Bifunctional aromatic alkylamine inhibitor for ferroptosis and / or sigma receptors, and preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a bifunctional aromatic alkylamine inhibitor for ferroptosis and / or Sigma receptors, and a preparation method therefor and the use thereof. The bifunctional inhibitor has the chemical structural formula of: (I). The bifunctional inhibitor is an arylalkylamine compound, can inhibit ferroptosis caused by a ferroptosis inducer, and can reduce the intracellular levels of reactive oxygen species. The arylalkylamine compound can also be used as a Sigma receptor inhibitor, showing relatively strong affinity for Sigma receptors. Moreover, it is verified by means of experiments that the arylalkylamine compound can inhibit monomeric amyloid Aβ1-42 from forming aggregates thereof. Therefore, the arylalkylamine compound provided in the present invention has great application value in the treatment, by means of multiple effects, of neurological diseases associated with ferroptosis and Sigma receptors.

Description

Aromatic amine ferroptosis and / or sigma receptor bifunctional inhibitor and preparation method and application thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and specifically relates to an aromatic amine ferroptosis and / or sigma receptor bifunctional inhibitor and a preparation method and application thereof. BACKGROUND

[0002] Cell death can be roughly divided into two categories: uncontrolled cell death caused by excessive cell damage and regulated cell death dependent on strictly controlled molecular pathways. Apoptosis is the most typical form of regulated cell death, which initiates cell death by activating caspases. Ferroptosis is a new cell death mode discovered in recent years, which is oxidative cell death induced by various reasons, and has iron ion dependence. Its occurrence is caused by the imbalance between the generation and degradation of intracellular lipid reactive oxygen species (ROS). The control mechanism of ferroptosis elucidated in the first few years of its discovery mainly revolves around cysteine and glutathione metabolism, and phospholipid peroxidase GPX4 prevents the accumulation of peroxidized lipids. Ferroptosis inducers directly or indirectly act on glutathione peroxidase (GPXs) through different pathways, leading to decreased cellular antioxidant capacity, ROS accumulation, and ultimately causing oxidative cell death. The complex interaction between lipid, iron and cysteine metabolism has become an important regulatory factor for this cell death pathway. Recently, the regulation of ferroptosis has become an attractive strategy for intervention in human diseases, including cancer, neurodegenerative diseases and ischemic diseases.

[0003] Ferroptosis can be inhibited by iron chelators, lipophilic antioxidants and / or ferrostatin-1 (fer-1). Fer-1 is an arylalkylamine with antioxidant properties, and is one of the earliest identified ferroptosis inhibitors. As a lipid peroxidation reductant, fer-1 intercepts and eliminates lipid free radicals through hydrogen atom transfer or direct reduction.

[0004] Sigma (σ) receptors are integral membrane proteins that are widely expressed in the central nervous system and peripheral tissues. Two subtypes, σ1 and σ2, are distinguished by differences in tissue distribution and pharmacological profile, although the two proteins are not related in sequence. The σ1 receptor was cloned in 1996 and has no close homolog in the human genome; its closest known functional homolog is the yeast Δ8,7 sterol isomerase ERG2. The σ2 receptor structure was only confirmed in recent years, also called TMEM97, a resident membrane protein of the endoplasmic reticulum (ER)-regulated sterol transporters NPC1. TMEM97 is predicted to be a four-helix bundle protein with its amino and carboxy termini facing the cytoplasm. The σ2 receptor is overexpressed in proliferating cells and many tumors, and labeled σ2 ligands have been proposed as tools for cancer diagnosis and treatment. Consistent with its high expression in the central nervous system (CNS), the σ2 receptor is also considered a target for the treatment of CNS diseases. σ2 receptor ligands can alleviate alcohol withdrawal symptoms and have neuroprotective effects in brain injury. SUMMARY

[0005] In view of the deficiencies of the prior art, the purpose of the present application is to provide a kind of aromatic amine class of ferroptosis and / or Sigma receptor bifunctional inhibitor and its preparation method and application.The aromatic amine class of compound is strong in anti-ferroptosis activity, can be used to prepare the drug for treating ferroptosis related diseases;And have Sigma receptor affinity, suitable for the treatment of Sigma receptor related diseases, such compounds have the advantages of both can be used to prepare the drug for treating both related diseases.In order to achieve the above-mentioned application purposes, the technical solutions adopted by the present application are as follows:

[0006] The present application provides a kind of aromatic amine class of ferroptosis and / or Sigma receptor bifunctional inhibitor, it has the structural formula as shown in the following:

