C-17 carbonyl substituted oleanane triterpenoid derivative as well as preparation method and application thereof

Abstract

The invention discloses a C-17 carbonyl substituted oleanane triterpenoid derivative as well as a preparation method and application thereof. The invention provides a C-17 carbonyl substituted oleanane triterpenoid derivative and a preparation method thereof. The compound provided by the invention is used for preparing an NRF2 activator. The medicine for treating / preventing diseases is prepared; diseases include small vascular diseases, mitochondrial brain myopathy, autism spectrum disorder, Rett syndrome, Friedel's ataxia, cerebral apoplexy, hemorrhagic cerebral apoplexy, ischemic cerebral apoplexy, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, cognitive impairment of schizophrenia, Parkinson's disease, cognitive impairment of Parkinson's disease, Alzheimer's disease, vascular dementia, and epilepsy. The present invention relates to compounds of formula (I), such as Huntington's disease, heart failure, myocardial infarction, renal failure, renal ischemia, or the like, or other disease states and conditions that are apparent to those skilled in the art, or prodrugs thereof, or pharmaceutically acceptable salts thereof, or hydrates or solvates thereof, and pharmaceutical compositions containing the same.

Description

Technical Field

[0001] The present invention generally relates to the field of biology and pharmaceutical technology, and specifically relates to C-17 carbonyl-substituted oleanane triterpene derivatives and preparation methods and uses thereof, and more specifically relates to compounds of formula (I) and their use in the preparation of drugs for preventing or treating diseases mediated by the NRF2-Leap1 system. Specifically, it relates to compounds represented by the general formula, their tautomers or pharmaceutically acceptable compositions thereof. Background Art

[0002] A large number of studies have found that oxidative stress can directly or indirectly induce the occurrence and development of a variety of diseases, so in-depth research on oxidative stress is particularly important. Oxidative stress is caused by a serious imbalance between the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) and the antioxidant defense system, enzymatic and non-enzymatic. In order to resist these injuries, the body has formed a complex oxidative stress response system to alleviate the damage to cells and maintain homeostasis by upregulating cytoprotective factors.

[0003] Nrf2 belongs to the leucine (CNC) regulatory protein family, including Neh1 to Neh6. It is an important transcription factor for the body to regulate the anti-oxidative stress response. Keap1 is the specific receptor of Nrf2. Under normal conditions, Nrf2 and Keap1 recognize and form a polymer, and its action is inhibited by Keap1. Under conditions such as oxidative stress, Nrf2 dissociates from Keap1 and is activated, activating the antioxidant response element (ARE), regulating the expression of phase II detoxification enzyme genes and antioxidant enzyme genes, and enhancing the cell's antibodies to substances such as oxidative stress and electrophilic biomacromolecules. Considering that the current molecules only focus on inflammatory treatment through Nrf2 agonism, they have the characteristics of poor inhibitory activity and poor specificity. Therefore, it is particularly important to develop new NRF2 activators that have both anti-inflammatory and antioxidant effects. Summary of the invention

[0004] In order to solve the above technical problems, the present invention provides a C-17 carbonyl substituted oleanane triterpene derivative and a preparation method and use thereof.

[0005] A compound represented by formula (I) or a pharmaceutically acceptable salt thereof:

[0006]

[0007] in:

[0008] R 1independently selected from: -C(=O)-alkane, -C(=O)-substituted alkane, -C(=O)-alkene, -C(=O)-substituted alkene, -C(=O)-alkyne, -C(=O)-substituted alkyne, -C(=O)-heteroarene, -C(=O)-heteroarene-R 1 ', -C(=O)-heteroarene diyl-R 1 '-C(=O)-NH-OH, -C(=O)-N(OH)-alkane, -C(=O)-N(OH)-substituted alkane, -C(=O)-N(OH)-alkene, -C(=O)-N(OH)-substituted alkene, -C(=O)-N(OH)-alkyne, -C(=O)-N(OH)-substituted alkyne, -C(=O)-N(OH)-arene, -C(=O)-N(OH)-arenediyl-R 1 ', -C(=O)-N(OH)-heteroarene, -C(=O)-N(OH)-heteroarene diyl-R 1 ', -C(=O)-NH-arene, -C(=O)-NH-arenediyl-R 1 ', -C(=O)-NH-heteroarene, -C(=O)-NH-heteroarene diyl-R 1 ', -C(=O)-O-arene, -C(=O)-O-arenediyl-R 1 ', -C(=O)-O-heteroarene, -C(=O)-O-heteroarene diyl-R 1 ', -C(=O)-CH 2 -heteroarene, -C(=O)-CH 2 -heteroarene diyl-R 1 ', -C(=O)-CR 2 'R 3 '-heteroarene, -C(=O)-CR 2 'R 3 '-heteroarene diyl-R 1 ', -C(=O)-L-amino acid-NH-heteroarene, -C(=O)-L-amino acid-NH-heteroarene diyl-R 1 ';

[0009] And R 2 : hydrogen or methyl; R 3 : Hydrogen or methyl.

[0010] The compound is further defined as:

[0011]

[0012] in:

[0013] R 1independently selected from: -C(=O)-alkane, -C(=O)-substituted alkane, -C(=O)-alkene, -C(=O)-substituted alkene, -C(=O)-alkyne, -C(=O)-substituted alkyne, -C(=O)-heteroarene, -C(=O)-heteroarene-R 1 ', -C(=O)-heteroarene diyl-R 1 '-C(=O)-NH-OH, -C(=O)-N(OH)-alkane, -C(=O)-N(OH)-substituted alkane, -C(=O)-N(OH)-alkene, -C(=O)-N(OH)-substituted alkene, -C(=O)-N(OH)-alkyne, -C(=O)-N(OH)-substituted alkyne, -C(=O)-N(OH)-arene, -C(=O)-N(OH)-arenediyl-R 1 ', -C(=O)-N(OH)-heteroarene, -C(=O)-N(OH)-heteroarene diyl-R 1 ', -C(=O)-NH-arene, -C(=O)-NH-arenediyl-R 1 ', -C(=O)-NH-heteroarene, -C(=O)-NH-heteroarene diyl-R 1 ', -C(=O)-O-arene, -C(=O)-O-arenediyl-R 1 ', -C(=O)-O-heteroarene, -C(=O)-O-heteroarene diyl-R 1 ', -C(=O)-CH 2 -heteroarene, -C(=O)-CH 2 -heteroarene diyl-R 1 ', -C(=O)-CR 2 'R 3 '-heteroarene, -C(=O)-CR 2 'R 3 '-heteroarene diyl-R 1 ', -C(=O)-L-amino acid-NH-heteroarene, -C(=O)-L-amino acid-NH-heteroarene diyl-R 1 ';

[0014] And R 2 : methyl; R 3 :methyl.

[0015] Preferably, R 1'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, wait.

[0016] Preferably, the aromatic hydrocarbon group is selected from:

[0017] wait.

[0018] Preferably, the length of the C chain of substituted alkanes, substituted alkenes, substituted alkynes, alkanes, alkenes, and alkynes is ≤ 6, and may be straight chain, branched chain, or cyclic. The substituent is selected from 1 to 5 -SO 3 H, -OH, -F, -Br, -Cl, -OH, methyl, ethyl, propyl replacement (and / or).

[0019] Further preferably, the compound, its pharmaceutically acceptable salt or stereoisomer is as follows:

[0020]

[0021]

[0022] The present invention also provides the use of the above-mentioned compounds, their pharmaceutically acceptable salts, and stereoisomers for preparing NRF2 activators. The biological experiments of the present invention show that many of the above-mentioned compounds of the present invention exhibit single-digit nanomolar human Nrf2 receptor agonist activity, and are significantly better than the existing NRF2 activator control drug Omaveloxolone. At the same time, the experiment also shows that the compounds of the present invention also have the activity of scavenging DPPH free radicals, inhibiting the production of lipid peroxide MDA, intervening in ferroptosis, etc. to exert antioxidant effects.

[0023] The present invention also provides the use of the above-mentioned compound, its pharmaceutically acceptable salt, stereoisomer, or pharmaceutical composition to prepare a drug for treating and / or preventing a patient's disease, and the use of the above-mentioned compound or its pharmaceutically acceptable salt or stereoisomer or pharmaceutical composition in preparing a drug. In addition, the prepared drug is used to prevent or treat diseases, including cerebral small vessel disease, mitochondrial encephalopathy, autism spectrum disorder, Rett syndrome, Friedreich's ataxia, stroke, hemorrhagic stroke, ischemic stroke, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, schizophrenia cognitive impairment, Parkinson's disease, Parkinson's cognitive impairment, Alzheimer's disease, vascular dementia, epilepsy, Huntington's disease, heart failure, myocardial infarction, renal failure, renal ischemia, etc. According to effective test data, the application of the drug prepared by the above-mentioned compound of the present invention, its pharmaceutically acceptable salt, and stereoisomer, especially for stroke, multiple sclerosis, amyotrophic lateral sclerosis has a significant effect.

[0024] Preferably, the compound, its pharmaceutically acceptable salt, stereoisomer is used to prepare a drug for preventing or treating stroke, multiple sclerosis, and amyotrophic lateral sclerosis.

[0025] Beneficial effects:

[0026] The present invention synthesizes for the first time a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, namely a C-17 carbonyl-substituted oleanane triterpene derivative.

[0027] At the same time, the present invention provides a method for preparing the compound represented by formula (I) through specific examples.

[0028] Furthermore, the present invention provides the use of a compound represented by formula (I) or a pharmaceutically acceptable salt thereof for preparing an NRF2 activator. The C-17 carbonyl-substituted oleanane triterpene derivative disclosed in the present invention can exert an antioxidant effect by scavenging DPPH free radicals, inhibiting the generation of lipid peroxide MDA, or inhibiting ferroptosis while maintaining a strong Nrf2 agonist effect.

[0029] Finally, the present invention also provides a pharmaceutical composition, including 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, etc. The pharmaceutical composition can be administered orally or parenterally (such as intravenously, subcutaneously or topically). The dosage can be appropriately adjusted according to the patient's age, gender and disease type. Diseases that can be used for prevention or treatment in the future include 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 drugs are effective for diseases such as stroke, multiple sclerosis, and amyotrophic lateral sclerosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Effects of compounds 19 and 28 on mNSS scores of rats with ischemic stroke: compared with the sham operation group: ###p<0.001; compared with the model group: ***p<0.001.

[0031] Figure 2 .Effects of compounds 19 and 28 on cerebral infarction area in rats with ischemic stroke, compared with the sham operation group: ###p<0.001; compared with the model group: ***p<0.001.

[0032] Figure 3 .The effect of compound 24 on the neurological function score of EAE model mice, compared with the sham operation group: ***p<0.001; compared with the model group: #p<0.05, ##p<0.01, ###p<0.001; compared with the Omaveloxolone 15mg / kg group: &&&p<0.001.

[0033] Figure 4 Effect of compound 24 on the 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.

[0034] Figure 5 . Effects of compounds 26 and 31 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 Omaveloxolone 3mg / kg group: & p<0.05.

[0035] Figure 6 Effects of compounds 26 and 31 on the motor coordination ability of SOD1 G93A mice, compared with the control group: * p<0.05, *** p<0.001; compared with the model group: ### p<0.001; compared with Omaveloxolone 3mg / kg group: & p<0.05.

[0036] Figure 7 Effects of compounds 26 and 31 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 Omaveloxolone 3mg / kg group: & p<0.05.

[0037] <0.05. DETAILED DESCRIPTION

[0038] The present invention is further described below in conjunction with specific embodiments and test examples, but the scope of the present invention is not limited in any form.

[0039] The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS).

[0040] NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using an AVANCE III 600 NMR spectrometer and the solvent used was deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated methanol (CD 3 OD) and deuterated chloroform (CDCl 3 ), and the internal standard was tetramethylsilane (TMS).

[0041] Liquid chromatography-mass spectrometry (LC-MS) was performed using a Shimadzu LCMS2020 mass spectrometer, and HPLC was performed using a Shimadzu LC20A liquid chromatograph.

[0042] 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.

[0043] Unless otherwise specified in the examples, SFC separation conditions are as follows: column model: DAICEL CHIRALPAK IC (250 mm*30 mm, 10 um); mobile phase: [CO2-i-PrOH / ACN]; B%: 40%, isocratic elution mode.

[0044] In the present invention, if the specific experimental conditions are not indicated, the conventional experimental conditions or the conditions recommended by the manufacturer shall be followed. If the manufacturer of the reagents or instruments is not indicated, conventional products can be obtained by commercial purchase.

[0045] In the present invention, the test results are expressed as average values.

[0046] The detection indexes in the present invention are: human Nrf2 receptor function test (agonist detection); DPPH free radical scavenging ability test; MDA anti-lipid peroxidation ability test.

[0047] ■Example 1

[0048] Compound 1:

[0049]

[0050] Synthesis route:

[0051]

[0052] Step 1: Synthesis of intermediate 2'

[0053] The raw material 1' (300 mg, 610.18 μmol) was dissolved in anhydrous dichloromethane (3 mL), and then N, N-dimethylformamide (22.30 mg, 309.09 μmol) and oxalyl chloride (309.79 mg, 2.44 mmol, 213.65 μL) were added and stirred at 25 ° C for 1 hour. After the reaction was completed by LCMS, the reaction solution was concentrated to obtain the intermediate 2' (300 mg, crude product, white solid). LCMS (methyl ester): rt = 0.660 min, 506.3 [M + H] + .

