C17-site nitrogen-substituted and methylene-substituted oleanane triterpene derivative, preparation method therefor and use thereof

Abstract

A C17-site nitrogen-substituted and methylene-substituted oleanane triterpene derivative, a preparation method therefor, and a use thereof. Provided are a C17-site nitrogen-substituted and methylene-substituted oleanane triterpene derivative, a use of the compound in the preparation of an NRF2-Leap1 decoupling agent, and a use of the compound in the preparation of a medicament for preventing and / or treating diseases in a patient, the diseases comprising cerebral small vessel disease, mitochondrial encephalomyopathy, autism spectrum disorder, Rett syndrome, Friedreich ataxia, stroke, hemorrhagic cerebral apoplexy, ischemic cerebral apoplexy, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, cognitive impairment in schizophrenia, Parkinson's disease, cognitive impairment in Parkinson's disease, Alzheimer's disease, vascular dementia, epilepsy, Huntington's disease, heart failure, myocardial infarction, renal failure, kidney ischemia, etc.

Description

C17 nitrogen-substituted and methylene-substituted oleanane triterpene derivatives and their preparation method and use Technical Field

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

[0002] [Corrected 10.03.2025 according to Rule 26] Current research shows 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 with 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 the C17 position, as well as preparation methods and uses thereof.

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

[0005] wherein: R1 is independently selected from: -NH-heteroarene, -NH-heteroarenediyl-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)-arenediyl-R1', -NH-C(=O)-alkyne 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)-arene-diyl-R1', -N(OH)-C(=O)-heteroarene, -N(OH)-C(=O)-heteroarene-diyl-R1', -NH-C(=O)-L-type amino acid-NH-heteroarene, -NH-C(=O)-L-type amino acid-NH-heteroarene-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: hydrogen or methyl; R3: hydrogen or methyl.

[0006] Preferably, the compound is further defined as:

[0007] wherein: R1 is independently selected from: -NH-heteroarene, -NH-heteroarenediyl-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)-arenediyl-R1', -NH-C(=O)- 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(OH)-C(=O)-arene-diyl-R1', -N(OH)-C(=O)-heteroarene, -N(OH)-C(=O)-heteroarene-diyl-R1', -NH-C(=O)-L-type amino acid-NH-heteroarene, -NH-C(=O)-L-type amino acid-NH-heteroarene-diyl-R1', -CH2-N(OH)-C(=O)-alkane, -CH2-N(OH)-C(=O) -substituted alkanes, -CH2-N(OH)-C(=O)-alkenes, -CH2-N(OH)-C(=O)-substituted alkenes, -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.

[0008] 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.

[0009] Preferably, the aromatic hydrocarbon group is selected from:

[0010] wait.

[0011] Preferably, the substituted alkane, substituted olefin, substituted alkyne, alkane, olefin, alkyne carbon 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).

[0012] Preferably, the compound, its pharmaceutically acceptable salt or stereoisomer is as follows:

[0013] The present invention also provides the use of the above-mentioned compounds, their pharmaceutically acceptable salts, and stereoisomers for preparing NRF2-Leap1 uncouplers. Biological experiments of the present invention have shown that many of the above-mentioned compounds exhibit single-digit nanomolar human Nrf2 receptor agonist activity, significantly outperforming the marketed Nrf2 agonist omaveloxolone. Some preferred compounds also exhibit antioxidant activities such as scavenging DPPH free radicals, inhibiting the production of lipid peroxides (MDA), and intervening in ferroptosis.

[0014] The present invention also provides the use of the above-mentioned compounds, pharmaceutically acceptable salts, and stereoisomers thereof for preparing drugs for treating and / or preventing diseases in patients. 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, and renal ischemia. Valid experimental data indicate that the use of drugs prepared from the above-mentioned compounds, pharmaceutically acceptable salts, and stereoisomers thereof is particularly effective in treating stroke, multiple sclerosis, and amyotrophic lateral sclerosis.

[0015] Preferably, the compound, its pharmaceutically acceptable salt, and stereoisomer are used to prepare drugs for preventing or treating stroke, multiple sclerosis, and amyotrophic lateral sclerosis. Beneficial effects:

[0016] The present invention synthesizes for the first time a compound represented by 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 the compound represented by formula (I) through a specific example. Furthermore, the present invention provides the use of the compound represented by 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. In addition to maintaining strong agonist activity on the Nrf2 target, they also have the activity of scavenging DPPH free radicals, inhibiting the formation of lipid peroxides 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, sex, and disease type. The present invention can be used to prevent or treat various 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

[0017] Figure 1. Effects of compound 28 on mNSS scores in rats with ischemic stroke, compared with the sham operation group: ### p<0.001; compared with the model group: *** p<0.001.

[0018] Figure 2. Effect of compound 28 on cerebral infarct area in rats with ischemic stroke, compared with the sham operation group: ### p<0.001; compared with the model group: *** p<0.001.

[0019] Figure 3. Effects of compound 28 on neurological function scores in 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 the Omaveloxolone 15mg / kg group: &&& p<0.001.

[0020] Figure 4. Effect 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 the Omaveloxolone 15mg / kg group: &&& p<0.001.

[0021] Figure 5. Effect of compound 28 on 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 the Omaveloxolone 3mg / kg group: & p<0.05.

[0022] Figure 6. Effects of compound 28 on motor coordination in SOD1 G93A mice compared with the control group: * p<0.05, *** p<0.001; compared with the model group: ### p<0.001; compared with the Omaveloxolone 3mg / kg group: & p<0.05.

[0023] Figure 7. Effects of compound 28 on muscle endurance in SOD1 G93A mice, compared with the control group: * p<0.05, *** p<0.001; compared with the model group: ### p<0.001; compared with the Omaveloxolone 3mg / kg group: & p<0.05. DETAILED DESCRIPTION

[0024] The present invention is further described below with reference to specific embodiments and test examples, but they are not intended to limit the scope of the present invention in any form.

[0025] The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS).

[0026] NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed on an AVANCE III 600 NMR spectrometer using deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3) as the solvents, with tetramethylsilane (TMS) as the internal standard.

[0027] Liquid chromatography-mass spectrometry (LC-MS) was performed using a Shimadzu LCMS2020 mass spectrometer, and HPLC was performed using a Shimadzu LC20A liquid chromatograph.

[0028] 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.

[0029] Unless otherwise specified in the examples, the SFC separation conditions were as follows: column model: DAICEL CHIRALPAK IC (250 mm*30 mm, 10 μm); mobile phase: [CO 2 -i -PrOH / ACN]; B%: 40%, isocratic elution mode.

[0030] In the present invention, if the specific experimental conditions are not indicated, conventional experimental conditions or conditions recommended by the manufacturer shall be followed. If the manufacturer of the reagents or instruments is not indicated, conventional products can be obtained through commercial purchase.

[0031] In the present invention, the test results are expressed as average values.

[0032] The detection indicators in the present invention are: human Nrf2 receptor function test (agonist test); DPPH free radical scavenging ability test; MDA anti-lipid peroxidation ability test.

[0033] ■Example 1

[0034] Compound 1:

[0035] Synthesis route:

[0036] Step 1: Synthesis of Intermediate 3

[0037] Raw material 1' (25 g, 244.78 mmol) was dissolved in anhydrous methanol (400 mL), followed by the addition of intermediate 2 (35.57 g, 269.25 mmol, 30.85 mL) and a 5.4 M sodium methoxide methanol solution (113.32 mL). The mixture was stirred at 70°C for 12 h. After completion of the reaction, concentrated hydrochloric acid was added to the reaction mixture at 0°C to adjust the pH to less than 7, as determined by LCMS. The mixture was then concentrated under reduced pressure to obtain a residue. Purification by column chromatography (silica gel, petroleum ether / ethyl acetate = 1 / 0 to 7 / 3) afforded intermediate 3 (31 g, 70.70% yield, yellow oil). LCMS: rt = 0.100 min, 169.2 [MH] + ; 1H 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).

[0038] Step 2: Synthesis of intermediate 4

[0039] Intermediate 3 (31 g, 182.18 mmol) was dissolved in anhydrous acetonitrile (310 mL). Phosphorus oxychloride (30.73 g, 200.39 mmol, 18.68 mL) was then added under nitrogen and stirred at 65°C for 12 hours. After completion of the reaction, LCMS analysis confirmed that the reaction mixture was quenched by adding room-temperature water (300 mL) at room temperature. After stirring for 1 hour, the mixture was concentrated and filtered to yield Intermediate 4 (21.4 g, 62.28% yield, as a 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).

[0040] Step 3: Synthesis of compound 1

[0041] 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. 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 thin layer chromatography (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 lyophilized to give compound 1 (4.2 mg, white solid, yield <1%). LCMS: rt = 1.502 min, 615.4 [M+H] + Purity: 97.48% 1H 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).