[0007] In the formula, R 1 selected from hydrogen, alkyl, aryl, C1-C6 alkyl-aryl, C1-C6 alkyl-phenol or C3-C 10 cycloalkyl;R 2 selected from C0-C8 alkyl, C3-C 12 cycloalkyl, adamantyl or polyacetylene group;

[0008] Linker is selected from a direct bond, C1-C6alkyl or C1-C6alkyl containing from 1 to 3 independent substituents, C2-C6alkenyl or C2-C6alkenyl containing from 1 to 3 independent substituents, (C0-C6alkyl)-(C3-C6cycloalkyl)-(C0-C6alkyl) or (C0-C6alkyl)-(C3-C6cycloalkyl)-(C0-C6alkyl) containing from 1 to 3 independent substituents, (C0-C6alkyl)-Z-(C0-C6alkyl) or (C0-C6alkyl)-Z-(C0-C6alkyl) containing from 1 to 3 independent substituents; wherein Z is selected from N(R a ), -SO2-, OC(=O) or C(=O)O;

[0009] when the A ring is present as an aromatic ring, R 3 , R 4 , R 5 and R 6 are 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, arahetero, 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), C(O)NH(C1-C4alkyl); wherein R 3 , R 4 , R 5 and R 6 may individually be the above substituents, or two, three or four of R 3 , R 4 , R 5 , R 6 are simultaneously the above substituents;

[0010] X is selected from CH2, O, S, NH, NH(C1-C4alkyl), N(C1-C4alkyl)2, NH(C3-C7cycloalkyl), NHC(O)(C1-C4alkyl), NC(O)R a 1 / 2 a ;

[0011] m, n are the number of carbon atoms and can be 0, 1, 2;

[0012] 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 groups are selected from C1-C6alkyl or C2-C7propenoic acid groups;

[0013] wherein C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C3-C8cycloalkyl, C3-C8cycloalkoxy, phenyl, benzyl, naphthyl, C5-C 10 aromatic heterocyclyl, C3-C7saturated heterocyclyl, C3-C 12 cycloalkyl, polyalkynyl, aryl, C1-C6alkyl-aryl, C1-C6alkyl-phenol or C3-C 10 cycloalkyl can be substituted by one or more atoms or groups.

[0014] Further, the inhibitors are compounds I-1 to I-22, II-1 to II-17, whose structural formulae are specifically as follows:

[0015] The present application also provides a preparation method of the aromatic alkylamine ferroptosis and / or Sigma receptor bifunctional inhibitor, and the preparation method comprises the following steps:

[0016] ​​The synthesis of the compounds I-1~I-22 is as follows: 4-chloro-3-nitrobenzaldehyde is used as a raw material, sodium borohydride is used for reduction to obtain a hydroxyl compound 2, then substitution reaction with a corresponding amine to obtain compound 3; compound 3 is oxidized by PCC to obtain an aldehyde compound 4, aldol condensation reaction of 4 with acetone under alkaline conditions to obtain compound 5; sodium borohydride is used for reduction of the double bond and the carbonyl group of compound 5 to obtain compound 6, hydrogen reduction of the nitro group to obtain a key intermediate 7; (Boc)2O is used for protection of the amino group to obtain a corresponding compound 8; halogenation reaction of 8 with CBr4 in the presence of PPh3 to obtain a halogenated compound 9; compound 10 is obtained by substitution reaction of 9 with a corresponding aromatic amine or aliphatic amine, then removal of the Boc protecting group to obtain compounds I-1~I-22;

[0017] Reagents and reaction conditions: (a) sodium borohydride, methanol; (b) cyclohexylamine, potassium carbonate, DMSO, 150°C; (c) PCC, chloroform; (d) acetone, sodium hydroxide; (e) sodium borohydride, methanol; (f) Pd / C, hydrogen, methanol; (g) (Boc)2O, triethylamine, tetrahydrofuran; (h) carbon tetrabromide, triphenylphosphine, dichloromethane; (i) corresponding amine, potassium carbonate, potassium iodide, DMF; (j) i: HCl / CH3CH2OH; ii: corresponding aldehyde, sodium triacetoxyborohydride, dichloromethane.