[0054] Step 2: Synthesis of intermediate 3

[0055] Ammonia (30.05 mg, 1.76 mmol) was introduced into toluene (4 mL) at -70 °C for 10 minutes, and then intermediate 2' (0.3 g, 588.11 μmol) was added and stirred at -70 °C for 1 hour. After the reaction was completed by LCMS, the reaction mixture was 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 freeze-drying, intermediate 6 (250 mg, yield 86.6%, yellow solid) was obtained. LCMS: rt = 0.557 min, 491.3 [M+H] + ; Purity: 100%.

[0056] Step 3: Synthesis of intermediate 4

[0057] Dissolve intermediate 4A (5 g, 29.38 mmol) in anhydrous acetonitrile (50 mL), and add phosphorus oxybromide (12.64 g, 44.07 mmol) under nitrogen. The reaction solution was stirred at 80 ° C for 0.5 hours. After the reaction was completed, the reaction mixture was added to normal temperature water (50 mL) at room temperature for quenching, and then filtered and concentrated after stirring for 1 hour to obtain a crude product. After purification by column chromatography (silica gel column, petroleum ether / ethyl acetate = 2 / 1 to 1 / 1), intermediate 4 (3.3 g, yield 45.92%, yellow solid) was obtained. LCMS: rt = 0.381, 0.749 min, 233.0 / 235.0 [M+H] + ; Purity 95.34% 1 H NMR (400MHz, DMSO-d6) δ = 5.93 (s, 1H), 5.00-4.86 (m, 1H), 1.33 (d, J = 7.0Hz, 6H).

[0058] Step 4: Synthesis of compound 1

[0059] Intermediate 3 (50 mg, 101.90 μmol) and intermediate 4 (35.62 mg, 152.85 μmol) were added to dioxane (4 mL) and dissolved. After nitrogen replacement three times, PEPPSIPd (5.99 mg, 7.13 μmol) and sodium tert-butoxide (29.38 mg, 305.70 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 prepared by chromatography (C 18 Column, 0.1% formic acid solution) and freeze-dried to obtain compound 1 (4.2 mg, white solid, yield 4.06%). LCMS: rt = 1.560 min, 643.4 [M+H] + ; Purity 94.25%; 1H NMR (400MHz, DMSO-d6)δ=10.81-10.41(m,1H),9.65-9.28(m,1H),8.65(s,1H),6.22(s,1H),6.03(br s,1H),5.04-4.92(m,1H),3.05(br d,J=4.2Hz,1H),2.84(br d,J=13.6Hz,1H),2.08-1.98(m,1H),1.87-1.73(m,3H),1.67-1.57(m,4H),1.44-1.40(m,4H),1.34(d,J=7 .0Hz,6H),1.28-1.22(m,6H),1.19-1.12(m,5H),1.08-1.00(m,4H),0.96(s,3H),0.89(brd,J=11.6Hz,6H).

[0060] ■Example 2

[0061] Compound 2:

[0062]

[0063] Synthesis route:

[0064]

[0065] ■Example 3

[0066] Compound 3:

[0067]

[0068] Synthesis route:

[0069]

[0070] Step 1: Synthesis of intermediate 3'

[0071] Raw material 1 (1 g, 5.30 mmol) and raw material 2 (1.57 g, 6.89 mmol) were added to dioxane (10 mL) and dissolved, and then diisopropylethylamine (2.06 g, 15.91 mmol, 2.77 mL) was added and microwaved at 120 ° C for 3 h. After the reaction was completed by LCMS detection, the reaction solution was concentrated to obtain a crude product. The crude product was purified by column chromatography (silica gel column, petroleum ether / ethyl acetate = 1 / 0 ~ dichloromethane / anhydrous methanol = 1 / 1) to obtain a crude product, and the crude product was slurried and filtered with methanol (10 mL) to obtain intermediate 3' (0.9 g, yield 45.95%, gray solid). LCMS: rt = 0.506 min, 380.1 [M+H] + ; Purity 96.28%; 1H NMR (400 MHz, DMSO-d 6 )δ=9.75(br s,1H),7.46-7.30(m,5H),7.25(d,J=8.6Hz,2H),6.99(d,J=8.8Hz,2H),6.44(br d,J=6.6Hz,1H),5.08(s,2H),4.96-4.84(m,1H),4.49-4.38(m,1H),4.34(s,1H),1.36(d,J=6.8Hz,3H),1.29-1.23(m,6H).

[0072] Step 2: Synthesis of intermediate 4

[0073] The intermediate 3' (450 mg, 1.19 mmol) was dissolved in anhydrous methanol (200 mL), and then wet palladium carbon (0.5 g, 10%) was added under nitrogen protection. After hydrogen was replaced three times, the mixture was stirred at 25°C for 2 hours. After the reaction was completed by LCMS, the reaction solution was filtered and concentrated 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 extraction, intermediate 4 (120 mg, yield 34.27%, yellow solid) was obtained. LCMS: rt = 0.397 min, 290.1 ​​[M + H] + ,purity: 98.23%.

[0074] Step 3: Synthesis of compound 3

[0075] Intermediate 5 (50 mg, 101.70 μmol) and intermediate 4 (35.31 mg, 122.04 μmol) were dissolved in dichloromethane (0.5 mL), and EDCI (23.39 mg, 122.04 μmol) and 4-dimethylaminopyridine (12.42 mg, 101.70 μmol) were added, and stirred at 25 ° C for 12 hours, and then stirred at 40 ° C for 2 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 freeze-dried to obtain compound 3 (28.6 mg, white solid, yield 36.02%). LCMS: rt = 1.746 min, 763.4 [M+H] + ; Purity 97.73%; 1H NMR (400MHz, DMSO-d6) δ = 9.81 (s, 1H), 8.67 (s, 1H), 7.39 (d, J = 8.4Hz, 2H), 7.07 (d, J = 8.4Hz, 2H), 6.54 (br d,J=6.6Hz,1H),6.26(s,1H),4.98-4.84(m,1H),4.53(br t,J=6.6Hz,1H),4.34(d,J=2.0Hz,1H),2.94(br d,J=3.4Hz,2H),2.08-1.93(m,2H),1.89-1.64(m,7H),1.52-1.43(m,6H),1.39(br d,J=6.8Hz,3H),1.36-1.30(m,4H),1.27(br d,J=6.8Hz,8H),1.18(s,3H),1.07(s,3H),0.99(s,3H),0.95(s,3H),0.91(s,3H).

[0076] ■Example 4

[0077] Compound 4:

[0078]

[0079] Synthetic route 1:

[0080]

[0081] Step 1: Synthesis of intermediate 2

[0082] The raw material 1 (500 mg) was dissolved in anhydrous dichloromethane (5 mL), and then N, N-dimethylformamide (7.82 μL) was added and ventilated with a nitrogen balloon. Subsequently, oxalyl chloride (356.09 μL) was slowly added to the reaction bottle and reacted at 25°C for 1 hour. After LC-MS detection of the reaction of the raw material, the reaction solution was concentrated under reduced pressure to obtain a residue. The obtained product was directly used in the next step without purification. Intermediate 2 (555 mg, crude product) was a light yellow solid powder.

[0083] Step 2: Synthesis of compound 4

[0084] 3 (46.16 mg) was dissolved in pyridine (1 mL), cooled to 0°C in an ice-water bath under nitrogen protection, and intermediate 2 (160 mg) was dissolved in pyridine (1 mL) and slowly added to the reaction bottle. The reaction was allowed to react in an ice-water bath for 5 hours. After the reaction of the raw materials was completed by LCMS, the reaction solution was poured into water (3 mL), extracted with ethyl acetate (2 mL*3), and the organic phase was washed with saturated brine (2.5 mL*2), dried over anhydrous sodium sulfate, and concentrated. The crude product was prepared by chromatography (C 18Column, 0.1% formic acid solution) and then freeze-dried to obtain compound 4 (27 mg, yield 14.45%). LCMS: Rt = 0.631 min, 572.5 [M+H] + ; HPLC: Rt = 2.098 min; 1 H NMR (400MHz, CHLOROFORM-d) δ = 8.05 (s, 1H), 5.98 (s, 1H), 5.92 (s, 1H), 3.13 (br s, 2H), 2.36-2.32 (m, 3H), 2.07-1.99 (m, 3H), 1.79 (br s,11H),1.49(s,3H),1.38(s,3H),1.27(s,5H),1.18(s,3H),1.05(d,J=4.0Hz,6H),0.94(s,3H).

[0085] Synthesis route 2:

[0086]

[0087] Step 1: Synthesis of intermediate 2

[0088] The raw material 1 (1g) was dissolved in anhydrous dichloromethane (10mL), N,N-dimethylformamide (15.65μL) was added and then ventilated with a nitrogen balloon, and then oxalyl chloride (712.16μL) was slowly added to the reaction bottle, and reacted at 25°C for 1 hour. After LCMS (quenched with methanol) detected that the raw material had reacted, the reaction solution was concentrated under reduced pressure to obtain a solid. The obtained product was directly used in the next step without purification. Intermediate 2 (1.1g, crude product is a light yellow solid powder.

[0089] Step 2: Synthesis of intermediate 3

[0090] Hydrazine hydrate (1.05 mL, 98%) was dissolved in anhydrous dichloromethane (5 mL), and the mixture was cooled to 0°C in an ice-water bath under nitrogen protection. Intermediate 2 (1.1 g) was dissolved in anhydrous dichloromethane (5 mL) and slowly added to the reaction flask. The reaction was allowed to react in an ice-water bath for 1 hour. After the reaction of the raw materials was completed by LCMS, the reaction solution was concentrated. The crude product was prepared by chromatography (C 18 Column, 0.1% formic acid solution) separation, acetonitrile was concentrated and then extracted with ethyl acetate (20mL*2), the organic phase was washed with saturated brine (30mL) and then dried over anhydrous sodium sulfate, and concentrated to obtain 3 (580mg, 1.15mmol, yield 53.19%). LCMS: rt = 0.502min, 506.3 [M+H] + .

[0091] Step 3: Synthesis of intermediate 5

[0092] Dissolve intermediate 3 (580 mg) and material 4' (145.20 μL) in anhydrous ethanol (7 mL) and then add acetic acid (6.57 μL) and react at 50°C for 3 hours. LCMS monitoring shows that 40% of the raw material remains and 35% of the product is generated. The reaction solution is concentrated under reduced pressure and dried and used directly in the next step. The crude product 5 (600 mg, yield 84.67%) is a tan solid. LCMS: rt = 0.599 min, 618.5 [M+H] + .

[0093] Step 4: Synthesis of compound 4

[0094] The intermediate 5 (500 mg) was dissolved in 6 (10 mL) and the temperature was raised to 145°C for 12 hours. The product was generated after the raw material was consumed by LCMS monitoring. The reaction solution was concentrated under reduced pressure and then silica gel was added to mix the sample. The crude product was separated by normal phase column chromatography (petroleum ether: ethyl acetate = 0 / 0-3 / 1). The chromatographic preparation (C 18 Column, 0.1% formic acid solution) to separate and purify compound 4 (39.9 mg, 69.30 μmol, yield 8.56%). LCMS: rt = 1.655 min, 572.4 [M+H] + ; HPLC: rt = 2.223min; 1 H NMR(400MHz,CHLOROFORM-d)δ=8.05(s,1H),5.98(s,1H),5.88(s,1H),3.14(br d,J=4.0Hz,2H),2.33(s,3H),2.02(s,3H),1.95-1.80(m,3H),1.75-1.54(m,4H),1.49(s,3 H), 1.43-1.33 (m, 5H), 1.31-1.23 (m, 6H), 1.17 (s, 3H), 1.05 (d, J = 4.4Hz, 6H), 0.94 (s, 3H).

[0095] ■Example 5

[0096] Compound 5:

[0097]

[0098] Synthesis route:

[0099]

[0100] Step 1: Synthesis of intermediate 2

[0101] The raw material 1 (500 mg, 1.02 mmol, 1 eq) was dissolved in anhydrous dichloromethane (5 mL), and then N, N-dimethylformamide (7.43 mg, 101.70 μmol, 7.82 μL, 0.1 eq) was added and protected with nitrogen. Then oxalyl chloride (516.33 mg, 4.07 mmol, 356.09 μL, 4 eq) was slowly added to the reaction bottle, and reacted at 25 ° C for 1 hour. After the reaction of the raw material was completed by LCMS, the reaction solution was concentrated under reduced pressure to obtain a residue. The obtained product was directly used in the next step without purification. Compound 2 (555 mg, crude product) is a light yellow solid powder.

[0102] Step 2: Synthesis of compound 5

[0103] Material 3 (66.78 mg, 439.12 μmol, 0.8 eq) was dissolved in pyridine (2 mL), ventilated with a nitrogen balloon, and then cooled to 0°C in an ice-water bath. Intermediate 2 (280 mg, 548.91 μmol, 1 eq) was dissolved in pyridine (2 mL) and slowly added to the reaction bottle. The reaction was allowed to react in an ice-water bath for 5 hours. After the reaction of the raw materials was completed by LCMS, the reaction solution was poured into water (5 mL), extracted with ethyl acetate (5 mL*3), and the organic phase was washed with saturated brine (5 mL*2), dried over anhydrous sodium sulfate, and concentrated. The crude product was prepared by chromatography (C 18 Column, 0.1% formic acid solution) and then freeze-dried to obtain compound 5 (80 mg, 122.10 μmol, yield 22.24%). LCMS: Rt = 1.802 min, 626.4 [M+H] +; HPLC: Rt = 2.388 min; 1 H NMR (400MHz, DMSO-d6) δ = 14.30-13.93 (m, 1H), 8.67 (s, 1H), 6.65 (br s, 1H), 6.26 (s, 1H), 2.91 (br s,2H),2.09-1.84(m,4H),1.72-1.61(m,5H),1.50-1.43(m,6H),1.30-1.2 2(m,6H),1.18(s,3H),1.07(s,3H),0.98(s,3H),0.94(s,3H),0.91(s,3H).

