Arylalkylamine ferroptosis inhibitor based on edaravone structure, and preparation method therefor and use thereof

Abstract

An arylalkylamine ferroptosis inhibitor based on an edaravone structure, and a preparation method therefor and a use thereof. The structural formula of the ferroptosis inhibitor is as shown in the following formula I, formula II, or formula III: formula I, formula II, formula III. The ferroptosis inhibitor has strong ROS scavenging ability and an inhibitory effect on lipid peroxidation, can inhibit ferroptosis caused by an ferroptosis inducer, and can reduce cerebral ischemia and nerve damage caused by cerebral ischemia, and alleviate symptoms of neurological diseases such as Parkinson's syndrome. On the basis of the ferrostain-1 / edaravone structure, the provided arylalkylamine ferroptosis inhibitor can exert a synergistic effect by means of multiple effects to treat ferroptosis-related diseases and can also expand the use of edaravone, thereby providing a candidate compound for subsequent drug research and development.

Description

An aromatic alkylamine ferroptosis inhibitor based on edaravone structure, its preparation method and application Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to an aromatic alkylamine ferroptosis inhibitor based on the edaravone structure, its preparation method, and its application. Background Technology

[0002] Ferroptosis, a novel cell death pathway discovered in recent years, is an oxidative cell death induced by various factors. It is iron-dependent and occurs due to an imbalance between the generation and degradation of intracellular lipid reactive oxygen species (ROS). In the early years after its discovery, the mechanisms controlling ferroptosis primarily revolved around cysteine ​​and glutathione metabolism, and the phospholipid peroxidase GPX4's prevention of lipid peroxidation. Ferroptosis inducers act directly or indirectly on glutathione peroxidases (GPXs) through different pathways, leading to decreased cellular antioxidant capacity, ROS accumulation, and ultimately oxidative cell death. The complex interactions between lipid, iron, and cysteine ​​metabolism have become important regulators of this cell death pathway. Recently, the regulation of ferroptosis has become an attractive strategy for intervening in human diseases, including cancer, neurodegenerative diseases, and ischemic diseases. Ferroptosis can be inhibited by iron chelators and lipophilic antioxidants such as ferrostatin-1 (Fer-1). Fer-1 is an aryl alkylamine with antioxidant properties and was one of the first inhibitors of ferroptosis. As a lipid peroxidation agent, Fer-1 intercepts and scavenges lipid free radicals through hydrogen atom transfer or direct reduction. The specific structural formula of Fer-1 is as follows:

[0003] Edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) is a neuroprotective agent (free radical scavenger) developed by Takeda Pharmaceutical Company of Japan. Mechanistic studies suggest that edaravone can scavenge free radicals and inhibit lipid peroxidation, thereby inhibiting oxidative damage to brain cells, vascular endothelial cells, and nerve cells. Clinical studies indicate that N-acetylaspartate (NAA) is a specific marker of surviving nerve cells, and its content decreases sharply in the early stages of cerebral infarction. Administration of edaravone to patients in the acute phase of cerebral infarction can inhibit the reduction of local cerebral blood flow around the infarction, resulting in a significantly higher NAA content in the brain on day 28 after onset compared to the glycerol control group. Preclinical studies suggest that intravenous administration of edaravone to rats after ischemia / ischemia-reperfusion can prevent the progression of cerebral edema and cerebral infarction, alleviate accompanying neurological symptoms, and inhibit delayed neuronal death. The specific structure of edaravone is as follows: Summary of the Invention

[0004] To address the shortcomings of Fer-1's poor efficacy and edaravone's inability to inhibit ferroptosis despite its free radical scavenging ability, this invention provides an aromatic alkylamine ferroptosis inhibitor based on the edaravone structure, its preparation method, and its applications. This invention utilizes a rational structural design-drug twinning strategy to fuse two components into a single molecule, resulting in a bifunctional ferroptosis inhibitor. This bifunctional inhibitor exhibits strong ROS scavenging ability, inhibits lipid peroxidation, reverses cell damage induced by ferroptosis inducers (RSL3 / Erastin), alleviates cerebral ischemia and its associated neurological damage, and relieves symptoms of neurological diseases such as Parkinson's syndrome, providing a novel compound entity for clinical research.

[0005] To achieve the above-mentioned objectives, the present invention employs the following technical solution:

[0006] This invention provides an aromatic alkylamine ferroptosis inhibitor based on the edaravone structure, the structural formula of which is shown in Formula I, Formula II or Formula III below:

[0007] In the formula, R 1 Selected from hydrogen, alkyl, aryl, C1 to C6 alkyl-aryl, C1 to C6 alkyl-phenolic or C3 to C4 10 cycloalkyl;

[0008] R 2 Selected from C0-C8 alkyl, C3-C 12 cycloalkyl, adamantyl, or polyacetylenic;

[0009] R 3 Selected from hydrogen, alkyl, aryl, C1 to C6 alkyl-aryl, C1 to C6 alkyl-phenolic or C3 to C4 10 cycloalkyl;

[0010] When ring A is an aromatic ring, R 4 R 5 R 6 R 7 and R 8 All are selected from H, C1-C6 alkyl, OH, OCH3, OCH(CH3)2, OCH2CH(CH3)2, OC(CH3)3, O(C1-C6 alkyl), OCF3, OCH2CH2OH, O(C1-C6 alkyl)OH, F, Cl, Br, I, CF3, CN, NO2, NH2, C1-C6 heteroalkyl, C1-C6 hydroxyalkyl, C i -C6 alkoxy, C1-C6 alkyl, aryl, aromatic heteroyl, C3-C7 cycloalkyl, heterocyclic alkyl, alkylaryl, CO2Ra C(O)R a NH (C1-C4 alkyl), N (C1-C4 alkyl)2, NH (C3-C7 cycloalkyl), NHC(O)(C1-C4 alkyl), CONR a ,NC(O)R a NS(O) 1 / 2 R a S(O) 1 / 2 NR a S(O) 1 / 2 R, C(O)O (C1-C4 alkyl), OC(O)N (R) a 2. C(O) (C1-C4 alkyl) or C(O)NH (C1-C4 alkyl); wherein R 4 R 5 R 6 R 7 and R 8 It is one of the above substituents, or R 4 R 5 R 6 R 7 and R 8 Two, three, or four of the substituents are selected from the above-mentioned substituents;

[0011] Among them, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, phenyl, benzyl, naphthyl, C5-C 10 Aromatic heterocyclic groups, C3-C7 saturated heterocyclic groups, C3-C 12 Cycloalkyl, polyynyl, aryl, C1-C6 alkyl-aryl, C1-C6 alkyl-phenolic or C3-C 10 Cycloalkyl groups can be substituted by one or more atoms or groups;

[0012] Where R a The group is selected from H, CH3, CH2CH3, C3-C6 alkyl, C1-C6 haloalkyl or optionally substituted aryl, alkylaryl, piperazinyl, piperidinyl, morpholinyl, heterocyclic alkyl, heteroaryl, C1-C6 alkoxy, NH (C1-C4 alkyl) and N (C1-C4 alkyl)2, wherein the optionally substituted group is selected from C1-C6 alkyl or C2-C7 acrylate.

[0013] Furthermore, the inhibitors are compounds I-1 to I-40, II-1 to II-3, and III-1 to III-3, whose structural formulas are as follows:

[0014] This invention also provides a method for preparing the aforementioned aromatic alkylamine ferroptosis inhibitor based on the edaravone structure, the method comprising the following steps:

[0015] The synthesis of compounds I-1 to I-40 is as follows: Compound 1 and Compound 2 are reacted in glacial acetic acid solution at 100°C to give edaravone analog 3; Compound 4 and the corresponding amine undergo a substitution reaction to give Compound 5, which is then subjected to hydrolysis and condensation to give the key intermediate compound 7; Compounds I-1 to I-23 are obtained by reducing Compound 7 on palladium on carbon and hydrogen, and then undergoing a reductive amination reaction with the corresponding amine aldehyde or ketone in the presence of sodium triacetoxyborohydride to give the target compounds I-24 to I-40.

[0016] Reagents and reaction conditions: (a) Glacial acetic acid, 100℃, 18h.

[0017] Reagents and reaction conditions: (b) the corresponding amine, potassium carbonate, dimethyl sulfoxide, 80 °C, 8 h; (c) sodium hydroxide, ethanol, room temperature, 6 h; (d) HATU, N,N-diisopropylethylamine, compound 3, dichloromethane, room temperature, 4 h; (e) hydrogen, 10% palladium on carbon, methanol, room temperature, 8 h; (f) the corresponding aldehyde / ketone, sodium triacetoxyborohydride, dichloromethane, room temperature, 8 h.

[0018] The synthesis of compounds II-1 to II-3 is as follows: Compound 8, sodium nitrate, and stannous chloride undergo a diazotization reaction in hydrochloric acid solution to generate compound 9, which then undergoes a cyclization reaction with ethyl acetoacetate or its analogue to generate compound 10. Subsequently, compound 10 undergoes a substitution reaction and a reduction reaction to give the target compound II-1; II-2 and II-3 are obtained by reductive amination reaction of II-1 with the corresponding aldehyde and ketone.

[0019] Reagents and reaction conditions: (a) Sodium nitrate, stannous chloride, hydrochloric acid, 0℃–room temperature, 4 h; (b) Ethyl acetoacetate or its analogue, glacial acetic acid, 100℃, 18 h; (c) The corresponding amine, potassium carbonate, dimethyl sulfoxide, 80℃, 8 h; (d) Hydrogen, 10% palladium on carbon, methanol, room temperature, 8 h; (e) The corresponding aldehyde / ketone, sodium triacetoxyborohydride, dichloromethane, room temperature, 8 h.

[0020] The synthesis of compounds III-1 to III-3 is as follows: 4-chloro-3-nitrobenzaldehyde is used as the starting material, and its corresponding hydroxyl-containing compound 13 is obtained by reduction with sodium borohydride; compound 13 undergoes a substitution reaction with the corresponding amine to obtain compound 14, which is then oxidized in chloroform solution by pyridinium chlorochromate (PCC) to obtain compound 15; in acetic acid solution, compound 15 undergoes a condensation reaction with an edaravone analog to obtain the key intermediate compound 16; target compound III-1 is obtained by reduction of compound 16 with hydrogen, and compounds III-2 and III-3 are obtained by reductive amination reaction of III-1 with the corresponding aldehyde and ketone.

