white solid. Prep-HPLC: column: YMC-Actus Triart C18150*30mm*7um; mobile phase: [water (FA)-ACN]; gradient: 62%-92% B over 10min. LCMS: rt = 1.593min, 663.4 [M + H] +. HPLC: 2.274min. 1H NMR: (400MHz, CHLOROFORM-d)δ=8.05(s,1H),7.99(d,J=8.8Hz,2H),7.76(d,J=8.8Hz,2H) ,6.02(s,1H),5.67(s,1H),3.47(s,2H),3.22(d,J=4.6Hz,1H),2.84-2.76(m,1H),2.43(br d,J=14.0Hz,1H),2.23(s,3H),2.18(br t,J=3.2Hz,1H),2.08-1.98(m,2H),1.93-1.84(m,1H),1.82-1.78(m,3H),1.63(br d,J=13.0Hz,2H),1.49(s,3H),1.42(s,3H),1.39-1.31(m,3H),1.28-1.22(m,4H),1.18(s,4H),1.08(d,J=3.4Hz,6H),0.94(s,3H).

[0131] ■Example 11

[0132] Compound 11:

[0133]

[0134] Synthesis route:

[0135]

[0136] Step 1: Synthesis of intermediate 3

[0137] Add material 2 (163.57 mg, 648.42 μmol, 1 eq), HATU (369.82 mg, 972.63 μmol, 1.5 eq) and N,N-diisopropylethylamine (251.41 mg, 1.95 mmol, 338.83 μL, 3 eq) to a solution of material 1 (300 mg, 648.42 μmol, 1 eq) in N,N-dimethylformamide (5 mL). The reaction solution was reacted at 20 ° C for 12 hours. LCMS monitoring showed that the reaction raw materials disappeared completely and the main peak was the target product. The reaction solution was poured into 30 mL of water and extracted 3 times with ethyl acetate (10 mL*3). The combined organic phase was washed with saturated sodium chloride solution (10 mL), then dried with anhydrous sodium sulfate, filtered, concentrated and mixed. The crude product was purified by normal phase column chromatography (petroleum ether / ethyl acetate = 0-50%) to obtain intermediate 3 (376 mg, 501.75 μmol, 87.42% yield) as a light yellow solid. LCMS: Rt = 0.624 min, 641.5 [M-56+H] + ESI pos.

[0138] Step 2: Synthesis of intermediate 4

[0139] Trifluoroacetic acid (2.5 mL) was added to a solution of intermediate 3 (370 mg, 530.91 μmol, 1 eq) in dichloromethane (5 mL), and the reaction solution was reacted at 30°C for 1 hour. LCMS monitoring showed that the raw material disappeared completely and the main peak was the target product. The reaction solution was concentrated under reduced pressure to obtain compound 4 (315 mg, 527.81 μmol, 99.42% yield) as a light yellow solid. LCMS: Rt = 0.515 min, 597.4 [M + H] + ESI pos.

[0140] Step 3: Synthesis of compound 11

[0141] To the acetonitrile (4mL) solution of intermediate 4 (315mg, 527.81μmol, 1eq) was added intermediate 5 (68.69mg, 527.81μmol, 66.82μL, 1eq), and the reaction solution was heated to 60℃ for 1 hour. LCMS monitoring showed that the raw material disappeared completely and the main peak was the target product. The reaction solution was filtered and concentrated, and the crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] and then freeze-dried to obtain compound 11 (147.2mg, 210.08μmol, yield 39.80%, purity 94.6%). Prep-HPLC::column:Phenomenex luna C18150*40mm*15um;mobile phase:[water(FA)-ACN];gradient:35%-65%B over 22min.LCMS:Rt=1.606min,663.4[M+H] + ESI pos. HPLC: Rt = 1.937 min. 1H NMR(400MHz,CHLOROFORM-d)δ=8.23(s,1H),8.04(d,J=6.8Hz,2H),7.55(d,J=7.8Hz,1H),7 .45(t,J=7.8Hz,1H),6.01(s,1H),5.80(s,1H),3.46(s,2H),3.25(d,J=4.6Hz,1H),2.84(br d,J=13.2Hz,1H),2.45-2.30(m,1H),2.20(s,3H),2.18-2.15(m,2H),2.12-1.86(m,1H),1.83- 1.75(m,4H),1.66-1.54(m,2H),1.48(d,J=8.4Hz,6H),1.41-1.29(m,4H),1.26(s,3H),1.21(br d,J=13.2Hz,1H),1.17(s,3H),1.07(d,J=2.8Hz,6H),0.93(s,3H).

[0142] ■Example 12

[0143] Compound 12:

[0144]

[0145] Synthesis route:

[0146]

[0147] ■Example 13

[0148] Compound 13:

[0149]

[0150] Synthesis route:

[0151]

[0152] Step 1: Synthesis of intermediate 2

[0153] Add thionyl chloride (6.55 g, 55.07 mmol, 4 mL, 42.71 eq) to a toluene (4 mL) solution of material 1 (200 mg, 1.29 mmol, 1 eq) and replace with nitrogen once, then stir at 70°C for 2 hours. After the reaction of the raw material was completed by LCMS, the reaction solution was concentrated under reduced pressure to obtain compound 2 (200 mg, crude) as a green solid.

[0154] Step 2: Synthesis of compound 13

[0155] Triethylamine (349.93 mg, 3.46 mmol, 481.34 μL, 8 eq) was added to a dichloromethane (2 mL) solution of material 3 (200 mg, 432.28 μmol, 1 eq), and the dichloromethane solution of intermediate 2 (150.05 mg, 864.56 μmol, 2 eq) was slowly added after nitrogen replacement 3 times, and stirred at 25 ° C for 1 hour. After the reaction of the raw material was completed by LCMS, the reaction solution was concentrated and then freeze-dried by reverse phase preparation to obtain compound 13 (67.2 mg, 110.48 μmol, 25.56% yield, 98.6% purity) as an off-white solid. Prep-HPLC (column: YMC-Actus Triart C18 150*30mm*7um; mobilephase: [water(FA)-ACN]; gradient: 45%-75% B over 15min). LCMS: Retention time=0.530min, 600.4[M+H]+. HPLC: retention time=2.438min, purity 98.77%. 1H NMR (400MHz, DMSO-d6) δ = 8.65 (s, 1H), 8.47 (br s,1H),7.36(dd,J=7.2,8.8Hz,1H),6.52(dd,J=1.6,9.2Hz,1H),6.24-6.16(m,2H),3.18-3.16(m,1H), 3.00-2.94(m,1H),2.19-2.09(m,1H),1.96-1.90(m,2H),1.90-1.78(m,4H),1.75-1.63(m,2H),1.43(br d,J=12.4Hz,8H),1.39-1.25(m,3H),1.25-1.21(m,1H),1.18(s,3H),1.07(s,3H),1.05-1.02(m,1H),1.00(s,3H),0.95(s,3H),0.87(s,3H).

[0156] ■Example 14

[0157] Compound 15:

[0158]

[0159] ■Synthesis route:

[0160]

[0161] ■Example 15

[0162] Compound 16:

[0163]

[0164] Synthesis route:

[0165]

[0166] ■Example 16

[0167] Compound 17:

[0168]

[0169] Synthesis route:

[0170] ■ Example 17 Compound 18:

[0171]

[0172] Synthesis route:

[0173] ■ Example 18 Compound 19:

[0174]

[0175] Synthesis route:

[0176] Step 1: Synthesis of intermediate 2

[0177] Add benzyl bromide (23.88 g, 136.85 mmol, 16.59 mL, purity 98%, 1.25 eq) and potassium carbonate (22.70 g, 164.22 mmol, 1.5 eq) to a solution of material 1 (50 g, 109.48 mmol, 1 eq) in N, N-dimethylformamide (1 L), and stir at 20 ° C for 12 hours. TLC (petroleum ether / ethyl acetate = 3 / 1) monitors that the raw material reaction is complete and new spots are generated. Water (4 L) is added to the reaction solution and extracted with ethyl acetate (2 L) 3 times. The organic phase is washed twice with saturated brine (2 L), dried and concentrated under reduced pressure to obtain a crude product, which is purified by column chromatography (ethyl acetate / petroleum ether = 10-30%) to obtain intermediate 2 (50 g, 91.44 mmol, yield 83.52%, purity 90%) as a light yellow solid. 1H NMR (400MHz, CHLOROFORM-d) δ = 7.35 (s, 5H), 5.32-5.28 (m, 1H), 5.14-5.02 (m, 2H), 3.26-3.18 (m, 1H), 2.91 (br dd,J=4.0,13.6Hz,1H),2.04-1.93(m,1H),1.86(dd,J=3.6,8.8Hz,2H),1.77-1.65(m,3 H),1.62-1.57(m,4H),1.55-1.49(m,3H),1.49-1.40(m,2H),1.39-1.30(m,3H),1.25(br s,3H),1.13(s,3H),1.08-1.02(m,1H),0.99(s,3H),0.93(s,3H),0.90(s,3H),0.89(s,3H),0.78(s,3H),0.72(br d,J=11.6Hz,1H),0.62(s,3H).