[0104] ■Example 6

[0105] Compound 6:

[0106]

[0107] Synthesis route:

[0108] ■Example 7

[0109] Compound 7:

[0110]

[0111] Synthesis route:

[0112] ■Example 8

[0113] Compound 8:

[0114]

[0115] ■Example 9

[0116] Compound 9:

[0117]

[0118] Synthesis route:

[0119] ■Example 10

[0120] Compound 10:

[0121]

[0122] Synthesis route:

[0123] ■Example 11

[0124] Compound 11:

[0125]

[0126] Synthesis route:

[0127]

[0128] Step 1: Synthesis of intermediate 2

[0129] Compound 1 (1 g, 5.52 mmol, 1 eq), triethylamine (614.32 mg, 6.07 mmol, 845.01 μL, 1.1 eq), Boc2O (1.32 g, 6.07 mmol, 1.39 mL, 1.1 eq) and DMAP (67.43 mg, 551.91 μmol, 0.1 eq) were added to anhydrous tetrahydrofuran (20 mL), and the reaction solution was stirred at 20 ° C for 14 hours. LCMS showed that the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (silica, petroleum ether: ethyl acetate = 1: 0 to 4: 1) to obtain intermediate 2 (1.1 g, 3.44 mmol, yield 62.35%, purity 88%) as a yellow oil. LCMS: rt = 0.524 min, 304.1 [M + Na] +, purity: 87.557%.

[0130] Step 2: Synthesis of intermediate 3

[0131] Compound 2 (1.1 g, 3.91 mmol, 1 eq) and hydrazine hydrate (4.27 g, 83.59 mmol, 4.14 mL, 98% purity, 21.38 eq) were added to anhydrous ethanol (15 mL). The reaction solution was heated at 80 ° C for 16 hours under a nitrogen atmosphere. LCMS showed that the reaction was complete. The reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain the product (900 mg, 3.61 mmol, yield 92.33%, purity 95%) as a yellow oil. LCMS: rt = 0.303 min, 150.0 [M-C5H9O2+H] - ,Purity: 94.705%. 1 H NMR (400MHz, DMSO-d6) δ = 11.21 (s, 1H), 6.70 (s, 1H), 6.23 (br d, J = 2.0Hz, 1H), 6.00 (t, J = 2.8Hz, 1H), 3.41 (br s, 2H), 1.36 (s, 9H).

[0132] Step 3: Synthesis of intermediate 5

[0133] Compound 4 (100 mg, 203.39 μmol, 1 eq), DMF (7.43 mg, 101.70 μmol, 7.82 μL, 0.5 eq) and oxalyl chloride (103.26 mg, 813.57 μmol, 71.22 μL, 4 eq) were added to anhydrous dichloromethane (1 mL). The reaction solution was stirred at 20 ° C for 1 hour. LCMS (quenched with methanol) showed that the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain intermediate 5 (100 mg, crude product) as a white solid. LCMS: rt = 0.717 min, 506.3 [M + H] + (Mass of methyl ester), purity: 82.696%.

[0134] Step 4: Synthesis of compound 11

[0135] Intermediate 5 (100 mg, 196.04 μmol, 1 eq) was added to pyridine (0.5 mL), and then a mixed solution of compound 3 (73.30 mg, 294.06 μmol, 1.5 eq) dissolved in pyridine (0.5 mL) was added, and the reaction solution was stirred at 0°C for 3 hours. LCMS showed that the reaction was complete. The reaction solution was concentrated under reduced pressure and purified by high performance liquid chromatography: column: Phenomenex lunaC18150*25mm*10um, mobile phase: [water (FA)-acetonitrile], gradient: 63%-93% B, 18 min to obtain compound 11 (9.2 mg, 14.45 μmol, yield 7.37%, purity 97.8%) as an off-white solid. LCMS (39): rt = 0.625 min, 623.4 [M + H] +, purity: 97.813%. HPLC (39): rt = 2.305 min, 97.949% purity. 1 H NMR (400MHz, DMSO-d6) δ = 12.63 (s, 1H), 11.26 (br s, 1H), 8.67 (s, 1H), 6.86 (br d, J = 1.6Hz, 1H), 6.49 (br s,1H),6.27(s,1H),6.20(d,J=2.0Hz,1H),6.10(br d,J=2.8Hz,1H),3.02-2.90(m,2H),2.09-1.99(m,1H),1.90-1.81(m,2H),1.79-1.61(m,5H),1.44 (s,5H),1.35-1.23(m,7H),1.18(s,3H),1.14-1.03(m,4H),1.01-0.89(m,8H),0.89-0.83(m,1H).

[0136] ■Example 12

[0137] Compound 12:

[0138]

[0139] Synthesis route:

[0140]

[0141] ■Example 13

[0142] Compound 13:

[0143]

[0144] Synthesis route:

[0145]

[0146] Step 1: Synthesis of intermediate a-1

[0147] To the material 6343-98-2 solution (2.00 g, 10.6 mmol, 1.00 eq) was added ethyl 3-oxobutanoate (1.37 g, 10.6 mmol, 1.34 mL, 1.00 eq). The mixture was stirred at 120 ° C for 2 h. LC-MS (EB11687-1-P1A1) showed a new peak on LC-MS, and about 71.0% of the desired compound was detected. The reaction mixture was concentrated under reduced pressure to obtain a residue. The crude product was triturated with ethanol (3V) at 25 ° C for 1 h, filtered and the solid was collected to obtain intermediate a-1 (1.70 g, 7.58 mmol, yield 72.0%, purity 98.2%) as a white solid. LCMS: (EB11687-1-P1A1_LCMS_SH), RT = 1.220 min, MS (ESI) m / z = 220 [M] + .LCMS:(EB11687-1-P1C1_LCMS_SH),RT=1.198min,MS(ESI)m / z=220[M] + .

[0148] Step 2: Synthesis of intermediate a

[0149] Under nitrogen atmosphere, Pd / C (246 mg, 232 μmol, 238 μL, 10% purity, 0.04 eq) was added to a MeOH solution (15.0 mL) of intermediate a-1 (1.50 g, 6.81 mmol, 1.00 eq). The suspension was degassed and heated with H 2 Purge 3 times. At 25℃H 2 The mixture was stirred for 2.5 h under the condition of (40.0 psi.). LC-MS (EB11687-2-P1A2) showed the appearance of a new peak, and about 85.9% of the expected compound was detected. The reaction mixture was filtered through a diatomaceous earth bed to remove Pd-C, and the diatomaceous earth bed was thoroughly washed with methanol containing 5.00% acetic acid. The filtrates were combined and the solvent was evaporated under vacuum. The residual syrup was suspended in ethyl acetate (10.0 mL) and diluted with hexane (50.0 mL). A yellow crystalline solid suspension was obtained. Stir for 10 min, filter the solid, wash with hexane and dry to obtain a yellow intermediate a solid (1.25 g, 6.57 mmol, yield 96.4%). LCMS: (EB11687-2-P1A2_LCMS_SH), RT = 0.431 min, MS (ESI) m / z = 190 [M] + .

[0150] Step 3: Synthesis of compound 13

[0151] The material 218600-44-3 (500 mg, 1.02 mmol, 1.00 eq), intermediate a (193 mg, 1.02 mmol, 1.00 eq) and NMI (292 mg, 3.56 mmol, 284 μL, 3.50 eq) in MeCN (8.00 mL) were mixed, and then TCFH (343 mg, 1.22 mmol, 1.20 eq) was added, and then the mixture was stirred 12 times at 25 ° C under nitrogen. LC-MS (EB11687-29-P1A1) showed that about 35.6% of the expected compound was detected. The residue was concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase HPLC column: Welch Xtimate C1840*200mm 7μm; mobile phase: [water (NH 4 HCO 3 )-ACN]; Gradient: 56.0%-96.0% B, 25 min to obtain compound 13, white solid (273 mg, 262 μmol, yield 39.3%). .LCMS: (EB11687-29-P1A1_LCMS_SH) RT = 1.855 min, MS (ESI) m / z = 663.9 [M+1] +.LCMS: (EB11687-35-P2C1_LCMS_SH) RT = 2.484 min, MS (ESI) m / z = 663.3 [M] +.HPLC: (EB11687-35-P1C4) RT = 4.362 min.1HNMR: (EB11687-35-P1N2) (CDCl3, 400 MHz) δ ppm 0.92 ( s,3H)0.97-1.01(m,8H)1.08-1.14(m,1H)1.16(s,3H)1.17-1.27(m,5H)1.27-1.45(m,4H)1.54-1.77(m,10H)1.90-2.00(m,3 H)2.31(s,3H)2.75(d,J=4.80Hz,1H)3.02-3.11(m,1H)5.94(s,2H)6.65(d,J=8.40Hz,2H)7.22(d,J=8.80Hz,2H)8.05(s,1H).

[0152] ■Example 14

[0153] Compound 14:

[0154]

[0155] Synthesis route:

[0156]

[0157] Step 1: Synthesis of intermediate 6B

[0158] p-TsOH (124 mg, 718 μmol, 0.05 eq) was added to a solution of material 6A (2.0 g, 13.1 mmol, 1.00 eq) and ethyl 3-oxobutanoate (1.95 g, 15.0 mmol, 1.90 mL, 1.15 eq) in EtOH (48.0 mL). The mixture was stirred at 80 ° C for 6 h. LC-MS (EB11687-6-P1 C3) showed a new peak on LC-MS, and about 96.4% of the expected compound was detected. The mixture was filtered and the filtrate was concentrated to obtain a crude product. The crude product was ground in ethanol (4.50 mL) at 25 ° C to obtain intermediate 6B (1.50 g, 6.60 mmol, yield 50.5%, purity 96.4%) as a red solid. LCMS: (EB11687-6-P1C3_LCMS_SH): RT=0.848min, MS(ESI)m / z=220[M+1] + .

[0159] Step 2: Synthesis of intermediate 6C

[0160] Pd / C (124 mg, 116 μmol, 10.0% purity, 0.04 eq) was added to a MeOH solution (1.00 mL) of intermediate 6B (750 mg, 3.42 mmol, 1.00 eq) under nitrogen atmosphere. The suspension was degassed and purged with H2 three times. The mixture was heated at 25 °C and H 2 (40.0psi) for 2.5h. LC-MS (EB11687-14-P1A1) showed that about 96.0% of the desired compound was detected. The reaction mixture was filtered through a diatomaceous earth bed to remove Pd-C, and the diatomaceous earth bed was thoroughly washed with methanol containing 5% acetic acid. The filtrates were combined and the solvent was evaporated under vacuum. The residual syrup was suspended in ethyl acetate (10.0mL) and diluted with hexane (50.0mL). A yellow crystalline solid suspension was obtained. Stir for 10min, filter the solid, wash with hexane and dry to obtain a red intermediate 6C solid (500mg, 2.64mmol, yield 77.2%). LCMS: (EB11687-14-P1A1_LCMS_SH): RT=0.154min, MS (ESI) m / z=189.9[M+1] + .

[0161] Step 3: Synthesis of compound 14

[0162] To a solution of material 218600-44-3 (500 mg, 1.02 mmol, 1.00 eq) and intermediate 6C (192 mg, 1.02 mmol, 1.00 eq) in acetonitrile (5.00 mL) was added CMPI (338 mg, 1.32 mmol, 1.30 eq) and Et 3 N (309 mg, 3.05 mmol, 425 μL, 3.00 eq). The mixture was stirred at 25 °C for 3 h. LC-MS (EB11687-40-P1A3) showed the appearance of a new peak and about 20% of the desired compound was detected. The mixture was concentrated to obtain a crude product. The crude product was purified by reverse phase HPLC (chromatographic column: Welch Xtimate C18 40*200mm 7μm; mobile phase: [water (NH3H2O+NH4HCO3)-ACN]; gradient: 54.0%-94.0% B in 25 min) to obtain compound 14 (74.0 mg, 112 μmol, 11.0% yield) as a brown solid. LCMS: (EB11687-40-P1A3_LCMS_SH): RT=1.834 min, MS (ESI) m / z=663[M+1] + ;LCMS: (EB11687-41-P1C1_LCMS_SH): RT=2.576min, MS(ESI)m / z=663[M+1] + ; HPLC: (EB11687-41-P1C3): RT=4.377min. 1 H NMR:(EB11687-41-P1N1)(CDCl 3 ,400MHz)δppm 0.95(s,3H)1.00-1.04(m,9H)1.18(s,3H)1.27(s,3H)1.33(br d,J=2.38Hz,2H)1.45(d,J=13.51Hz,2H)1.50(s,3H)1.72-1.82(m,10H)1.92-2.02(m,3H)2.34(s,3H)2. 77(d,J=4.63Hz,1H)3.03-3.17(m,1H)5.97(s,2H)6.71(d,J=8.25Hz,2H)7.27(s,2H)8.06-8.10(m,1H).