[0021] Reagents and reaction conditions: (a) Sodium borohydride, methanol, 0°C–room temperature, 2 h; (b) The corresponding amine, potassium carbonate, dimethyl sulfoxide, 150°C, 8 h; (c) Pyridinium chlorochromate, chloroform, room temperature, 12 h; (d) Compound 3, acetic acid, room temperature, 8 h; (e) Hydrogen, 10% palladium on carbon, methanol, room temperature, 8 h; (f) The corresponding aldehyde / ketone, sodium triacetoxyborohydride, dichloromethane, room temperature, 8 h.

[0022] The present invention also provides the use of the described inhibitor in the preparation of ferroptosis inhibitors and / or ferroptosis inhibitor-edaravone twins.

[0023] The present invention also provides the use of the described inhibitor in the preparation of treatments for ferroptosis-related diseases and / or diseases that can be treated with edaravone.

[0024] Furthermore, the iron death-related diseases include neurodegenerative diseases (including but not limited to Alzheimer's disease, Parkinson's syndrome, depression, etc.), tissue ischemia-reperfusion injury, stroke, cardiovascular disease, liver and kidney failure, inflammation, diabetic complications, and cardiovascular diseases.

[0025] Furthermore, the diseases that edaravone can treat include neurological diseases, including but not limited to stroke, chronic neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Parkinson's syndrome, cerebrovascular diseases, and lung diseases such as chronic obstructive pulmonary disease (COPD) and sepsis.

[0026] The present invention also provides a medicament for treating ferroptosis-related diseases and / or diseases treatable by edaravone, said medicament comprising the aforementioned aromatic alkylamine ferroptosis inhibitor and a pharmaceutically acceptable salt, carrier, or adjuvant.

[0027] Furthermore, the compounds treat ferroptosis because they act as free radical scavengers, reducing intracellular reactive oxygen species (ROS) and lipid peroxides, thereby rescuing ferroptosis caused by ROS; wherein ROS include, but are not limited to, H₂O₂, t-BuOOH, and O₂.2- ,OH·,ONOO - ,ClO - .

[0028] Furthermore, the treatment of ferroptosis with the compounds is that these compounds act as free radical scavengers to rescue ferroptosis caused by ferroptosis inducers; wherein ferroptosis inducers include, but are not limited to, RSL3 and its derivatives, Erastin and its derivatives, ML162 and its derivatives.

[0029] Furthermore, the compound can be used to treat both ferroptosis-related diseases and edaravone-treatable related diseases simultaneously.

[0030] Compared with the prior art, the advantages and beneficial technical effects of the present invention are as follows: the edaravone structure has a keto-enol tautomerism, as shown below. The present invention utilizes rational drug design to combine Fer-1 and edaravone structures to form a twin drug, increasing the drug-like properties of Fer-1 while endowing edaravone with anti-ferroptosis ability, for the treatment of ferroptosis-related diseases.

[0031] This invention synthesizes a novel ferroptosis inhibitor containing both a ferroptosis inhibitor pharmacophore and an edaravone core, enriching the structure of existing ferroptosis inhibitors. Experimental verification shows that the bifunctional ferroptosis inhibitor based on the ferrostain-1 / edaravone structure can effectively inhibit ferroptosis induced by ferroptosis inducers; experimental verification shows that the bifunctional inhibitor can reduce the content of intracellular reactive oxygen species and lipid peroxides; experimental verification shows that the bifunctional inhibitor exhibits good safety; and experimental verification shows that the bifunctional inhibitor can scavenge free radicals under cell-free conditions.

[0032] Therefore, the bifunctional ferroptosis inhibitor based on the ferrostain-1 / edaravone structure provided by this invention can exert a synergistic effect through multiple actions to treat ferroptosis-related diseases, and can also expand the application of edaravone, providing candidate compounds for subsequent drug development. Attached Figure Description

[0033] Figure 1 shows the experimental results of the free radical scavenging ability of the compound.

[0034] Figure 2 shows the toxicity test results of different compounds on HT22 cells, where the compound concentrations are 100 μM, 30 μM, and 10 μM.

[0035] Figure 3 shows the toxicity results of different compounds on HUVEC cells, with compound concentrations of 100 μM, 50 μM, and 25 μM. Detailed Implementation

[0036] The embodiments of the present invention are described in detail below. These embodiments are implemented based on the technical solution of the present invention, and provide detailed implementation methods and specific operation processes. However, the scope of protection of the present invention is not limited to the following embodiments.

[0037] Example 1

[0038] 1. 1-(4-Chlorophenyl)-3-methyl-1H-pyrazole-5-ol (Compound 3)

[0039] 4-Chlorophenylhydrazine (0.30 g, 1.67 mmol) and ethyl 3-oxobutyrate (0.19 g, 1.67 mmol) were added to a 25 mL round-bottom flask. 10 mL of glacial acetic acid was added as a solvent, and the mixture was reacted at 100 °C for 18 hours under N2 protection. After the reaction was complete, the reaction mixture was cooled to room temperature and poured into a conical flask containing 100 mL of water. The aqueous layer was extracted three times with ethyl acetate, the organic layers were combined and dried over saturated brine, and the organic layer was concentrated under vacuum. The resulting product was then separated by column chromatography to obtain the target compound 3 as a white solid (0.21 g, 60.25%).

[0040] 2. Methyl 4-(cyclohexylamino)-3-nitrobenzene (Compound 5)

[0041] Methyl 4-chloro-3-nitrobenzene (2.00 g, 9.30 mmol), cyclohexylamine (4.61 g, 46.50 mmol), and potassium carbonate (2.57 g, 18.60 mmol) were added to a 25 mL reaction flask, followed by the addition of 3 mL of DMSO as a solvent. The reaction mixture was reacted at 80 °C for 8 hours. After the reaction was complete, the reaction mixture was poured into an Erlenmeyer flask containing 50 mL of water. The aqueous layer was extracted with DCM (50 mL × 3), the organic layers were combined and concentrated to obtain a yellow oily substance, which was used directly in the next reaction without further treatment.

[0042] 3,4-(cyclohexylamino)-3-nitrobenzoic acid (compound 6)

[0043] The mixture obtained in the previous step was transferred to a solution containing 20 mL of ethanol, followed by the addition of 5 mL of sodium hydroxide aqueous solution (0.74 g, 18.60 mmol). The reaction mixture was stirred at room temperature for 6 hours. After the reaction was complete, the ethanol was distilled off, and the pH of the remaining aqueous solution containing the mixture was adjusted to 7 with 1 N hydrochloric acid, resulting in the precipitation of a yellow solid. The mixture was filtered, the filter cake was washed with water, and dried. After drying, the target compound 6 was obtained as a yellow solid.

[0044] 4. 1-(4-chlorophenyl)-3-methyl-1H-pyrazole-5-yl-4-(cyclohexylamino)-3-nitrobenzene ester (compound 7)

[0045] 4-(cyclohexylamino)-3-nitrobenzoic acid (0.10 g, 0.38 mmol), 1-(4-chlorophenyl)-3-methyl-1H-pyrazole-5-ol (0.08 g, 0.38 mmol), HATU (0.16 g, 0.42 mmol), and DIEPA (0.054 g, 0.42 mmol) were added to a reaction flask, with DCM (15 mL) as the solvent. The mixture was stirred at room temperature for 4 hours. After the reaction was complete, the solvent was evaporated under vacuum, and the residue was separated by column chromatography to obtain the target compound 7 (0.10 g, 52.90%) as a gray solid.

[0046] 5. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-1)

[0047] Compound 3-methyl-1-phenyl-1H-pyrazole-5-yl 4-(cyclohexylamino)-3-nitrobenzene ester (0.10 g, 0.23 mmol) was added to a reaction flask containing a methanol solution, followed by the addition of Pd / C (0.01 g). The reaction was carried out under hydrogen atmosphere until completion. Pd / C was filtered through filter paper, and the solvent was concentrated to obtain compound I-1 as a pale yellow solid (0.08 g, 86.20%). 1 H NMR (400MHz, DMSO-d6) δ7.55(d,J=7.6Hz,2H),7.45(t,J=7.9Hz,2H),7.31(td,J=6.3,5.8,2.5Hz,2H),7.20(d,J=2.0Hz,1H),6.52(d,J=8.6Hz, 1H),6.17(s,1H),5.26(d,J=7.4Hz,1H),4.91(s,2H),2.24(s,3H),1.96 (m,2H),1.74(m,2H),1.66–1.59(m,1H),1.36(m,2H),1.26–1.19(m,3H). 13 C NMR (101MHz, DMSO-d6) δ162.92,148.55,145.29,141.43,138.35,134.76,129.72,127. 30,122.76,122.51,114.94,113.01,108.60,97.08,51.16,32.91,25.95,25.13,14.72.

[0048] 6. 3-Methyl-1-(p-Tolyl)-1H-pyrazole-5-yl3-amino-4-(cyclohexylamino)benzoate (I-2)

[0049] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 80.12%.1 H NMR (400MHz, DMSO-d6) δ7.40–7.37(m,2H),7.26(dd,J=8.4,2.1Hz,1H),7.21(d,J= 8.2Hz,2H),7.16(d,J=2.1Hz,1H),6.48(d,J=8.8Hz,1H),6.10(s,1H),5.24(d,J=7 .5Hz,1H),4.88(s,2H),3.61–3.36(m,1H),2.26(s,3H),2.19(s,3H),1.95–1.89(m ,2H),1.73–1.67(m,2H),1.62–1.56(m,1H),1.37–1.29(m,2H),1.24–1.20(m,3H). 13 C NMR (101MHz, DMSO-d6) δ162.98,148.23,145.16,141.40,136.71,135.98,130.10,122. 71,122.49,114.92,113.04,108.57,96.88,51.17,32.96,25.95,25.14,20.96,14.70.