[0042] ■Example 2

[0043] Compound 2:

[0044] Synthesis route:

[0045] Step 1: Synthesis of intermediate 2'

[0046] Raw material 1 (3 g, 6.10 mmol) was dissolved in anhydrous toluene (60 mL). Triethylamine (12.35 g, 122.04 mmol, 16.99 mL) and DPPA (5.04 g, 18.31 mmol, 3.95 mL) were then added at 0°C and stirred at 25°C for 2 h. After completion of the reaction, the reaction solution was concentrated to obtain the crude product. Purification by column chromatography (silica gel, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1) afforded intermediate 2' (2.8 g, 79.93% yield, colorless oil). LCMS: rt = 0.677 min, 517.4 [M+H] + ; Purity 90.74%.

[0047] Step 2: Synthesis of intermediate 3'

[0048] 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 complete, the reaction solution was concentrated to give Intermediate 3' (2.6 g, white solid). LCMS: rt = 0.693 min, 489.3 [M+H] + .

[0049] Step 3: Synthesis of Intermediate 4

[0050] Intermediate 3' (2.9 g, 5.93 mmol) was dissolved in anhydrous acetonitrile (30 mL), and concentrated hydrochloric acid (12 M, 5 mL) was added. The reaction mixture was stirred at 25°C for 0.5 h. After completion of the reaction, 20% sodium hydroxide solution (50 mL) was added to the reaction mixture at 0°C to adjust the pH to >8, followed by extraction with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, and filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography (silica gel column, petroleum ether / ethyl acetate = 1 / 1 to dichloromethane / anhydrous methanol = 10 / 1). Intermediate 4 (2.2 g, 79.32% yield, white solid) was obtained. LCMS: rt = 0.481, 0.749 min, 485.3 [M+Na] + ;Purity 99.18%; 1 H 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).

[0051] Step 4: Synthesis of Intermediate 6

[0052] 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%.

[0053] Step 5: Synthesis of Intermediate 7

[0054] Intermediate 6 (360 mg, 567.95 μmol) was dissolved in anhydrous dichloromethane (4 mL), trifluoroacetic acid (0.4 mL) was added, and the mixture was stirred at 25°C for 3 hours. The reaction solution was concentrated under reduced pressure to give Intermediate 7 (360 mg, yellow oil). LCMS: rt = 0.477 min, 534.3 [M+H] + .

[0055] Step 6: Synthesis of Compound 2

[0056] 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 microwave-reacted 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 18 Column, 0.1% formic acid solution) and lyophilized to give 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(br d,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).

[0057] ■Example 3

[0058] Compound 3:

[0059] Synthesis route:

[0060] ■Example 4

[0061] Compound 4:

[0062] Synthesis route:

[0063] Step 1: Synthesis of Intermediate 2

[0064] Intermediate 1 (150 mg, 1.18 mmol, 1 eq) was dissolved in anhydrous toluene (1.5 mL). Thionyl chloride (702.04 mg, 5.90 mmol, 428.59 μL, 5 eq) was then added and stirred at 70°C for 1 hour. After completion of the reaction as determined by LCMS, the reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product (150 mg) was used directly in the next reaction.

[0065] Step 2: Synthesis of compound 4

[0066] 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 mixture was cooled to 0°C and slowly added with Intermediate 2 (125 mg, 858.85 μmol, 1 eq). The reaction mixture was stirred at 25°C for 1 h. After completion of the reaction, the reaction mixture was concentrated to obtain the crude product. The crude product was purified by preparative chromatography (C18 column, 0.1% formic acid solution) and lyophilized to obtain compound 4 (73.4 mg, off-white solid, 14.85% yield). 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).

[0067] ■Example 5

[0068] Compound 5:

[0069] Synthesis route:

[0070] ■Example 6

[0071] Compound 6:

[0072] Synthesis route:

[0073] Step 1: Synthesis of Intermediate 1

[0074] 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 approximately 84.0% of the product was detected. The mixture was concentrated to give the crude product. The residue was purified by column chromatography (SiO2, 0-10% EtOAc in CH2C l2 The product was purified by TLC (CH2Cl2:EtOH = 10:1, Rf = 0.5) to afford Intermediate 1 (1.40 g, 2.71 mmol, 67.0% yield) as a white solid. LCMS: (EB11687-32-P1A1_LCMS_SH), RT = 1.236 min, MS (ESI) m / z = 517 [M+1]+.

[0075] Step 2: Synthesis of Intermediate 2

[0076] The mixture was added to a toluene solution (104 mL) of intermediate 1 (1.40 g, 2.71 mmol, 1.00 eq) and stirred at 110°C for 2 h. LC-MS (EB11687-34-P1 C1) showed approximately 97.0% detection of the desired compound. The mixture was concentrated to give a crude product. The crude compound 2 (1.20 g, 2.46 mmol, 90.6% yield) was obtained as 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] + .

[0077] Step 3: Synthesis of Intermediate 3

[0078] To a MeCN solution (7.70 mL) of intermediate 2 (1.20 g, 2.46 mmol, 1.00 eq) was added HCl (12 M, 13.00 mL, 58.6 eq) dropwise. After stirring 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 NaHCO 3 (4.60 mL) were added and stirred for 5 min. LC-MS (EB11687-45-P1A1) showed that approximately 96.6% of the desired compound was detected. The organic phase was separated, washed with brine, dried over NaSO 4, and concentrated. The crude intermediate 3 (1.10 g, 2.38 mmol, 93.3% yield) was obtained as a yellow solid. It was used in the next step without further purification. LCMS: (EB11687-45-P1A1_LCMS_SH)RT=1.730min, MS(ESI)m / z=463[M+1] + .

[0079] Step 4: Synthesis of compound 6

[0080] To a solution of intermediate 3 (200 mg, 432 μmol, 1.00 eq) and TME (131 mg, 1.30 mmol, 181 μL, 3.00 eq) in DCM (2 mL) 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) indicated that intermediate 3 had been consumed and the desired MS peak was detected. The mixture was concentrated at 25°C to obtain a residue. The residue was purified by prep-HPLC using Xtimate C18 150*40mm*10μm; mobile phase: [water(NH4HCO3)-ACN]; gradient: 52%-92% B / 25 to afford 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-P1C2). LCMS: EB11739-8-P1A1, RT = 1.681 min, 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=4.788 min, 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). .

[0081] ■Example 7

[0082] Compound 7:

[0083] Synthesis route:

[0084] Step 1: Synthesis of Intermediate 1

[0085] 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 approximately 84.0% of the product was detected. The mixture was concentrated to give the crude product. The residue was purified by column chromatography (SiO2, 0-10% EtOAc in CH2C l2 The product was purified by TLC (CH2Cl2:EtOH = 10:1, Rf = 0.5) to afford Intermediate 1 (1.40 g, 2.71 mmol, 67.0% yield) as a white solid. LCMS: (EB11687-32-P1A1_LCMS_SH), RT = 1.236 min, MS (ESI) m / z = 517 [M+1]+.

[0086] Step 2: Synthesis of Intermediate 2

[0087] The mixture was added to a toluene solution (104 mL) of intermediate 1 (1.40 g, 2.71 mmol, 1.00 eq) and stirred at 110°C for 2 h. LC-MS (EB11687-34-P1 C1) showed approximately 97.0% detection of the desired compound. The mixture was concentrated to give a crude product. The crude compound 2 (1.20 g, 2.46 mmol, 90.6% yield) was obtained as 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] + .

[0088] Step 3: Synthesis of Compound 7: To a THF solution (12.0 mL) of intermediate 2 (650 mg, 1.33 mmol, 1.00 eq) was added pyrazole (453 mg, 6.65 mmol, 5.00 eq). The mixture was stirred at 25°C for 20 h. LC-MS (EB11687-36-P1A1) indicated an approximately 88.0% yield. The crude product was purified by reverse-phase HPLC (column: Welch Xtimate C1840*200 mm 7 μm; mobile phase: [water (NH3H2O ​​+ NH4HCO3)-ACN]; gradient: 58% to 98% B over 25 min). Compound 7 (450 mg, 771 μmol, 58.0% yield, 95.4% purity) 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).δppm 0.94(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).