[0178] Step 2: Synthesis of intermediate 3

[0179] To a pyridine (500 mL) solution of intermediate 2 (50 g, 91.44 mmol, 1 eq), 4-dimethylaminopyridine (1.12 g, 9.14 mmol, 0.1 eq) was added and the mixture was replaced with nitrogen three times, then cooled to 0°C, acetic anhydride was slowly added at 0°C, and stirred at 20°C for 12 hours. TLC (petroleum ether / ethyl acetate = 3 / 1) showed that new spots were generated but the raw material was not completely reacted. 4-dimethylaminopyridine (1.12 g, 9.14 mmol, 0.1 eq) was added to the reaction solution, the mixture was replaced with nitrogen three times, then cooled to 0°C, acetic anhydride was slowly added at 0°C, and stirred at 20°C for 24 hours. TLC (petroleum ether / ethyl acetate = 3 / 1) showed that the raw material was completely reacted and new spots became concentrated. The reaction solution was concentrated under reduced pressure to obtain a solid, water (1 L) was added to the solid, and the solid was extracted with ethyl acetate (1 L) three times. The organic phase was dried and concentrated under reduced pressure to obtain intermediate 3 (48 g, crude product) as a white solid, which was used directly in the next step. 1H NMR (400MHz, CHLOROFORM-d) δ = 7.35 (s, 5H), 5.29 (t, J = 3.6Hz, 1H), 5.13-5.02 (m, 2H), 4.52-4.47 (m, 1H), 2.91 (br dd,J=4.0,13.6Hz,1H),2.05(s,3H),1.86(dd,J=3.6,8.8Hz,2H),1.75-1.60(m,7H),1.59-1.45(m,4H),1.44-1.29 (m,3H),1.28-1.16(m,3H),1.13(s,3H),1.08-1.01(m,2H),0.94-0.89(m,9H),0.86(d,J=4.0Hz,7H),0.62(s,3H).

[0180] Step 3: Synthesis of intermediate 4

[0181] To a solution of intermediate 3 (48 g, 81.50 mmol, 1 eq) in dichloromethane (2 L) was added meta-chloroperbenzoic acid (49.65 g, 244.55 mmol, 85% purity, 3 eq) in batches. The reaction solution was stirred at 40 ° C for 12 hours. TLC (petroleum ether / ethyl acetate = 5 / 1) monitored that the raw material reaction was complete and new spots were generated. At 0 ° C, sodium sulfite aqueous solution was slowly added to the reaction solution until the starch potassium iodide test paper did not turn blue, then water (2 L) was added and extracted with dichloromethane (1 L) 3 times. The organic phase was dried and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (ethyl acetate / petroleum ether = 1-5%) to obtain intermediate 4 (50 g, 70.26 mmol, yield 86.20%, purity 85%) as a white solid. 1 H NMR(400MHz,CHLOROFORM-d)δ=7.40-7.33(m,5H),5.26-5.20(m,1H),5.14-5.07(m,1H),4.49(dd,J=5.2,11 .6Hz,1H),2.92-2.81(m,1H),2.48(d,J=4.4Hz,1H),2.26-2.18(m,1H),2.08(s,3H),1.97-1.83(m,3H),1.75

[0182] -1.43(m,10H),1.43-1.32(m,3H),1.28-1.20(m,3H),1.06(br d,J=4.0Hz,1H),1.02(s,3H),1.01-0.95(m,2H),0.93(s,6H),0.88(d,J=1.2Hz,6H),0.85(s,3H),0.68-0.61(m,3H).

[0183] Step 4: Synthesis of intermediate 5

[0184] Dissolve intermediate 4 (20 g, 33.07 mmol, 1 eq) in acetic acid (100 mL) solution, add hydrobromic acid acetic acid solution (3.24 g, 13.23 mmol, 2.18 mL, 33% purity, 0.4 eq) dropwise at room temperature, then heat to 50°C, slowly add bromine (6.34 g, 39.68 mmol, 2.04 mL, 1.2 eq) dropwise under heating conditions. After addition, stir at 50°C for 12 hours. LCMS monitoring shows that the raw material reaction is complete and there is product. The reaction solution was diluted with water (500 mL) and then quenched with 20% sodium thiosulfate aqueous solution (50 mL), followed by extraction with dichloromethane (200 mL*3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate / petroleum ether=0-26%) to obtain intermediate 5 (7.89 g, 11.52 mmol, yield 34.83%) as a yellow solid. LCMS: rt=0.869 min, 603.3 [M+H] + .

[0185] Step 5: Synthesis of intermediate 6

[0186] Dissolve intermediate 5 (35.6 g, 59.05 mmol, 1 eq) in anhydrous methanol (300 mL) and add potassium hydroxide (23.19 g, 413.38 mmol, 7 eq). Heat the reaction mixture to 70 °C and stir for 4 hours. LCMS monitors that the raw material reaction is complete and there is product. The reaction mixture is adjusted to pH 2-3 with 6N HCl in an ice-water bath, then diluted and filtered with water (400 mL). The filter cake is washed with water (300 mL*3) and then dried under reduced pressure to obtain intermediate 6 (30.9 g, crude product) as a yellow solid. LCMS: rt = 0.789 min, 561.4 [M+H] + .

[0187] Step 6: Synthesis of intermediate 7

[0188] Dissolve intermediate 6 (30.9 g, 55.10 mmol, 1 eq) in acetonitrile (600 mL) and add 2-iodobenzoic acid (30.86 g, 110.20 mmol, 2 eq). Heat the reaction mixture to 80 °C and stir for 1 hour. LCMS monitors that the reaction of the raw material is complete and there is product. The reaction mixture is filtered, the filter cake is washed with ethyl acetate (400 mL*3), and the filtrate is concentrated to obtain intermediate 7 (30.79 g, crude product) as a yellow solid. LCMS: rt = 0.816 min, 559.4 [M+H] + .

[0189] Step 7: Synthesis of Intermediate 8

[0190] Dissolve intermediate 7 (30.79 g, 55.10 mmol, 1 eq) in anhydrous dichloromethane (300 mL), and add ethyl formate (16.33 g, 220.41 mmol, 17.73 mL, 4 eq) and sodium methoxide (17.86 g, 330.61 mmol, 6 eq) in turn. The reaction solution was stirred at room temperature for 2 hours. LCMS monitored that the raw material reaction was complete and there was product. The reaction solution was filtered, the filter cake was washed with dichloromethane (500 mL*3), and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether=0-15%) to obtain intermediate 8 (30.47 g, crude product) as a yellow solid. LCMS: rt=0.873 min, 587.3 [M+H] + .

[0191] Step 8: Synthesis of Intermediate 9

[0192] Dissolve intermediate 8 (1g, 1.70mmol, 1eq) in ethanol (10mL) and water (1mL), add hydroxylamine hydrochloride (1.18g, 17.04mmol, 10eq), raise the reaction solution to 100°C and stir for 1 hour. LCMS monitors that the raw material reaction is complete and there is product. The reaction solution is concentrated, diluted with water (20mL) and then extracted with dichloromethane (20mL*3). The organic layers are combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product is purified by column chromatography (dichloromethane / petroleum ether=0-88%) to obtain intermediate 9 (580mg, 993.49μmol, yield 58.30%) as a white solid. LCMS: rt=0.822min, 584.4[M+H] + .1H NMR (400MHz, CHLOROFORM-d) δ = 8.07 (s, 1H), 7.40-7.28 (m, 5H), 5.86 (s, 1H), 5.16 (d,J=4.8Hz,2H),3.15-3.05(m,1H),2.83(d,J=4.6Hz,1H),2.77(d,J=15.0Hz,1H ),2.38(d,J=15.0Hz,1H),1.96-1.84(m,2H),1.78-1.52(m,9H),1.50-1.32(m,6H ),1.26(s,3H),1.13(s,4H),1.02(s,3H),1.00(s,3H),0.96(s,3H),0.90(s,3H).

[0193] Step 9: Synthesis of intermediate 10

[0194] Dissolve intermediate 9 (20.08 g, 34.40 mmol, 1 eq) in anhydrous tetrahydrofuran (300 mL) and add wet palladium carbon (4.09 g, 3.84 mmol, purity 10%, 1.12 e-1 eq). Stir the reaction solution at room temperature for 2 hours under the protection of a hydrogen balloon. LCMS monitored that the raw material reaction was complete and there was a product. The reaction solution was filtered, the filter cake was washed with dichloromethane (500 mL*3), and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 0-31%) to obtain intermediate 10 (9.8 g, 17.07 mmol, yield 49.63%) as a white solid. LCMS: rt = 0.684 min, 494.3 [M+H] + .1H NMR (400MHz, CHLOROFORM-d) δ = 8.07 (s, 1H), 5.90 (s, 1H), 3.09-2.98 (m, 2H), 2.79 (d, J = 15.0Hz, 1H), 2.42 (d, J = 14.8Hz, 1H), 2.00-1.90 (m, 2H), 1.83-1.69(m,5H),1.67-1.60(m,2H),1.59-1.48(m,3H),1.46-1.39(m, 1H),1.36(s,3H),1.32(s,3H),1.29-1.28(m,1H),1.27(s,3H),1.23(br t,J=4.4Hz,1H),1.17(s,3H),1.05(s,3H),1.03(s,3H),0.92(s,3H).

[0195] Step 10: Synthesis of intermediate 11

[0196] Dissolve the intermediate 10 (7 g, 14.18 mmol, 1 eq) in acetone (70 mL), add potassium carbonate (5.88 g, 42.54 mmol, 3 eq) and iodomethane (6.04 g, 42.54 mmol, 2.65 mL, 3 eq), and stir the reaction solution at room temperature for 12 hours. LCMS monitored that the raw material reaction was complete and there was product. The reaction solution was filtered, the filter cake was washed with dichloromethane (100 mL*3), and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 0-30%) to obtain the intermediate 11 (6.3 g, 11.42 mmol, yield 80.51%) as a white solid. LCMS: rt = 0.774 min, 508.2 [M+H] +.1H NMR(400MHz,CHLOROFORM-d)δ=8.07(s,1H),5.90(s,1H),3.71(s,3H),3.06(t d,J=3.6,13.4Hz,1H),2.95(d,J=4.8Hz,1H),2.79(d,J=15.0Hz,1H),2.41(d,J =15.0Hz,1H),1.97-1.82(m,2H),1.81-1.66(m,5H),1.66-1.60(m,1H),1.57(s ,3H),1.55-1.47(m,3H),1.39-1.34(m,3H),1.30(s,3H),1.28(s,3H),1.23(br d,J=2.6Hz,1H),1.17(s,3H),1.04(s,3H),1.02(s,3H),0.91(s,3H).