[0050] 7. 1-(4-Ethylphenyl)-3-methyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-3)

[0051] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 76.58%. 1 H NMR(400MHz,DMSO-d6)δ7.45–7.39(m,2H),7.29–7.22(m,3H),7.17(d,J=2.1Hz,1H),6 .49(d,J=8.8Hz,1H),6.10(s,1H),5.23(d,J=7.6Hz,1H),4.87(s,2H),3.34(dt,J=9.8, 3.3Hz,1H),2.57(q,J=7.6Hz,2H),2.20(s,3H),1.96–1.89(m,2H),1.75–1.67(m,2H), 1.62–1.56(m,1H),1.32(ddd,J=14.9,7.6,3.3Hz,2H),1.24–1.18(m,3H),1.14(s,3H). 13C NMR(101MHz,DMSO-d6)δ163.04,148.25,145.15,142.87,141.41,136.19,134.78,128.92,122 .73,122.44,114.92,113.03,108.59,96.96,51.16,32.96,28.03,25.95,25.13,15.80,14.72.

[0052] 8. 1-(4-methoxyphenyl)-3-methyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-4)

[0053] The synthesis method is the same as I-1. The product is a pale yellow solid with a yield of 72.05%. 1 H NMR(400MHz, DMSO-d6)δ7.43–7.36(m,2H),7.24(dd,J=8.4,2.1Hz,1H),7.15(d,J=2.1 Hz,1H),6.99–6.94(m,2H),6.48(d,J=8.8Hz,1H),6.08(s,1H),5.21(d,J=7.4Hz,1H),4 .86(s,2H),3.72(s,3H),3.42–3.35(m,1H),2.19(s,3H),1.96–1.88(m,2H),1.74–1.66 (m,2H),1.62–1.55(m,1H),1.32(ddd,J=12.5,7.2,2.8Hz,2H),1.19(d,J=10.2Hz,3H). 13 C NMR (101MHz, DMSO-d6) δ163.01,158.43,147.92,145.07,141.37,134.74,131.45,124. 39,122.72,114.76,113.06,108.59,96.47,55.81,51.16,32.95,25.95,25.13,14.69.

[0054] 9. 1-(3-methoxyphenyl)-3-methyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-5)

[0055] The synthesis method is the same as I-1. The product is a pale yellow solid with a yield of 74.30%. 1H NMR (400MHz, DMSO-d6) δ7.34–7.27(m,2H),7.18(d,J=2.1Hz,1H),7.13–7.06(m ,2H),6.86–6.80(m,1H),6.49(d,J=8.7Hz,1H),6.12(s,1H),5.24(d,J=7.5Hz,1 H),4.87(s,2H),3.67(s,3H),2.20(s,3H),1.96–1.89(m,2H),1.74–1.67(m,2H) ,1.63–1.56(m,1H),1.31(dd,J=17.3,7.7Hz,2H),1.20(dd,J=22.7,2.9Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ163.00,160.10,148.57,145.34,141.44,139.45,134.79,130.55,122.78, 114.90,114.35,113.22,112.96,108.56,107.61,97.41,55.67,51.16,32.93,25.95,25.13,14.72.

[0056] 10. 1-(4-fluorophenyl)-3-methyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-6)

[0057] The synthesis method is the same as I-1. The solid is pale gray and the yield is 80.03%. 1 H NMR (400MHz, DMSO-d6) δ7.56–7.52(m,2H),7.31–7.24(m,3H),7.15(d,J=2. 2Hz,1H),6.48(d,J=8.9Hz,1H),6.15–6.11(m,1H),5.23(d,J=7.5Hz,1H),4 .86(s,2H),3.34(s,1H),2.20(s,3H),1.96–1.89(m,2H),1.73–1.66(m,2H) ,1.62–1.56(m,1H),1.32(ddd,J=15.4,7.4,2.9Hz,2H),1.24–1.15(m,3H). 13C NMR (101MHz, DMSO-d6) δ162.79,148.61,145.31,141.44,134.76,124.89,124.80,122. 79,116.68,116.46,114.88,112.90,108.60,96.84,51.16,32.95,25.95,25.13,14.69.

[0058] 11. 1-(4-aminophenyl)-3-methyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (1-7)

[0059] The synthesis method is the same as I-1. The solid is pale gray and the yield is 69.20%. 1 H NMR(400MHz, DMSO-d6)δ7.21(dd,J=8.4,2.1Hz,1H),7.14(d,J=2.1Hz,1H),7.09–7.0 5(m,2H),6.54–6.50(m,2H),6.46(d,J=8.8Hz,1H),6.00(s,1H),5.18(d,J=8.4Hz,3H ),4.83(s,2H),3.44–3.40(m,1H),2.16(s,3H),1.95–1.88(m,2H),1.70(dt,J=12.7, 2.6Hz, 2H), 1.62–1.54 (m, 1H), 1.31 (ddd, J=15.6, 8.9, 2.5Hz, 2H), 1.23–1.17 (m, 3H). 13 C NMR(101MHz,DMSO-d6)δ163.12,148.43,147.03,144.80,141.28,134.71,127.15,124.51,122 .58,114.98,114.04,113.29,108.53,95.85,56.50,51.15,32.95,25.96,25.12,19.02,14.68.

[0060] 12. 1-(4-chlorophenyl)-3-methyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-8)

[0061] The synthesis method is the same as I-1. The solid is pale gray and the yield is 76.40%. 1H NMR (400MHz, DMSO-d6) δ7.67–7.48(m,4H),7.34(d,J=7.2Hz,1H),7.23(s,1H),6.54(d,J=7.2Hz,1H),6.22(s,1H),5.37–5.21(m ,1H),4.95(s,2H),2.25(s,3H),2.03–1.91(m,2H),1.81–1.69(m,2H),1.69–1.57(m,1H),1.43–1.31(m,2H),1.29–1.14(m,3H). 13 C NMR (101MHz, DMSO-d6) δ162.71,149.02,145.44,141.50,137.21,134.80,131.48,129. 74,124.08,122.85,114.91,112.87,108.62,97.21,51.19,32.96,25.96,25.14,14.70.

[0062] 13. 1-(4-cyanophenyl)-3-methyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-9)

[0063] The synthesis method is the same as I-1. The product is a gray solid with a yield of 83.30%. 1 H NMR (400MHz, DMSO-d6) δ7.92(d,J=8.5Hz,2H),7.77(d,J=8.8Hz,2H),7.32(dd, J=8.4,1.8Hz,1H),7.19–7.16(m,1H),6.52(d,J=8.6Hz,1H),6.24(s,1H),5.28 (d,J=7.5Hz,1H),4.92(s,2H),2.23(s,3H),1.97–1.90(m,2H),1.74–1.67(m,2 H),1.63–1.56(m,1H),1.32(td,J=10.9,9.2,3.1Hz,2H),1.22(t,J=6.0Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ162.52,150.21,145.99,141.79,141.61,134.82,134.20,122.99,1 22.17,118.87,114.88,112.69,109.22,108.68,98.10,51.20,32.95,25.95,25.14,14.75.

[0064] 14. 3-Methyl-1-(4-(trifluoromethyl)phenyl)-1H-pyrazole-5-yl3-amino-4-(cyclohexylamino)benzoate (I-10)

[0065] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 76.70%. 1 H NMR (400MHz, DMSO-d6) δ7.84–7.77(m,4H),7.32(dd,J=8.4,2.1Hz,1H),7.19(d,J=2.2Hz,1H),6.51(d,J=8.8Hz,1H),6.22(s,1H),5.28(d,J=7 .5Hz,1H),4.90(s,2H),2.23(s,3H),1.96–1.90(m,2H),1.71(dt,J=12. 0,2.9Hz,2H),1.63–1.55(m,1H),1.39–1.28(m,2H),1.25–1.16(m,3H). 13 C NMR (101MHz, DMSO-d6) δ162.67,149.79,145.84,141.59,134.84,127.12,122. 93,122.20,114.89,112.73,108.66,97.92,51.19,32.95,25.95,25.14,14.73.

[0066] 15. 3-Methyl-1-(naphth-1-yl)-1H-pyrazole-5-yl3-amino-4-(cyclohexylamino)benzoate (I-11)

[0067] The synthesis method is the same as I-1. The product is a gray solid with a yield of 79.10%. 1 H NMR (400MHz, DMSO-d6) δ8.07(d,J=2.0Hz,1H),7.97(d,J=8.9Hz,1H),7.92–7.87(m,2H),7.72(dd,J =8.8,2.2Hz,1H),7.52–7.46(m,2H),7.31(dd,J=8.4,2.1Hz,1H),7.21(d,J=2.1Hz,1H),6.48(d,J= 8.6Hz,1H),6.19(s,1H),5.23(d,J=7.5Hz,1H),4.86(s,2H),3.50–3.33(m,1H),2.25(s,3H),1.94– 1.87(m,2H),1.73–1.66(m,2H),1.58(dt,J=11.8,2.9Hz,1H),1.37–1.27(m,2H),1.24–1.19(m,3H). 13C NMR (101MHz, DMSO-d6) δ162.92,148.85,145.56,141.44,135.83,134.78,133.35,131.77,129.50,128.38,128. 11,127.47,126.77,122.80,121.36,120.05,114.97,113.01,108.58,97.36,51.15,32.94,25.95,25.12,14.77.

[0068] 16. 3-Ethyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-12)

[0069] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 82.23%. 1 H NMR(400MHz,DMSO-d6)δ7.52(dt,J=8.6,1.7Hz,2H),7.45–7.39(m,2H),7.31–7.24 (m,2H),7.17(d,J=2.1Hz,1H),6.48(d,J=8.8Hz,1H),6.18(s,1H),5.26(d,J=7.5Hz ,1H),4.89(s,2H),3.28(d,J=7.0Hz,1H),2.57(q,J=7.6Hz,2H),1.92(dd,J=8.7,3. 2Hz,2H),1.74–1.67(m,2H),1.58(s,1H),1.37–1.27(m,2H),1.19(t,J=7.6Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ162.93,154.25,145.24,141.43,138.41,134.79,129.70,127.28, 122.74,122.55,114.92,113.04,108.58,95.71,51.17,32.95,25.96,25.13,22.26,13.74.