[0163] ■Example 15

[0164] Compound 15:

[0165]

[0166] Synthesis route:

[0167]

[0168] Step 1: Synthesis of intermediate 3

[0169] Compound 1 (5 g, 40.60 mmol, 4.81 mL, 1 eq) was dissolved in anhydrous tetrahydrofuran (50 mL) and sodium hydrogen (1.95 g, 48.72 mmol, 60% purity, 1.2 eq) was added in batches at 20 ° C under nitrogen. The reaction was carried out at 0 ° C for 0.5 hours, and then compound 2 (5.28 g, 44.66 mmol, 5.41 mL, 1.1 eq) was added dropwise into the reaction system under the protection of nitrogen and reacted at 20 ° C for 2 hours. TLC detected that most of the raw materials had reacted and a new point was generated. The reaction solution was slowly poured into a saturated aqueous ammonium chloride solution (200 mL), and then extracted three times with ethyl acetate (100 mL*3). The combined organic phase was washed once with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by normal phase column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to obtain intermediate 3 (4 g, 20.49 mmol, yield 50.47%) as a yellow oil. 1 H NMR(400MHz,CHLOROFORM-d)δ=6.97(dd,J=1.6,4.2Hz,1H),6.85(s,1H),6.15(dd,J=2 .4, 4.2Hz, 1H), 4.21 (q, J = 7.2Hz, 2H), 3.95 (s, 3H), 3.81 (s, 2H), 1.28 (t, J = 7.2Hz, 3H).

[0170] Step 2: Synthesis of intermediate 4

[0171] After the intermediate 3 (2g, 10.25mmol, 1eq) was dissolved in anhydrous ethanol (20mL), hydrazine hydrate (5.23g, 102.45mmol, 5.07mL, purity 98% 10eq) was added at 20°C, and then reacted at 80°C for 2 hours. After the reaction of the raw materials was completed, the reaction solution was concentrated and separated and purified by a reverse phase column [220g, water (formic acid)-acetonitrile system] and then freeze-dried to obtain the intermediate 4 (900mg, 5.52mmol, yield 53.84%, purity 100%) as a yellow oil. LCMS: rt = 0.418min, 164.1 [M+H] +. 1 H NMR (400MHz, DMSO-d6) δ = 8.45 (s, 1H), 6.77 (s, 1H), 6.27 (dd, J = 1.8, 3.6Hz, 1H), 6.00 (dd, J = 2.8, 3.6Hz, 1H), 5.56 (s, 1H), 3.68 (s, 3H).

[0172] Step 3: Synthesis of intermediate 6

[0173] Compound 5 (300 mg, 610.18 μmol, 1 eq) was dissolved in anhydrous dichloromethane (5 mL), and N, N-dimethylformamide (4.46 mg, 61.02 μmol, 4.69 μL, 0.1 eq) and oxalyl chloride (232.34 mg, 1.83 mmol, 160.24 μL, 3 eq) were added at 20°C, and nitrogen was replaced three times, and then reacted at 20°C for 1 hour. After the reaction of the raw materials was completed by LCMS (a small sample was dissolved in methanol for detection), the reaction solution was concentrated to obtain intermediate 6 (300 mg, 588.11 μmol, yield 96.38%) as a yellow oil. LCMS: rt = 0.608 min, 506.4 [M + H] + (Ms of methyl ester).

[0174] Step 4: Synthesis of compound 15

[0175] Intermediate 4 (95.97 mg, 588.11 μmol, 1 eq) and intermediate 6 (300 mg, 588.11 μmol, 1 eq) were dissolved in dichloromethane (5 mL), and pyridine (69.78 mg, 882.17 μmol, 71.20 μL, 1.5 eq) was added, and the mixture was reacted at 20°C for 16 hours. LCMS monitored that the raw material was consumed and the product was generated. The reaction solution was spin-dried and separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] and then freeze-dried to obtain compound 15 (225.1 mg, 355.90 μmol, yield 60.70%, purity 99.78%) as a white solid. Prep-HPLC:column:Phenomenex luna C18 150*40mm*15um;mobile phase:[water(FA)-ACN];gradient:62%-92%B over 15min.LCMS:rt=1.741min,637.4[M+H] + HPLC: rt = 2.144 min. 1H NMR (400MHz, CHLOROFORM-d) δ = 8.05 (s, 1H), 7.27 (s, 1H), 6.78-6.73 (m, 1H), 6.38 (dd, J = 1.4, 3.4Hz, 1H), 6.22-6.15 ( m,2H),5.98(s,1H),3.72(s,3H),3.20-3.13(m,2H),2.13-2.02(m,2H),1.98-1.86(m,2H),1.85-1.75(m,5H),1.67(br d,J=13.2Hz,1H),1.60-1.54(m,1H),1.51-1.47(m,3H),1.43-1.37(m,4H) ,1.35-1.24(m,6H),1.18(s,3H),1.06(d,J=1.6Hz,6H),0.97-0.92(m,3H).

[0176] ■Example 16

[0177] Compound 16:

[0178]

[0179] Synthesis route:

[0180]

[0181] Step 1: Synthesis of intermediate 3

[0182] Compound 1 (5 g, 40.60 mmol, 4.81 mL, 1 eq) was dissolved in anhydrous tetrahydrofuran (50 mL) and sodium hydrogen (1.95 g, 48.72 mmol, 60% purity, 1.2 eq) was added in batches at 20 ° C under nitrogen. The reaction was carried out at 0 ° C for 0.5 hours, and then compound 2 (5.28 g, 44.66 mmol, 5.41 mL, 1.1 eq) was added dropwise into the reaction system under the protection of nitrogen and reacted at 20 ° C for 2 hours. TLC detected that most of the raw materials had reacted and a new point was generated. The reaction solution was slowly poured into a saturated aqueous ammonium chloride solution (200 mL), and then extracted three times with ethyl acetate (100 mL*3). The combined organic phase was washed once with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by normal phase column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to obtain intermediate 3 (4 g, 20.49 mmol, yield 50.47%) as a yellow oil. 1H NMR(400MHz,CHLOROFORM-d)δ=6.97(dd,J=1.6,4.2Hz,1H),6.85(s,1H),6.15(dd,J=2 .4, 4.2Hz, 1H), 4.21 (q, J = 7.2Hz, 2H), 3.95 (s, 3H), 3.81 (s, 2H), 1.28 (t, J = 7.2Hz, 3H).

[0183] Step 2: Synthesis of intermediate 4

[0184] After the intermediate 3 (2g, 10.25mmol, 1eq) was dissolved in anhydrous ethanol (20mL), hydrazine hydrate (5.23g, 102.45mmol, 5.07mL, purity 98% 10eq) was added at 20°C, and then reacted at 80°C for 2 hours. After the reaction of the raw materials was completed, the reaction solution was concentrated and separated and purified by a reverse phase column [220g, water (formic acid)-acetonitrile system] and then freeze-dried to obtain the intermediate 4 (900mg, 5.52mmol, yield 53.84%, purity 100%) as a yellow oil. LCMS: rt = 0.418min, 164.1 [M+H] +. 1 H NMR (400MHz, DMSO-d6) δ = 8.45 (s, 1H), 6.77 (s, 1H), 6.27 (dd, J = 1.8, 3.6Hz, 1H), 6.00 (dd, J = 2.8, 3.6Hz, 1H), 5.56 (s, 1H), 3.68 (s, 3H).

[0185] Step 3: Synthesis of intermediate 6

[0186] Compound 5 (300 mg, 610.18 μmol, 1 eq) was dissolved in anhydrous dichloromethane (5 mL), and N, N-dimethylformamide (4.46 mg, 61.02 μmol, 4.69 μL, 0.1 eq) and oxalyl chloride (232.34 mg, 1.83 mmol, 160.24 μL, 3 eq) were added at 20°C, and nitrogen was replaced three times, and then reacted at 20°C for 1 hour. After the reaction of the raw materials was completed by LCMS (a small sample was dissolved in methanol for detection), the reaction solution was concentrated to obtain intermediate 6 (300 mg, 588.11 μmol, yield 96.38%) as a yellow oil. LCMS: rt = 0.608 min, 506.4 [M + H] + (Ms of methyl ester).

[0187] Step 4: Synthesis of compound 16

[0188] Intermediate 4 (95.97 mg, 588.11 μmol, 1 eq) and intermediate 6 (300 mg, 588.11 μmol, 1 eq) were dissolved in dichloromethane (5 mL), and pyridine (69.78 mg, 882.17 μmol, 71.20 μL, 1.5 eq) was added, and the mixture was reacted at 20°C for 16 hours. LCMS monitored that the raw material was consumed and the product was generated. The reaction solution was spin-dried and separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] and then freeze-dried to obtain compound 16 (227.1 mg, 355.83 μmol, yield 60.50%, purity 99.78%) as a white solid. Prep-HPLC:column:Phenomenex luna C18 150*40mm*15um;mobile phase:[water(FA)-ACN];gradient:62%-92%B over 15min.LCMS:rt=1.741min,637.4[M+H] + HPLC: rt = 2.144 min. 1 H NMR (400MHz, CHLOROFORM-d) δ = 8.05 (s, 1H), 7.27 (s, 1H), 6.78-6.73 (m, 1H), 6.38 (dd, J = 1.4, 3.4Hz, 1H), 6.22-6.15 ( m,2H),5.98(s,1H),3.72(s,3H),3.20-3.13(m,2H),2.13-2.02(m,2H),1.98-1.86(m,2H),1.85-1.75(m,5H),1.67(br d,J=13.2Hz,1H),1.60-1.54(m,1H),1.51-1.47(m,3H),1.43-1.37(m,4H) ,1.35-1.24(m,6H),1.18(s,3H),1.06(d,J=1.6Hz,6H),0.97-0.92(m,3H).

[0189] ■Example 17

[0190] Compound 17:

[0191]

[0192] Synthesis route:

[0193]

[0194] Step 1: Synthesis of intermediate 2

[0195] Compound 1 (1 g, 5.52 mmol, 1 eq), triethylamine (614.32 mg, 6.07 mmol, 845.01 μL, 1.1 eq), Boc2O (1.32 g, 6.07 mmol, 1.39 mL, 1.1 eq) and DMAP (67.43 mg, 551.91 μmol, 0.1 eq) were added to anhydrous tetrahydrofuran (20 mL), and the reaction solution was stirred at 20 ° C for 14 hours. LCMS showed that the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (silica, petroleum ether: ethyl acetate = 1: 0 to 4: 1) to obtain intermediate 2 (1.1 g, 3.44 mmol, yield 62.35%, purity 88%) as a yellow oil. LCMS: rt = 0.524 min, 304.1 [M + Na] +, purity 87.557%.

[0196] Step 2: Synthesis of intermediate 3

[0197] Compound 2 (1.1 g, 3.91 mmol, 1 eq) and hydrazine hydrate (4.27 g, 83.59 mmol, 4.14 mL, purity 98%, 21.38 eq) were added to anhydrous ethanol (15 mL). The reaction solution was heated at 80 ° C for 16 hours under a nitrogen atmosphere. LCMS showed that the reaction was complete. The reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain intermediate 3 (900 mg, 3.61 mmol, yield 92.33%, purity 95%) as a yellow oil. LCMS: rt = 0.303 min, 150.0 [M-C5H9O2+H] - , purity 94.705%. 1 H NMR (400MHz, DMSO-d6) δ = 11.21 (s, 1H), 6.70 (s, 1H), 6.23 (br d, J = 2.0Hz, 1H), 6.00 (t, J = 2.8Hz, 1H), 3.41 (br s, 2H), 1.36 (s, 9H).

[0198] Step 3: Synthesis of intermediate 4

[0199] After dissolving the intermediate 3 (1.7 g, 6.82 mmol, 1 eq) in anhydrous N,N-dimethylformamide (25 mL), lithium tert-butoxide (818.92 mg, 10.23 mmol, 922.20 μL, 1.5 eq) was added at 0°C and reacted for 0.5 hours. Then 2-(trimethylsilyl)ethoxymethyl chloride (1.36 g, 8.18 mmol, 1.45 mL, 1.2 eq) was added at 0°C and reacted at 20°C for 1.5 hours. After the reaction of the raw materials was completed by LCMS detection, the reaction solution was poured into a saturated aqueous ammonium chloride solution (100 mL), and then extracted three times with ethyl acetate (70 mL*3). The combined organic phase was washed three times with saturated brine (70 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by normal phase column chromatography (petroleum ether / ethyl acetate = 0-50%) to obtain intermediate 4 (610 mg, 1.45 mmol, yield 21.21%, purity 90%) as a yellow solid. LCMS: rt = 0.557 min, 280.3 [M-Boc+H] + .

[0200] Step 4: Synthesis of intermediate 6

[0201] Intermediate 4 (368.29 mg, 970.39 μmol, 1 eq) and intermediate 5 (495 mg, 970.39 μmol, 1 eq) were dissolved in dichloromethane (10 mL), and pyridine (230.27 mg, 2.91 mmol, 234.97 μL, 3 eq) was added, and the mixture was reacted at 20°C for 16 hours. LCMS monitoring showed that the raw materials were consumed and the product was generated. The reaction solution was dried and purified by normal phase column chromatography (petroleum ether / ethyl acetate = 0-50%) to obtain compound 6 (437 mg, 580.30 μmol, yield 59.80%) as a yellow solid. LCMS: rt = 0.734 min, 753.6 [M+H] + .