[0197] Step 11: Synthesis of Intermediate 12

[0198] The intermediate 11 (957 mg, 1.88 mmol, 1 eq) was dissolved in anhydrous tetrahydrofuran (10 mL) and diisobutylaluminum hydride (1 M, 9.42 mL, 5 eq) was slowly added dropwise in an ice-water bath. After the addition was complete, the mixture was stirred in an ice-water bath for 0.5 hours, and then the reaction solution was slowly warmed to room temperature and stirred for 2 hours. LCMS monitored that the raw material reaction was complete and there was a product. The reaction solution was slowly added with water (10 mL) in an ice-water bath to quench the reaction, and then 1N HCl (50 mL) was added to acidify and extracted with ethyl acetate (50 mL*4). The organic layers were combined, washed with water and saturated brine in turn, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the intermediate 12 (970 mg, crude product) as a white solid. LCMS: rt = 0.713 min, 464.2 [M-OH] + .

[0199] Step 12: Synthesis of intermediate 13

[0200] The intermediate 12 (970 mg, 2.01 mmol, 1 eq) was dissolved in anhydrous dichloromethane (20 mL), 4A molecular sieves (2 g) and N-methylmorpholine oxide (518.96 mg, 4.43 mmol, 467.54 μL, 2.2 eq) were added, and the reaction solution was reacted at room temperature for 10 min under nitrogen protection, and then tetrapropylammonium perruthenate (70.77 mg, 201.37 μmol, 0.1 eq) was added to the reaction solution, and the reaction solution was stirred at room temperature for 1.5 hours. LCMS monitoring showed that the raw material reaction was complete and the product was produced. The reaction solution was quenched with saturated aqueous sodium carbonate solution (50 mL), then extracted with dichloromethane (50 mL*3), the organic layers were combined and washed with water and saturated brine in turn, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate / petroleum ether=0-22%) to obtain intermediate 13 (647 mg, 1.19 mmol, 59.19% yield) as a white solid. LCMS: rt=0.743 min, 478.3 [M+H] + .

[0201] Step 13: Synthesis of Intermediate 14

[0202] The intermediate 13 (300 mg, 628.04 μmol, 1 eq) was dissolved in anhydrous ethanol (3 mL) and water (0.2 mL), and p-methoxybenzyloxyamine hydrochloride (154.83 mg, 816.45 μmol, 1.3 eq) and sodium acetate (92.74 mg, 1.13 mmol, 1.8 eq) were added. The reaction solution was stirred at room temperature for 12 hours. LCMS monitored that the raw material reaction was complete and there was a product. The reaction solution was diluted with water (20 mL), then extracted with dichloromethane (20 mL*3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the intermediate 14 (384 mg, crude) as a white solid. LCMS: rt = 0.846 min, 613.3 [M + H] + .

[0203] Step 14: Synthesis of Intermediate 15

[0204] Intermediate 14 (384 mg, 626.59 μmol, 1 eq) was dissolved in anhydrous ethanol (10 mL) and pyridine borane (174.69 mg, 1.88 mmol, 188.04 μL, 3 eq) and 10% aqueous hydrochloric acid solution (2.19 g, 6.02 mmol, 2.15 mL, 10% purity, 9.6 eq) were slowly added dropwise under an ice-water bath. After the addition was complete, the reaction solution was warmed to room temperature and stirred for 12 hours. LCMS monitored the remaining raw materials and the presence of products. The reaction solution was adjusted to alkaline pH with saturated sodium bicarbonate aqueous solution, then extracted with dichloromethane (20 mL*3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate / petroleum ether = 0-15%) to obtain intermediate 15 (280 mg, 318.77 μmol, 50.87% yield) as a colorless jelly. LCMS: rt=0.754min,615.3[M+H] + .

[0205] Step 15: Synthesis of Intermediate 16

[0206] The intermediate 15 (280 mg, 455.39 μmol, 1 eq) was dissolved in anhydrous dichloromethane (4 mL) and material 2 (150.36 mg, 1.37 mmol, 3 eq), N,N-diisopropylethylamine (588.55 mg, 4.55 mmol, 793.19 μL, 10 eq) and n-butylphosphonic anhydride (50% ethyl acetate solution) (984.36 mg, 1.37 mmol, 50% purity, 3 eq) were added in sequence. The reaction solution was stirred at room temperature for 12 hours. LCMS monitored that a small amount of raw materials remained and there was product. The reaction solution was concentrated to obtain a crude product, which was purified by column chromatography (petroleum ether / ethyl acetate = 0-20%) to obtain the intermediate 16 (179 mg, 230.43 μmol, 50.60% yield) as a colorless colloid. LCMS: rt = 0.786 min, 707.3 [M+H] + .

[0207] Step 16: Synthesis of Intermediate 17

[0208] Dissolve intermediate 16 (179 mg, 253.22 μmol, 1 eq) in anhydrous methanol (1 mL) and tetrahydrofuran (1 mL). Add 30% sodium methoxide methanol solution (227.98 mg, 1.27 mmol, 30% purity, 5 eq) under ice-water bath. Warm the reaction solution to room temperature and stir for 1 hour. LCMS monitors that a small amount of raw materials remain and there is product. Dilute the reaction solution with 1N hydrochloric acid aqueous solution (10 mL), then extract with dichloromethane (10 mL*3), wash the organic layers with saturated brine, dry with anhydrous sodium sulfate, filter and concentrate to obtain intermediate 17 (179 mg, crude) as a colorless jelly. LCMS: rt = 0.733 min, 729.4 [M+Na] + .

[0209] Step 17: Synthesis of Intermediate 18

[0210] The intermediate 17 (179 mg, 253.22 μmol, 1 eq) was dissolved in anhydrous N, N-dimethylformamide (2 mL) and material 3 (36.20 mg, 126.61 μmol, 0.5 eq) was added under an ice-water bath, and then the reaction solution was kept under an ice-water bath for 2 hours. Pyridine (80.12 mg, 1.01 mmol, 81.75 μL, 4 eq) was then added, and the reaction solution was heated to 55 ° C and stirred for 4 hours. LCMS monitored that a small amount of raw materials remained and there was product. The reaction solution was diluted with water (10 mL), then extracted with ethyl acetate (10 mL*3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate / petroleum ether = 0-20%) to obtain intermediate 18 (130 mg, 184.43 μmol, 72.83% yield) as a white solid. LCMS: rt=0.743min,727.2[M+Na] + ..

[0211] Step 18: Synthesis of compound 19

[0212] Intermediate 18 (125 mg, 177.33 μmol, 1 eq) was dissolved in hexafluoroisopropanol (1.5 mL), and methanesulfonic acid (170.44 mg, 1.77 mmol, 126.72 μL, 10 eq) was added, and the temperature was raised to 60 ° C for 1 hour. LCMS monitored that the raw material reaction was complete and there was product. The reaction solution was quenched with saturated sodium bicarbonate (10 mL), then extracted with dichloromethane (10 mL*3), the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was then prepared by reverse phase freeze drying to obtain compound 19 (58.01 mg, 95.22 μmol, 53.69% yield) as a white solid. Prep-HPLC:column:Phenomenex Luna C18 150*25mm*10um; mobile phase:[H2O(0.225%FA)-ACN]; gradient:48%-78%B over 15.0min.LCMS: rt=0.654min,585.3[M+H] + .HPLC: rt = 3.863 min. 1 H NMR (400MHz, DMSO-d6) δ = 10.17 (s, 1H), 8.66 (s, 1H), 6.22 (s, 1H), 3.77 (br d, J = 14.4Hz, 1H), 3.51 (br d, J = 14.8Hz, 1H), 3.19 (br d,J=4.6Hz,1H),2.31-2.22(m,1H),1.99-1.77(m,8H),1.76-1.60(m,2H),1.53-1.39(m,7H),1.38-1.20(m ,4H),1.18(s,3H),1.16-1.10(m,2H),1.08(s,3H),1.06-0.99(m,1H),0.93(s,3H),0.84(d,J=8.4Hz,6H).

[0213] ■Example 19

[0214] Compound 20:

[0215]

[0216] Synthesis route:

[0217]

[0218] ■Example 20

[0219] Compound 21:

[0220]

[0221] Synthesis route:

[0222]

[0223] ■Example 21

[0224] Compound 22:

[0225]

[0226] Synthesis route:

[0227] ■Example 22

[0228] Compound 23:

[0229]

[0230] Synthesis route:

[0231] Step 1: Synthesis of intermediate 3

[0232] Material 1 (916 mg, 1.57 mmol, 1 eq) was dissolved in anhydrous dichloromethane (10 mL) and material 2 (583.06 mg, 4.70 mmol, 3 eq), N,N-diisopropylethylamine (2.02 g, 15.66 mmol, 2.73 mL, 10 eq) and n-butylphosphonic anhydride (50% ethyl acetate solution) (3.39 g, 4.70 mmol, 50% purity, 3 eq) were added in sequence. The reaction solution was stirred at room temperature for 2 hours. LCMS monitored that the raw material reaction was complete and there was product. The reaction solution was concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate / petroleum ether = 0-21%) to obtain compound 3 (980 mg, 1.38 mmol, 87.84% yield) as a white solid. LCMS: rt = 0.834 min, 691.3 [M+H] + . 1H NMR (400MHz, CHLOROFORM-d) δ = 8.09 (s, 1H), 7.47-7.33 (m, 5H), 5.91 (s, 1H), 4.90 (s, 2H), 3.22 (br s,1H),2.81(d,J=15.0Hz,1H),2.47-2.32(m,2H),2.31-2.15(m,2H),2.15-2.07(m,1H),2.05-1.90(m, 2H),1.85-1.70(m,3H),1.65-1.59(m,2H),1.57-1.47(m,5H),1.41-1.34(m,4H),1.31(s,4H),1.25(br d,J=7.0Hz,1H),1.21(s,3H),1.19-1.07(m,5H),1.07-1.00(m,5H),0.93(s,3H),0.88(s,3H).