[0070] 17. 1-Phenylacetyl-3-propyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-13)

[0071] The synthesis method is the same as I-1. The product is a gray solid with a yield of 74.03%. 1H NMR (400MHz, DMSO-d6) δ7.55–7.50(m,2H),7.45–7.39(m,2H),7.30–7.24(m,2H),7.17( d,J=2.1Hz,1H),6.48(d,J=8.9Hz,1H),6.16(s,1H),5.24(d,J=7.5Hz,1H),4.89(s,2H), 3.28(dd,J=10.8,3.5Hz,1H),2.55–2.50(m,2H),1.92(dd,J=8.8,3.4Hz,2H),1.74–1.67 (m,2H),1.65–1.55(m,3H),1.38–1.27(m,2H),1.22–1.12(m,3H),0.92(t,J=7.4Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ162.92,152.97,145.20,141.42,138.41,134.77,129.70,127.25,122 .75,122.50,114.94,113.06,108.59,96.09,51.17,32.96,31.01,25.96,25.13,22.39,14.30.

[0072] 18. 3-Isopropyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-14)

[0073] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 70.84%. 1 H NMR(400MHz,DMSO-d6)δ7.53(dt,J=8.7,1.7Hz,2H),7.45–7.40(m,2H),7.30–7.2 5(m,2H),7.16(d,J=2.1Hz,1H),6.48(d,J=8.8Hz,1H),6.20(s,1H),5.25(d,J=7.5 Hz,1H),4.89(s,2H),3.30–3.23(m,1H),2.93–2.83(m,1H),1.96–1.88(m,2H),1.7 5–1.65(m,2H),1.59(dd,J=9.3,3.4Hz,1H),1.38–1.28(m,2H),1.23–1.17(m,9H). 13C NMR(101MHz,DMSO-d6)δ162.94,158.51,145.17,141.43,138.48,134.80,129.69,127.28, 122.75,122.58,114.93,113.08,108.59,94.46,51.19,32.97,28.53,25.96,25.15,22.63.

[0074] 19. 3-Cyclopropyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-15)

[0075] The synthesis method is the same as I-1. The product is a pale yellow solid with a yield of 73.15%. 1 H NMR(400MHz, DMSO-d6)δ7.51(dd,J=8.6,1.1Hz,2H),7.44–7.38(m,2H),7.30–7.24( m,2H),7.16(d,J=2.1Hz,1H),6.48(d,J=8.8Hz,1H),6.05(s,1H),5.28(d,J=7.5Hz, 1H),4.91(s,2H),1.90(ddt,J=13.4,8.4,4.2Hz,3H),1.74–1.66(m,2H),1.62–1.55 (m,1H),1.36–1.25(m,2H),1.23–1.14(m,3H),0.92–0.86(m,2H),0.73–0.68(m,2H). 3 C NMR(101MHz,DMSO-d6)δ162.93,154.89,145.21,141.46,138.38,134.80,129.69, 127.23,122.75,114.91,108.57,93.98,51.18,32.95,25.95,25.13,10.22,8.30.

[0076] 20. 3-Butyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(cyclohexylamino)benzoate (I-16)

[0077] The synthesis method is the same as I-1. The product is a pale yellow solid with a yield of 75.46%. 1H NMR(400MHz,DMSO-d6)δ7.55–7.51(m,2H),7.45–7.39(m,2H),7.29–7.24(m,2H), 7.17(d,J=2.1Hz,1H),6.49(d,J=8.9Hz,1H),6.24(s,1H),5.25(d,J=7.5Hz,1H),4 .89(s,2H),3.29(dd,J=8.9,5.5Hz,1H),1.96–1.88(m,2H),1.74–1.65(m,2H),1. 62–1.55(m,1H),1.38–1.28(m,2H),1.26(s,9H),1.19(td,J=8.3,7.7,4.2Hz,3H). 13 C NMR (101MHz, DMSO-d6) δ162.96,161.25,145.08,141.42,138.53,134.77,129. 68,127.22,122.55,114.93,108.59,93.99,51.18,32.95,30.44,25.95,25.13.

[0078] 21. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(isopropylamino)benzoate (I-17)

[0079] The synthesis method is the same as I-1. The product is a pale yellow solid with a yield of 70.63%. 1 H NMR (400MHz, DMSO-d6) δ7.54–7.50(m,2H),7.44–7.40(m,2H),7.31–7.25(m,2H),7.18(d,J=2.1Hz,1H),6.46(d,J=8.9Hz ,1H),6.14(s,1H),5.27(d,J=6.4Hz,1H),4.92(s,2H),3.66(dq,J=11.6,5.6Hz,1H),2.21(s,3H),1.15(d,J=6.3Hz,6H). 13 C NMR(101MHz,DMSO-d6)δ162.96,148.56,145.31,141.57,138.36,134.80,12 9.71,127.30,122.79,114.87,113.12,108.73,97.07,43.68,22.78,14.71.

[0080] 22. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(cyclopropylamino)benzoate (I-18)

[0081] The synthesis method is the same as I-1. The product is a pale yellow solid with a yield of 75.40%. 1 H NMR (400MHz, DMSO-d6) δ7.56–7.50(m,2H),7.47–7.39(m,2H),7.34–7.25(m,2H),7.17(d,J=2.1Hz,1H),6.80(d,J=8.4Hz,1 H),6.15(s,1H),6.02(s,1H),4.84(s,2H),2.45–2.37(m,1H),2.21(s,3H),0.74(td,J=6.7,4.6Hz,2H),0.45–0.37(m,2H). 13 C NMR (101MHz, DMSO-d6) δ163.04,148.57,145.25,138.34,134.89,129.72,127.32,122.55,122.26,114.49,109.83,97.07,24.94,14.71,7.38.

[0082] 23. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(isobutylamino)benzoate (I-19)

[0083] The synthesis method is the same as I-1. The product is a pale yellow solid with a yield of 70.86%. 1 H NMR (400MHz, DMSO-d6) δ7.54–7.50(m,2H),7.45–7.40(m,2H),7.30–7.25(m,2H),7.21(d,J=2.2Hz,1H), 6.73(d,J=8.6Hz,1H),6.13(s,1H),4.86(d,J=21.2Hz,3H),3.46–3.35(m,2H),2.21(s,3H),1.35(s,7H). 13 C NMR(101MHz,DMSO-d6)δ162.99,148.56,145.27,141.41,138.35,135.73,12 9.73,127.31,122.52,115.66,113.18,111.11,97.08,51.23,29.68,14.71.

[0084] 24. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(cyclopentanamino)benzoate (I-20)

[0085] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 76.20%. 1H NMR (400MHz, DMSO-d6) δ7.53(d,J=7.2Hz,2H),7.42(t,J=7.1Hz,2H),7.29(s,2H),7.18(s,1H),6.48(d,J=8.2Hz,1H),6.14(s, 1H),5.40–5.32(m,1H),4.92(s,2H),3.86–3.72(m,1H),2.21(s,3H),2.01–1.89(m,2H),1.72–1.62(m,2H),1.56–1.44(m,4H). 13 C NMR(101MHz,DMSO-d6)δ163.00,148.57,145.32,141.95,138.37,134.94,129.7 0,127.29,122.61,114.55,113.33,109.16,97.06,54.12,32.95,24.31,14.71.

[0086] 25. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-3-amino-4-(cycloheptaneamino)benzoate (I-21)

[0087] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 70.36%. 1 H NMR (400MHz, DMSO-d6) δ7.54–7.50(m,2H),7.44–7.39(m,2H),7.31–7.25(m,2H),7.17(d,J=2.2Hz,1H),6.38(d,J=8.9Hz,1H),6.13(s,1H ),5.28(d,J=7.4Hz,1H),4.93(s,2H),3.50(ddq,J=12.3,8.4,4.4Hz,1H),2.20(s,3H),1.87(dq,J=10.0,3.9Hz,2H),1.64–1.44(m,10H). 13 C NMR (101MHz, DMSO-d6) δ162.97,148.56,141.26,129.71,127.30,122.52,114.81,108.81,97.08,53.07,34.28,28.47,24.35,14.71.

[0088] 26. 3-Methyl-1-phenyl-1H-pyrazole-5-yl4-(adamantane-1-ylamino)-3-aminobenzoate (I-22)

[0089] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 66.82%. 1H NMR (400MHz, DMSO-d6) δ7.55–7.51(m,2H),7.45–7.40(m,2H),7.30–7.24(m,2H),7.23(d,J=2.2Hz,1H),6.83(d,J= 8.5Hz,1H),6.13(s,1H),4.87(s,2H),4.63(s,1H),2.21(s,3H),2.05(s,3H),1.95(s,6H),1.66(d,J=14.4Hz,6H). 13 C NMR (101MHz, DMSO-d6) δ162.93,148.55,145.27,138.36,135.79,129.73,127. 29,122.51,116.00,113.34,112.14,97.07,52.02,42.12,36.37,29.52,14.71.

[0090] 27. 3-Methyl-1-phenyl-1H-pyrazol-5-yl-3-amino-4-((tetrahydro-2H-pyran-4-yl)amino)benzoate (I-23)

[0091] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 75.63%. 1 H NMR (400MHz, DMSO-d6) δ7.60–7.53 (m, 2H), 7.45 (t, J = 7.9Hz, 2H), 7.34–7. 27(m,2H),7.22(d,J=2.1Hz,1H),6.60(d,J=8.7Hz,1H),6.18(s,1H),5.37( d,J=7.5Hz,1H),4.95(s,2H),3.93–3.85(m,2H),3.66–3.56(m,1H),3.47– 3.42(m,2H),2.24(s,3H),1.94–1.86(m,2H),1.46(qd,J=12.7,4.3Hz,2H). 13 C NMR(101MHz,DMSO-d6)δ162.90,148.57,145.27,138.35,135.00,129.71,1 27.32,122.55,115.00,113.53,108.93,97.04,66.47,48.51,33.11,14.71.