[0018] The application also provides a preparation method of the aromatic alkylamine ferroptosis and / or Sigma receptor bifunctional inhibitor, and the preparation method comprises the following steps:

[0019] The synthesis of the compounds II-1~II-5, II-16 and II-17 is as follows: compound 11 is subjected to radical substitution reaction with NBS under the catalysis of AIBN to obtain compound 12, then affinity substitution reaction with 2-methylbut-3-yn-2-amine to obtain a key intermediate 13; compound 13 is subjected to coupling reaction with a corresponding compound, such as 4-bromo-N-cyclohexyl-2-nitroaniline, under the catalysis of Pd(PPh3)2Cl2 and CuI to obtain a key intermediate compound 14; compound II-16 / 17 is obtained by reduction of compound 14 with zinc powder; and compounds II-1~II-5 need to be further reduced under hydrogen conditions to obtain;

[0020] Reagents and reaction conditions: (k) NBS, AIBN, chloroform, reflux; (l) 2-methylbut-3-yn-2-amine, potassium carbonate, tetrahydrofuran, 60°C; (m) 4-bromo-N-cyclohexyl-2-nitroaniline, Pd(PPh3)2Cl2, cuprous iodide, triethylamine, 40°C; (n) i: zinc powder, hydrochloric acid, methanol; ii: corresponding aldehyde, sodium triacetoxyborohydride, dichloromethane; (o)

[0021] Pd / C, hydrogen, methanol.

[0022] The application also provides a preparation method of the aromatic alkylamine ferroptosis and / or Sigma receptor bifunctional inhibitor, and the preparation method comprises the following steps:

[0023] The synthesis of the compounds II-4~II-15 is as follows: 4-chloro-3-nitrobenzoic acid is used as a starting material, and a condensation reaction is performed with a corresponding amine group segment to obtain compound 16, then a nucleophilic substitution reaction is performed with an amine group derivative to obtain compound 17, and after the Boc protecting group is removed, an intermediate 18 is obtained. The nucleophilic substitution is performed between compound 18 and a corresponding bromide compound to obtain a key intermediate 19, and further reduction with zinc powder obtains the target compounds II-4~II-15.

[0024] Reagents and reaction conditions: (p) i: sulfurous acid chloride, dichloromethane, DMF; ii: (2-aminoethyl) tert-butyl carbamate, triethylamine, dichloromethane; (q) cyclohexylamine, potassium carbonate, DMF, 80 DEG C; (r) trifluoroacetic acid, dichloromethane; (s) 1,2-bis(bromomethyl)benzene, potassium carbonate, tetrahydrofuran, 60 DEG C; (t) i: zinc powder, hydrochloric acid, methanol; ii: corresponding aldehyde, sodium triacetoxyborohydride, dichloromethane.

[0025] The application also provides application of the inhibitor in preparation of a ferroptosis inhibitor and / or a Sigma receptor inhibitor.

[0026] The application also provides application of the inhibitor in preparation of a drug for treating a ferroptosis related disease and / or a Sigma receptor related disease.

[0027] The application also provides application of the inhibitor in preparation of a related neurological disease caused by Aβ amyloid protein aggregation.

[0028] Further, the ferroptosis related disease includes neurodegeneration, tissue ischemia-reperfusion injury, stroke, cardiovascular, liver and kidney failure, inflammation, diabetic complications.

[0029] Further, the Sigma receptor related disease includes neurodegeneration (Alzheimer's disease, Parkinson's disease), stroke, drug addiction, neuropathic pain, schizophrenia and depression.

[0030] Further, the related neurological disease caused by Aβ amyloid protein aggregation includes but is not limited to Alzheimer's disease, Parkinson's syndrome and the like.

[0031] Further, the drug further comprises a pharmaceutically acceptable salt, a carrier or an auxiliary agent.

[0032] Further, the treatment of ferroptosis by the aromatic alkylamine compound is that the compound can act as a free radical scavenger to reduce the level of active oxygen in cells, lipid peroxide, and thus rescue ferroptosis caused by active oxygen, wherein the active oxygen includes but is not limited to H2O2, t-BuOOH, OH·, ONOO 2- - -

[0033] Further, the treatment of ferroptosis by the aromatic alkylamine compound is that the compound can act as a free radical scavenger to rescue ferroptosis caused by ferroptosis inducers, wherein the ferroptosis inducers include but are not limited to RSL3 and its derivatives, Erastin and its derivatives, and ML162 and its derivatives.

[0034] Further, the treatment of Sigma receptor related diseases by the aromatic alkylamine compound is that the compound can act as a Sigma receptor inhibitor to inhibit related diseases caused by incorrect or abnormal expression of Sigma.