[0202] Step 5: Synthesis of compound 17

[0203] After dissolving intermediate 6 (100 mg, 132.79 μmol, 1 eq) in hydrochloric acid / 1,4-dioxane (5 mL, 2 M), the mixture was heated to 50 °C and reacted for 2 hours. LCMS monitoring showed that the raw material was consumed and the product was generated. After the reaction solution was dried, it was adjusted to alkalinity with sodium bicarbonate aqueous solution, and then extracted three times with ethyl acetate (10 mL*3). The combined organic phases were washed once with saturated brine (15 mL), dried over anhydrous sodium sulfate, and dried to obtain a crude product. The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] and then freeze-dried to obtain compound 17 (6.2 mg, 9.75 μmol, yield 7.34%, purity 97.96%) as a white solid. Prep-HPLC: column: Phenomenex luna C18 150*25mm*10um; mobile phase: [water(FA)-ACN]; gradient: 55%-85%B over 10min. LCMS: rt=1.664min, 623.4[M+H]+. HPLC: rt=2.043min. 1 H NMR (400MHz, CHLOROFORM-d) δ = 8.78-8.63 (m, 1H), 8.04 (s, 1H), 6.89 (br s, 1H), 6.46 (br s, 1H), 6.29 (br d,J=2.8Hz,1H),6.17(s,1H),5.98(s,1H),3.19-3.10(m,2H),2.64(s,1H),2.06-2.01(m,2H),1.95-1.88(m,2H),1.78(br s,5H),1.67(br d,J=13.2Hz,1H),1.55(brd,J=12.8Hz,1H),1.48(s,3H),1.36(s,4H),1.32-1.26(m,6H),1.18(s,3H),1.05(d,J=4.2Hz,6H),0.95(s,3H).

[0204] ■Example 18

[0205] Compound 18:

[0206]

[0207] Synthesis route:

[0208]

[0209] ■Example 19

[0210] Compound 19:

[0211]

[0212] Synthesis route:

[0213]

[0214] Step 1: Synthesis of intermediate 2

[0215] To a solution of material 1 (500 mg, 1.02 mmol, 1 eq) in dichloromethane (8 mL) was added N, N-dimethylformamide (7.43 mg, 101.70 μmol, 7.82 μL, 0.1 eq) and oxalyl chloride (387.25 mg, 3.05 mmol, 267.07 μL, 3 eq). The reaction solution was reacted at 25 ° C for 1 hour. LCMS monitoring showed that the reaction raw material completely disappeared and the main peak was the target product. The reaction solution was concentrated under reduced pressure to obtain intermediate 2 (500 mg, 980.19 μmol, 96.38% yield) as a light yellow solid. LCMS: Rt = 0.917 min, 510.4 [M + H] + ESI pos.

[0216] Step 2: Synthesis of compound 19

[0217] At 0°C, a solution of intermediate 2 (500 mg, 980.19 μmol, 1 eq) in dichloromethane (5 mL) was added to a solution of hydroxylamine hydrochloride (81.74 mg, 1.18 mmol, 1.2 eq) and N,N-diisopropylethylamine (380.04 mg, 2.94 mmol, 512.18 μL, 3 eq) in dichloromethane (5 mL), and the reaction solution was heated to 20°C for 2 hours. LCMS monitoring showed that the raw material was consumed and the product was generated. The reaction solution was filtered and concentrated, and the crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] and then freeze-dried to obtain compound 19 (136.20 mg, 266.39 μmol, 27.18% yield, 99.1% purity). Prep-HPLC:column:Phenomenex luna C18 150*40mm*15um;mobile phase:[water(FA)-ACN];gradient:45%-75%B over 15min.LCMS:Rt=1.339min,507.3[M+H]+ESI pos.HPLC:Rt=1.816min. 1HNMR(400MHz,CHLOROFORM-d)δ=9.39-9.07(m,1H),8.03(s,1H),6.04(s,1H),2.97(d,J=4.6Hz,1H),2.92-2.84(m,1H) ,2.01-1.92(m,1H),1.85-1.79(m,1H),1.78-1.71(m,4H),1.70-1.62(m,3H),1.57-1.45(m,4H),1.42(s,3H),1.31(br d,J=4.0Hz,1H),1.28(s,3H),1.26-1.23(m,1H),1.21(s,3H),1.12(s,3H),0.97(d,J=5.8Hz,6H),0.88(s,3H).

[0218] ■Example 20

[0219] Compound 21:

[0220]

[0221] Synthesis route:

[0222]

[0223] Step 1: Synthesis of compound 21

[0224] Material 2 (24.53 mg, 196.04 μmol, 24.44 μL, 1 eq) was added to anhydrous dichloromethane (1 mL), and then N, N-diisopropylethylamine (126.68 mg, 980.19 μmol, 170.73 μL, 5 eq) was added. The reaction solution was cooled to 0 ° C and material 1 (100.00 mg, 196.04 μmol, 1 eq) dissolved in anhydrous dichloromethane (1 mL) was slowly added. The reaction solution was stirred at 25 ° C for 12 h. After the reaction was completed by LCMS detection, the reaction solution was concentrated to obtain a crude product. The crude product was purified by reverse phase preparation (C18 column, 0.1% trifluoroacetic acid solution) and lyophilized to obtain a crude product. The crude product was purified by reverse phase preparation (C18 column, 0.1% hydrogen chloride solution) and lyophilized to obtain compound 21 (5.0 mg, off-white solid, yield 4.26%). Prep-HPLC (column: Phenomenexluna C18 150*25mm*10um; mobile phase: [water (TFA)-ACN]; gradient: 40%-70%B over10min).LCMS: rt=1.225min,599.3[M+H] +, purity 100%. HPLC: retention time = 1.724min, purity 97.989%. 1 H NMR:1H NMR (400MHz, DMSO-d6) δ = 8.65 (s, 1H), 7.88-7.76 (m, 1H), 6.19 (s, 1H), 3.47-3.36 (m, 1H), 3.35-3 .24(m,1H),3.03-2.97(m,1H),2.90-2.80(m,1H),2.57-2.53(m,2H),1.98-1.79(m,3H),1.64(br s,3H),1.63-1.47(m,3H),1.44-1.43(m,3H),1.42-1.36(m,2H),1.36-1.27(m,2H),1.27-1.24(m ,3H),1.19-1.16(m,3H),1.16-1.08(m,2H),1.06(s,3H),0.93(s,3H),0.89(s,3H),0.86(s,3H).

[0225] ■Example 21

[0226] Compound 22:

[0227]

[0228] Synthesis route:

[0229]

[0230] Step 1: Synthesis of intermediate 2

[0231] Add material 1 (200 mg, 406.79 μmol, 1 eq) and N,N-dimethylformamide (2.97 mg, 40.68 μmol, 3.13 μL, 0.1 eq) to anhydrous dichloromethane (1.2 mL) and dissolve. Then replace with nitrogen three times, cool to 0 ° C and add oxalyl chloride (154.89 mg, 1.22 mmol, 106.82 μL, 3 eq) dissolved in anhydrous dichloromethane (0.8 mL), and stir at 25 ° C for 1 h. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product of intermediate 2 (200 mg, crude) was directly used for the next step reaction.

[0232] Step 2: Synthesis of compound 22

[0233] 3-Amino-1-propanesulfonic acid (27.28 mg, 196.04 μmol, 1 eq) was added to anhydrous dichloromethane (1 mL), and then N, N-diisopropylethylamine (126.68 mg, 980.19 μmol, 170.73 μL, 5 eq) was added. The reaction solution was cooled to 0 ° C and the intermediate 2 (100 mg, 196.04 μmol, 1 eq) dissolved in anhydrous dichloromethane (1 mL) was slowly added. The reaction solution was stirred at 25 ° C for 12 h. After the reaction was completed by LCMS detection, the reaction solution was concentrated to obtain a crude product. The crude product was purified by reverse phase preparation (C18 column, 0.1% sodium bicarbonate solution) and lyophilized to obtain a crude product. The crude product was purified by reverse phase preparation (C18 column, 0.1% hydrogen chloride solution) and lyophilized to obtain compound 22 (9.5 mg, off-white solid, yield 7.91%). Prep-HPLC (column: WatersXbridge 150*25mm*5um; mobile phase: [water (NH4HCO3)-ACN]; gradient: 22%-52% Bover 9min). over 18min). LCMS: rt=1.219min, 613.4[M+H]+, 100% purity. HPLC: retention time=1.708min, purity 99.318%. 1 H NMR:1H NMR (400MHz, DMSO-d6) δ = 8.64 (s, 1H), 7.89-7.73 (m, 1H), 6.18 (s, 1H), 3.22-3.12 (m, 1H), 3. 12-3.01(m,2H),2.90-2.80(m,1H),2.44-2.39(m,2H),1.92-1.77(m,3H),1.76-1.68(m,3H), 1.67-1.48(m,5H),1.47-1.39(m,6H),1.36-1.30(m,1H),1.29-1.25(m,1H),1.24-1.22(m,3H ),1.18-1.15(m,3H),1.15-1.08(m,2H),1.05(s,3H),0.92(s,3H),0.88(s,3H),0.85(s,3H).

[0234] ■Example 22

[0235] Compound 23:

[0236]

[0237] Synthesis route:

[0238]

[0239] Step 1: Synthesis of intermediate 2

[0240] Dissolve material 1 (300 mg, 610.18 μmol, 1 eq) in anhydrous dichloromethane (2 mL) and add a drop of N, N-dimethylformamide. Cool the reaction solution with an ice-water bath. Dissolve oxalyl chloride (309.79 mg, 2.44 mmol, 213.65 μL, 4 eq) in anhydrous dichloromethane (2 mL) and slowly dropwise add it to the stirred reaction solution. Slowly warm the reaction solution to 20 ° C and stir for 1 hour. LCMS (quenched with methanol) monitors the complete reaction of the raw materials. The reaction solution is concentrated by rotary evaporation to obtain intermediate 2 (311 mg, crude) as a white solid. LCMS (quenched with methanol): rt = 0.648 min, 506.3 [M + H] +.

[0241] Step 2: Synthesis of intermediate 4

[0242] The material 3 (179.86 mg, 914.52 μmol, 177.03 μL, 1.5 eq, HCl) was dissolved in anhydrous dichloromethane (3 mL), and then triethylamine (308.46 mg, 3.05 mmol, 424.30 μL, 5 eq) was added. The reaction solution was cooled with an ice-water bath, and then a solution of compound 2 (311 mg, 609.68 μmol, 1 eq) dissolved in anhydrous dichloromethane (2 mL) was slowly dripped. The reaction solution was slowly heated to 20 ° C and stirred for 3 hours. LCMS monitored the complete reaction of the raw materials. After the reaction solution was concentrated, it was purified by column chromatography (ethyl acetate / petroleum ether = 0-51%) to obtain intermediate 4 (370 mg, 583.73 μmol, yield 95.74%) as a colorless colloid. LCMS: rt = 0.619 min, 634.5 [M + H] + .

[0243] Step 3: Synthesis of intermediate 5

[0244] Intermediate 4 (370 mg, 583.73 μmol, 1 eq) was dissolved in a prepared dichloromethane / trifluoroacetic acid solution = 5 / 1 (6 mL) and stirred at 20°C for 1 hour. LCMS monitored the complete reaction of the raw material. The reaction solution was concentrated to obtain intermediate 5 (378 mg, crude, TFA) as a light yellow jelly. LCMS: rt = 0.494 min, 534.3 [M+H] +.

[0245] Step 4: Synthesis of compound 23

[0246] Anhydrous dichloromethane (3mL) and triethylamine (472.39mg, 4.67mmol, 649.77μL, 8eq) were added to intermediate 5 (378mg, 583.54μmol, 1eq, TFA) in turn, and the reaction solution was cooled in an ice-water bath. Material 6 (202.55mg, 1.17mmol, 2eq) was dissolved in anhydrous dichloromethane (2mL) and then slowly added dropwise to the stirred reaction solution. The reaction solution was slowly heated to 20°C and stirred for 3 hours. LCMS monitored the complete reaction of the raw materials. After the reaction solution was concentrated, the crude product was obtained by reverse phase preparative lyophilization, which was then purified by prep-TLC (dichloromethane / methanol = 15:1) and then lyophilized to obtain compound 23 (62.8mg, 89.28μmol, yield 15.30%, purity 93.92%) as a yellow solid. HPLC: (1) column: Phenomenex luna C18150*40mm*15um; mobile phase: [water (FA)-ACN]; gradient: 45%-75%B over 15min.LCMS: rt=0.630min, 671.4[M+H]+.HPLC: rt=1.761min.1H NMR (400MHz, METHANOL-d4) δ = 8.37 (s, 1H), 7.92-7.75 (m, 1H), 7.53-7.33 (m, 2H), 6.97-6.79 (m, 1H), 6.07 (s, 1H), 3.64 (br d, J = 1.0Hz, 3H), 3.10 (br s,1H),3.06-2.97(m,1H),1.90-1.78(m,2H),1.70(br d,J=1.2Hz,4H),1.62-1.51(m,4H),1.46-1.37(m,4H),1.31(br s,2H),1.24(br s,4H),1.14(br s,6H),1.09(br s,2H),0.94(br s,6H),0.88(br s,3H).