[0233] Step 2: Synthesis of intermediate 4

[0234] Dissolve intermediate 3 (880 mg, 1.27 mmol, 1 eq) in anhydrous methanol (5 mL) and tetrahydrofuran (5 mL) and add 30% sodium methoxide methanol solution (1.15 g, 6.37 mmol, 30% purity, 5 eq) under an ice-water bath. Warm the reaction solution to room temperature and stir for 1 hour. LCMS monitors that a small amount of raw materials remain and there is product. The reaction solution is diluted with 1N hydrochloric acid aqueous solution (30 mL), then extracted with dichloromethane (30 mL*3), the organic layers are combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain intermediate 4 (880 mg, crude) as a white solid. LCMS: rt = 0.759 min, 691.4 [M+H] + .

[0235] Step 3: Synthesis of intermediate 6

[0236] The intermediate 4 (880 mg, 1.27 mmol, 1 eq) was dissolved in anhydrous N, N-dimethylformamide (10 mL) and material 5 (182.09 mg, 636.85 μmol, 0.5 eq) was added under an ice-water bath, and then the reaction solution was kept in an ice-water bath for 2 hours. Pyridine (403.00 mg, 5.09 mmol, 411.22 μL, 4 eq) was then added, and the reaction solution was heated to 55 ° C and stirred for 4 hours. LCMS monitored the remaining raw materials and the product. The reaction solution was diluted with water (20 mL), then extracted with ethyl acetate (20 mL * 3), the organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate / petroleum ether = 0-19%) to obtain intermediate 6 (796 mg, 1.11 mmol, 87.09% yield) as a white solid. LCMS: rt=0.769min,689.3[M+H] + .

[0237] Step 4: Synthesis of compound 23

[0238] The intermediate 6 (640 mg, 929.04 μmol, 1 eq) was dissolved in hexafluoroisopropanol (10 mL), and methanesulfonic acid (1.34 g, 13.94 mmol, 995.80 μL, 15 eq) was added, and the temperature was raised to 60 ° C for 2 hours. LCMS monitored the remaining raw materials and the product. The reaction solution was slowly added to a stirred saturated sodium bicarbonate aqueous solution (20 mL), and then extracted with dichloromethane (20 mL * 3). The organic layers were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by column chromatography (ethyl acetate / petroleum ether = 0-28%) to obtain a crude product, which was then prepared by reverse phase freeze drying to obtain compound 23 (65.2 mg, 106.62 μmol, yield 11.48%) as a white solid. Prep-HPLC:column:Phenomenex LunaC18150*25mm*10um;mobile phase:[H2O(0.225%FA)-ACN];gradient:58%-88%B over15.0min.LCMS:rt=0.678min,599.3[M+H] + .HPLC: rt = 4.015 min. 1H NMR (400MHz, DMSO-d6) δ = 10.14 (s, 1H), 8.66 (s, 1H), 6.21 (s, 1H), 3.82 (br d, J = 14.4Hz, 1H), 3.47 (br d, J = 14.4Hz, 1H), 3.23 (br d,J=4.4Hz,1H),2.29-2.12(m,3H),1.97-1.79(m,5H),1.76-1.62(m,2H),1.47(d,J=8.6Hz,7H),1.38-1.20(m,4H),1.18(s,3H),1.14(br s,1H),1.08(s,3H),1.07-0.98(m,2H),0.97-0.91(m,6H),0.84(d,J=7.8Hz,6H).

[0239] ■Example 23

[0240] Compound 24:

[0241]

[0242] Synthesis route:

[0243]

[0244] ■Example 24

[0245] Compound 25:

[0246]

[0247] Synthesis route:

[0248]

[0249] ■Example 25

[0250] Compound 26:

[0251]

[0252] Synthesis route:

[0253]

[0254] Step 1: Synthesis of intermediate 3

[0255] N,N-diisopropylethylamine (1.33 g, 10.26 mmol, 1.79 mL, 3 eq) was added to a solution of material 1 (2 g, 3.42 mmol, 1 eq) in dichloromethane (20 mL), and then a solution of material 2 (402.67 mg, 5.13 mmol, 364.74 μL, 1.5 eq) in dichloromethane (20 mL) was added at 0°C. The reaction solution was reacted at 20°C for 2 hours. LCMS monitoring showed that the reaction raw material completely disappeared and the main peak was the target product. The reaction solution was concentrated under reduced pressure, and the crude product was purified by normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 1:0 to 3:1) and then concentrated under reduced pressure to obtain intermediate 3 (1.3 g, 1.85 mmol, 53.97% yield, 89% purity) as a white solid. LCMS: Rt = 0.766 min, 627.5 [M+H] + ESI.

[0256] Step 2: Synthesis of intermediate 4

[0257] To a solution of intermediate 3 (1.3 g, 2.07 mmol, 1 eq) in anhydrous methanol (20 mL) was slowly added sodium methoxide solution (1.87 g, 10.37 mmol, 30% purity, 5 eq) at 0 ° C, and the reaction solution was reacted at 20 ° C for 1 hour. The main peak monitored by LCMS is the target product. The reaction solution was slowly poured into 1N HCl solution in turn to adjust the pH value to 3, and extracted with ethyl acetate (20 mL*2). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain intermediate 4 (1.3 g, crude product) as a white solid. LCMS: Rt = 0.706 min, 627.5 [M + H] + ESI.

[0258] Step 3: Synthesis of intermediate 6

[0259] To the solution of intermediate 4 (1.3 g, 2.07 mmol, 1 eq) in N, N-dimethylformamide (15 mL) was slowly added material 5 (296.47 mg, 1.04 mmol, 0.5 eq) at 0 ° C, the reaction solution was reacted at 0 ° C for 2 hours, and then pyridine (656.15 mg, 8.30 mmol, 669.54 μL, 4 eq) was slowly added, and the reaction solution was heated to 55 ° C for 4 hours. The main peak monitored by LCMS is the target product. The reaction solution was poured into water (20 mL) and extracted with ethyl acetate (20 mL*2). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 1:0 to 3:1) and then concentrated under reduced pressure to obtain intermediate 6 (1 g, 1.50 mmol, 72.54% yield, 94% purity) as a white solid. LCMS: Rt=0.718min,625.5[M+H] + ESI.

[0260] Step 4: Synthesis of compound 26

[0261] Boron trichloride solution (1M, 7.20mL, 5eq) was slowly added to a dichloromethane (20mL) solution of intermediate 6 (900mg, 1.44mmol, 1eq) at -70°, and the reaction solution was reacted at 35°C for 4 hours. LCMS monitoring showed that the reaction of the raw materials was complete and the main peak was the target product. The reaction solution was slowly poured into a saturated sodium bicarbonate aqueous solution (20mL) in turn to adjust the pH value to 8, and extracted with dichloromethane (20mL*2). The combined organic phase was washed with saturated brine (20mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] to obtain compound 26 (96.37mg, 164.00μmol, 11.39% yield, 91% purity) as a yellow solid. Prep-HPLC column: Phenomenex lunaC18150*40mm*15um; mobile phase: [H2O(0.225%FA)-ACN]; gradient: 50%-80%B over15.0min.LCMS: rt=0.634min, 535.5[M+H]+.HPLC: rt=2.803min; 1H NMR (400MHz, DMSO-d6) δ = 9.69 (s, 1H), 8.66 (s, 1H), 6.21 (s, 1H), 3.66-3.55 (br m,1H),3.46-3.35(m,1H),3.30-3.20(m,1H),2.24-2.16(m,1H),2.06-1.91(m,5H),1.89-1.7 9(m,4H),1.76-1.61(m,2H),1.50-1.42(m,6H),1.38-1.26(m,3H),1.20-1.16(m,4H),1.12(br s,1H),1.08(s,3H),1.04-0.96(m,2H),0.93(s,3H),0.85(d,J=12.0Hz,6H).

[0262] ■Example 26

[0263] Compound 27:

[0264]

[0265] Synthesis route:

[0266]

[0267] Step 1: Synthesis of intermediate 3

[0268] N,N-diisopropylethylamine (4.20 g, 32.49 mmol, 5.66 mL, 10 eq) and T4P (7.02 g, 9.75 mmol, 50% purity, 3 eq) were added to a solution of material 1 (1.9 g, 3.25 mmol, 1 eq) and material 2 (722.00 mg, 9.75 mmol, 727.09 μL, 3 eq) in dichloromethane (50 mL). The reaction solution was reacted at 25 ° C for 12 hours. LCMS monitoring showed that the reaction raw material disappeared completely and the main peak was the target product. The reaction solution was concentrated under reduced pressure, and the crude product was purified by normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 1:0 to 4:1) and then concentrated under reduced pressure to obtain intermediate 3 (1.8 g, 2.67 mmol, 82.13% yield, 95% purity) as a white solid. LCMS: Rt = 0.786 min, 641.5 [M+H] + ESI.

[0269] Step 2: Synthesis of intermediate 4

[0270] To a solution of intermediate 3 (1.8 g, 2.81 mmol, 1 eq) in anhydrous methanol (20 mL) was slowly added sodium methoxide solution (2.53 g, 14.04 mmol, 30% purity, 5 eq) at 0°C, and the reaction solution was reacted at 25°C for 1.5 hours. The main peak monitored by LCMS is the target product. The reaction solution was slowly poured into 1N HCl solution in turn to adjust the pH value to 3, and extracted with ethyl acetate (20 mL*2). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain intermediate 4 (1.78 g, 2.53 mmol, 89.99% yield, 91% purity) as a white solid. LCMS: Rt = 0.735 min, 641.5 [M+H] + ESI.

[0271] Step 3: Synthesis of intermediate 6

[0272] To the solution of intermediate 4 (1.78 g, 2.78 mmol, 1 eq) in N, N-dimethylformamide (20 mL) was slowly added material 5 (397.06 mg, 1.39 mmol, 0.5 eq) at 0 ° C, the reaction solution was reacted at 0 ° C for 2 hours, and then pyridine (878.76 mg, 11.11 mmol, 896.69 μL, 4 eq) was slowly added, and the reaction solution was heated to 55 ° C for 4 hours. The main peak monitored by LCMS is the target product. The reaction solution was poured into water (20 mL) and extracted with ethyl acetate (20 mL*2). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 1:0 to 3:1) and then concentrated under reduced pressure to obtain intermediate 6 (1.3 g, 1.99 mmol, 71.80% yield, 98% purity) as a white solid. LCMS: Rt=0.746min,639.5[M+H] + ESI.