[0092] 28. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-4-(cyclohexylamino)-3-(cyclopentylamino)benzoate (I-24)

[0093] Compound I-1 (0.10 g, 0.25 mmol) and cyclopentanone (0.10 g, 1.28 mmol) were added to a reaction flask containing 15 mL of dichloromethane. Then, sodium triacetoxyborohydride (0.16 g, 0.75 mmol) was slowly added, and the mixture was stirred at room temperature until the reaction was complete. 1 mL of saturated ammonium chloride aqueous solution was added to the reaction mixture, followed by extraction with dichloromethane. The organic layers were combined and dried. The organic layers were concentrated under vacuum. The mixture was then separated by column chromatography to obtain the target compound I-24, a pale yellow solid, in a yield of 50.34%. 1 HNMR(400MHz,Chloroform-d)δ7.61(ddd,J=8.4,4.1,1.6Hz,3H),7.43–7.37(m,3H),7. 29–7.25(m,1H),6.59(d,J=8.5Hz,1H),6.24(s,1H),3.73(p,J=6.2Hz,1H),3.32(ddt,J= 10.1,7.2,3.6Hz,1H),2.34(s,3H),2.08–2.00(m,2H),1.96(ddd,J=12.7,7.7,3.4Hz,2 H),1.76(ddt,J=21.3,11.0,5.3Hz,5H),1.60(tt,J=7.3,4.5Hz,2H),1.50–1.31(m,5H). 13 C NMR(101MHz,Chloroform-d)δ162.41,149.04,143.78,138.44,134.43,128.98,126.78, 123.20,116.14,109.03,95.59,55.22,51.39,33.52,33.22,25.82,24.93,24.36,14.59.

[0094] 29. 3-Methyl-1-phenyl-1H-pyrazole-5-yl 3,4-bis(cyclohexylamino)benzoate (I-25)

[0095] The synthesis method is the same as that for I-24. The product is a pale yellow solid with a yield of 53.34%. 1 H NMR(400MHz,Chloroform-d)δ7.62(s,2H),7.42(s,2H),7.26(s,2H),6.60(s,1H),6.22(s,1H ),3.49–2.87(m,2H),2.35(s,3H),2.11–1.92(m,4H),1.83–1.62(m,6H),1.43–1.16(m,10H). 13C NMR(101MHz,Chloroform-d)δ162.38,149.04,145.20,138.41,129.00,126.79, 123.10,109.21,95.70,52.90,51.30,33.59,33.16,25.97,25.82,24.82,14.60.

[0096] 30. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-4-(cyclohexylamino)-3-((cyclohexylmethyl)amino)benzoate (I-26)

[0097] The synthesis method is the same as that for I-24. The solid is grayish-white and the yield is 80.42%. 1 H NMR(400MHz,Chloroform-d)δ7.62(dt,J=8.7,1.7Hz,3H),7.43–7.38(m,2H),7.33 (s,1H),7.30–7.25(m,1H),6.60(d,J=8.4Hz,1H),6.25(s,1H),3.37–3.28(m,1H),2 .90–2.82(m,2H),2.34(s,3H),2.11–2.03(m,2H),1.75(ddt,J=34.1,19.9,9.1Hz, 9H), 1.38 (dd, J=17.4, 7.6Hz, 2H), 1.30–1.19 (m, 6H), 1.01 (qd, J=12.2, 2.8Hz, 2H). 13 C NMR(101MHz,Chloroform-d)δ162.43,149.05,145.27,143.39,129.00,126.82,124.92, 123.18,114.92,109.07,95.58,51.63,33.25,31.56,26.59,26.00,25.81,24.93,14.59.

[0098] 31. 3-Methyl-1-phenyl-1H-pyrazole-5-yl 3-(benzylamino)-4-(cyclohexylamino)benzoate (I-27)

[0099] The synthesis method is the same as that for I-24. The product is a pale yellow solid with a yield of 76.60%. 1H NMR(400MHz,Chloroform-d)δ7.64(dd,J=8.4,2.0Hz,1H),7.63–7.59(m,2H),7.43(d,J =1.9Hz,1H),7.40(d,J=7.5Hz,2H),7.37(dd,J=3.3,1.2Hz,4H),7.28–7.25(m,1H),6.6 2(d,J=8.6Hz,1H),6.23(s,1H),4.23(s,2H),3.35(tt,J=10.4,3.6Hz,1H),2.34(s,3H) ,2.08–2.03(m,2H),1.77(dt,J=13.1,3.7Hz,2H),1.71–1.51(m,4H),1.43–1.36(m,2H). 13 C NMR(101MHz,Chloroform-d)δ162.31,149.03,145.18,138.73,138.39,128.97,128.74,128.17,127. 63,126.81,125.35,123.17,115.14,109.10,95.58,51.43,49.35,33.21,29.70,25.75,24.93,14.57.

[0100] 32. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-4-(cyclohexylamino)-3-(phenethylamino)benzoate (I-28)

[0101] The synthesis method is the same as that for I-24. The product is a pale yellow solid with a yield of 82.62%. 1 H NMR(400MHz,Chloroform-d)δ7.66–7.57(m,3H),7.40–7.21(m,9H),7.18(t,J=7.4Hz,1H),6.58(d,J=8.5Hz,1H),6.25(s,1H),4.04(s,1H),3.31(q ,J=7.3Hz,3H),2.96(t,J=7.0Hz,2H),2.35(s,3H),2.05–1.95(m,2H),1. 79–1.71(m,2H),1.69–1.64(m,1H),1.44–1.34(m,2H),1.27–1.15(m,3H). 13C NMR(101MHz,Chloroform-d)δ162.32,149.04,145.24,143.49,139.21,138.39,134.57,128.95,128.71,126. 84,126.62,125.28,123.18,115.25,115.08,109.08,95.55,51.15,46.08,35.85,33.08,25.80,24.77,14.60.

[0102] 33. 3-Methyl-1-phenyl-1H-pyrazole-5-yl(E)-4-(cyclohexylamino)-3-((pyridin-4-ylmethylene)amino)benzoate (I-29)

[0103] The synthesis method is the same as that for I-24. The product is a yellow solid with a yield of 66.32%. 1 HNMR(400MHz,Chloroform-d)δ8.78(d,J=5.4Hz,2H),8.55(s,1H),7.88(dd,J=8.7,1.9Hz,1 H),7.77(d,J=1.9Hz,1H),7.73(d,J=5.9Hz,2H),7.61(d,J=7.7Hz,2H),7.44–7.40(m,2H),7. 35–7.27(m,2H),6.66(d,J=8.8Hz,1H),6.24(s,1H),3.44(dq,J=13.7,9.7,7.0Hz,1H),2.35( s,3H),2.11–2.04(m,2H),1.80(dt,J=12.3,3.4Hz,2H),1.66(ddd,J=24.6,14.2,5.0Hz,6H).

[0104] 34. 3-Methyl-1-phenyl-1H-pyrazol-5-yl 4-(cyclohexylamino)-3-((pyridin-4-ylmethyl)amino)benzoate (I-30)

[0105] The synthesis method is the same as that for I-24. The product is a yellow solid with a yield of 70.67%. 1HNMR(400MHz,Chloroform-d)δ8.56–8.53(m,2H),7.64(dd,J=8.5,2.0Hz,1H),7.59–7.55(m,2H), 7.41–7.36(m,2H),7.29(ddd,J=7.4,5.8,2.6Hz,3H),7.25–7.24(m,1H),6.64(d,J=8.5Hz,1H),6.2 0(s,1H),4.30(s,2H),3.36(ddt,J=10.3,7.3,3.8Hz,1H),2.33(s,3H),2.09–2.03(m,2H),1.79(dt ,J=12.8,3.5Hz,2H),1.68(dt,J=12.6,3.4Hz,1H),1.45–1.35(m,2H),1.27(dd,J=8.1,5.0Hz,3H).

[0106] 35. 3-Methyl-1-phenyl-1H-pyrazole-5-yl 4-(cyclohexylamino)-3-((4-fluorobenzyl)amino)benzoate (I-31)

[0107] The synthesis method is the same as that for I-24. The product is a yellow solid with a yield of 74.36%. 1 HNMR(400MHz,Chloroform-d)δ7.67–7.63(m,1H),7.62–7.59(m,2H),7.42–7.37(m,3H),7.36–7.26(m,4H),7.03(td,J=8.7,1.4Hz,3H),6.62(d,J= 8.7Hz,1H),6.22(s,1H),4.20(s,2H),3.39–3.31(m,1H),2.34(s,3H),2. 09–2.03(m,2H),1.81–1.74(m,2H),1.70–1.65(m,1H),1.47–1.27(m,5H). 13 C NMR(101MHz,Chloroform-d)δ162.32,149.07,145.18,143.47,138.41,134.34,129.70,128.98,126. 85,125.47,123.20,115.70,115.29,109.19,95.62,64.66,51.43,48.58,33.24,25.77,24.92,14.58.

[0108] 36. 3-Methyl-1-phenyl-1H-pyrazole-5-yl 3-((4-bromobenzyl)amino)-4-(cyclohexylamino)benzoate (I-32)

[0109] The synthesis method is the same as that for I-24. The product is a yellow solid with a yield of 72.10%. 1 HNMR(400MHz,Chloroform-d)δ7.64(dd,J=8.5,1.5Hz,1H),7.62–7.56(m,2H),7.48– 7.44(m,2H),7.42–7.36(m,3H),7.29–7.26(m,1H),7.25–7.21(m,2H),6.63(d,J=8.5H z,1H),6.22(s,1H),4.19(s,2H),3.40–3.26(m,1H),2.34(s,3H),2.09–2.01(m,2H),1 .82–1.74(m,2H),1.71–1.65(m,1H),1.44–1.32(m,2H),1.26(dt,J=10.8,4.5Hz,4H). 13 C NMR(101MHz,Chloroform-d)δ162.28,149.07,145.17,138.40,137.74,131.83,129.70,128.99,126. 86,125.50,123.19,121.41,115.31,109.26,95.63,64.50,51.45,48.59,33.22,25.77,24.92,14.59.

[0110] 37. 3-Methyl-1-phenyl-1H-pyrazole-5-yl 4-(cyclohexylamino)-3-((4-methylbenzyl)amino)benzoate (I-33)

[0111] The synthesis method is the same as that for I-24. The product is a yellow solid with a yield of 64.80%. 1 HNMR(400MHz,Chloroform-d)δ7.70(dd,J=8.6,2.0Hz,1H),7.66(d,J=2.0Hz,1H),7.60–7.5 6(m,2H),7.39(tt,J=8.8,2.0Hz,2H),7.29(dt,J=8.3,1.6Hz,1H),7.09–7.04(m,3H),6.47(d ,J=8.8Hz,1H),6.25(s,1H),5.68(d,J=8.0Hz,1H),3.89(s,2H),3.36–3.20(m,1H),2.35(s,3 H),2.29(s,3H),1.95–1.86(m,2H),1.76–1.68(m,2H),1.66–1.61(m,1H),1.45–1.25(m,5H). 13C NMR(101MHz,Chloroform-d)δ161.93,149.06,148.69,138.46,136.83,135.61,134.87,129.76,128.98,1 28.81,126.82,126.10,123.25,113.10,108.83,95.43,57.20,32.82,29.73,25.77,24.62,21.12,14.60.