[0247] ■Example 23

[0248] Compound 24:

[0249]

[0250] Synthesis route:

[0251]

[0252] Step 1: Synthesis of compound 24

[0253] N,N-diisopropylethylamine (633.41 mg, 4.90 mmol, 853.65 μL, 5 eq) was added to a solution of N-methylhydroxylamine hydrochloric acid (98.24 mg, 1.18 mmol, 1.2 eq) in dichloromethane (7.5 mL). After replacing with nitrogen three times, the temperature was lowered to 0°C and a solution of material 1 (500 mg, 980.19 μmol, 1 eq) in dichloromethane (7.5 mL) was slowly added. The mixture was stirred at 25°C for 2 h. LCMS monitoring showed that the reaction raw material completely disappeared and the main peak was the target product. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] and lyophilized to obtain compound 24 (267.5 mg, 500.48 μmol, 51.06% yield, 97.42% purity) as a white solid. Prep-HPLC (column: Phenomenex luna C18 150*40mm*15um; mobile phase: [water (FA)-ACN]; gradient: 55%-85% B over 15min). LCMS: Retention time = 1.461 min, 521.3 [M + H] +. HPLC: retention time = 1.831 min, purity 97.42%. 1H NMR(400MHz,CHLOROFORM-d)δ=8.31(s,1H),8.08(s,1H),5.99(s,1H),3.46(s,3H),3.25(brd,J=12.8Hz,1H),3.17(br d,J=4.0Hz,1H),2.11-1.94(m,2H),1.85-1.78(m,5H),1.76-1.69(m,2H),1.61-1.53(m,3H),1.50(s,3H),1.39(br s,5H),1.30-1.26(m,4H),1.19(s,3H),1.04(d,J=6.4Hz,6H),0.93(s,3H).

[0254] ■Example 24

[0255] Compound 25:

[0256]

[0257] Synthesis route:

[0258]

[0259] ■Example 25

[0260] Compound 26:

[0261]

[0262] Synthesis route:

[0263]

[0264] Step 1: Synthesis of compound 26

[0265] Anhydrous tetrahydrofuran (2mL) and sodium bicarbonate (204.87mg, 2.44mmol, 94.89μL, 4eq) were added to material 2 (204.07mg, 1.83mmol, 3eq) in sequence, and the reaction solution was cooled in an ice-water bath. Material 1 (311mg, 609.68μmol, 1eq) was dissolved in anhydrous tetrahydrofuran (3mL) and then slowly added dropwise to the stirred reaction solution. The reaction solution was slowly warmed to room temperature and stirred for 5 hours. LCMS monitored the complete reaction of the raw materials. The reaction solution was filtered, the filter cake was washed with dichloromethane (20mL*3), and the filtrate was concentrated and separated by plate (petroleum ether / ethyl acetate = 2:1) to obtain a crude product. The crude product was then freeze-dried by reverse phase preparation to obtain compound 26 (13.6mg, 24.63μmol, 4.04% yield) as an off-white solid. Prep-HPLC:column:Phenomenex Luna C18 150*25mm*10um;mobile phase:[H2O(0.225%FA)-ACN];gradient:55%-85%B over 10.0min.LCMS:rt=0.669min,549.3[M+H] + .HPLC: rt = 3.810 min. 1 H NMR (400MHz, DMSO-d6) δ = 9.06 (s, 1H), 8.65 (s, 1H), 6.17 (s, 1H), 4.60 (quin, J = 6.6Hz, 1H), 3.19 (br d,J=3.2Hz,1H),3.10-2.98(m,1H),2.23-2.09(m,1H),1.89-1.74(m,4H),1.73-1.56(m,4H),1.48-1.36(m,5H) ,1.32-1.22(m,5H),1.17(s,3H),1.15-1.12(m,1H),1.11-1.02(m,10H),0.94(s,3H),0.90(s,3H),0.85(s,3H).

[0266] ■Example 26

[0267] Compound 27:

[0268]

[0269] Synthesis route:

[0270]

[0271] ■Example 27

[0272] Compound 28:

[0273]

[0274] Synthesis route:

[0275]

[0276] Step 1: Synthesis of compound 28

[0277] N, N-dimethylethylamine (126.68 mg, 980.19 μmol, 170.73 μL, 10 eq) was added to a solution of material 2 (28.68 mg, 294.06 μmol, 3 eq, HCl) in dichloromethane (0.5 mL), and then a solution of material 1 (50 mg, 98.02 μmol, 1 eq) in dichloromethane (0.5 mL) was added at 0 ° C. The reaction solution was reacted at 25 ° C for 1 hour. LCMS monitoring showed that the reaction raw material completely disappeared and the main peak was the target product. The reaction solution was concentrated and separated by a large plate (petroleum ether: ethyl acetate = 1: 1) to obtain a crude product. The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] to obtain compound 28 (4.0 mg, 7.27 μmol, yield 7.42%, purity 97.25%) as a white solid. Prep-HPLC:column:Phenomenex Luna C18 150*25mm*10um; mobile phase:[H 2 O(0.225%FA)-ACN]; gradient: 50%-80%B over 10.0minLCMS: rt=0.671min, 535.4[M+H]+. HPLC: rt=2.253min. 1H NMR (400MHz, DMSO-d6)δ=9.50(s,1H),8.65(s,1H),6.18(s,1H),3.66-3.54(m,1H),3.51-3.40(m,1H),3.18(br s,1H),3.11-2.98(m,1H),2.16-2.07(m,1H),1.87-1.74(m,4H),1.71-1.58(m,4H),1.48-1.37(m,5H), 1.34-1.25(m,2H),1.24(s,3H),1.17(s,3H),1.13-1.04(m,8H),0.94(s,3H),0.90(s,3H),0.85(s,3H).

[0278] ■Example 28

[0279] Compound 29:

[0280]

[0281] Synthesis route:

[0282]

[0283] Step 1: Synthesis of compound 29

[0284] N, N-dimethylethylamine (380.04 mg, 2.94 mmol, 512.18 μL, 5 eq) was added to a dichloromethane (5 mL) solution of material 2 (267.53 mg, 1.76 mmol, 3 eq, HCl), and then a dichloromethane (2 mL) solution of material 1 (300 mg, 588.11 μmol, 1 eq) was added at 0 ° C. The reaction solution was reacted at 25 ° C for 1 hour. LCMS monitoring showed that the reaction raw material completely disappeared and the peak was the target product. The reaction solution was concentrated and a crude product was obtained by prep-TLC (petroleum ether: ethyl acetate = 1: 1). The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] to obtain compound 29 (10.70 mg, 18.00 μmol, yield 3.06%, purity 99.04%) as a white solid. Prep-HPLC:column:Phenomenex Luna C18 150*25mm*10um; mobile phase:[H 2 O(0.225%FA)-ACN]; gradient:60%-90%B over 10.0min.LCMS: rt=0.703min,589.4[M+H] + .HPLC: rt = 2.583 min. 1H NMR (400MHz, DMSO-d6) δ = 9.09 (s, 1H), 8.65 (s, 1H), 6.17 (s, 1H), 4.27-4.14 (m, 1H), 3.19 (br d,J=3.8Hz,1H),3.11-2.96(m,1H),2.23-2.07(m,1H),1.83(br s,3H),1.78-1.63(m,6H),1.60-1.48(m,6H),1.48-1.40(m,5H),1.40-1.35(m,1H),1.31-1.2 1(m,7H),1.17(s,3H),1.13-1.08(m,2H),1.06(s,3H),0.94(s,3H),0.89(s,3H),0.85(s,3H).

[0285] ■Example 29

[0286] Compound 30:

[0287]

[0288] Synthesis route:

[0289]

[0290] Step 1: Synthesis of compound 30

[0291] N,N-dimethylethylamine (126.68 mg, 980.19 μmol, 170.73 μL, 5 eq) was added to a dichloromethane (2 mL) solution of compound 2 (93.87 mg, 588.11 μmol, 3 eq, HCl), and then a dichloromethane (1 mL) solution of compound 1 (100 mg, 196.04 μmol, 1 eq) was added at 0 ° C. The reaction solution was reacted at 25 ° C for 1 hour. LCMS monitoring showed that the reaction raw material completely disappeared and the main peak was the target product. The reaction solution was concentrated and a crude product was obtained by prep-TLC (petroleum ether: ethyl acetate = 1:1). The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] to obtain compound 30 (3.20 mg, 5.14 μmol, yield 2.62%, purity 95.93%) as a white solid. Prep-HPLC:column:Phenomenex Luna C18 150*25mm*10μm; mobile phase:[H 2 O(0.225%FA)-ACN]; gradient: 55%-85%B over 10.0min. LCMS: rt=0.725min, 597.3[M+H]+. HPLC: rt=2.419min. 1H NMR (400MHz, DMSO-d6) δ = 9.80 (s, 1H), 8.64 (s, 1H), 7.35-7.22 (m, 5H), 6.18 (s, 1H), 4.84-4.58 (m, 2H), 3.16 (br s,1H),3.12-3.01(m,1H),2.19-2.08(m,1H),1.89-1.76(m,4H),1.75-1.56(m,4H),1.47-1.40(m ,4H),1.37-1.25(m,3H),1.20-1.10(m,7H),1.06(s,4H),0.94(s,3H),0.91(s,3H),0.86(s,3H).

[0292] ■Example 30

[0293] Compound 31:

[0294]

[0295] Synthesis route:

[0296]

[0297] Step 1: Synthesis of intermediate 2

[0298] Triethylamine (1.67 g, 16.52 mmol, 2.30 mL, 1.1 eq) and tert-butyldiphenylsilyl chloride (4.13 g, 15.02 mmol, 3.84 mL, 1 eq) were added to a solution of material 1 (2 g, 15.02 mmol, 1 eq) in dichloromethane (20 mL). The reaction solution was reacted at 25 ° C for 12 hours. LCMS monitoring showed that the reaction raw material completely disappeared and the main peak was the target product. The reaction solution was concentrated under reduced pressure, and the crude product was purified by normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 1: 0 to 100: 1) and then concentrated under reduced pressure to obtain intermediate 2 (5.5 g, 14.06 mmol, yield 93.62%, purity 95%), which was a colorless oil. LCMS: Rt = 0.711 min, 394.2 [M + Na] + ESI 1 HNMR(400MHz,CHLOROFORM-d)δ=7.80-7.71(m,4H),7.49-7.37(m,6H),6.71(s,1H),1.38(s,9H),1.15(s,9H).

[0299] Step 2: Synthesis of intermediate 4

[0300] To a tetrahydrofuran (30 mL) solution of intermediate 2 (2 g, 5.38 mmol, 1 eq), sodium hydrogen (322.98 mg, 8.07 mmol, 60% purity, 1.5 eq) was slowly added at 0 ° C. The reaction solution was reacted at 0 ° C for 30 minutes, and then material 3 (3.12 g, 13.46 mmol, 2.5 eq) was slowly added. The reaction solution was reacted at 25 ° C for 12 hours. LCMS monitoring showed that the reaction raw materials disappeared completely and the main peak was the target product. The reaction solution was slowly poured into a saturated aqueous ammonium chloride solution (20 mL) and extracted with ethyl acetate (20 mL*2). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by normal phase silica gel column chromatography (petroleum ether: ethyl acetate = 1:0) and concentrated under reduced pressure to obtain intermediate 4 (1.87 g, 3.71 mmol, yield 68.93%, purity 90%), which was a colorless oil. LCMS: Rt=0.801min,476.2[M+Na] +. ESI 1 H NMR (400MHz, CHLOROFORM-d)δ=7.74-7.69(m,4H),7.48-7.43(m,2H),7.41-7.37(m,4H),3.93-3.82(m,2H),1.26(s,9H),1.17(s,9H).

[0301] Step 3: Synthesis of intermediate 5

[0302] To intermediate 4 (500 mg, 1.10 mmol, 1 eq) was added ethyl acetate hydrochloride solution (2 M, 5.00 mL, 9.07 eq), and the reaction solution was reacted at 25 ° C for 12 hours. TLC monitoring showed that the reaction raw material completely disappeared. The reaction solution was concentrated under reduced pressure, and the filter cake was filtered and concentrated with petroleum ether (2 mL). Intermediate 5 (50 mg, 330.00 μmol, 29.94% yield, HCl) was obtained as a white solid. 1 HNMR (400MHz, DMSO-d6) δ = 3.94-3.81 (m, 2H).

[0303] Step 4: Synthesis of compound 31

[0304] N,N-dimethylethylamine (63.34 mg, 490.09 μmol, 85.36 μL, 5 eq) was added to a solution of intermediate 5 (44.55 mg, 294.06 μmol, 3 eq, HCl) in dichloromethane (0.5 mL), and then a solution of material 6 (50 mg, 98.02 μmol, 1 eq) in dichloromethane (0.5 mL) was added at 0 ° C. The reaction solution was reacted at 25 ° C for 1 hour. LCMS monitoring showed that the reaction raw material completely disappeared and the main peak was the target product. The reaction solution was concentrated and a crude product was obtained by prep-TLC (petroleum ether: ethyl acetate = 2: 1). The crude product was separated and purified by reverse phase preparation [water (formic acid)-acetonitrile system] to obtain compound 31 (7.48 mg, 12.58 μmol, yield 12.84%, purity 99.04%) as a white solid. Prep-HPLC:column:Phenomenex Luna C18 150*25mm*10um; mobile phase:[H 2 O(0.225%FA)-ACN]; gradient:53%-83%B over10.0min.LCMS: rt=0.661min,589.4[M+H] + .HPLC: rt = 2.969 min. 1 H NMR (400MHz, DMSO-d6) δ = 10.20 (s, 1H), 8.65 (s, 1H), 6.19 (s, 1H), 4.63-4.4 4(m,1H),4.28-4.10(m,1H),3.14-2.97(m,2H),2.19-2.07(m,1H),1.93-1.7 6(m,4H),1.74-1.59(m,4H),1.46-1.41(m,4H),1.40-1.26(m,3H),1.22(s, 3H),1.19-1.10(m,5H),1.06(s,3H),0.95(s,3H),0.90(s,3H),0.86(s,3H).