[0273] Step 4: Synthesis of compound 27

[0274] Boron trichloride solution (1M, 7.83mL, 5eq) was slowly added to a dichloromethane (20mL) solution of intermediate 6 (1g, 1.57mmol, 1eq) at -70°C, and the reaction solution was reacted at 30°C for 12 hours. LCMS monitored the remaining reaction raw materials and the main peak was the target product. The reaction solution was slowly poured into a saturated sodium bicarbonate aqueous solution (20mL) to adjust the pH value to 8, and extracted with dichloromethane (20mL*2). The combined organic phase was washed with saturated brine (20mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 1:0 to 1:1) to obtain a crude product. The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] to obtain compound 27 (113.80mg, 199.29μmol, yield 12.73%, purity 96.10%), as a white solid. Prep-HPLC:column:Phenomenex Luna C18150*25mm*10um; mobile phase:[H2O(0.225%FA)-ACN]; gradient:50%-80%B over 10.0min; LCMS: rt=0.646min,549.4[M+H] + .HPLC: rt = 2.928 min; 1 H NMR (400MHz, DMSO-d6) δ = 9.54 (s, 1H), 8.66 (s, 1H), 6.20 (s, 1H), 3.71-3.42 (m, 2H), 3.27 (br d, J = 3.6Hz, 1H), 2.43-2.36 (m, 2H), 2.20 (br d,J=12.8Hz,1H),2.03-1.92(m,1H),1.89-1.77(m,4H),1.75-1.62(m,2H),1.52-1.42(m,7H),1.38-1.31(m,1H),1.30 -1.21(m,2H),1.18(s,3H),1.15-1.11(m,1H),1.08(s,4H),1.03-0.94(m,5H),0.93(s,3H),0.86(s,3H),0.83(s,3H).

[0275] ■Example 27

[0276] Compound 28:

[0277]

[0278] Synthesis route:

[0279]

[0280] Step 1: Synthesis of intermediate 3

[0281] To a solution of material 1 (1 g, 1.71 mmol, 1 eq) in dichloromethane (20 mL), material 2 (441.61 mg, 5.13 mmol, 405.89 μL, 3 eq), N,N-diisopropylethylamine (1.33 g, 10.26 mmol, 1.79 mL, 3 eq) and then T4P (3.70 g, 5.13 mmol, 50% purity, 3 eq) were added. The reaction solution was reacted at 20 ° C for 2 hours. LCMS monitoring showed that the reaction raw materials disappeared completely and the main peak was the target product. The reaction solution was concentrated under reduced pressure, and the crude product was purified by normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 1:0 to 3:1) and then concentrated under reduced pressure to obtain intermediate 3 (900 mg, 1.20 mmol, 70.14% yield, 87% purity) as a white solid. LCMS: Rt = 0.808 min, 653.5 [M + H] + ESI.

[0282] Step 2: Synthesis of intermediate 4

[0283] To a solution of intermediate 3 (740 mg, 1.13 mmol, 1 eq) in anhydrous methanol (10 mL) was slowly added sodium methoxide solution (1.02 g, 5.67 mmol, 30% purity, 5 eq) at 0°C, and the reaction solution was reacted at 20°C for 1 hour. The main peak monitored by LCMS is the target product. The reaction solution was slowly poured into 1N HCl solution in turn to adjust the pH value to 3, and extracted with ethyl acetate (20 mL*2). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain intermediate 4 (740 mg, crude product) as a white solid. LCMS: Rt = 0.738 min, 653.5 [M+H] + ESI.

[0284] Step 3: Synthesis of intermediate 6

[0285] To the solution of intermediate 4 (740 mg, 1.13 mmol, 1 eq) in N,N-dimethylformamide (10 mL) was slowly added material 5 (162.03 mg, 566.70 μmol, 0.5 eq) at 0°C, the reaction solution was reacted at 0°C for 2 hours, then pyridine (358.61 mg, 4.53 mmol, 365.92 μL, 4 eq) was slowly added, and the reaction solution was heated to 55°C for 4 hours. The main peak monitored by LCMS was the target product. The reaction solution was poured into water (20 mL) and extracted with ethyl acetate (20 mL*2). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 1:0 to 3:1) and then concentrated under reduced pressure to obtain intermediate 6 (500 mg, 736.69 μmol, 65.00% yield, 95.9% purity) as a white solid. LCMS: Rt = 0.752 min, 651.5 [M+H] + ESI.

[0286] Step 4: Synthesis of compound 28

[0287] Boron trichloride solution (1M, 3.07mL, 5eq) was slowly added to a dichloromethane (8mL) solution of intermediate 6 (400mg, 614.55μmol, 1eq) at -70°, and the reaction solution was reacted at 35°C for 2 hours. LCMS monitoring showed that the reaction raw materials reacted completely and the main peak was the target product. The reaction solution was slowly poured into a saturated sodium bicarbonate aqueous solution (20mL) in turn to adjust the pH value to 8, and extracted with dichloromethane (20mL*2). The combined organic phase was washed with saturated brine (20mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] to obtain compound 28 (48.49mg, 84.41μmol, 13.74% yield, 97.618% purity) as a yellow solid. Prep-HPLC column: Phenomenexluna C18150*40mm*15um; mobile phase: [H2O(0.225%FA)-ACN]; gradient:55%-85%Bover 15.0min.LCMS: rt=0.655min,561.5[M+H] + .HPLC: rt = 1.939 min; 1H NMR (400MHz, DMSO-d6) δ = 9.80 (s, 1H), 8.66 (s, 1H), 6.20 (s, 1H), 3.69-3.62 (m, 1H), 3.59-3.52 (m, 1H),3.29-3.22(m,1H),2.33-2.28(m,1H),2.23-2.16(m,1H),2.02-1.94(m,1H),1.88-1.82(m,3H ),1.74-1.63(m,2H),1.46(d,J=2.0Hz,6H),1.38-1.22(m,4H),1.21-1.13(m,5H),1.11-1.03(m,5 H),1.01-0.96(m,1H),0.93(s,3H),0.85(d,J=12.8Hz,6H),0.77-0.73(m,2H),0.72-0.68(m,2H).

[0288] ■Example 28

[0289] Compound 29:

[0290]

[0291] Synthesis route:

[0292]

[0293] Step 1: Synthesis of intermediate 3

[0294] Add material 2 (858.71 mg, 9.75 mmol, 903.90 μL, 3 eq), N,N-diisopropylethylamine (4.20 g, 32.49 mmol, 5.66 mL, 10 eq) and T4P (7.02 g, 9.75 mmol, 50% purity, 3 eq) to a solution of material 1 (1.9 g, 3.25 mmol, 1 eq) in dichloromethane (50 mL). Stir at 25 °C for 12 hours. LCMS monitored that the raw material reacted completely. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (ethyl acetate / petroleum ether = 20-25%) to obtain intermediate 3 (1.6 g, 2.20 mmol, 67.68% yield, 90% purity) as a white solid. LCMS: rt = 0.821 min, 655.3 [M+H] + .

[0295] Step 2: Synthesis of intermediate 4

[0296] Sodium methoxide (2.20 g, 12.22 mmol, 30% purity, 5 eq) was added to a solution of intermediate 3 (1.6 g, 2.44 mmol, 1 eq) in methanol (20 mL) at 0°C and stirred at 25°C for 2 hours under a nitrogen atmosphere. LCMS monitored that the reaction of the raw material was complete. The reaction solution was adjusted to pH 3-4 with 1N hydrochloric acid, extracted with dichloromethane, washed and dried, and then concentrated under reduced pressure to obtain intermediate 4 (1.5 g, crude product, white solid. LCMS: rt = 0.757 min, 655.3 [M+H] + ..

[0297] Step 3: Synthesis of intermediate 6

[0298] Add material 5 (327.43 mg, 1.15 mmol, 0.5 eq) to a solution of intermediate 4 (820 mg, 3.01 mmol, 1 eq) in N, N-dimethylformamide (15 mL) at 0 ° C and stir at 0 ° C for 2 hours under a nitrogen atmosphere. Add pyridine (724.67 mg, 9.16 mmol, 739.46 μL, 4 eq) to the reaction solution and stir at 55 ° C for 4 hours. LCMS monitored that the raw material reaction was complete. The reaction solution was decompressed and poured into water and filtered. The filtered solid was slurried with water to obtain intermediate 6 (1.4 g, 1.93 mmol, yield 84.26%, purity 90%) as a light yellow solid. LCMS: rt = 0.767 min, 653.5 [M + H] + ..

[0299] Step 4: Synthesis of compound 29

[0300] A solution of compound 6 (1 g, 1.53 mmol, 1 eq) in dichloromethane (10 mL) was replaced with nitrogen three times and then cooled to -70 ° C. Boron trichloride (1 M, 7.66 mL, 5 eq) was slowly added, and the reaction solution was reacted at 35 ° C for 4 hours. LCMS monitored that the raw material reaction was complete. The reaction solution was poured into a saturated sodium bicarbonate solution, extracted with dichloromethane, washed, dried and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] and then freeze-dried to obtain compound 29 (150.65 mg, 267.69 μmol, 17.48% yield) as a white solid. Prep-HPLC:column:Phenomenex luna C18150*25mm*10um; mobile phase:[H2O(0.225%FA)-ACN]; gradient:60%-90%B over 10.0min.LCMS:Retention time=4.177min,563.4[M+H] +.HPLC: retention time = 2.396 min, purity 98.87%; 1 HNMR (400MHz, DMSO-d6) δ = 9.56 (s, 1H), 8.67 (s, 1H), 6.21 (s, 1H), 3.74 (br d, J = 14.4Hz, 1H), 3.41 (br d, J = 14.4Hz, 1H), 3.29 (br d,J=4.8Hz,1H),3.10(quin,J=6.8Hz,1H),2.27-2.17(m,1H),2.02-1.93(m,1H),1.92-1.79(m,4H),1.75-1.6 3(m,2H),1.48(d,J=8.4Hz,6H),1.46-1.39(m,1H),1.37-1.31(m,1H),1.31-1.21(m,2H),1.19(s,3H),1.15(br d,J=3.2Hz,1H),1.12(br s,1H),1.09(s,3H),1.04(br s,1H),1.00(dd,J=5.6,6.8Hz,7H),0.93(s,3H),0.86(s,3H),0.83(s,3H).