[0089] ■Example 8

[0090] Compound 8:

[0091] Synthesis route:

[0092] ■Example 9

[0093] Compound 9:

[0094] Synthesis route:

[0095] ■Example 10

[0096] Compound 10:

[0097] Synthesis route:

[0098] Step 1: Synthesis of Intermediate 3

[0099] Material 1 (250 mg, 540.35 μmol, 1 eq) and material 2 (136.31 mg, 540.35 μmol, 1 eq) were dissolved in anhydrous N,N-dimethylformamide (3 mL). HATU (246.55 mg, 648.42 μmol, 1.2 eq) and diisopropylethylamine (209.50 mg, 1.62 mmol, 282.35 μL, 3 eq) were added, and nitrogen was applied. The mixture was reacted at 25°C for 12 hours. After the reaction of the raw materials was completed, the reaction solution was poured into water (10 mL) and extracted with ethyl acetate (10 mL*3). The organic phase was washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by normal phase column chromatography (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.

[0100] Step 2: Synthesis of intermediate 4

[0101] Intermediate 3 (445 mg, 638.53 μmol, 1 eq) was dissolved in anhydrous dichloromethane (5 mL) and trifluoroacetic acid (1.54 g, 13.46 mmol, 1 mL, 21.08 eq) was added and allowed to react at room temperature for 1 hour. After the reaction was complete, as determined by LCMS, the reaction solution was concentrated to afford Intermediate 4 (380 mg, 636.73 μmol, 99.72% yield) as a yellow solid, which was used directly in the next step. LCMS: rt = 0.530 min, 597.4 [M+H] + .

[0102] Step 3: Synthesis of compound 10

[0103] Intermediate 4 (380 mg, 636.73 μmol, 1 eq) was dissolved in anhydrous acetonitrile (5 mL), and ethyl acetoacetate (82.86 mg, 636.73 μmol, 80.61 μL, 1 eq) was added. The mixture was then heated to 60°C for 1 hour. After the reaction was complete, the solvent was partially concentrated and the mixture was lyophilized on a reverse phase basis to afford compound 10 (147.8 mg, 215.17 μmol, 33.79% yield) as a white solid. Prep-HPLC: column: YMC-Actus Triart C18 150*30mm*7um; mobile phase: [water(FA)-ACN]; gradient: 62%-92% B over 10 min. LCMS: rt = 1.593 min, 663.4 [M+H]+. HPLC: 2.274 min. 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).

[0104] ■Example 11

[0105] Compound 11:

[0106] Synthesis route:

[0107] Step 1: Synthesis of Intermediate 3

[0108] To a solution of material 1 (300 mg, 648.42 μmol, 1 eq) in N,N-dimethylformamide (5 mL) were added 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). The reaction mixture was incubated at 20°C for 12 hours. LCMS monitoring confirmed the complete disappearance of the starting material and the main peak was the desired product. The reaction mixture was poured into 30 mL of water and extracted three times with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and mixed. The crude product was purified by normal phase column chromatography (petroleum ether / ethyl acetate = 0-50%) to give 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.

[0109] Step 2: Synthesis of intermediate 4

[0110] To a solution of intermediate 3 (370 mg, 530.91 μmol, 1 eq) in dichloromethane (5 mL) was added trifluoroacetic acid (2.5 mL), and the reaction mixture was allowed to react at 30°C for 1 hour. LCMS analysis indicated the complete disappearance of the starting material and the main peak was the desired product. The reaction mixture was concentrated under reduced pressure to afford 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.

[0111] Step 3: Synthesis of compound 11

[0112] To a solution of intermediate 4 (315 mg, 527.81 μmol, 1 eq) in acetonitrile (4 mL) was added intermediate 5 (68.69 mg, 527.81 μmol, 66.82 μL, 1 eq). The reaction was heated to 60°C for 1 hour. LCMS analysis revealed the complete disappearance of the starting material and the main peak was the desired product. The reaction mixture was filtered and concentrated. The crude product was purified by reverse phase preparative chromatography using water (formic acid)-acetonitrile and lyophilized to afford compound 11 (147.2 mg, 210.08 μmol, 39.80% yield, 94.6% purity). Prep-HPLC: column: Phenomenex luna C18 150*40mm*15um; mobile phase: [water (FA)-ACN]; gradient: 35%-65% B over 22 min. LCMS: Rt = 1.606 min, 663.4 [M+H] + ESI pos. HPLC: Rt = 1.937 min. 1 H 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).

[0113] ■Example 12

[0114] Compound 12:

[0115] Synthesis route:

[0116] ■Example 13

[0117] Compound 13:

[0118] Synthesis route:

[0119] Step 1: Synthesis of Intermediate 2

[0120] To a toluene (4 mL) solution of material 1 (200 mg, 1.29 mmol, 1 eq) was added thionyl chloride (6.55 g, 55.07 mmol, 4 mL, 42.71 eq). The atmosphere was purged with nitrogen and stirred at 70°C for 2 hours. After the reaction was complete as determined by LCMS, the reaction solution was concentrated under reduced pressure to afford compound 2 (200 mg, crude) as a green solid.

[0121] Step 2: Synthesis of compound 13

[0122] To a solution of material 3 (200 mg, 432.28 μmol, 1 eq) in dichloromethane (2 mL) was added triethylamine (349.93 mg, 3.46 mmol, 481.34 μL, 8 eq). After nitrogen substitution three times, a solution of intermediate 2 (150.05 mg, 864.56 μmol, 2 eq) in dichloromethane was slowly added and stirred at 25°C for 1 hour. After the reaction was complete, as determined by LCMS, the reaction solution was concentrated and lyophilized by reverse phase preparative lyophilization to afford 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*30 mm*7 um; mobile phase: [water(FA)-ACN]; gradient: 45%-75% B over 15 min) was performed. 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).

[0123] ■Example 14

[0124] Compound 15:

[0125] ■Synthesis route:

[0126] ■Example 15

[0127] Compound 16:

[0128] Synthesis route:

[0129] ■Example 16

[0130] Compound 17:

[0131] Synthesis route:

[0132] ■Example 17

[0133] Compound 18:

[0134] Synthesis route:

[0135] ■Example 18

[0136] Compound 19:

[0137] Synthesis route:

[0138] Step 1: Synthesis of Intermediate 2

[0139] To a solution of material 1 (50 g, 109.48 mmol, 1 eq) in N,N-dimethylformamide (1 L) were added benzyl bromide (23.88 g, 136.85 mmol, 16.59 mL, 98% purity, 1.25 eq) and potassium carbonate (22.70 g, 164.22 mmol, 1.5 eq). The mixture was stirred at 20°C for 12 hours. TLC (petroleum ether / ethyl acetate = 3 / 1) confirmed complete reaction and the formation of new spots. Water (4 L) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (2 L). The organic phase was washed twice with saturated brine (2 L), dried, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether = 10-30%) to afford intermediate 2 (50 g, 91.44 mmol, 83.52% yield, 90% purity) as a pale 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).

[0140] Step 2: Synthesis of Intermediate 3

[0141] To a solution of Intermediate 2 (50 g, 91.44 mmol, 1 eq) in pyridine (500 mL) was added 4-dimethylaminopyridine (1.12 g, 9.14 mmol, 0.1 eq). The atmosphere was purged with nitrogen three times, then the temperature was lowered to 0°C. Acetic anhydride was slowly added at 0°C, and the mixture was stirred at 20°C for 12 hours. TLC (petroleum ether / ethyl acetate = 3 / 1) revealed the formation of a new spot, but the starting material was not completely reacted. 4-dimethylaminopyridine (1.12 g, 9.14 mmol, 0.1 eq) was added to the reaction solution, the atmosphere was purged with nitrogen three times, the temperature was lowered to 0°C, acetic anhydride was slowly added at 0°C, and the mixture was stirred at 20°C for 24 hours. TLC (petroleum ether / ethyl acetate = 3 / 1) confirmed the complete reaction of the starting material and the concentration of the new spot. The reaction solution was concentrated under reduced pressure to obtain a solid. Water (1 L) was added to the solid, and the solution was extracted three times with ethyl acetate (1 L). The organic phase was dried and concentrated under reduced pressure to give 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).

[0142] Step 3: Synthesis of Intermediate 4

[0143] To a solution of Intermediate 3 (48 g, 81.50 mmol, 1 eq) in dichloromethane (2 L) was added portionwise m-chloroperbenzoic acid (49.65 g, 244.55 mmol, 85% purity, 3 eq). The reaction was stirred at 40°C for 12 hours. TLC (petroleum ether / ethyl acetate = 5 / 1) monitored the complete reaction of the starting material and the formation of new spots. Aqueous sodium sulfite solution was slowly added to the reaction solution at 0°C until the color did not turn blue on potassium iodide paper. Water (2 L) was then added, and the mixture was extracted three times with dichloromethane (1 L). The organic phase was dried and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (ethyl acetate / petroleum ether = 1-5%) to afford Intermediate 4 (50 g, 70.26 mmol, 86.20% yield, 85% purity) 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),1.75-1.43(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).