[0305] ■Example 31

[0306] Compound 32:

[0307]

[0308] Synthesis route:

[0309]

[0310] ■Example 32

[0311] Compound 33:

[0312]

[0313] Synthesis route:

[0314]

[0315] ■Example 33

[0316] Compound 34:

[0317]

[0318] Synthesis route:

[0319]

[0320] ■Example 34

[0321] Compound 35:

[0322]

[0323] Synthesis route:

[0324]

[0325] ■Example 35

[0326] Biological evaluation

[0327] Test Example 1 In vitro human Nrf2 receptor agonist activity assay

[0328] Reagents: Opti-MEM serum-free medium; PEI 40000 transfection reagent; DMEM complete medium.

[0329] Equipment: Cx7Pro high-content rapid imaging platform, etc.

[0330] For each well of cells, use 10 μL Opti-MEM serum-free medium (Cienry) to dilute 360 ​​ng of Nrf2 phase change probe plasmid, mix thoroughly to make DNA dilution solution, let stand for 5 minutes, use 10 μL Opti-MEM serum-free medium to dilute 0.6 μL of PEI 40000 transfection reagent (YEASEN), mix thoroughly to make PEI 40000 dilution solution, let stand for 5 minutes. After mixing the DNA dilution solution and PEI 40000 dilution solution, incubate at room temperature for 20 minutes to form a DNA-PEI cationic nucleic acid transfection reagent complex. Remove 20 μL of cell growth medium and add 20 μL of DNA-PEI cationic nucleic acid transfection reagent complex to each well. Shake the culture plate and mix gently. 37°C, 5% CO 2Cultured in an incubator, 1 hour after transfection, remove 75 μL of cell growth medium, add 75 μL of fresh preheated DMEM complete medium (Meilunbio) to each well, and maintain the volume of each well at 100 μL. 18 hours after transfection, the gradient dilution of drugs (0.0625nM, 0.125nM, 0.25nM, 0.5nM, 1nM, 2nM, 4nM, 8nM, 16nM, 32nM, 128Nm) was first mixed into the preheated DMEM complete medium, and 100 μL of medium containing gradient dilution of drugs was added to the sample wells, and the final volume of each sample well was 200 μL. 6 hours after adding the drug, the Cx7Pro high-content rapid imaging platform (Thermo Fisher) was used with a 20x high-definition lens to randomly select 16 fields of view from each well for photography. The position of the cell nucleus was determined by the NLS-mTagBFP2 independently expressed in the probe, thereby determining the number of transfected cells. Calculate the total fluorescence intensity of the Nrf2 phase change probe phase transition. Count the total fluorescence intensity of the Nrf2 phase change probe phase change "droplet" in each cell. Compare the total fluorescence intensity of the Nrf2 phase change probe phase transition in each cell in the drug group with the total fluorescence intensity of the Nrf2 phase change probe phase transition in each cell in the DMSO group. Use the log(agonist) vs.response--Variable slope(fourparameters) 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:

[0331]

[0332] Table 1 Test results of representative compounds of the present invention on in vitro human Nrf2 receptor agonist activity

[0333]

[0334]

[0335] The biological experimental data are shown in Table 1. Moreover, the test experimental data show that, in particular, multiple molecules such as compounds 1, 4, 5, 11, 16, 17, 19, 24, 26, 28, 29, 30, 31 prepared by the present invention all exhibit single-digit nanomolar human Nrf2 receptor agonist activity, and are significantly better than 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 activators.

[0336] In addition, control experiments with the control drug Omaveloxolone showed that both the compound of the present invention and the control drug Omaveloxolone have human Nrf2 receptor agonist activity, but there are significant differences in the activity of the compound in terms of DPPH free radical scavenging and MDA anti-lipid peroxidation, as shown in Test Examples 2-4.

[0337] Test Example 2 DPPH free radical scavenging ability test

[0338] Purpose of the experiment: To determine the DPPH free radical scavenging ability of the compounds of the present invention.

[0339] Test Materials:

[0340] Material factory Part Number Dimethyl sulfoxide Sigma D2650 D4Plus Sample Head Carrier Tecan 30097371 T8Plus Sample Head Carrier Tecan 30097370 96-well plate Corning 3599 Anhydrous ethanol Bioengineering A500737 2,2-Diphenyl-1-picrylhydrazyl(DPPH) MCE HY-112053

[0341] Test equipment:

[0342]

[0343]

[0344] Test method:

[0345] First, add 20 μL of the compound to be tested to a 96-well plate and dilute it continuously in 1:2 with DMSO. Then add 200 μL of 200 μM DPPH prepared with anhydrous ethanol as solvent to each well, shake it slightly and incubate it at room temperature in the dark for 30 minutes. Detect the absorbance value at 517 nm with an ELISA reader. Calculate the DPPH clearance rate using the following formula: DPPH clearance rate % = (1-Ai / A0) * 100%, Ai refers to the sample absorbance value, and A0 refers to the DMSO control group absorbance value. Use XLfit 5.3.1.3 software to process the data, and use the nonlinear fitting formula to obtain the IC50 value of the compound. The results are shown in Table 2.

[0346] Table 2 DPPH free radical scavenging ability test results of representative compounds of the present invention

[0347]

[0348] The results showed that the representative compounds 19 and 24 prepared by the present invention had the ability to scavenge DPPH free radicals, while Omaveloxolone had no such activity.

[0349] Test Example 3 MDA Anti-lipid Peroxidation Ability Test

[0350] Experimental purpose: To test the MDA anti-lipid peroxidation ability of the compounds of the present invention.

[0351] Experimental Materials:

[0352] Material factory Part Number DPBS (1×) Corning 21-031-CVC Dimethyl sulfoxide Sigma D2650 D4Plus Sample Head Carrier Tecan 30097371 T8Plus Sample Head Carrier Tecan 30097370 96-well plate Corning 3599 ascorbic acid ST1434 Blue Sky Lipid oxidation (MDA) detection kit S0131M Blue Sky

[0353] Test equipment:

[0354]

[0355]

[0356] Test method:

[0357] First, extract brain tissue homogenate: an adult male SD rat was anesthetized with isoflurane and then killed by dislocating the cervical vertebrae. The whole brain was removed and washed twice in DPBS. The meninges were peeled off and transferred to a 50mL centrifuge tube containing 10mL DPBS. After being cut with scissors, they were divided into 10 1.5mL centrifuge tubes. Three grinding beads were added to each tube and ground at 90Hz for 60min for 3 times. The ground tissue homogenate was transferred to the same new 50mL centrifuge tube, and DPBS was added to a total volume of 30mL and mixed. Next, 20μL of the test compound was added to the 96-well plate and diluted continuously 1:3 with DMSO. Then 100μL of brain tissue homogenate, 50μL DPBS and 50μL of 200μg / ml vitamin C were added in sequence, and a series of concentration standards were prepared as standard curves. After shaking, incubate at 37℃ for 1 hour, then add 400μL MDA working solution and heat at 100℃ for 15 minutes. After cooling to room temperature, centrifuge at 1000g for 10 minutes, aspirate 200μL of supernatant into another new plate, and detect the absorbance at 532nm using an enzyme reader. The clearance rate of the compound on MDA was calculated using the following formula: MDA clearance rate % = [((A1-A0)-(A2-A0)) / ((A1-A0)-(A3-A0))]*100%, where A1 refers to the absorbance value of the high control group, A2 refers to the sample absorbance value, A3 refers to the absorbance value of the low control group, and A0 refers to the absorbance value of the blank group. The data were processed using XLfit5.3.1.3 software, and the IC value of the compound was obtained using the nonlinear fitting formula. 50 The results are shown in Table 3.

[0358] Table 3 Test results of MDA anti-lipid peroxidation ability of representative compounds of the present invention

[0359]

[0360] The results show that the compounds 19, 24, 26 and 28 prepared in the present invention have anti-lipid peroxidation ability, and the activity is better than that of Edaravone, while Omaveloxolone has no such activity.

[0361] Experimental Example 4: Intervention Effect of Representative Compounds on Ferroptosis Process (qPCR)

[0362] 1) Experimental purpose: To test the intervention effect of a series of compounds on the ferroptosis process by detecting changes in the expression levels of key genes in the ferroptosis signaling pathway at the cellular level. The specific indicator is the detection of the mRNA level of PTGS2, a key gene in the ferroptosis process (fluorescence real-time quantitative qRT-PCR).

[0363] 2) Experimental methods:

[0364] HT-1080 cells (Cat. No.: CCL-121, Shanghai Cell Bank, introduced from ATCC) were plated in six-well plates at 4×105 / well for 22h; HT-1080 cells were treated with different concentration gradient compounds (1.37nM to 333nM, a total of 6 concentration gradients) for 1h in advance; HT-1080 cells were treated with 200nM of the classic ferroptosis inducer RSL3 (Cat. No.: HY-100218A, MCE, USA) for 16h; mRNA was extracted and reverse transcribed into cDNA, and PTGS2 mRNA levels were detected by qRT-PCR (Model: ABI7500, Thermo Fisher, USA). The ferroptosis inhibitor Fer-1 (Cat. No.: HY-100579, MCE, USA) was used as a positive control.

[0365] 3) Experimental results:

[0366] The IC of each drug was calculated using the log(agonist) vs. response--Variable slope(four parameters) analysis method in GraphPad Prism. 50 , the results are shown in Table 4.

[0367] Table 4 Intervention effect of the compounds of the present invention on the ferroptosis process

[0368]

[0369] The results showed that the representative compounds 26 and 28 prepared in the present invention can inhibit ferroptosis, while Omaveloxolone has no such activity.

[0370] The above-mentioned pharmacological experiments prove that the preferred NRF2-Keap1 compounds prepared by the present invention, such as 19, 24, 26, 28, etc., in addition to maintaining Nrf2 agonist activity similar to the marketed Nrf2 agonist Omaveloxolone, also have the effects of scavenging DPPH free radicals, inhibiting the production of lipid peroxide MDA or intervening in ferroptosis.

[0371] At the same time, the present invention has been verified by biological in vivo tests to show that the compounds of the present invention have significant effects on a variety of diseases, including: cerebral small vessel disease, mitochondrial encephalopathy, autism spectrum disorder, Rett syndrome, Friedreich's ataxia, stroke, hemorrhagic stroke, ischemic stroke, multiple sclerosis, amyotrophic lateral sclerosis, schizophrenia, schizophrenia cognitive impairment, Parkinson's disease, Parkinson's cognitive impairment, Alzheimer's disease, vascular dementia, epilepsy, Huntington's disease, heart failure, myocardial infarction, renal failure, renal ischemia. In particular, it has significant effects on diseases such as stroke, multiple sclerosis, and amyotrophic lateral sclerosis, and can be used as a drug for the preparation of drugs for preventing or treating stroke, multiple sclerosis, and amyotrophic lateral sclerosis.

[0372] Experimental Example 5: Tissue distribution experiment of representative compounds in SD rats

[0373] Test animals: SD rats, male, weighing about 250±20g, were randomly divided into groups, 3 rats in each group.

[0374] Vehicle: DMSO: 10% solution (5%: 95%).

[0375] Test method:

[0376] The drug was administered by gavage at a dose of 20 mg / kg (30 mg / kg for compound 30). The animals were fasted for 12 hours before administration and had free access to water. Plasma was collected at a specific time after administration and brain tissue was collected after cardiac perfusion. After pretreatment, LC-MS / MS was used for analysis to determine the concentration of the analyte in plasma and brain. The test results are shown in Table 4:

[0377] Table 4: Plasma and brain concentrations in SD rats after administration

[0378]

[0379] Experimental Example 6 Protective Effects of Representative Compounds on Ischemic Stroke

[0380] 1) Reagents: Omaveloxolone, MedChemexpress Biotechnology, USA; Transient Middle Cerebral Artery Occlusion Model (tMCAO) suture, Beijing Reward Life Science Co., Ltd.; DMAO, Bio-Technology Co., Ltd.; Solutal, Sigma-Aldrich (Shanghai) Trading Co., Ltd.; Normal saline, Sinopharm Chemical Reagent Co., Ltd.; TTC, Sigma-Aldrich (Shanghai) Trading Co., Ltd.

[0381] 2) Establishment of cerebral ischemia rat model by tMCAO method:

[0382] Rats were anesthetized with 10% chloral hydrate (350 mg / kg), fixed on the operating table in the supine position, and the neck was disinfected. A midline incision was made in the neck, and the intermuscular space between the left sternocleidomastoid muscle and the sterno-glossus muscle was bluntly separated to expose the common carotid artery. The common carotid artery was picked out by ophthalmic curved forceps and threaded with silk thread for later use. The external carotid artery was separated under the right digastric muscle and hyoid bone, and the thread was threaded for later use. The suture was passed through and tied to the common carotid artery, and a small incision was made at the free end of the common carotid artery and a loose knot was tied. The thread plug was inserted along the common carotid artery through the incision, and the thread plug was slowly pushed forward. When resistance was felt, it stopped, and the pre-tied loose knot was tied. The body temperature was maintained at 37°C during the operation, and the thread plug was pulled out after 1.5 hours. In the sham operation group, the thread plug was not inserted after the vascular separation, and the other operation steps were the same.

[0383] 3) Experimental grouping and drug administration:

[0384] 48 SD male rats, weighing 230-250g, were randomly divided into sham operation group, model group, Omaveloxolone 3mg / kg group, Omaveloxolone 9mg / kg group, compound 192.74mg / kg group, and compound 282.89mg / kg group. After 3 days of adaptive feeding, the rats were injected with drugs through the tail vein when the plug was inserted into the model. The sham operation group and the model group were injected with an equal volume of solvent (10% DMSO + 10% solutal + 80% saline).

[0385] 4) Index detection:

[0386] mNSS score: Neurobehavioral score was performed 24 hours after modeling, mainly to evaluate the neurological function of rats with ischemic stroke from the two aspects of sensation and movement.