[0112] 38. 3-Methyl-1-phenyl-1H-pyrazole-5-yl 4-(cyclohexylamino)-3-((4-methoxybenzyl)amino)benzoate (I-34)

[0113] The synthesis method is the same as that for I-24. The product is a yellow solid with a yield of 70.42%. 1 HNMR(400MHz,Chloroform-d)δ7.63(ddd,J=9.8,8.4,1.4Hz,3H),7.44–7.37(m,3H),7.2 8(t,J=8.1Hz,3H),6.93–6.86(m,2H),6.62(d,J=8.6Hz,1H),6.23(s,1H),4.16(s,2H),3 .81(s,3H),3.34(ddd,J=13.2,10.0,3.3Hz,1H),2.35(s,3H),2.10–2.01(m,2H),1.77(d t,J=13.2,3.3Hz,2H),1.68(dd,J=8.6,4.2Hz,1H),1.43–1.32(m,2H),1.26–1.18(m,3H). 13 C NMR(101MHz,Chloroform-d)δ162.37,159.11,149.06,145.24,143.36,138.42,134.67,130.80,129.44,128.99,1 26.84,125.27,123.20,115.25,115.02,114.11,109.05,95.61,55.33,51.45,48.79,33.23,25.78,24.96,14.59.

[0114] 39. 3-Methyl-1-phenyl-1H-pyrazole-5-yl 4-(cyclohexylamino)-3-((4-nitrobenzyl)amino)benzoate (I-35)

[0115] The synthesis method is the same as that for I-24. The product is a yellow solid with a yield of 75.52%. 1HNMR(400MHz,Chloroform-d)δ8.20–8.15(m,2H),7.65(dd,J=8.5,2.0Hz,1H),7.59–7.55 (m,2H),7.48–7.45(m,2H),7.41–7.36(m,2H),7.31–7.26(m,2H),6.65(d,J=8.8Hz,1H),6 .19(s,1H),4.38(s,2H),3.36(ddt,J=9.8,7.3,3.7Hz,1H),2.33(s,3H),2.11–2.04(m,2H ),1.79(dt,J=13.1,3.6Hz,2H),1.71–1.65(m,1H),1.46–1.35(m,2H),1.31–1.24(m,3H).

[0116] 40. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-4-(cyclohexylamino)-3-((thiophen-3-ylmethyl)amino)benzoate (I-36)

[0117] The synthesis method is the same as that for I-24. The product is a yellow solid with a yield of 64.38%. 1 HNMR(400MHz,Chloroform-d)δ7.65(dd,J=8.4,1.9Hz,1H),7.62–7.59(m,2H),7.43–7.37(m,3H),7 .33(dd,J=4.9,3.0Hz,1H),7.29–7.25(m,1H),7.20–7.17(m,1H),7.10(dd,J=5.0,1.1Hz,1H),6.62 (d,J=8.6Hz,1H),6.23(s,1H),4.25(s,2H),3.34(ddd,J=13.3,9.5,2.7Hz,1H),2.34(s,3H),2.09– 2.02(m,2H),1.78(dt,J=12.3,3.1Hz,2H),1.70–1.65(m,1H),1.43–1.34(m,2H),1.28–1.21(m,3H). 13 C NMR(101MHz,Chloroform-d)δ162.33,149.06,145.22,143.55,139.77,138.42,134.40,129.00,127.51,1 26.84,126.31,125.50,123.21,122.34,115.24,109.13,95.62,51.41,44.50,33.22,25.78,24.93,14.59.

[0118] 41. 3-Methyl-1-phenyl-1H-pyrazole-5-yl-4-(cyclohexylamino)-3-((naphth-1-ylmethyl)amino)benzoate (I-37)

[0119] The synthesis method is the same as that for I-24. The product is a yellow solid with a yield of 60.16%. 1 HNMR(400MHz,Chloroform-d)δ8.09(dd,J=6.2,3.5Hz,1H),7.91(dt,J=6.6,3.3Hz,1H),7.86(d,J=7.8Hz,1H),7. 69(dd,J=8.4,1.8Hz,1H),7.63(d,J=7.7Hz,2H),7.59(d,J=1.7Hz,1H),7.53(dt,J=6.4,3.3Hz,2H),7.50–7.44(m ,2H),7.39(t,J=7.9Hz,2H),7.22(d,J=7.5Hz,1H),6.63(d,J=8.5Hz,1H),6.26(s,1H),4.65(s,2H),3.36–3.25(m ,1H),2.36(s,3H),2.01–1.92(m,2H),1.75–1.68(m,2H),1.67–1.62(m,1H),1.40–1.29(m,2H),1.19–1.03(m,3H).

[0120] 42. 1-(4-cyanophenyl)-3-methyl-1H-pyrazole-5-yl-4-(cyclohexylamino)-3-(cyclopentylamino)benzoate (I-38)

[0121] The synthesis method is the same as that for I-24. The product is a gray solid with a yield of 62.47%. 1 HNMR(400MHz,Chloroform-d)δ7.86–7.81(m,2H),7.71–7.67(m,2H),7.61(dd,J=8.4,2.0Hz,1H),7.37(d,J=2.0Hz,1H),6.62(d,J=8.5Hz,1H),6.25(s ,1H),3.79–3.71(m,1H),3.35(ddd,J=13.9,9.9,3.4Hz,1H),2.34(s,3H),2 .12–2.04(m,2H),2.04–1.95(m,2H),1.81–1.60(m,10H),1.51–1.39(m,4H).

[0122] 43. 1-(4-cyanophenyl)-3-methyl-1H-pyrazole-5-yl-4-(cyclohexylamino)-3-((cyclohexylmethyl)amino)benzoate (I-39)

[0123] The synthesis method is the same as that for I-24. The product is a gray solid with a yield of 79.64%. 1 HNMR(400MHz,Chloroform-d)δ7.84–7.81(m,2H),7.71–7.68(m,2H),7.61(dd,J=8.4,1. 9Hz,1H),7.33(d,J=2.0Hz,1H),6.63(d,J=8.6Hz,1H),6.27(s,1H),3.37(td,J=10.0,5.0 Hz,1H),2.89(d,J=6.6Hz,2H),2.34(s,3H),2.12–2.05(m,2H),1.87–1.75(m,6H),1.72– 1.66(m,3H),1.63–1.58(m,1H),1.47–1.37(m,3H),1.32–1.25(m,5H),1.07–0.98(m,2H).

[0124] 44. 1-(4-cyanophenyl)-3-methyl-1H-pyrazole-5-yl-4-(cyclohexylamino)-3-(phenethylamino)benzoate (I-40)

[0125] The synthesis method is the same as that for I-24. The product is a pale yellow solid with a yield of 74.62%. 1 H NMR(400MHz,Chloroform-d)δ7.80(d,J=8.7Hz,2H),7.69–7.58(m,3H),7.40–7. 30(m,3H),7.24(d,J=7.4Hz,3H),6.61(d,J=8.5Hz,1H),6.24(s,1H),3.34(t,J=6 .9Hz,3H),2.98(t,J=6.9Hz,2H),2.34(s,3H),2.08–1.93(m,2H),1.76(dd,J=9.4 ,3.9Hz,2H),1.67(dt,J=12.1,3.4Hz,1H),1.47–1.35(m,2H),1.27–1.18(m,3H).

[0126] Example 2

[0127] 1. (4-Bromo-3-nitrophenyl)hydrazine (compound 9)

[0128] Compound 4-bromo-3-nitroaniline (1.00 g, 5.81 mmol) and concentrated hydrochloric acid (12 N, 3 mL) were added to a reaction flask. Sodium nitrite aqueous solution (0.48 g, 6.97 mmol) was slowly added dropwise at 0 °C, and the mixture was stirred at 0 °C for 15 minutes after the addition was complete. Stannous chloride (3.30 g, 17.43 mmol) was then dissolved in 5 mL of concentrated hydrochloric acid and added dropwise to the reaction mixture at 0 °C, with stirring until complete. The pH was adjusted to 9 by slowly adding saturated sodium bicarbonate to the reaction mixture. The mixture was extracted with ethyl acetate (30 × 3), the organic layers were combined and dried, the solvent was concentrated under vacuum, and the product was separated by column chromatography to obtain compound 9 as a brown solid, with a yield of 71.30%.

[0129] 2. 2-(4-bromo-3-nitrophenyl)-5-methyl-2,4-dihydro-3H-pyrazole-3-one (Compound 10)

[0130] The synthesis method is the same as that for compound 3. The product is a pale yellow solid with a yield of 62.74%. 1 H NMR (400MHz, DMSO-d6) δ12.21(s,1H),8.39(s,1H),8.06(d,J=13.0Hz,1H),7.79(d,J=13.1Hz,1H),5.39(s,1H),2.10(s,3H).

[0131] 3. 2-(4-(cyclohexylamino)-3-nitrophenyl)-5-methyl-2,4-dihydro-3H-pyrazole-3-one (compound 11), synthesized by the same method as compound 6, yellow solid.

[0132] 4. 2-(3-amino-4-(cyclohexylamino)phenyl)-5-methyl-2,4-dihydro-3H-pyrazole-3-one (II-1)

[0133] The synthesis method is the same as I-1, the product is a gray solid, and the yield is 82.42%. 1 HNMR (400MHz, DMSO-d6) δ7.12(d,J=2.0Hz,1H),7.08(dd,J=8.2,2.0Hz,1H),6.49(d,J=8.4Hz,1H),4.86(s,2H),4.80(d,J=7.5Hz,1H),3.28 –3.20(m,1H),3.07(s,2H),2.30(s,3H),1.94(d,J=10.2Hz,2H),1.74– 1.65(m,2H),1.62–1.54(m,1H),1.36–1.26(m,2H),1.23–1.11(m,3H).