[0144] Step 4: Synthesis of Intermediate 5

[0145] Intermediate 4 (20 g, 33.07 mmol, 1 eq) was dissolved in acetic acid (100 mL). A solution of hydrobromic acid in acetic acid (3.24 g, 13.23 mmol, 2.18 mL, 33% purity, 0.4 eq) was added dropwise at room temperature. The mixture was then heated to 50°C and bromine (6.34 g, 39.68 mmol, 2.04 mL, 1.2 eq) was slowly added dropwise. After the addition, the mixture was stirred at 50°C for 12 hours. LCMS confirmed the complete reaction of the starting material and the presence of product. The reaction mixture was diluted with water (500 mL) and quenched with 20% aqueous sodium thiosulfate (50 mL). The mixture was then extracted with dichloromethane (200 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether = 0-26%) to afford Intermediate 5 (7.89 g, 11.52 mmol, 34.83% yield) as a yellow solid. LCMS: rt = 0.869 min, 603.3 [M+H] + .

[0146] Step 5: Synthesis of Intermediate 6

[0147] Intermediate 5 (35.6 g, 59.05 mmol, 1 eq) was dissolved in anhydrous methanol (300 mL) and potassium hydroxide (23.19 g, 413.38 mmol, 7 eq) was added. The reaction mixture was heated to 70°C and stirred for 4 hours. LCMS confirmed the complete reaction of the starting material and the presence of product. The reaction mixture was adjusted to pH 2-3 with 6N HCl in an ice-water bath, then diluted with water (400 mL) and filtered. The filter cake was washed with water (300 mL x 3) and then dried under reduced pressure to afford Intermediate 6 (30.9 g, crude) as a yellow solid. LCMS: rt = 0.789 min, 561.4 [M+H] + .

[0148] Step 6: Synthesis of Intermediate 7

[0149] Intermediate 6 (30.9 g, 55.10 mmol, 1 eq) was dissolved in acetonitrile (600 mL) and 2-iodoacylbenzoic acid (30.86 g, 110.20 mmol, 2 eq) was added. The reaction mixture was heated to 80°C and stirred for 1 hour. LCMS confirmed complete reaction and the presence of product. The reaction mixture was filtered, and the filter cake was washed with ethyl acetate (400 mL x 3). The filtrate was concentrated to afford Intermediate 7 (30.79 g, crude) as a yellow solid. LCMS: rt = 0.816 min, 559.4 [M+H] + .

[0150] Step 7: Synthesis of Intermediate 8

[0151] Intermediate 7 (30.79 g, 55.10 mmol, 1 eq) was dissolved in anhydrous dichloromethane (300 mL), and ethyl formate (16.33 g, 220.41 mmol, 17.73 mL, 4 eq) and sodium methoxide (17.86 g, 330.61 mmol, 6 eq) were added sequentially. The reaction mixture was stirred at room temperature for 2 hours. LCMS confirmed the complete reaction of the starting material and the presence of product. The reaction mixture was filtered, and the filter cake was washed with dichloromethane (500 mL x 3). The filtrate was concentrated to obtain the crude product, which was purified by column chromatography (ethyl acetate / petroleum ether = 0-15%) to obtain Intermediate 8 (30.47 g, crude) as a yellow solid. LCMS: rt = 0.873 min, 587.3 [M+H] + .

[0152] Step 8: Synthesis of Intermediate 9

[0153] Intermediate 8 (1 g, 1.70 mmol, 1 eq) was dissolved in ethanol (10 mL) and water (1 mL). Hydroxylamine hydrochloride (1.18 g, 17.04 mmol, 10 eq) was added, and the reaction mixture was heated to 100°C and stirred for 1 hour. LCMS confirmed the complete reaction and the presence of product. The reaction mixture was concentrated, diluted with water (20 mL), and extracted with dichloromethane (20 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product. The crude product was purified by column chromatography (dichloromethane / petroleum ether = 0-88%) to afford Intermediate 9 (580 mg, 993.49 μmol, 58.30% yield) as a white solid. LCMS: rt = 0.822 min, 584.4 [M+H] + . 1 H 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).

[0154] Step 9: Synthesis of Intermediate 10

[0155] Intermediate 9 (20.08 g, 34.40 mmol, 1 eq) was dissolved in anhydrous tetrahydrofuran (300 mL) and wet palladium on carbon (4.09 g, 3.84 mmol, 10% purity, 1.12 e-1 eq) was added. The reaction was stirred at room temperature for 2 hours under a hydrogen balloon. LCMS confirmed the complete reaction and the presence of product. The reaction mixture was filtered, the filter cake was washed with dichloromethane (500 mL x 3), and the filtrate was concentrated to obtain the 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, 49.63% yield) 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).

[0156] Step 10: Synthesis of Intermediate 11

[0157] Intermediate 10 (7 g, 14.18 mmol, 1 eq) was dissolved in acetone (70 mL) and potassium carbonate (5.88 g, 42.54 mmol, 3 eq) and iodomethane (6.04 g, 42.54 mmol, 2.65 mL, 3 eq) were added. The reaction was stirred at room temperature for 12 hours. LCMS confirmed the complete reaction and the presence of product. The reaction mixture was filtered, the filter cake was washed with dichloromethane (100 mL x 3), and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 0-30%) to obtain 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).

[0158] Step 11: Synthesis of Intermediate 12

[0159] 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 complete addition, the mixture was stirred in an ice-water bath for 0.5 hours. The reaction mixture was then slowly warmed to room temperature and stirred for 2 hours. LCMS confirmed the complete reaction of the starting material and the presence of product. The reaction mixture was quenched by the slow addition of water (10 mL) in an ice-water bath. 1N HCl (50 mL) was then added to acidify the mixture and extracted with ethyl acetate (50 mL*4). The organic layers were combined, washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to afford Intermediate 12 (970 mg, crude) as a white solid. LCMS: rt = 0.713 min, 464.2 [M-OH] + .

[0160] Step 12: Synthesis of Intermediate 13

[0161] 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. The reaction was allowed to react at room temperature under nitrogen for 10 min. Tetrapropylammonium perruthenate (70.77 mg, 201.37 μmol, 0.1 eq) was then added to the reaction solution, which was stirred at room temperature for 1.5 hours. LCMS analysis confirmed the complete reaction of the starting material and the presence of product. The reaction mixture was quenched with saturated aqueous sodium carbonate (50 mL) and subsequently extracted with dichloromethane (50 mL x 3). The combined organic layers were washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether = 0-22%) to afford Intermediate 13 (647 mg, 1.19 mmol, 59.19% yield) as a white solid. LCMS: rt = 0.743 min, 478.3 [M+H] + .

[0162] Step 13: Synthesis of Intermediate 14

[0163] Intermediate 13 (300 mg, 628.04 μmol, 1 eq) was dissolved in anhydrous ethanol (3 mL) and water (0.2 mL). 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, and the reaction mixture was stirred at room temperature for 12 hours. LCMS monitoring showed that the reaction of the starting material was complete and the product was present. The reaction mixture was diluted with water (20 mL) and 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 give Intermediate 14 (384 mg, crude) as a white solid. LCMS: rt = 0.846 min, 613.3 [M+H] + .

[0164] Step 14: Synthesis of Intermediate 15

[0165] Intermediate 14 (384 mg, 626.59 μmol, 1 eq) was dissolved in anhydrous ethanol (10 mL). Pyridine borane (174.69 mg, 1.88 mmol, 188.04 μL, 3 eq) and 10% aqueous hydrochloric acid (2.19 g, 6.02 mmol, 2.15 mL, 10% purity, 9.6 eq) were slowly added dropwise under an ice-water bath. After complete addition, the reaction mixture was warmed to room temperature and stirred for 12 hours. LCMS confirmed the presence of product and the presence of starting material. The pH of the reaction mixture was adjusted to alkaline with saturated aqueous sodium bicarbonate solution, followed by extraction with dichloromethane (20 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether = 0-15%) to afford Intermediate 15 (280 mg, 318.77 μmol, 50.87% yield) as a colorless gum. LCMS: rt=0.754min,615.3[M+H] + .

[0166] Step 15: Synthesis of Intermediate 16

[0167] Intermediate 15 (280 mg, 455.39 μmol, 1 eq) was dissolved in anhydrous dichloromethane (4 mL) and then added sequentially with 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). The reaction was stirred at room temperature for 12 hours. LCMS analysis indicated a small amount of starting material remaining and the presence of product. The reaction solution was concentrated to obtain the crude product, which was purified by column chromatography (petroleum ether / ethyl acetate = 0-20%) to afford Intermediate 16 (179 mg, 230.43 μmol, 50.60% yield) as a colorless gum. LCMS: rt = 0.786 min, 707.3 [M+H] + .