[0035] Compared with the prior art, the present application has the advantages and beneficial effects that the present application obtains an aromatic alkylamine compound capable of acting on ferroptosis and Sigma receptor at the same time, and enriches the structure of ferroptosis and Sigma receptor inhibitors. The aromatic alkylamine compound can inhibit ferroptosis caused by ferroptosis inducers, and can reduce the level of active oxygen in cells, which is verified by experiments. The aromatic alkylamine compound can act as a Sigma receptor inhibitor, and shows strong affinity with Sigma receptor, which is also verified by experiments. The aromatic alkylamine compound can inhibit the formation of monomeric amyloid protein Aβ 1-42 into its polymer, and thus the aromatic alkylamine compound provided by the present application can play a good application value in the treatment of neurological diseases related to ferroptosis and Sigma receptor through multiple actions. BRIEF DESCRIPTION OF DRAWINGS

[0036] Figure 1 shows the inhibitory effect of compound I-7 on Aβ 1-42 . DETAILED DESCRIPTION

[0037] The embodiments of the present application are described in detail below, and the embodiments are implemented on the premise of the technical scheme of the present application, and detailed implementation modes and specific operation processes are given, but the protection scope of the present application is not limited to the following embodiments.

[0038] Embodiment 1

[0039] 1, (4-chloro-3-nitrophenyl) methanol (compound 2)​​​

[0040] To a flask containing 40 mL of methanol was added 4-chloro-3-nitrobenzaldehyde (2.19 g, 11.8 mmol) and NaBH4(0.22 g, 5.9 mmol) was added portionwise at room temperature. The reaction was monitored by TLC until completion. The NaBH4was quenched with 1 N HCl and the solvent was evaporated. The residue was extracted with ethyl acetate and the organic layers were combined. The organic layer was washed with saturated brine and dried over anhydrous Na2SO4. The organic layer was concentrated under vacuum to give compound 2 as a light yellow solid (2 g, 90%). 1 H NMR (400 MHz, Chloroform-d) δ 7.88 (d, J = 8.5 Hz, 1H), 7.51 (d, J = 8.7 Hz, 2H), 4.76 (s, 2H).

[0041] 2, (4-(cyclohexylamino)-3-nitrophenyl)methanol (compound 3)

[0042] To a flask containing 40 mL of methanol was added 4-chloro-3-nitrobenzaldehyde (2.19 g, 11.8 mmol) and NaBH4(0.22 g, 5.9 mmol) was added portionwise at room temperature. The reaction was monitored by TLC until completion. The NaBH4was quenched with 1 N HCl and the solvent was evaporated. The residue was extracted with ethyl acetate and the organic layers were combined. The organic layer was washed with saturated brine and dried over anhydrous Na2SO4. The organic layer was concentrated under vacuum to give compound 2 as a light yellow solid (2 g, 90%). 1 H NMR (400 MHz, Chloroform-d) δ 7.88 (d, J = 8.5 Hz, 1H), 7.51 (d, J = 8.7 Hz, 2H), 4.76 (s, 2H).

[0043] 3, 4-(cyclohexylamino)-3-nitrobenzaldehyde (compound 4)

[0044] To a flask containing 40 mL of methanol was added 4-chloro-3-nitrobenzaldehyde (2.19 g, 11.8 mmol) and NaBH4(0.22 g, 5.9 mmol) was added portionwise at room temperature. The reaction was monitored by TLC until completion. The NaBH4was quenched with 1 N HCl and the solvent was evaporated. The residue was extracted with ethyl acetate and the organic layers were combined. The organic layer was washed with saturated brine and dried over anhydrous Na2SO4. The organic layer was concentrated under vacuum to give compound 2 as a light yellow solid (2 g, 90%).

[0045] 4, 4-(4-(cyclohexylamino)-3-nitrophenyl)but-3-en-2-one (Compound 5)

[0046] To a solution of 4-(cyclohexylamino)-3-nitrobenzaldehyde (0.4 g, 1.6 mmol) in acetone / ethanol (1.6 mL / 0.2 mL) was added 10% aqueous sodium hydroxide solution (7 mL) at room temperature under stirring condition. TLC was monitored until the reaction was complete. The organic solvent was evaporated under vacuum and the residual water was extracted with DCM. The organic layer was dried and concentrated. The mixture was separated by column chromatography to obtain the target compound 5 as a dark red solid in 79.3% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.38 (d, J = 7.1 Hz, 1H), 8.33 (s, 1H), 7.61 (d, J = 9.0 Hz, 1H), 7.40 (d, J = 16.2 Hz, 1H), 6.90 (d, J = 9.0 Hz, 1H), 6.57 (d, J = 16.2 Hz, 1H), 3.61 - 3.49 (m, 1H), 2.35 (s, 3H), 2.05 (dd, J = 9.3, 3.5 Hz, 2H), 1.75 (dd, J = 58.5, 12.6 Hz, 3H), 1.48 - 1.28 (m, 5H).