[0134] 5. 2-(3,4-bis(cyclohexylamino)phenyl)-5-methyl-2,4-dihydro-3H-pyrazole-3-one (II-2)

[0135] The synthesis method is the same as that for I-24. The product is a pale yellow solid with a yield of 56.73%. 1 H NMR (400MHz, Chloroform-d) δ7.36 (s, 1H), 7.25–7.19 (m, 1H), 6.62 (d, J = 7.3Hz, 1H), 3.35–3.18 (m, 2H), 3. 08(s,2H),2.34(s,3H),2.09–2.01(m,4H),1.82–1.72(m,4H),1.69–1.63(m,2H),1.37(m,5H),1.28(m,5H).

[0136] 6. 2-(3-(benzylamino)-4-(cyclohexylamino)phenyl)-5-methyl-2,4-dihydro-3H-pyrazole-3-one (II-3)

[0137] The synthesis method is the same as that for I-24. The product is a pale yellow solid with a yield of 72.64%. 1 H NMR(400MHz,Chloroform-d)δ7.52–7.23(m,5H),7.02-6.94(m,2H),6.68(d,J=8.0Hz,1H),4.32(s,2H),3.34(t,J=10.0Hz,1H),3 .10(s,2H),2.33(s,3H),2.08(d,J=11.1Hz,2H),1.79(d,J=13.1Hz,2H),1.68(d,J=12.6Hz,1H),1.40(m,2H),1.28–1.20(m,3H).

[0138] Example 3

[0139] 1. (4-Chloro-3-nitrobenzene)methanol (Compound 13)

[0140] 4-Chloro-3-nitrobenzaldehyde (2.00 g, 10.81 mmol) was added to a round-bottom flask containing 40 mL of methanol. Sodium borohydride (0.41 g, 10.81 mmol) was added in small portions under ice bath conditions, followed by stirring at room temperature until the reaction was complete. The sodium borohydride was quenched with 1 N hydrochloric acid solution (1 mL), the solvent was evaporated, the residual mixture was extracted with ethyl acetate, the organic layers were combined, the organic layer was extracted with saturated brine and dried with anhydrous Na₂SO₄, and the organic layer was concentrated under vacuum to obtain compound 13, a pale yellow solid (1.80 g, 89.06%). 1H NMR (400MHz, Chloroform-d) δ7.88 (d, J = 8.5 Hz, 1H), 7.51 (d, J = 8.7 Hz, 2H), 4.76 (s, 2H).

[0141] 2. (4-(cyclohexylamino)-3-nitrophenyl)methanol (compound 14)

[0142] (4-chloro-3-nitrophenyl)methanol (1.00 g, 5.34 mmol), cyclohexylamine (2.64 g, 26.73 mmol), and potassium carbonate (1.47 g, 10.68 mmol) were added to a 25 mL reaction flask, and DMSO (5 mL) was added as a solvent. The reaction mixture was reacted at 150 °C for 8 hours. After the reaction was completed, the reaction solution was cooled to room temperature and then poured into 50 mL of water. The aqueous layer was extracted with DCM (50 mL × 3), and the organic layers were combined and concentrated. The mixture was separated by column chromatography (PE:EA = 10:1) to give a yellow solid compound 14 (1.02 g, 76.03%). 1 H NMR(400MHz,Chloroform-d)δ8.14(s,2H),7.45(dd,J=6.5,2.2Hz,1H),6.92–6.82(m,1H),4.57(d,J= 5.7Hz,2H),3.57–3.42(m,1H),2.09–1.98(m,2H),1.79(s,2H),1.73–1.61(m,3H),1.45–1.37(m,3H).

[0143] 3,4-(cyclohexylamino)-3-nitrobenzaldehyde (compound 15)

[0144] (4-(cyclohexylamino)-3-nitrophenyl)methanol (1.00 g, 3.99 mmol) and pyridinium chlorochromate (1.72 g, 7.99 mmol) were added to a round-bottom flask containing 100 mL of chloroform and stirred at room temperature until the reaction was complete. The reaction solution was filtered through diatomaceous earth, the filtrate was dried and concentrated, and the mixture was separated by column chromatography to give compound 15 as a yellow solid, with a yield of 75.30%.

[0145] 4. (E)-4-(4-(cyclohexylamino)-3-nitrobenzylidene)-5-methyl-2-phenyl-2,4-dihydro-3H-pyrazole-3-one (Compound 16)

[0146] Compound 4-(cyclohexylamino)-3-nitrobenzaldehyde (0.16 g, 0.63 mmol) and 5-methyl-2-phenyl-2,4-dihydro-3H-pyrazole-3-one (0.10 g, 0.57 mmol) were added to a reaction flask, with acetic acid (5 mL) as the solvent. The mixture was stirred at room temperature until the reaction was complete. After the reaction was complete, the reaction solution was poured into 50 mL of water, and the aqueous layer was extracted with dichloromethane (30 mL × 3). The organic layers were combined and dried. The organic layer was concentrated under vacuum, and the residue was separated by column chromatography to obtain the target compound 16 as a yellow solid with a yield of 52.40%. 1 HNMR(400MHz,DMSO-d6)δ9.65(d,J=2.0Hz,1H),8.78(dd,J=9.3,1.9Hz,1H),8.4 8(d,J=7.9Hz,1H),7.93–7.85(m,2H),7.75(s,1H),7.44–7.37(m,2H),7.26(d,J =9.4Hz,1H),7.19–7.12(m,1H),3.83–3.74(m,1H),2.28(s,3H),1.95(dt,J=8.8 ,5.2Hz,2H),1.68(dt,J=9.8,5.4Hz,2H),1.62–1.54(m,1H),1.46–1.37(m,5H).

[0147] 5. 4-(3-amino-4-(cyclohexylamino)benzyl)-5-methyl-2-phenyl-2,4-dihydro-3H-pyrazole-3-one (III-1)

[0148] The synthesis method is the same as I-1, and the product is a pale yellow solid with a yield of 64.21%. 1 H NMR(400MHz,Chloroform-d)δ7.35–7.26(m,5H),6.60(s,2H),6.53(s,1H),3.85(s,2H),3.32(td,J=10.0,5.0Hz,1H),2.8 6(s,2H),2.09–2.03(m,2H),1.81–1.74(m,3H),1.64(ddd,J=14.1,10.3,7.2Hz,2H),1.41–1.33(m,2H),1.24–1.16(m,3H).

[0149] 6. 4-(3,4-bis(cyclohexylamino)benzyl)-5-methyl-2-phenyl-2,4-dihydro-3H-pyrazole-3-one (III-2)

[0150] The synthesis method is the same as that for I-24, and the product is a pale yellow solid with a yield of 52.83%. 1H NMR(400MHz,Chloroform-d)δ7.36(d,J=2.0Hz,1H),7.34–7.30(m,4H),7.30–7.27(m,1H),7.25–7.22(m,1H),6.59(d,J=8.3 Hz,1H),3.39-3.16(m,4H),2.09–2.01(m,4H),1.92(s,3H),1.82–1.72(m,4H),1.69–1.63(m,2H),1.37(m,5H),1.28(m,5H).

[0151] 7. 4-(3-(benzylamino)-4-(cyclohexylamino)benzyl)-5-methyl-2-phenyl-2,4-dihydro-3H-pyrazole-3-one (III-3)

[0152] The synthesis method is the same as that for I-24, and the product is a pale yellow solid with a yield of 70.65%. 1 H NMR(400MHz,Chloroform-d)δ7.64(d,J=3.4Hz,2H),7.44–7.35(m,5H),7.33–7.28(m,2H),6.66(d,J=8.2Hz,1H ),3.35–3.26(m,3H),2.10–2.02(m,2H),1.92(s,3H),1.80–1.73(m,2H),1.72–1.61(m,2H),1.45–1.33(m,4H).

[0153] Example 4: The compound can effectively inhibit ferroptosis.

[0154] Ferroplasmosis inducers, such as RSL3 and erastin, can increase intracellular ROS and lipid peroxides, thereby inducing ferroptosis. Ferroplasmosis can be inhibited by free radical scavengers (Fer-1, liproxstatin), iron ion chelators, etc. Therefore, the ability of ferroptosis inhibitors to alleviate ferroptosis can be indicated by the reversal of ferroptosis induced by ferroptosis inducers.

[0155] Cell lines: human fibrosarcoma cells HT1080 and mouse hippocampal neurons HT22, purchased from Wuhan Pronosei Life Science Technology Co., Ltd.

[0156] Methods: The MTT assay was used, specifically as follows: Human renal cell carcinoma lines HT1080 or HT22 in logarithmic growth phase were collected by digestion and diluted. Approximately 4000-5000 cells per well were seeded into 96-well plates, with three replicates and 80 μL per well. The plates were incubated overnight at 37°C and 5% CO2. The experiment included a DMSO control group and nine different concentrations of the compound treatment groups. Each treatment group received 10 μL of a different concentration of the compound, and a DMSO control group (diluted to the highest concentration compound) was also included. After incubation for 1 hour at 37°C and 5% CO2, ferroptosis was induced by adding 2 μM / 200 nM RSL3 to each compound concentration. RSL3 and DMSO control groups were also included. The plates were incubated for another 48 hours at 37°C and 5% CO2. Add 20 μL of 5 mg / mL MTT solution to each well and incubate the cells at 37°C for 2 hours. Then add 100 μL of DMSO to each well, shake gently for 10 minutes, and mix thoroughly. Measure the optical density (OD value) of each well at 570 nm using an ELISA reader. Repeat the experiment three times. Calculate the survival rate (%) using the following formula: Survival rate % = (OD value of experimental group / OD value of DMSO control group) × 100%.

[0157] The results are shown in Table 1, indicating that the various compounds of the present invention can significantly inhibit ferroptosis and have good ferroptosis inhibitory activity.

[0158] Table 1. Inhibitory activity of compounds against RSL3-induced ferroptosis in HT1080 cells.

[0159] In the table: "A" indicates IC 50 ≤0.1μM, “B” indicates IC50>0.1μM and ≤0.5μM, “C” indicates IC50>0.5μM.

[0160] Table 2. Inhibitory activity of compounds against RSL3-induced ferroptosis in HT22 cells.

[0161] In the table: "A" indicates IC 50 ≤0.1μM, “B” indicates IC50>0.1μM and ≤0.5μM, “C” indicates IC50>0.5μM.