[0301] ■Example 29

[0302] Biological evaluation

[0303] Test Example 1 In vitro human Nrf2 receptor agonist activity assay

[0304] Reagents: Opti-MEM serum-free medium; PEI 40000 transfection reagent; DMEM complete medium.

[0305] Equipment: Cx7Pro high-content rapid imaging platform, etc.

[0306] For each well of cells, use 10 μL Opti-MEM serum-free medium (Cienry) to dilute 360 ​​ng of Nrf2 phase change probe plasmid, mix thoroughly to make DNA dilution solution, let stand for 5 minutes, use 10 μL Opti-MEM serum-free medium to dilute 0.6 μL of PEI 40000 transfection reagent (YEASEN), mix thoroughly to make PEI 40000 dilution solution, and let stand for 5 minutes. After mixing the DNA dilution solution and the PEI 40000 dilution solution, incubate at room temperature for 20 minutes to form a DNA-PEI cationic nucleic acid transfection reagent complex. Remove 20 μL of cell growth medium and add 20 μL of DNA-PEI cationic nucleic acid transfection reagent complex to each well. Shake the culture plate and mix gently. 37 ℃, 5% CO2 incubator culture, 1 hour after transfection, remove 75 μL of cell growth medium, add 75 μL of fresh preheated DMEM complete medium (Meilunbio) to each well, and maintain the volume of each well medium at 100 μL. 18 hours after transfection, the gradient dilution of drugs (0.0625nM, 0.125nM, 0.25nM, 0.5nM, 1nM, 2nM, 4nM, 8nM, 16nM, 32nM, 128nM) was first mixed into the preheated DMEM complete medium, and 100 μL of medium containing gradient dilution of drugs was added to the sample wells, and the final volume of each sample well was 200 μL. 6 hours after adding the drug, the Cx7Pro high-content rapid imaging platform (Thermo Fisher) was used with a 20x high-definition lens to randomly select 16 fields of view from each well for photography. The position of the cell nucleus was determined by the NLS-mTagBFP2 independently expressed in the probe, thereby determining the number of transfected cells. Calculate the total fluorescence intensity of the Nrf2 phase change probe phase transition. Count the total fluorescence intensity of the Nrf2 phase change probe phase change "droplet" in each cell. Compare the total fluorescence intensity of the Nrf2 phase change probe phase transition in each cell in the drug group with the total fluorescence intensity of the Nrf2 phase change probe phase transition in each cell in the DMSO group. Use the log(agonist) vs.response--Variable slope(four parameters) analysis method in GraphPad Prism to calculate the EC value of each drug. 50 , the results are shown in Table 1. The structural formula of Omaveloxolone is as follows:

[0307]

[0308] Table 1 Test results of representative compounds of the present invention on in vitro human Nrf2 receptor agonist activity

[0309]

[0310]

[0311] The biological experimental data are shown in Table 1. Moreover, the test experimental data show that compounds 2, 4, 6, 7, 11, 13, 19, 23, 26, 27, 28 and 29 prepared by the present invention all exhibit Nrf2 receptor agonist activity, especially compounds 4, 19, 23, 26, 27, 28 and 29 prepared by the present invention exhibit single-digit nanomolar human Nrf2 receptor agonist activity, and are superior to the control drug Omaveloxolone. The above in vitro human Nrf2 receptor agonist activity data show that the compounds of the present invention, their pharmaceutically acceptable salts, and stereoisomers can be used to prepare NRF2-Leap1 uncouplers. Moreover, compared with the control drug Omaveloxolone, the compounds of the present invention have significant differences in biological activities such as DPPH free radical scavenging and MDA anti-lipid peroxidation, as shown in Test Examples 2-4.

[0312] Test Example 2 DPPH free radical scavenging ability test

[0313] Purpose of the experiment: To determine the DPPH free radical scavenging ability of the compounds of the present invention.

[0314] Test Materials:

[0315] Material factory Part Number Dimethyl sulfoxide Sigma D2650 D4Plus Sample Head Carrier Tecan 30097371 T8Plus Sample Head Carrier Tecan 30097370 96-well plate Corning 3599 Anhydrous ethanol Bioengineering A500737 2,2-Diphenyl-1-picrylhydrazyl(DPPH) MCE HY-112053

[0316] Test equipment:

[0317]

[0318]

[0319] Test method:

[0320] First, add 20 μL of the compound to be tested to a 96-well plate and dilute it continuously in 1:2 with DMSO. Then add 200 μL of 200 μM DPPH prepared with anhydrous ethanol as solvent to each well, shake it slightly and incubate it at room temperature in the dark for 30 minutes. Detect the absorbance value at 517 nm with an ELISA reader. Calculate the DPPH clearance rate using the following formula: DPPH clearance rate % = (1-Ai / A0) * 100%, Ai refers to the sample absorbance value, and A0 refers to the DMSO control group absorbance value. Use XLfit 5.3.1.3 software to process the data, and use the nonlinear fitting formula to obtain the IC50 value of the compound. The results are shown in Table 2.

[0321] Table 2 DPPH free radical scavenging ability test results of representative compounds of the present invention

[0322]

[0323] The results showed that the compounds 27 and 28 prepared in the present invention had the ability to scavenge DPPH free radicals, while Omaveloxolone had no such activity.

[0324] Test Example 3 MDA Anti-lipid Peroxidation Ability Test

[0325] Experimental purpose: To test the MDA anti-lipid peroxidation ability of the compounds of the present invention.

[0326] Experimental Materials:

[0327] Material factory Part Number DPBS (1×) Corning 21-031-CVC Dimethyl sulfoxide Sigma D2650 D4Plus Sample Head Carrier Tecan 30097371 T8Plus Sample Head Carrier Tecan 30097370 96-well plate Corning 3599 ascorbic acid ST1434 Blue Sky Lipid oxidation (MDA) detection kit S0131M Blue Sky

[0328] Test equipment:

[0329]

[0330]

[0331] Test method:

[0332] First, extract brain tissue homogenate: an adult male SD rat was anesthetized with isoflurane and then killed by dislocating the cervical vertebrae. The whole brain was removed and washed twice in DPBS. The meninges were peeled off and transferred to a 50mL centrifuge tube containing 10mL DPBS. After being cut with scissors, they were divided into 10 1.5mL centrifuge tubes. Three grinding beads were added to each tube and ground at 90Hz for 60min for 3 times. The ground tissue homogenate was transferred to the same new 50mL centrifuge tube, and DPBS was added to a total volume of 30mL and mixed. Next, 20μL of the test compound was added to the 96-well plate and diluted continuously 1:3 with DMSO. Then 100μL of brain tissue homogenate, 50μL DPBS and 50μL of 200μg / ml vitamin C were added in sequence, and a series of concentration standards were prepared as standard curves. After shaking, incubate at 37℃ for 1 hour, then add 400μL MDA working solution and heat at 100℃ for 15 minutes. After cooling to room temperature, centrifuge at 1000g for 10 minutes, aspirate 200μL of supernatant into another new plate, and detect the absorbance at 532nm using an enzyme reader. The clearance rate of the compound on MDA was calculated using the following formula: MDA clearance rate % = [((A1-A0)-(A2-A0)) / ((A1-A0)-(A3-A0))]*100%, where A1 refers to the absorbance value of the high control group, A2 refers to the sample absorbance value, A3 refers to the absorbance value of the low control group, and A0 refers to the absorbance value of the blank group. The data were processed using XLfit5.3.1.3 software, and the IC value of the compound was obtained using the nonlinear fitting formula. 50 The results are shown in Table 3.

[0333] Table 3 MDA anti-lipid peroxidation ability test results of the compounds of the present invention

[0334]

[0335] The results show that the compounds 23, 26, 27, 28 and 29 prepared in the present invention all have anti-lipid peroxidation ability, and their activity is better than that of Edaravone, while Omaveloxolone has no such activity.

[0336] Experimental Example 4: Intervention Effect of Representative Compounds on Ferroptosis Process (qPCR)

[0337] 1) Experimental purpose: To test the intervention effect of a series of compounds on the ferroptosis process by detecting changes in the expression levels of key genes in the ferroptosis signaling pathway at the cellular level. The specific indicator is the detection of the mRNA level of PTGS2, a key gene in the ferroptosis process (fluorescence real-time quantitative qRT-PCR).

[0338] 2) Experimental methods:

[0339] HT-1080 cells (Cat. No.: CCL-121, Shanghai Cell Bank, introduced from ATCC) were plated in six-well plates at 4×105 / well for 22h; HT-1080 cells were treated with different concentration gradient compounds (1.37nM to 333nM, a total of 6 concentration gradients) for 1h in advance; HT-1080 cells were treated with 200nM of the classic ferroptosis inducer RSL3 (Cat. No.: HY-100218A, MCE, USA) for 16h; mRNA was extracted and reverse transcribed into cDNA, and PTGS2 mRNA levels were detected by qRT-PCR (Model: ABI7500, Thermo Fisher, USA). The ferroptosis inhibitor Fer-1 (Cat. No.: HY-100579, MCE, USA) was used as a positive control.

[0340] 3) Experimental results:

[0341] The IC of each drug was calculated using the log(agonist) vs. response--Variable slope(four parameters) analysis method in GraphPad Prism. 50 , the results are shown in Table 4.