[0168] Step 16: Synthesis of Intermediate 17

[0169] Intermediate 16 (179 mg, 253.22 μmol, 1 eq) was dissolved in anhydrous methanol (1 mL) and tetrahydrofuran (1 mL). A 30% sodium methoxide solution in methanol (227.98 mg, 1.27 mmol, 30% purity, 5 eq) was added under an ice-water bath. The reaction mixture was warmed to room temperature and stirred for 1 hour. LCMS analysis indicated a small amount of starting material remaining and the presence of product. The reaction mixture was diluted with 1N aqueous hydrochloric acid (10 mL) and then extracted with dichloromethane (10 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to afford Intermediate 17 (179 mg, crude) as a colorless gum. LCMS: rt = 0.733 min, 729.4 [M+Na] + .

[0170] Step 17: Synthesis of Intermediate 18

[0171] Intermediate 17 (179 mg, 253.22 μmol, 1 eq) was dissolved in anhydrous N,N-dimethylformamide (2 mL). Material 3 (36.20 mg, 126.61 μmol, 0.5 eq) was added under ice-water bath. The reaction mixture was then kept under ice-water bath for 2 hours. Pyridine (80.12 mg, 1.01 mmol, 81.75 μL, 4 eq) was then added, and the reaction mixture was heated to 55°C and stirred for 4 hours. LCMS analysis indicated a small amount of starting material remaining and the presence of product. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether = 0-20%) to afford Intermediate 18 (130 mg, 184.43 μmol, 72.83% yield) as a white solid. LCMS: rt=0.743min,727.2[M+Na] + ..

[0172] Step 18: Synthesis of compound 19

[0173] 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. The reaction temperature was raised to 60°C for 1 hour. LCMS confirmed the complete reaction and the presence of product. The reaction solution was quenched with saturated sodium bicarbonate (10 mL) and extracted with dichloromethane (10 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product. The crude product was then lyophilized by reverse phase preparative to afford 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).

[0174] ■Example 19

[0175] Compound 20:

[0176] Synthesis route:

[0177] ■Example 20

[0178] Compound 21:

[0179] Synthesis route:

[0180] ■Example 21

[0181] Compound 22:

[0182] Synthesis route:

[0183] ■Example 22

[0184] Compound 23:

[0185] Synthesis route:

[0186] Step 1: Synthesis of Intermediate 3

[0187] 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 sequentially. The reaction solution was stirred at room temperature for 2 hours. LCMS analysis confirmed the complete reaction of the starting material and the presence of 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] + . 1 H 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).

[0188] Step 2: Synthesis of intermediate 4

[0189] Intermediate 3 (880 mg, 1.27 mmol, 1 eq) was dissolved in anhydrous methanol (5 mL) and tetrahydrofuran (5 mL). A 30% sodium methoxide solution in methanol (1.15 g, 6.37 mmol, 30% purity, 5 eq) was added under an ice-water bath. The reaction mixture was warmed to room temperature and stirred for 1 hour. LCMS analysis indicated a small amount of starting material remaining and the presence of product. The reaction mixture was diluted with 1N aqueous hydrochloric acid (30 mL) and then extracted with dichloromethane (30 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to afford Intermediate 4 (880 mg, crude) as a white solid. LCMS: rt = 0.759 min, 691.4 [M+H] + .

[0190] Step 3: Synthesis of Intermediate 6

[0191] Intermediate 4 (880 mg, 1.27 mmol, 1 eq) was dissolved in anhydrous N,N-dimethylformamide (10 mL). Material 5 (182.09 mg, 636.85 μmol, 0.5 eq) was added under ice-water bath. The reaction mixture was then kept under ice-water bath for 2 hours. Pyridine (403.00 mg, 5.09 mmol, 411.22 μL, 4 eq) was then added, and the reaction mixture was heated to 55°C and stirred for 4 hours. LCMS confirmed the presence of product and the presence of starting material. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to yield the crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether = 0-19%) to yield Intermediate 6 (796 mg, 1.11 mmol, 87.09% yield) as a white solid. LCMS: rt=0.769min,689.3[M+H] + .

[0192] Step 4: Synthesis of compound 23

[0193] 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. The reaction temperature was raised to 60°C for 2 hours. LCMS confirmed the presence of product and the presence of residual starting material. The reaction solution was slowly added to a stirred saturated aqueous sodium bicarbonate solution (20 mL), followed by extraction with dichloromethane (20 mL x 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (ethyl acetate / petroleum ether = 0-28%) to obtain the crude product, which was then lyophilized by reverse phase preparative lyophilization to afford compound 23 (65.2 mg, 106.62 μmol, 11.48% yield) as a white solid. Prep-HPLC:column:Phenomenex Luna C18 150*25mm*10um;mobile phase:[H2O(0.225%FA)-ACN];gradient:58%-88%B over 15.0min.LCMS:rt=0.678min,599.3[M+H] + .HPLC: rt = 4.015 min. 1 H 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).

[0194] ■Example 23

[0195] Compound 24:

[0196] Synthesis route:

[0197] ■Example 24

[0198] Compound 25:

[0199] Synthesis route:

[0200] ■Example 25

[0201] Compound 26:

[0202] Synthesis route:

[0203] Step 1: Synthesis of Intermediate 3

[0204] To a solution of material 1 (2 g, 3.42 mmol, 1 eq) in dichloromethane (20 mL) was added N,N-diisopropylethylamine (1.33 g, 10.26 mmol, 1.79 mL, 3 eq), followed by a solution of material 2 (402.67 mg, 5.13 mmol, 364.74 μL, 1.5 eq) in dichloromethane (20 mL) at 0°C. The reaction mixture was allowed to react at 20°C for 2 hours. LCMS analysis indicated the complete disappearance of the starting material and the main peak was the desired product. The reaction mixture was concentrated under reduced pressure, and the resulting 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 afford 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.

[0205] Step 2: Synthesis of intermediate 4

[0206] 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. The reaction mixture was allowed to react at 20°C for 1 hour. LCMS analysis indicated the main peak to be the desired product. The reaction mixture was slowly poured into 1N HCl solution to adjust the pH to 3, and extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield intermediate 4 (1.3 g, crude) as a white solid. LCMS: Rt = 0.706 min, 627.5 [M+H] + ESI.

[0207] Step 3: Synthesis of Intermediate 6

[0208] To a 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 mixture was allowed to react at 0°C for 2 hours, followed by the slow addition of pyridine (656.15 mg, 8.30 mmol, 669.54 μL, 4 eq). The reaction mixture was then heated to 55°C and allowed to react for 4 hours. LCMS analysis revealed the main peak to be the desired product. The reaction mixture was poured into water (20 mL) and extracted with ethyl acetate (20 mL x 2). The combined organic phases were 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 afford 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.

[0209] Step 4: Synthesis of compound 26

[0210] To a solution of intermediate 6 (900 mg, 1.44 mmol, 1 eq) in dichloromethane (20 mL) was slowly added boron trichloride solution (1 M, 7.20 mL, 5 eq) at -70°C, and the reaction mixture was allowed to react at 35°C for 4 hours. LCMS monitoring confirmed complete reaction of the starting materials, with the main peak being the desired product. The reaction mixture was slowly poured into saturated aqueous sodium bicarbonate (20 mL) to adjust the pH to 8, and extracted with dichloromethane (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was isolated and purified via reverse phase preparative (water (formic acid)-acetonitrile) to afford compound 26 (96.37 mg, 164.00 μmol, 11.39% yield, 91% purity) as a yellow solid. Prep-HPLC column: Phenomenex luna C18 150*40mm*15um; mobile phase: [H2O (0.225% FA)-ACN]; gradient: 50%-80% B over 15.0min.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).

[0211] ■Example 26

[0212] Compound 27:

[0213] Synthesis route:

[0214] Step 1: Synthesis of Intermediate 3

[0215] 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) were added 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). The reaction mixture was allowed to react at 25°C for 12 hours. LCMS analysis indicated the complete disappearance of the starting material and the main peak was the desired product. The reaction mixture was concentrated under reduced pressure, and the resulting 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 afford 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.

[0216] Step 2: Synthesis of intermediate 4

[0217] 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. The reaction mixture was allowed to react at 25°C for 1.5 hours. LCMS analysis indicated the main peak was the desired product. The reaction mixture was slowly poured into 1N HCl solution to adjust the pH to 3, and extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford 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.

[0218] Step 3: Synthesis of Intermediate 6

[0219] To a 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 mixture was allowed to react at 0°C for 2 hours, followed by the slow addition of pyridine (878.76 mg, 11.11 mmol, 896.69 μL, 4 eq). The reaction mixture was then heated to 55°C and allowed to react for 4 hours. LCMS analysis revealed the main peak to be the desired product. The reaction mixture was poured into water (20 mL) and extracted with ethyl acetate (20 mL x 2). The combined organic phases were 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 afford 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.