[0162] Example 5: The compound exhibits strong free radical scavenging ability in vitro.

[0163] Methods: Fer-1, Vc, and the compound were diluted in ethanol to prepare different concentrations, which were then added to 96-well plates. Five concentration gradients were set for all compounds: 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, and 0 μM. 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH) was dissolved in ethanol to prepare a reaction solution with a final concentration of 100 μM, which was then added to the 96-well plates containing the compound. The mixture was then incubated at room temperature in the dark for 30 minutes. The absorbance at 517 nm was measured. The results are shown in Figure 1. Taking compound I-39 as an example, it can scavenge free radicals in a dose-dependent manner, and it also exhibits superior free radical scavenging ability at low concentrations. Compared with Fer-1 and Vc, compound I-39 showed similar free radical scavenging ability, indicating that aromatic alkylamine ferroptosis inhibitors based on the edaravone structure have a strong ability to scavenge free radicals.

[0164] Example 6: The compound exhibits low cytotoxicity and high safety, making it suitable for in vivo efficacy evaluation.

[0165] Cell lines: Mouse hippocampal neuron HT22 and human umbilical vein endothelial cells HUVEC were purchased from Wuhan Pronosei Life Science Technology Co., Ltd.

[0166] Methods: The MTT assay was used as follows: Human renal cell carcinoma line HT22 / HUVEC in logarithmic growth phase was collected by digestion and diluted. Approximately 4000-5000 cells per well were seeded into 96-well plates, with three replicates and 90 μL per well. Cells were incubated overnight at 37°C with 5% CO2. A DMSO control group and different concentrations of the compound were included in the incubation. Cells were incubated for another 48 hours at 37°C with 5% CO2. 20 μL of 5 mg / mL MTT solution was added to each well, and cells were incubated at 37°C for another 2 hours. Then, 100 μL of DMSO was added to each well, and the mixture was gently shaken for 10 minutes to mix. The optical density (OD) of each well was measured at 570 nm using an ELISA reader. The experiment was repeated three times. The survival rate (%) was calculated using the following formula: Survival rate % = (OD value of experimental group / OD value of DMSO control group) × 100%. The results are shown in Figures 2 and 3. Taking compound I-39 as an example, it did not show toxicity to mouse hippocampal neurons (HT22) and human umbilical vein endothelial cells (HUVEC) at 100 μM, which was significantly better than compound Fer-1. This demonstrates that aromatic alkylamine ferroptosis inhibitors based on the edaravone structure have strong safety at the cellular level and are suitable for further pharmacological and biological evaluation.

[0167] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. An aromatic alkylamine-based ferroptosis inhibitor based on the structure of edaravone, characterized in that, The structural formula of the aromatic alkylamine ferroptosis inhibitor is as shown in the following formula I, formula II or formula III: wherein R 1 is selected from hydrogen, alkyl, aryl, C1to C6alkyl-aryl, C1to C6alkyl-phenol, or C3to C6cycloalkyl; and 10 cycloalkyl; R 2 Selected from C0-C8 alkyl, C3-C 12 Cycloalkyl, adamantyl, or polyacetylenic; R 3 selected from hydrogen, alkyl, aryl, C1to C6alkyl-aryl, C1to C6alkyl-phenyl, or C3to C6cycloalkyl; and 10 cycloalkyl; R 4 , R 5 , R 6 , R 7 and R 8 are each selected from H, C1-C6alkyl, OH, OCH3, OCH(CH3)2, OCH2CH(CH3)2, OC(CH3)3, O(C1-C6alkyl), OCF3, OCH2CH2OH, O(C1-C6alkyl)OH, F, Cl, Br, I, CF3, CN, NO2, NH2, C1-C6heteroalkyl, C1-C6hydroxyalkyl, C i -C6alkoxy, C1-C6alkyl, aryl, heteroaryl, C3-C7cycloalkyl, heterocycloalkyl, alkylaryl, CO2R a , C(O)R a , NH(C1-C4alkyl), N(C1-C4alkyl)2, NH(C3-C7cycloalkyl), NHC(O)(C1-C4alkyl), CONR a , NC(O)R a , NS(O) 1 / 2 R a , S(O) 1 / 2 NR a , S(O) 1 / 2 R, C(O)O(C1-C4alkyl), OC(O)N(R a )2, C(O)(C1-C4alkyl), or C(O)NH(C1-C4alkyl); wherein R 4 , R 5 , R 6 , R 7 and R 8 are each one of the above substituents, or wherein two, three, or four of R 4 , R 5 , R 6 , R 7 and R 8 are simultaneously selected from the above substituents; Among them, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C3-C8 cycloalkoxy, phenyl, benzyl, naphthyl, C5-C 10 Aromatic heterocyclic groups, C3-C7 saturated heterocyclic groups, C3-C 12 Cycloalkyl, polyynyl, aryl, C1-C6 alkyl-aryl, C1-C6 alkyl-phenolic or C3-C 10 Cycloalkyl groups can be substituted by one or more atoms or groups; wherein R a is selected from H, CH3, CH2CH3, C3-C6alkyl, C1-C6haloalkyl, or optionally substituted aryl, alkylaryl, piperazinyl, piperidinyl, morpholinyl, heterocycloalkyl, heteroaryl, C1-C6alkoxy, NH(C1-C4alkyl), and N(C1-C4alkyl)2, wherein the optionally substituted group is selected from C1-C6alkyl or C2-C7propenoyl.

2. The edaravone structure-based aromatic alkylamine ferroptosis inhibitor according to claim 1, characterized in that, The inhibitors are compounds I-1 to I-40, II-1 to II-3, III-1 to III-3, the structural formula of which is as follows:

3. The process for the preparation of the idarubicin structure based aromatic alkylamine ferroptosis inhibitor as claimed in claim 2, wherein the process is characterized by, The preparation method comprises the following steps: The synthesis of the compounds I-1 to I-40 is as follows: compound 1 and compound 2 react in an ice acetic acid solution to obtain edaravone analogue 3; compound 4 and the corresponding amine undergo substitution reaction to obtain compound 5, which then undergoes hydrolysis reaction and condensation reaction to obtain key intermediate compound 7; compounds I-1 to I-23 are obtained by reduction of compound 7 with palladium-carbon and hydrogen; then, the corresponding aldehyde or ketone undergoes reductive amination reaction in the presence of sodium triacetoxyborohydride to obtain target compounds I-24 to I-40; Reagents and reaction conditions: (a) Glacial acetic acid, 100℃; Reagents and reaction conditions: (a) sodium nitrate, stannous chloride, hydrochloric acid, 0°C- room temperature; (b) ethyl acetoacetate or its analogues, glacial acetic acid; (c) corresponding amine, potassium carbonate, dimethyl sulfoxide; (d) hydrogen, 10% palladium on carbon, methanol, room temperature; (e) corresponding aldehyde / ketone, sodium triacetoxyborohydride, dichloromethane, room temperature.

4. The process for the preparation of the idarubicin structure based aromatic alkylamine ferroptosis inhibitor as claimed in claim 2, wherein the process is characterized by, The preparation method comprises the following steps: The synthesis of the compounds II-1 to II-3 is as follows: diazotization of compound 8, sodium nitrate and stannous chloride in hydrochloric acid solution to generate compound 9, followed by cyclization with ethyl acetoacetate or its analogs to generate compound 10; then substitution reaction, reduction reaction of compound 10 to obtain the target compound II-1; II-2 and II-3 are obtained from II-1 and the corresponding aldehyde, ketone by reductive amination reaction; Reagents and reaction conditions: (a) sodium nitrate, stannous chloride, hydrochloric acid, 0°C- room temperature; (b) ethyl acetoacetate or its analogues, glacial acetic acid; (c) corresponding amine, potassium carbonate, dimethyl sulfoxide; (d) hydrogen, 10% palladium on carbon, methanol, room temperature; (e) corresponding aldehyde / ketone, sodium triacetoxyborohydride, dichloromethane, room temperature.

5. The process for the preparation of the idarubicin structure based aromatic alkylamine ferroptosis inhibitor as claimed in claim 2, wherein the process is characterized by, The preparation method comprises the following steps: The synthesis of the compounds III-1~III-3 is as follows: 4-chloro-3-nitrobenzaldehyde is used as a starting material, and sodium borohydride is used for reduction to obtain the corresponding hydroxyl-containing compound 13; substitution reaction of the compound 13 and the corresponding amine obtains compound 14, and then pyridine chlorochromate is used for oxidation in chloroform solution to obtain compound 15; condensation reaction of compound 15 and an edaravone analogue in acetic acid solution obtains the key intermediate compound 16; the target compound III-1 is obtained by hydrogen reduction of compound 16, and compounds III-2 and III-3 are obtained by reductive amination reaction of III-1 and the corresponding aldehyde, ketone; Reagents and reaction conditions: (a) sodium nitrate, stannous chloride, hydrochloric acid, 0°C- room temperature; (b) ethyl acetoacetate or its analogues, glacial acetic acid; (c) corresponding amine, potassium carbonate, dimethyl sulfoxide; (d) hydrogen, 10% palladium on carbon, methanol, room temperature; (e) corresponding aldehyde / ketone, sodium triacetoxyborohydride, dichloromethane, room temperature.

6. Use of the inhibitor of claim 1 or 2 in the preparation of an inhibitor of ferroptosis and / or an inhibitor of ferroptosis-Edaravone.

7. Use of the inhibitor of claim 1 or 2 in the preparation of a treatment of a disease related to ferroptosis and / or a disease that can be treated by Edaravone.

8. Use according to claim 7, characterized in that, The disease related to ferroptosis includes neurodegenerative diseases, tissue ischemia-reperfusion injury, stroke, cardiovascular, liver and kidney failure, inflammation, diabetic complications and cardiovascular diseases.

9. Use according to claim 7, characterized in that, The disease that can be treated by Edaravone includes neurological diseases, chronic neurodegenerative diseases amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's syndrome, cerebrovascular disease, etc., pulmonary diseases, sepsis.

10. A medicament for treating a disease associated with ferroptosis and / or a disease capable of being treated by edaravone, characterized by, The drug contains the aromatic amine ferroptosis inhibitor of claim 1 or 2 and a pharmaceutically acceptable salt, carrier or adjuvant.

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