[0342] Table 4 Intervention effect of the compounds of the present invention on the ferroptosis process

[0343]

[0344] The results showed that compounds 26, 27, 28 and 29 prepared in the present invention can inhibit ferroptosis, while Omaveloxolone has no such activity.

[0345] The above-mentioned pharmacological experiments prove that the preferred NRF2-Keap1 compounds prepared by the present invention, such as 26, 27, 28, 29, etc., not only maintain the Nrf2 agonist activity similar to the marketed Nrf2 agonist Omaveloxolone, but also increase the effects of scavenging DPPH free radicals, inhibiting the production of lipid peroxide MDA or intervening in ferroptosis.

[0346] At the same time, biological in vivo tests have shown that the compounds of the present invention have significant effects on a variety of diseases, including: cerebral small vessel disease, mitochondrial encephalopathy, autism spectrum disorder, Rett syndrome, Friedreich's ataxia, stroke, hemorrhagic stroke, ischemic stroke, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, schizophrenia cognitive impairment, Parkinson's disease, Parkinson's cognitive impairment, Alzheimer's disease, vascular dementia, epilepsy, Huntington's disease, heart failure, myocardial infarction, renal failure, renal ischemia. In particular, the compounds of the present invention have significant effects on diseases such as stroke, multiple sclerosis, and amyotrophic lateral sclerosis, and can be used as a drug for the preparation of drugs for preventing or treating stroke, multiple sclerosis, and amyotrophic lateral sclerosis.

[0347] Experimental Example 5 Protective effect of representative compounds on ischemic stroke rats

[0348] 1) Reagents: Omaveloxolone, MedChemexpress Biotechnology, USA; Transient Middle Cerebral Artery Occlusion Model (tMCAO) suture, Beijing Reward Life Science Co., Ltd.; DMAO, Bio-Technology Co., Ltd.; Solutal, Sigma-Aldrich (Shanghai) Trading Co., Ltd.; Normal saline, Sinopharm Chemical Reagent Co., Ltd.; TTC, Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0349] 2) Establishment of cerebral ischemia rat model by tMCAO method:

[0350] Rats were anesthetized with 10% chloral hydrate (350 mg / kg), fixed on the operating table in the supine position, and the neck was disinfected. A midline incision was made in the neck, and the intermuscular space between the left sternocleidomastoid muscle and the sterno-glossus muscle was bluntly separated to expose the common carotid artery. The common carotid artery was picked out by ophthalmic curved forceps and threaded with silk thread for later use. The external carotid artery was separated under the right digastric muscle and hyoid bone, and the thread was threaded for later use. The suture was passed through and tied to the common carotid artery, and a small incision was made at the free end of the common carotid artery and a loose knot was tied. The thread plug was inserted along the common carotid artery through the incision, and the thread plug was slowly pushed forward. When resistance was felt, it stopped, and the pre-tied loose knot was tied. The body temperature was maintained at 37°C during the operation, and the thread plug was pulled out after 1.5 hours. In the sham operation group, the thread plug was not inserted after the vascular separation, and the other operation steps were the same.

[0351] 3) Experimental grouping and drug administration:

[0352] 40 SD male rats, weighing 230-250g, were randomly divided into sham operation group, model group, Omaveloxolone 3mg / kg group, Omaveloxolone 9mg / kg group, and compound 283.03mg / kg group. After 3 days of adaptive feeding, the rats were injected with drugs through the tail vein when the plug was inserted in the modeling. The rats in the sham operation group and the model group were injected with an equal volume of solvent (10% DMSO + 10% solutal + 80% saline).

[0353] 4) Index detection:

[0354] mNSS score: Neurobehavioral score was performed 24 hours after modeling, mainly to evaluate the neurological function of rats with ischemic stroke from the two aspects of sensation and movement.

[0355] Determination of cerebral infarction area: Rats were euthanized by overdose of chloral hydrate, and the brain was removed and placed in a -20°C refrigerator for 20 minutes. The brain was removed and placed in the brain trough, sliced, and the thickness of the brain slice was 2 mm. The brain slice was placed in a 2% TTC solution, shielded from light, and incubated in a 37°C constant temperature box for 15 minutes. After staining, pictures were taken for analysis of cerebral infarction area.

[0356] 5) Statistical methods:

[0357] All data in this study were expressed as mean ± standard deviation (Mean ± SD) and statistically analyzed using GraphPad Prism 7.0. Differences among multiple groups were evaluated by one-way ANOVAs followed by Tukey's test. Behavioral tests were tested by Krystal-Wallis test. P < 0.05 was considered statistically significant.

[0358] 6) Experimental results:

[0359] Effects of compound 28 on neurological function in rats with ischemic stroke:

[0360] like Figure 1 As shown, Omaveloxolone 3mg / kg and 9mg / kg had no effect on the mNSS score of rats with cerebral ischemia. At an equimolar dose to Omaveloxolone 3mg / kg, compound 28 (3.03mg / kg) could significantly reduce the mNSS score of rats with cerebral ischemia.

[0361] Effects of compound 28 on cerebral infarction area in rats with ischemic stroke:

[0362] like Figure 2As shown, Omaveloxolone 3mg / kg and 9mg / kg had no effect on the cerebral infarction area of ​​rats with cerebral ischemia. At an equimolar dose to Omaveloxolone 3mg / kg, compound 28 (3.03mg / kg) could significantly reduce the cerebral infarction area of ​​rats with cerebral ischemia.

[0363] 7) Conclusion: The Nrf2-Keap1 uncouplers of the present invention, such as compound 28, can reduce the cerebral infarction area in rats with ischemic stroke and have a protective effect on neurological damage.

[0364] Experimental Example 6 Effects of Representative Compounds on Neurobehavior in Multiple Sclerosis Model Mice

[0365] 1. Materials and Methods

[0366] 1) Main reagents

[0367]

[0368] 2) Experimental animals and grouping and drug administration

[0369] Female C57BL / 6J mice were randomly divided into 5 groups: sham operation group, model group, Omaveloxolone 5 mg / kg group, Omaveloxolone 15 mg / kg group, and compound 635.05 mg / kg, with 8 mice in each group. The mice in the sham operation group and the model group were given the same volume of solvent (10% solutol + 90% saline) twice a day for 42 consecutive days.

[0370] 3) Preparation of Multiple Sclerosis Model-EAE Model

[0371] The EAE model is a commonly used animal model of multiple sclerosis. The mixed emulsion containing myelin oligodendrocyte glycoprotein MOG35-55 peptide and complete Freund's adjuvant (CFA) containing Mycobacterium tuberculosis was injected subcutaneously on the back of mice, and pertussis toxin (PTX) was injected intraperitoneally on the day of immunization and 48 hours later to establish the EAE (experimental autoimmune encephalosporin) model.

[0372] 4) Weight and neurological function score (5-point scale)

[0373] Starting from the day of immune induction (Day 0), the rats were weighed and neurological function was scored daily. Neurological function scoring criteria (5-point method): 0 points, no clinical deficits; 1 point, partial tail paralysis; 2 points, complete tail paralysis; 3 points, partial hind limb paralysis; 4 points, complete hind limb paralysis; 5 points, forelimb paralysis; 6 points, death.

[0374] 5) Statistical processing

[0375] All data are presented as mean ± standard error (Mean ± SEM). Two-way ANOVA and Tukey's multiple comparison test were used to compare the differences in neurological function scores and body weight among the groups. All data were analyzed using GraphPad Prism 9.0.0 software. p < 0.05 indicated that the difference was statistically significant.

[0376] 2. Experimental results

[0377] like Figure 3 As shown, Omaveloxolone 5 and 15 mg / kg can significantly reduce the neurological function scores of EAE model mice, but there is no significant difference between the two groups, indicating that Omaveloxolone 5 mg / kg has the maximum effect. At the same molar dose as Omaveloxolone 5 mg / kg, compound 28 (5.05 mg / kg) can significantly reduce the neurological function scores of EAE model mice, and the effect intensity is better than Omaveloxolone 15 mg / kg, and the difference is statistically significant.

[0378] like Figure 4 As shown, Omaveloxolone 5 and 15 mg / kg can significantly increase the body weight of EAE model mice, but there is no significant difference between the two groups, indicating that Omaveloxolone 5 mg / kg has the maximum effect. At the same molar dose as Omaveloxolone 5 mg / kg, compound 28 (5.05 mg / kg) can significantly reduce the neurological function score of EAE model mice, and the effect intensity is better than Omaveloxolone 15 mg / kg, and the difference is statistically significant.

[0379] 3. Experimental conclusion

[0380] Compound 28 has a protective effect on the neurological function of multiple sclerosis model mice, and its effect intensity is better than that of Omaveloxolone.

[0381] Experimental Example 6 Protective effect of representative compounds on amyotrophic lateral sclerosis model mice

[0382] 1. Materials and methods

[0383] 1) Main reagents

[0384] Omaveloxolone was purchased from MCE Biotech; Solutal was purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.; and normal saline was purchased from Sinopharm Chemical Reagent Co., Ltd.

[0385] 2) Animals

[0386] Forty-eight B6SJL-Tg(SOD1 G93A)-1Gur / J transgenic mice (half male and half female) of the amyotrophic lateral sclerosis (ALS) model were purchased from Shanghai Model Organisms Technology Co., Ltd.

[0387] 3) Experimental grouping and drug administration

[0388] SOD1 G93A mice were randomly divided into model group, Omaveloxolone 1mg / kg group, Omaveloxolone 3mg / kg group, and compound 281.01mg / kg group, with 12 mice in each group. At the same time, 12 C57BL / 6J mice were set as the control group. The control group and the model group were intraperitoneally injected with an equal volume of solvent (1% DMSO + 4% solutal + 95% saline), once a day for 10 consecutive weeks.