[0220] Step 4: Synthesis of compound 27

[0221] To a solution of intermediate 6 (1 g, 1.57 mmol, 1 eq) in dichloromethane (20 mL) was slowly added boron trichloride solution (1 M, 7.83 mL, 5 eq) at -70°C. The reaction mixture was allowed to react at 30°C for 12 hours. LCMS analysis indicated residual starting material and the main peak was the desired product. The reaction mixture was slowly poured into saturated aqueous sodium bicarbonate (20 mL) to adjust the pH to 8 and extracted with dichloromethane (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was purified by normal-phase silica gel column chromatography (petroleum ether:ethyl acetate = 1:0 to 1:1) to afford the crude product. This crude product was then isolated and purified by reverse-phase preparative chromatography (water (formic acid)-acetonitrile) to afford compound 27 (113.80 mg, 199.29 μmol, 12.73% yield, 96.10% purity) as a white solid. Prep-HPLC:column:Phenomenex Luna C18 150*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).

[0222] ■Example 27

[0223] Compound 28:

[0224] Synthesis route:

[0225] Step 1: Synthesis of Intermediate 3

[0226] To a solution of material 1 (1 g, 1.71 mmol, 1 eq) in dichloromethane (20 mL) were added 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). The reaction mixture was allowed to react at 20°C for 2 hours. LCMS analysis indicated the complete disappearance of the starting material and the main peak was the desired product. The reaction mixture was concentrated under reduced pressure, and the resulting 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 afford 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.

[0227] Step 2: Synthesis of intermediate 4

[0228] 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. The reaction mixture was allowed to react at 20°C for 1 hour. LCMS analysis indicated the main peak was the desired product. The reaction mixture was slowly poured into 1N HCl solution to adjust the pH to 3 and extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield Intermediate 4 (740 mg, crude) as a white solid. LCMS: Rt = 0.738 min, 653.5 [M+H] + ESI.

[0229] Step 3: Synthesis of Intermediate 6

[0230] To a 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 mixture was allowed to react at 0°C for 2 hours. Pyridine (358.61 mg, 4.53 mmol, 365.92 μL, 4 eq) was then slowly added. The reaction mixture was heated to 55°C and reacted for 4 hours. LCMS analysis indicated the main peak was the desired product. The reaction mixture was poured into water (20 mL) and extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting 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 afford 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.

[0231] Step 4: Synthesis of compound 28

[0232] To a solution of intermediate 6 (400 mg, 614.55 μmol, 1 eq) in dichloromethane (8 mL) was slowly added boron trichloride solution (1 M, 3.07 mL, 5 eq) at -70°C. The reaction mixture was allowed to react at 35°C for 2 hours. LCMS analysis confirmed complete reaction of the starting materials, with the main peak being the desired product. The reaction mixture was slowly poured into saturated aqueous sodium bicarbonate (20 mL) to adjust the pH to 8, and extracted with dichloromethane (20 mL x 2). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was isolated and purified via reverse phase preparative (water (formic acid)-acetonitrile) to afford compound 28 (48.49 mg, 84.41 μmol, 13.74% yield, 97.618% purity) as a yellow solid. Prep-HPLC column: Phenomenex luna C18 150*40mm*15um; mobile phase: [H2O(0.225%FA)-ACN]; gradient:55%-85%B over 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).

[0233] ■Example 28

[0234] Compound 29:

[0235] Synthesis route:

[0236] Step 1: Synthesis of Intermediate 3

[0237] To a solution of material 1 (1.9 g, 3.25 mmol, 1 eq) in dichloromethane (50 mL) were added 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). The mixture was stirred at 25°C for 12 hours. LCMS analysis indicated that the reaction was complete. 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 afford 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] + .

[0238] Step 2: Synthesis of intermediate 4

[0239] To a solution of intermediate 3 (1.6 g, 2.44 mmol, 1 eq) in methanol (20 mL) was added sodium methoxide (2.20 g, 12.22 mmol, 30% purity, 5 eq) at 0°C and stirred at 25°C under a nitrogen atmosphere for 2 hours. LCMS confirmed the complete reaction. The reaction solution was adjusted to pH 3-4 with 1N hydrochloric acid, extracted with dichloromethane, washed, dried, and concentrated under reduced pressure to afford intermediate 4 (1.5 g, crude product) as a white solid. LCMS: rt = 0.757 min, 655.3 [M+H] + ..

[0240] Step 3: Synthesis of Intermediate 6

[0241] To a solution of intermediate 4 (820 mg, 3.01 mmol, 1 eq) in N,N-dimethylformamide (15 mL) was added material 5 (327.43 mg, 1.15 mmol, 0.5 eq) at 0°C and stirred at 0°C for 2 hours under a nitrogen atmosphere. Pyridine (724.67 mg, 9.16 mmol, 739.46 μL, 4 eq) was added to the reaction solution and stirred at 55°C for 4 hours. LCMS monitoring showed that the reaction of the starting material was complete. The reaction solution was poured into water under reduced pressure 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] + ..

[0242] Step 4: Synthesis of compound 29

[0243] 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 mixture was allowed to react at 35°C for 4 hours. LCMS confirmed the complete reaction. The reaction mixture was poured into saturated sodium bicarbonate solution, extracted with dichloromethane, washed, dried, and concentrated under reduced pressure to obtain the crude product. The crude product was isolated and purified by reverse phase preparative reaction (water (formic acid)-acetonitrile system) and lyophilized to obtain compound 29 (150.65 mg, 267.69 μmol, 17.48% yield) as a white solid. Prep-HPLC:column:Phenomenex luna C18 150*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).

[0244] ■Example 29

[0245] Biological evaluation

[0246] Test Example 1 In vitro human Nrf2 receptor agonist activity assay

[0247] Reagents: Opti-MEM serum-free medium; PEI 40000 transfection reagent; DMEM complete medium.

[0248] Equipment: Cx7Pro high-content rapid imaging platform, etc.

[0249] For each well of cells, dilute 360 ​​ng of Nrf2 phase change probe plasmid with 10 μL of Opti-MEM serum-free medium (Cienry), mix thoroughly to form a DNA dilution solution, and let it stand for 5 minutes. Then, dilute 0.6 μL of PEI 40000 transfection reagent (YEASEN) with 10 μL of Opti-MEM serum-free medium, mix thoroughly to form a PEI 40000 dilution solution, and let it stand for 5 minutes. After mixing the DNA dilution solution and PEI 40000 dilution solution, incubate at room temperature for 20 minutes to allow the formation of the DNA-PEI cationic nucleic acid transfection reagent complex. Remove 20 μL of cell growth medium and add 20 μL of the DNA-PEI cationic nucleic acid transfection reagent complex to each well. Shake the culture plate gently to mix. Cells were cultured in a 37°C, 5% CO2 incubator. One hour after transfection, 75 μL of cell growth medium was removed and 75 μL of fresh, prewarmed DMEM (Meilunbio) was added to each well, maintaining a medium volume of 100 μL per well. Eighteen hours after transfection, serially diluted drug (0.0625 nM, 0.125 nM, 0.25 nM, 0.5 nM, 1 nM, 2 nM, 4 nM, 8 nM, 16 nM, 32 nM, 128 nM) was first mixed with prewarmed DMEM. 100 μL of this serially diluted drug medium was then added to each well, bringing the final volume to 200 μL. Six hours after drug addition, 16 randomly selected fields of view per well were imaged using a Cx7Pro high-content rapid imaging platform (Thermo Fisher) at a 20x magnification lens. The number of transfected cells was determined by determining the location of the cell nucleus using the independently expressed NLS-mTagBFP2 in the probe. Calculate the total fluorescence intensity of the Nrf2 phase transition probe phase transition. Count the total fluorescence intensity of the Nrf2 phase transition probe phase transition "droplets" in each cell. Compare the total fluorescence intensity of the Nrf2 phase transition probe phase transition in each cell in the drug group with the total fluorescence intensity of the Nrf2 phase transition 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:

[0250] Table 1 Test results of representative compounds of the present invention on the agonist activity of human Nrf2 receptor in vitro

[0251] 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.

[0252] Test Example 2 DPPH free radical scavenging ability test

[0253] Purpose of the experiment: To determine the DPPH free radical scavenging ability of the compounds of the present invention.