[0387] Determination of cerebral infarction area: Rats were euthanized by overdose of chloral hydrate, and the brain was removed and placed in a -20°C refrigerator for 20 minutes. The brain was removed and placed in the brain trough, sliced, and the thickness of the brain slice was 2 mm. The brain slice was placed in a 2% TTC solution, shielded from light, and incubated in a 37°C constant temperature box for 15 minutes. After staining, pictures were taken for analysis of cerebral infarction area.

[0388] 5) Statistical methods:

[0389] All data in this study were expressed as mean ± standard deviation (Mean ± SD) and statistically analyzed using GraphPad Prism 7.0. Differences among multiple groups were evaluated by one-way ANOVAs followed by Tukey's test. Behavioral tests were tested by Krystal-Wallis test. P < 0.05 was considered statistically significant.

[0390] 6) Experimental results:

[0391] Effects of compounds 19 and 28 on neurological function in rats with ischemic stroke:

[0392] like Figure 1 As shown, Omaveloxolone 3mg / kg and 9mg / kg had no effect on the mNSS score of rats with cerebral ischemia. At the same molar dose as Omaveloxolone 3mg / kg, Compound 19 (2.74mg / kg) and Compound 28 (2.89mg / kg) could significantly reduce the mNSS score of rats with cerebral ischemia.

[0393] Effects of compounds 19 and 28 on cerebral infarction area in rats with ischemic stroke:

[0394] like Figure 2 As shown, Omaveloxolone 3mg / kg and 9mg / kg had no effect on the cerebral infarction area of ​​rats with cerebral ischemia. At the same molar dose as Omaveloxolone 3mg / kg, compound 19 (2.74mg / kg) and compound 28 (2.89mg / kg) could significantly reduce the cerebral infarction area of ​​rats with cerebral ischemia.

[0395] 7) Conclusion: The novel Nrf2-Keap1 uncouplers of the present invention, such as compounds 19 and 28, can reduce the cerebral infarction area in rats with ischemic stroke and have a protective effect on neurological damage.

[0396] Experimental Example 7 Effects of Representative Compounds on Neurobehavior in Multiple Sclerosis Model Mice

[0397] 1. Materials and Methods

[0398] 1) Main reagents

[0399]

[0400] 2) Experimental animals and grouping and drug administration

[0401] Female C57BL / 6J mice were randomly divided into 5 groups: sham operation group, model group, Omaveloxolone 5 mg / kg group, Omaveloxolone 15 mg / kg group, and compound 244.69 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). Each group was given twice a day for 42 consecutive days.

[0402] 3) Preparation of Multiple Sclerosis Model-EAE Model

[0403] The mixed emulsion containing myelin oligodendrocyte glycoprotein MOG35-55 peptide and complete Freund's adjuvant (CFA) containing Mycobacterium tuberculosis was injected subcutaneously on the back of mice, and pertussis toxin (PTX) was injected intraperitoneally on the day of immunization and 48 hours later to establish the EAE (experimental autoimmune encephalosporin) model.

[0404] 4) Weight and neurological function score (5-point scale)

[0405] Starting from the day of immune induction (Day 0), the rats were weighed and neurological function was scored daily. Neurological function scoring criteria (5-point method): 0 points, no clinical deficits; 1 point, partial tail paralysis; 2 points, complete tail paralysis; 3 points, partial hind limb paralysis; 4 points, complete hind limb paralysis; 5 points, forelimb paralysis; 6 points, death.

[0406] 5) Statistical processing

[0407] All data are presented as mean ± standard error (Mean ± SEM). Two-way ANOVA and Tukey's multiple comparison test were used to compare the differences in neurological function scores and body weight among the groups. All data were analyzed using GraphPad Prism 9.0.0 software. p < 0.05 indicated that the difference was statistically significant.

[0408] 2. Experimental results

[0409] like Figure 3 As shown, Omaveloxolone 5mg / kg and 15mg / kg can significantly reduce the neurological function scores of EAE model mice, but there is no significant difference between the two groups, indicating that Omaveloxolone 5mg / kg has the maximum effect. At the same molar dose as Omaveloxolone 5mg / kg, compound 24 (4.69mg / kg) can significantly reduce the neurological function scores of EAE model mice, and the effect intensity is better than Omaveloxolone 15mg / kg, and the difference is statistically significant.

[0410] like Figure 4 As shown, Omaveloxolone 5mg / kg and 15mg / kg can significantly increase the body weight of EAE model mice, but there is no significant difference between the two groups, indicating that Omaveloxolone 5mg / kg has the maximum effect. At the same molar dose as Omaveloxolone 5mg / kg, compound 24 (4.69mg / kg) can significantly increase the body weight of EAE model mice, and the effect intensity is better than Omaveloxolone 15mg / kg, and the difference is statistically significant.

[0411] 3. Experimental conclusion

[0412] Compound 24 has a protective effect on the neurological function of multiple sclerosis model mice, and its effect intensity is better than that of Omaveloxolone.

[0413] Test Example 8 Protective effect of representative compounds on amyotrophic lateral sclerosis model mice

[0414] 1. Materials and methods

[0415] 1) Main reagents

[0416] Omaveloxolone was purchased from MCE Biotech; Solutal was purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd.; and normal saline was purchased from Sinopharm Chemical Reagent Co., Ltd.

[0417] 2) Animals

[0418] Sixty 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.

[0419] 3) Experimental grouping and drug administration

[0420] SOD1 G93A mice were randomly divided into model group, Omaveloxolone 1mg / kg group, Omaveloxolone 3mg / kg group, Compound 2 60.99mg / kg group, Compound 3 11.06mg / kg, 12 mice in each group. At the same time, 12 C57BL / 6J mice were set as the control group. The control group and the model group were intraperitoneally injected with an equal volume of solvent (1% DMSO + 4% solutal + 95% saline), once a day for 10 consecutive weeks.

[0421] 4) Index detection

[0422] Rotarod test

[0423] The rotarod test is a classic behavioral experiment to evaluate the motor coordination of mice. The rotarod test (Panlab rotarod instrument, purchased from Harvard Bioscience, USA) was performed twice a week, and the rotation speed of the rotarod ranged from 4 to 40 r / min. The experimental steps are as follows: 1. Before the formal experiment, the mice were trained to adapt to the rotation speed, and the rotation speed was set to 12 r / min, 5 min each time, twice a day, for 3 days; 2. In the formal experiment, 3 consecutive tests were performed (3 min / time, with an interval of 30 min each time); 3. The time the mouse stayed on the rotarod each time was recorded, and the longest time on the rotarod in the 3 tests was taken as the latent period of falling.

[0424] Onset time

[0425] 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.

[0426] Cage experiment

[0427] The hanging cage test is used to evaluate the grip strength and endurance of the mouse's limbs. The hanging cage test is performed twice a week. Each mouse is placed in the center of the iron net, and the iron net is gently shaken to make the mouse hold it tightly. Then the iron net is slowly inverted to a horizontal position, and the time the mouse hangs on the iron net is recorded. Each mouse is measured three times, with an interval of 30 minutes between each test, and the maximum value is taken as the fall latency.

[0428] 2. Experimental results

[0429] 1) Effects of compounds 26 and 31 on the onset of disease in SOD1 G93A mice

[0430] like Figure 5 As shown, both Omaveloxolone 1mg / kg and 3mg / kg can significantly delay the onset of SOD1 G93A mice, but there is no significant difference between the two groups, indicating that Omaveloxolone 1mg / kg reaches the maximum effect. At the same molar dose as Omaveloxolone 1mg / kg, compound 26 (0.99mg / kg) and compound 31 (1.06mg / kg) can significantly delay the onset of SOD1 G93A mice, and the intensity of action is better than Omaveloxolone 3mg / kg, and the difference is statistically significant.

[0431] 2) Effects of compounds 26 and 31 on motor coordination ability of SOD1 G93A mice

[0432] like Figure 6 As shown, both Omaveloxolone 1mg / kg and 3mg / kg can significantly improve the motor coordination ability of SOD1 G93A mice, which is manifested by a significant increase in the latency to fall, but there is no significant difference between the two groups, indicating that Omaveloxolone 1mg / kg has reached its maximum effect. At the same molar dose as Omaveloxolone 1mg / kg, compound 26 (0.99mg / kg) and compound 31 (1.06mg / kg) can significantly increase the latency to fall of mice, and the intensity of action is better than Omaveloxolone 3mg / kg, and the difference is statistically significant.

[0433] 3) Effects of compounds 26 and 31 on muscle endurance in SOD1 G93A mice

[0434] like Figure 7 As shown, both Omaveloxolone 1mg / kg and 3mg / kg can significantly improve the muscle endurance of SOD1 G93A mice, which is manifested by a significant increase in the latency to fall, but there is no significant difference between the two groups, indicating that Omaveloxolone 1mg / kg has reached its maximum effect. At equimolar doses with Omaveloxolone 1mg / kg, compound 26 (0.99mg / kg) and compound 31 (1.06mg / kg) can significantly increase the latency to fall of mice, and the intensity of action is better than Omaveloxolone 3mg / kg, and the difference is statistically significant.

[0435] 3. Experimental conclusion

[0436] Compounds 26 and 31 can improve the neurobehavior of ALS model mice, and the effect intensity is better than Omaveloxolone.

[0437] In summary, the compounds involved in the present invention, as novel NRF2 activators, have excellent Nrf2 agonism, and the compounds exert antioxidant effects through new mechanisms such as scavenging DPPH free radicals or inhibiting the generation of lipid peroxide MDA. Further, the in vivo experiments of the present invention also show that the compounds provided by the present invention can be more effectively used for the prevention or treatment of related 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, some compounds have obvious effects on the treatment and / or prevention of stroke, multiple sclerosis, and amyotrophic lateral sclerosis.

[0438] 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, various equivalent transformations can be made to the technical solutions of the present invention, and these equivalent transformations all belong to the protection scope of the present invention.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, characterized in that As shown below: in: R1 is independently selected from: -C(=O)-alkane, -C(=O)-substituted alkane, -C(=O)-alkene, -C(=O)-substituted alkene, -C(=O)-alkyne, -C(=O)-substituted alkyne, -C(=O)-heteroarene, -C(=O)-heteroarene-R1', -C(=O)-heteroarene-R1', -C(=O)-NH-OH, -C(=O)-N(OH )-alkane, -C(=O)-N(OH)-substituted alkane, -C(=O)-N(OH)-olefin, -C(=O)-N(OH)-substituted olefin, -C(=O)-N(OH)-alkyne, -C(=O)-N(OH)-substituted alkyne, -C(=O)-N(OH)-arene, -C(=O)-N(OH)-arenediyl-R1', -C(=O)-N(OH) -heteroarene, -C(=O)-N(OH)-heteroarene-diyl-R1', -C(=O)-NH-arene, -C(=O)-NH-arene-diyl-R1', -C(=O)-NH-heteroarene, -C(=O)-NH-heteroarene-diyl-R1', -C(=O)-O-arene, -C(=O)-O-arene-diyl-R1', -C(=O)-O-heteroarene, -C(= -C(=O)-O-heteroarene-diyl-R1', -C(=O)-CH2-heteroarene, -C(=O)-CH2-heteroarene-diyl-R1', -C(=O)-CR2'R3'-heteroarene, -C(=O)-CR2'R3'-heteroarene-diyl-R1', -C(=O)-L-type amino acid-NH-heteroarene, -C(=O)-L-type amino acid-NH-heteroarene-diyl-R1'; And R2: hydrogen or methyl; R3: hydrogen or methyl.

2. The compound according to claim 1, characterized in that The compound is: in: R1 is independently selected from: -C(=O)-alkane, -C(=O)-substituted alkane, -C(=O)-alkene, -C(=O)-substituted alkene, -C(=O)-alkyne, -C(=O)-substituted alkyne, -C(=O)-heteroarene, -C(=O)-heteroarene-R1', -C(=O)-heteroarene-R1', -C(=O)-NH-OH, -C(=O)-N(OH )-alkane, -C(=O)-N(OH)-substituted alkane, -C(=O)-N(OH)-olefin, -C(=O)-N(OH)-substituted olefin, -C(=O)-N(OH)-alkyne, -C(=O)-N(OH)-substituted alkyne, -C(=O)-N(OH)-arene, -C(=O)-N(OH)-arenediyl-R1', -C(=O)-N(OH) -heteroarene, -C(=O)-N(OH)-heteroarene-diyl-R1', -C(=O)-NH-arene, -C(=O)-NH-arene-diyl-R1', -C(=O)-NH-heteroarene, -C(=O)-NH-heteroarene-diyl-R1', -C(=O)-O-arene, -C(=O)-O-arene-diyl-R1', -C(=O)-O-heteroarene, -C(= -C(=O)-O-heteroarene-diyl-R1', -C(=O)-CH2-heteroarene, -C(=O)-CH2-heteroarene-diyl-R1', -C(=O)-CR2'R3'-heteroarene, -C(=O)-CR2'R3'-heteroarene-diyl-R1', -C(=O)-L-type amino acid-NH-heteroarene, -C(=O)-L-type amino acid-NH-heteroarene-diyl-R1'; And R2: methyl; R3: methyl.

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

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

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

7. Use of the compound according to claim 6 for preparing an NRF2 activator.

8. Use of the compound according to claim 6 for preparing a medicament for treating and / or preventing a patient's disease, characterized in that The prepared drugs are used to prevent or treat patient 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 or treating stroke, multiple sclerosis and amyotrophic lateral sclerosis.

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