[0389] 4) Index detection

[0390] Rotarod test

[0391] The rotarod test is a classic behavioral experiment to evaluate the motor coordination of mice. The rotarod test (Panlab rotarod instrument, purchased from Harvard Bioscience, USA) was performed twice a week, and the rotation speed of the rotarod ranged from 4 to 40 r / min. The experimental steps are as follows: 1. Before the formal experiment, the mice were trained to adapt to the rotation speed, and the rotation speed was set to 12 r / min, 5 min each time, twice a day, for 3 days; 2. In the formal experiment, 3 consecutive tests were performed (3 min / time, with an interval of 30 min each time); 3. The time the mouse stayed on the rotarod each time was recorded, and the longest time on the rotarod in the 3 tests was taken as the latent period of falling.

[0392] Onset time

[0393] The rotarod test was used to detect the onset time of mice, and the first time the mouse fell from the rotarod within 3 minutes was recorded as the onset date.

[0394] Cage experiment

[0395] The hanging cage test is used to evaluate the grip strength and endurance of the mouse's limbs. The hanging cage test is performed twice a week. Each mouse is placed in the center of the iron net, and the iron net is gently shaken to make the mouse hold it tightly. Then the iron net is slowly inverted to a horizontal position, and the time the mouse hangs on the iron net is recorded. Each mouse is measured three times, with an interval of 30 minutes between each test, and the maximum value is taken as the fall latency.

[0396] 2. Experimental results

[0397] 1) Effect of compound 28 on the onset time of SOD1 G93A mice

[0398] like Figure 5 As shown, both Omaveloxolone 1mg / kg and 3mg / kg can significantly delay the onset of SOD1 G93A mice, but there is no significant difference between the two groups, indicating that Omaveloxolone 1mg / kg has the maximum effect. At the same molar dose as Omaveloxolone 1mg / kg, compound 28 (1.01mg / kg) can significantly delay the onset of SOD1 G93A mice, and the intensity of action is better than Omaveloxolone 3mg / kg, and the difference is statistically significant.

[0399] 2) Effect of compound 28 on motor coordination ability of SOD1 G93A mice

[0400] like Figure 6 As shown, both Omaveloxolone 1mg / kg and 3mg / kg can significantly improve the motor coordination ability of SOD1 G93A mice, which is manifested by a significant increase in the latency to fall, but there is no significant difference between the two groups, indicating that Omaveloxolone 1mg / kg has reached its maximum effect. At an equimolar dose to Omaveloxolone 1mg / kg, compound 28 (1.01mg / kg) can significantly increase the latency to fall of mice, and the intensity of action is better than Omaveloxolone 3mg / kg, and the difference is statistically significant.

[0401] 3) Effect of compound 28 on muscle endurance of SOD1 G93A mice

[0402] like Figure 7 As shown, both Omaveloxolone 1mg / kg and 3mg / kg can significantly improve the motor coordination ability of SOD1 G93A mice, which is manifested by a significant increase in the latency to fall, but there is no significant difference between the two groups, indicating that Omaveloxolone 1mg / kg has reached its maximum effect. At an equimolar dose to Omaveloxolone 1mg / kg, compound 28 (1.01mg / kg) can significantly increase the latency to fall of mice, and the intensity of action is better than Omaveloxolone 3mg / kg, and the difference is statistically significant.

[0403] 3. Experimental conclusion

[0404] Compound 28 can improve the neurobehavior of ALS model mice, and its effect intensity is better than Omaveloxolone.

[0405] The above test results show that the C17 nitrogen-substituted and methylene-substituted oleanol triterpene derivatives disclosed in the present invention can be used as a new class of Nrf2-Keap1 uncouplers, have excellent Nrf2 agonism, and have the activity of scavenging DPPH free radicals, inhibiting the generation of lipid peroxide MDA, or intervening in ferroptosis to exert antioxidant effects. Moreover, the in vivo test of the present invention shows that the compounds of the present invention can be effectively used for the prevention or treatment of diseases including cerebral small vessel disease, mitochondrial encephalomyopathy, autism spectrum disorder, Rett syndrome, Friedreich's ataxia, stroke, hemorrhagic stroke, ischemic stroke, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, schizophrenia cognitive impairment, Parkinson's disease, Parkinson's cognitive impairment, Alzheimer's disease, vascular dementia, epilepsy, Huntington's disease, heart failure, myocardial infarction, renal failure, renal ischemia, etc. In particular, the in vivo experiments of the present invention also show that some compounds have significant effects on the treatment and / or prevention of stroke, multiple sclerosis, and amyotrophic lateral sclerosis.

[0406] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of equivalent transformations, and these equivalent transformations all belong to the protection scope of the present invention. It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further explain various possible combinations. In addition, the various different embodiments of the present invention can also be arbitrarily combined, as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, as shown below: in: R1 is independently selected from: -NH-heteroarene, -NH-heteroarene diyl-R1', -NH-C(=O)-alkane, -NH-C(=O)-substituted alkane, -NH-C(=O)-alkene, -NH-C(=O)-substituted alkene, -NH-C(=O)-alkyne, -NH-C(=O)-substituted alkyne, -NH-C(=O)-arene, -NH-C(=O)-arene diyl-R1', -NH- C(=O)-heteroarene, -NH-C(=O)-heteroarenediyl-R1', -N(OH)-C(=O)-alkane, -N(OH)-C(=O)-substituted alkane, -N(OH)-C(=O)-alkene, -N(OH)-C(=O)-substituted alkene, -N(OH)-C(=O)-alkyne, -N(OH)-C(=O)-substituted alkyne, -N(OH)-C(=O)-arene, -N( -NH-C(=O)-L-type amino acid-NH-heteroarene, -NH-C(=O)-L-type amino acid-NH-heteroarene, -CH2-N(OH)-C(=O)-alkane, -CH2-N(OH)-C(=O)-substituted alkane hydrocarbon, -CH2-N(OH)-C(=O)-olefin, -CH2-N(OH)-C(=O)-substituted olefin, -CH2-N(OH)-C(=O)-alkyne, -CH2-N(OH)-C(=O)-substituted alkyne, -CH2-N(OH)-C(=O)-heteroarene, -CH2-N(OH)-C(=O)-heteroarene diyl-R1'; and R2: hydrogen or methyl; R3: hydrogen or methyl.

2. The compound according to claim 1, characterized in that The structure is as follows: Wherein: R1 is independently selected from: -NH-heteroarene, -NH-heteroarene diyl-R1', -NH-C(=O)-alkane, -NH-C(=O)-substituted alkane, -NH-C(=O)-alkene, -NH-C(=O)-substituted alkene, -NH-C(=O)-alkyne, -NH-C(=O)-substituted alkyne, -NH-C(=O)-arene, -NH-C(=O)-arene diyl-R1', -NH-C(=O)-alkane ... NH-C(=O)-heteroarene, -NH-C(=O)-heteroarene diyl-R1', -N(OH)-C(=O)-alkane, -N(OH)-C(=O)-substituted alkane, -N(OH)-C(=O)-alkene, -N(OH)-C(=O)-substituted alkene, -N(OH)-C(=O)-alkyne, -N(OH)-C(=O)-substituted alkyne, -N(OH)-C(=O)-arene, -N(OH)-C(=O)-aromatic diyl-R1', -N(OH)-C(=O)-heteroaromatic, -N(OH)-C(=O)-heteroaromatic diyl-R1', -NH-C(=O)-L-type amino acid-NH-heteroaromatic, -NH-C(=O)-L-type amino acid-NH-heteroaromatic diyl-R1', -CH2-N(OH)-C(=O)-alkane, -CH2-N(OH)-C(=O) -substituted alkanes, -CH2-N(OH)-C(=O)-olefins, -CH2-N(OH)-C(=O)-substituted olefins, -CH2-N(OH)-C(=O)-alkynes, -CH2-N(OH)-C(=O)-substituted alkynes, -CH2-N(OH)-C(=O)-heteroarene, -CH2-N(OH)-C(=O)-heteroarenediyl-R1'; and R2: methyl; R3: methyl.

3. The compound according to claim 1 or 2, characterized in that In the aromatic hydrocarbon compound, R1' is independently selected from: -Cl, -F, -Br, -OH, isopropyl, straight-chain / branched alkyl (C≤6), straight-chain / branched alkyl (C≤6) substituted with 1 to 5 halogens, -OH, straight-chain / branched alkyl (C≤6) substituted with 1 to 5 -OHs, straight-chain / branched alkenyl (C≤6), straight-chain / branched alkenyl (C≤6) substituted with 1 to 5 halogens, straight-chain / branched alkenyl (C≤6) substituted with 1 to 5 -OHs, straight-chain / branched alkynyl (C≤6), straight-chain / branched alkynyl (C≤6) substituted with 1 to 5 halogens, straight-chain / branched alkynyl (C≤6) substituted with 1 to 5 -OHs, 4. The compound according to claim 3, characterized in that The aromatic hydrocarbon group is selected from:

5. The compound according to claim 3, characterized in that The substituted alkanes, substituted alkenes, substituted alkynes, alkanes, alkenes, alkynes have a C chain length of ≤6 and are independently selected from: linear, branched or cyclic.

6. The compound according to claim 1, characterized in that The compounds are as follows:

7. Use of the compound according to claim 6 for preparing an NRF2-Leap1 uncoupler.

8. Use of the compound according to claim 6 for preparing a medicament for preventing and / or treating a patient's disease, characterized in that The prepared drugs are used to prevent and / or treat diseases including cerebral small vessel disease, mitochondrial encephalomyopathy, autism spectrum disorder, Rett syndrome, Friedreich's ataxia, stroke, hemorrhagic stroke, ischemic stroke, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, schizophrenia cognitive impairment, Parkinson's disease, Parkinson's cognitive impairment, Alzheimer's disease, vascular dementia, epilepsy, Huntington's disease, heart failure, myocardial infarction, renal failure, and renal ischemia.

9. The use according to claim 8, characterized in that The compound is used in preparing drugs for preventing and / or treating stroke, multiple sclerosis and amyotrophic lateral sclerosis.

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