[0254] Test materials:

[0255] Test equipment:

[0256] Test method:

[0257] First, add 20 μL of the test compound to a 96-well plate and serially dilute it 1:2 with DMSO. Then, add 200 μL of 200 μM DPPH prepared in anhydrous ethanol to each well. After gentle shaking, incubate at room temperature in the dark for 30 minutes. Measure the absorbance at 517 nm using a microplate reader. Calculate the DPPH clearance using the following formula: DPPH clearance (%) = (1-Ai / A0) * 100%, where Ai refers to the sample absorbance and A0 refers to the DMSO control absorbance. Data were processed using XLfit 5.3.1.3 software, and the IC50 values ​​of the compounds were calculated using a nonlinear fitting formula. The results are shown in Table 2.

[0258] Table 2 DPPH radical scavenging ability test results of representative compounds of the present invention

[0259] The results showed that compounds 27 and 28 prepared in the present invention had the ability to scavenge DPPH free radicals, while Omaveloxolone had no such activity.

[0260] Test Example 3 MDA Anti-lipid Peroxidation Ability Test

[0261] Experimental purpose: To test the MDA anti-lipid peroxidation ability of the compounds of the present invention.

[0262] Experimental Materials:

[0263] Test equipment:

[0264] Test method:

[0265] First, brain tissue homogenate was prepared: an adult male Sprague-Dawley rat was anesthetized with isoflurane and sacrificed by cervical dislocation. The whole brain was removed and washed twice in DPBS. The meninges were stripped and transferred to a 50-mL centrifuge tube containing 10 mL of DPBS. The brain was minced with scissors and divided into ten 1.5-mL centrifuge tubes. Three grinding beads were added to each tube and the mixture was ground at 90 Hz for 60 minutes three times. The ground tissue homogenate was transferred to a fresh 50-mL centrifuge tube, DPBS was added to a total volume of 30 mL, and the mixture was mixed. Next, 20 μL of the test compound was added to a 96-well plate and serially diluted 1:3 in DMSO. Then, 100 μL of brain tissue homogenate, 50 μL of DPBS, and 50 μL of 200 μg / mL vitamin C were added, along with a series of concentration standards as a standard curve. After shaking, the mixture was incubated at 37°C for 1 hour. Then, 400 μL of MDA working solution was added and the mixture was heated at 100°C 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 a microplate reader. The MDA clearance rate of the compound was calculated by 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 low control group absorbance value, and A0 refers to the blank group absorbance value. The data were processed using XLfit5.3.1.3 software, and the IC value of the compound was obtained using a nonlinear fitting formula. 50 The results are shown in Table 3.

[0266] Table 3 MDA anti-lipid peroxidation ability test results of the compounds of the present invention

[0267] The results showed that compounds 23, 26, 27, 28, and 29 prepared in the present invention all had anti-lipid peroxidation activity, and their activity was better than that of edaravone, while omaveloxolone had no such activity.

[0268] Experimental Example 4: Intervention Effect of Representative Compounds on Ferroptosis (qPCR)

[0269] 1) Experimental Objective: To examine the effects of a series of compounds on ferroptosis by measuring changes in the expression of key genes in the ferroptosis signaling pathway at the cellular level. The specific indicator is the mRNA level of PTGS2, a key gene in ferroptosis (fluorescence real-time quantitative RT-PCR).

[0270] 2) Experimental methods:

[0271] HT-1080 cells (Cat. No. CCL-121, Shanghai Cell Bank, purchased from ATCC) were plated in six-well plates at 4 × 10⁵ / well for 22 hours. HT-1080 cells were treated with a gradient of compound concentrations (1.37 nM to 333 nM, a total of six concentrations) for 1 hour beforehand. The classic ferroptosis inducer RSL3 (Cat. No. HY-100218A, MCE, USA) was then treated with 200 nM for 16 hours. mRNA was extracted and reverse-transcribed into cDNA, and PTGS2 mRNA levels were measured by qRT-PCR (ABI7500, Thermo Fisher Scientific, USA). The ferroptosis inhibitor Fer-1 (Cat. No. HY-100579, MCE, USA) was used as a positive control.

[0272] 3) Experimental results:

[0273] The IC value of each drug was calculated using the log(agonist) vs. response--Variable slope (four parameters) analysis method in GraphPadPrism. 50 , the results are shown in Table 4.

[0274] Table 4 Interventional effects of the compounds of the present invention on the ferroptosis process

[0275] The results showed that compounds 26, 27, 28 and 29 prepared in the present invention can inhibit ferroptosis, while Omaveloxolone has no such activity.

[0276] The above-mentioned pharmacological experiments demonstrate that the preferred NRF2-Keap1 compounds prepared by the present invention, such as 26, 27, 28, 29, etc., not only maintain Nrf2 agonist activity similar to that of the marketed Nrf2 agonist Omaveloxolone, but also increase the effects of scavenging DPPH free radicals, inhibiting the production of lipid peroxides MDA, or intervening in ferroptosis.

[0277] 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, and renal ischemia. In particular, the compounds have significant effects on diseases such as stroke, multiple sclerosis, and amyotrophic lateral sclerosis, and can be used to prepare drugs for the prevention or treatment of stroke, multiple sclerosis, and amyotrophic lateral sclerosis.

[0278] Experimental Example 5: Protective Effects of Representative Compounds on Ischemic Stroke Rats

[0279] 1) Reagents: Omaveloxolone, MedChemexpress Biotechnology, USA; Transient Middle Cerebral Artery Occlusion (tMCAO) suture, Beijing Reward Life Sciences Co., Ltd.; DMAO, Beyotime Biotechnology Co., Ltd.; Solutal, Sigma-Aldrich (Shanghai) Trading Co., Ltd.; Normal saline, Sinopharm Chemical Reagent Co., Ltd.; TTC, Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0280] 2) Establishment of cerebral ischemia rat model using tMCAO method:

[0281] Rats were anesthetized with 10% chloral hydrate (350 mg / kg), fixed in the supine position on the operating table, and the neck was disinfected. A midline incision was made in the neck, and the intermuscular space between the left sternocleidomastoid and sternoglossi muscles was bluntly dissected to expose the common carotid artery. The common carotid artery was extracted with ophthalmic curved forceps and threaded with silk suture for later use. The external carotid artery was isolated between the right digastric muscle and the hyoid bone, and threaded with silk suture for later use. Suture was passed through and tied to the common carotid artery. A small incision was made at the free end of the common carotid artery and a loose knot was tied. A suture was inserted through the incision along the common carotid artery. The suture was slowly advanced until resistance was felt and the pre-tied loose knot was tightened. The body temperature was maintained at 37°C during the operation, and the suture was removed after 1.5 hours. Rats in the sham operation group did not have a suture inserted after vascular isolation, and the other procedures were the same.

[0282] 3) Experimental groups and drug administration:

[0283] Forty male Sprague-Dawley rats, weighing 230-250g, were randomly divided into a sham-operated group, a model group, an omaveloxolone 3mg / kg group, an omaveloxolone 9mg / kg group, and a compound 283.03mg / kg group. After three days of acclimatization, the rats were injected into the tail vein during modeling. Rats in the sham-operated and model groups received an equal volume of vehicle (10% DMSO + 10% solutal + 80% saline).

[0284] 4) Index detection:

[0285] mNSS score: Neurobehavioral scoring was performed 24 hours after model establishment, mainly to evaluate the neurological function of rats with ischemic stroke from the perspectives of sensation and movement.

[0286] Measurement of cerebral infarct area: Rats were euthanized with an overdose of chloral hydrate. The brains were removed and placed in a -20°C refrigerator for 20 minutes. The brains were then placed in the cerebral trough and sliced ​​to a thickness of 2 mm. The slices were then incubated in a 37°C incubator in a 2% TTC solution, shielded from light, for 15 minutes. After staining, images were taken and analyzed for cerebral infarct area.

[0287] 5) Statistical methods:

[0288] All data in this study are expressed as mean ± standard deviation (mean ± SD) and statistically analyzed using GraphPad Prism 7.0. Differences between groups were assessed using one-way ANOVAs followed by Tukey's test. Behavioral tests were evaluated using the Krystal-Wallis test. Data were considered statistically significant when P < 0.05.

[0289] 6) Experimental results:

[0290] Effects of compound 28 on neurological function in rats with ischemic stroke:

[0291] As shown in Figure 1, Omaveloxolone 3 mg / kg and 9 mg / kg had no effect on the mNSS score of rats with cerebral ischemia. At an equimolar dose to Omaveloxolone 3 mg / kg, Compound 28 (3.03 mg / kg) significantly reduced the mNSS score of rats with cerebral ischemia.

[0292] Effects of compound 28 on cerebral infarction area in rats with ischemic stroke:

[0293] As shown in Figure 2, Omaveloxolone 3 mg / kg and 9 mg / kg had no effect on the cerebral infarction area in rats with cerebral ischemia. At an equimolar dose to Omaveloxolone 3 mg / kg, Compound 28 (3.03 mg / kg) significantly reduced the cerebral infarction area in rats with cerebral ischemia.

[0294] 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.

[0295] Effects of the representative compounds of Experimental Example 6 on the neurobehavior of multiple sclerosis model mice

[0296] 1. Materials and Methods

[0297] 1) Main reagents

[0298] 2) Experimental animals and grouping and drug administration

[0299] Female C57BL / 6J mice were randomly divided into five 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.

[0300] 3) Preparation of Multiple Sclerosis Model - EAE Model

[0301] The EAE model is a commonly used animal model for multiple sclerosis. An EAE (experimental autoimmune encephalosporin) model is established by subcutaneously injecting a mixture of a peptide fragment of myelin oligodendrocyte glycoprotein MOG35-55 and complete Freund's adjuvant (CFA) containing Mycobacterium tuberculosis into the back of mice. Pertussis toxin (PTX) is then injected intraperitoneally on the day of immunization and 48 hours later.

[0302] 4) Weight and neurological function score (5-point scale)

[0303] Starting from the day of immunization induction (Day 0), the rats were weighed and neurological function was scored daily. Neurological function scoring criteria (5-point scale): 0, no clinical deficit; 1, partial tail paralysis; 2, complete tail paralysis; 3, partial hindlimb paralysis; 4, complete hindlimb paralysis; 5, forelimb paralysis; 6, death.

[0304] 5) Statistical processing

[0305] All data are presented as mean ± standard error (SEM). Two-way analysis of variance and Tukey's multiple comparison test were used to compare 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 statistical significance.

[0306] 2. Experimental Results

[0307] As shown in Figure 3, omaveloxolone at 5 and 15 mg / kg significantly reduced neurological function scores in EAE model mice, but no significant difference was observed between the two groups, indicating that omaveloxolone 5 mg / kg achieved its maximum effect. At an equimolar dose to omaveloxolone 5 mg / kg, compound 28 (5.05 mg / kg) significantly reduced neurological function scores in EAE model mice, with a greater potency than omaveloxolone 15 mg / kg, achieving statistically significant differences.

[0308] As shown in Figure 4, omaveloxolone at 5 and 15 mg / kg significantly increased the body weight of EAE model mice, but no significant difference was observed between the two groups, indicating that omaveloxolone 5 mg / kg achieved its maximum effect. At an equimolar dose to omaveloxolone 5 mg / kg, compound 28 (5.05 mg / kg) significantly reduced neurological function scores in EAE model mice, with a greater potency than omaveloxolone 15 mg / kg, achieving statistically significant differences.

[0309] 3. Experimental Conclusion

[0310] Compound 28 has a protective effect on the neurological function of multiple sclerosis model mice, and its effect is stronger than that of Omaveloxolone.

[0311] Protective effect of the representative compound of Experimental Example 6 on amyotrophic lateral sclerosis model mice

[0312] 1. Materials and Methods

[0313] 1) Main reagents

[0314] Omaveloxolone was purchased from MCE Biotechnology Co., Ltd.; Solutal was purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.; and normal saline was purchased from Sinopharm Chemical Reagent Co., Ltd.

[0315] 2) Animals

[0316] 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.

[0317] 3) Experimental grouping and drug administration

[0318] SOD1 G93A mice were randomly divided into a model group, an omaveloxolone 1 mg / kg group, an omaveloxolone 3 mg / kg group, and a compound 28 1.01 mg / kg group, with 12 mice in each group. Twelve C57BL / 6J mice were also assigned to the control group. The control and model groups were intraperitoneally injected with an equal volume of vehicle (1% DMSO + 4% solutal + 95% saline) once daily for 10 weeks.

[0319] 4) Index detection

[0320] Rotarod test

[0321] The rotarod test is a classic behavioral test for evaluating motor coordination in mice. The rotarod test (Panlab rotarod apparatus, purchased from Harvard Bioscience, USA) was performed twice weekly, with the rotation speed ranging from 4 to 40 rpm. The experimental procedures were as follows: 1. Before the formal experiment, the mice were acclimated to the rotation speed at 12 rpm for 5 minutes twice daily for 3 days. 2. During the formal experiment, three consecutive tests were performed (3 minutes each, with a 30-minute interval between each test). 3. The time the mouse remained on the rotarod was recorded each time, and the longest time spent on the rotarod across the three tests was used as the latency to fall.

[0322] Onset time

[0323] 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.

[0324] Cage experiment

[0325] The hanging cage test assesses the grip strength and endurance of mice's limbs. This test is performed twice weekly. Each mouse is placed in the center of a wire mesh. The mesh is gently shaken to ensure the mouse's grip is firm. The mesh is then slowly inverted to a horizontal position, and the time the mouse remains suspended is recorded. Each mouse is tested three times, with each test separated by 30 minutes. The maximum value is used as the fall latency.

[0326] 2. Experimental results

[0327] 1) Effect of compound 28 on the onset time of SOD1 G93A mice

[0328] As shown in Figure 5, both 1 mg / kg and 3 mg / kg of omaveloxolone significantly delayed the onset of SOD1 G93A mice, but no significant difference was observed between the two groups, indicating that omaveloxolone 1 mg / kg achieved its maximum effect. At an equimolar dose to omaveloxolone 1 mg / kg, compound 28 (1.01 mg / kg) significantly delayed the onset of SOD1 G93A mice, and its efficacy was superior to that of omaveloxolone 3 mg / kg, achieving statistically significant results.

[0329] 2) Effect of compound 28 on motor coordination ability of SOD1 G93A mice

[0330] As shown in Figure 6, both 1mg / kg and 3mg / kg of omaveloxolone significantly improved the motor coordination of SOD1 G93A mice, as evidenced by a significant increase in fall latency. However, no significant difference was observed between the two groups, indicating that omaveloxolone 1mg / kg achieved its maximum effect. At an equimolar dose to omaveloxolone 1mg / kg, compound 28 (1.01mg / kg) significantly increased the fall latency of mice, with a greater potency than omaveloxolone 3mg / kg, achieving statistically significant results.

[0331] 3) Effect of compound 28 on muscle endurance in SOD1 G93A mice

[0332] As shown in Figure 7, both 1mg / kg and 3mg / kg of omaveloxolone significantly improved the motor coordination of SOD1 G93A mice, as evidenced by a significant increase in fall latency. However, no significant difference was observed between the two groups, indicating that omaveloxolone 1mg / kg achieved its maximum effect. At an equimolar dose to omaveloxolone 1mg / kg, compound 28 (1.01mg / kg) significantly increased the fall latency of mice, with a greater potency than omaveloxolone 3mg / kg, achieving statistically significant results.

[0333] 3. Experimental Conclusion

[0334] Compound 28 can improve the neurobehavior of ALS model mice, and its effect is stronger than Omaveloxolone.

[0335] 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 type of Nrf2-Keap1 uncoupler, with excellent Nrf2 agonist effect, and at the same time 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 experiments of the present invention show that the compounds of the present invention can be effectively 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. In particular, the in vivo experiments of the present invention also show that some of the compounds have significant effects in treating and / or preventing stroke, multiple sclerosis, and amyotrophic lateral sclerosis.

[0336] [Corrected 19.02.2025 according to Rule 26] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, the technical solutions of the present invention can be subjected to a variety of equivalent transformations, and these equivalent transformations all fall within the scope of protection 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 unless there is any contradiction. In order to avoid unnecessary repetition, the present invention will not further describe 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. [Part of the accompanying drawings moved to the correct position]

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-heteroarenediyl-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)-arenediyl-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-R1', -CH2-N(OH)-C(=O)-alkane, -CH2-N(OH)-C(=O)-substituted alkane hydrocarbon, -CH2-N(OH)-C(=O)-alkene, -CH2-N(OH)-C(=O)-substituted alkene, -CH2-N(OH)-C(=O)-alkyne, -CH2-N(OH)-C(=O)-substituted alkyne, -CH2-N(OH)-C(=O)-heteroarene, -CH2-N(OH)-C(=O)-heteroarenediyl-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-heteroarenediyl-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)-arenediyl-R1', -NH-C(=O)- 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(OH)-C(=O)-arene-diyl-R1', -N(OH)-C(=O)-heteroarene, -N(OH)-C(=O)-heteroarene-diyl-R1', -NH-C(=O)-L-type amino acid-NH-heteroarene, -NH-C(=O)-L-type amino acid-NH-heteroarene-diyl-R1', -CH2-N(OH)-C(=O)-alkane, -CH2-N(OH)-C(=O) -substituted alkanes, -CH2-N(OH)-C(=O)-alkenes, -CH2-N(OH)-C(=O)-substituted alkenes, -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, and 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 in preparing an NRF2-Leap1 uncoupling agent.

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 medicines for preventing and / or treating stroke, multiple sclerosis and amyotrophic lateral sclerosis.

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