[0047] 5, 4-(4-(cyclohexylamino)-3-nitrophenyl)but-3-en-2-one (Compound 5)

[0048] Synthesis method is the same as compound 2, yield is 83%. 1 H NMR (400 MHz, Chloroform-d) δ 8.19 (d, J = 6.7 Hz, 1H), 8.12 (s, 1H), 7.48 (d, J = 9.0 Hz, 1H), 6.84 (d, J = 8.7 Hz, 1H), 6.43 (d, J = 15.8 Hz, 1H), 6.16 - 6.05 (m, 1H), 4.47 (p, J = 6.6 Hz, 1H), 3.56 - 3.46 (m, 1H), 2.07 - 2.01 (m, 2H), 1.84 - 1.76 (m, 2H), 1.66 (dd, J = 12.9, 2.8 Hz, 1H), 1.46 - 1.32 (m, 8H).

[0049] 6, 4-(3-amino-4-(cyclohexylamino)phenyl)butan-2-ol (Compound 7)

[0050] Compound 6 (0.3 g, 1.0 mmol) was added to a reaction vial containing a methanol solution, Pd / C (0.03 g) was added and the reaction was allowed to proceed to completion under hydrogen conditions. The Pd / C was filtered through filter paper and the solvent was concentrated to yield compound 7 as a light yellow solid (0.24 g, 90%). 1 H NMR (400 MHz, Chloroform-d) δ 6.59 (dd, J = 11.9, 4.2 Hz, 3H), 3.81 (dt, J = 12.1, 6.0 Hz, 1H), 3.22 - 3.09 (m, 1H), 2.66 - 2.49 (m, 2H), 2.05 (d, J = 4.6 Hz, 2H), 1.79 - 1.62 (m, 5H), 1.29 (dd, J = 22.0, 4.9 Hz, 3H), 1.22 - 1.18 (m, 5H).

[0051] 7, tert-Butyl (2-(cyclohexylamino)-5-(3-hydroxybutyl)phenyl)carbamate (Compound 8)

[0052] Compound 7 (1 g, 3.8 mmol) was dissolved in 30 mL THF, triethylamine (0.46 g, 4.56 mmol) was added, followed by slow dropwise addition of (Boc)20 under ice bath conditions. After the dropwise addition was complete, the reaction was allowed to proceed to completion at room temperature. The reaction was concentrated and the mixture was separated by column chromatography to yield compound 8 as a yellow solid in 90% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.31 (s, 1H), 6.86 (t, J = 8.0 Hz, 1H), 6.70 (t, J = 7.0 Hz, 1H), 3.88 - 3.74 (m, 1H), 3.17 - 3.02 (m, 1H), 2.60 (qd, J = 15.9, 15.1, 9.1 Hz, 2H), 2.05 - 1.92 (m, 2H), 1.81 - 1.57 (m, 7H), 1.51 (d, J = 9.0 Hz, 9H), 1.34 (dd, J = 12.4, 6.9 Hz, 2H), 1.23 - 1.18 (m, 4H).

[0053] 8, tert-Butyl (5-(3-bromobutyl)-2-(cyclohexylamino)phenyl)carbamate (Compound 9)

[0054] Compound 8 (1 g, 2.7 mmol) was added to a 40 mL vial, followed by the addition of PPh3 (0.8 g, 3.3 mmol) and CBr4 (1.1 g, 3.3 mmol), and the reaction was allowed to proceed for 1 h at room temperature. The reaction was concentrated and the mixture was separated by column chromatography to yield compound 9 (0.7 g, 60%) as a yellow oil. 1H NMR (400 MHz, Chloroform-d) δ 7.27 (s, 1H), 6.86 (d, J = 8.1 Hz, 1H), 6.72 - 6.66 (m, 1H), 6.31 (s, 1H), 4.07 (dq, J = 12.2, 7.1 Hz, 1H), 3.18 - 3.07 (m, 1H), 2.77 - 2.58 (m, 2H), 2.05 - 1.94 (m, 4H), 1.77 (d, J = 6.5 Hz, 2H), 1.70 (d, J = 6.6 Hz, 3H), 1.50 (s, 9H), 1.33 (d, J = 11.0 Hz, 2H), 1.25 - 1.13 (m, 4H).

[0055] 9. tert-Butyl (2-(cyclohexylamino)-5-(3-((4-(trifluoromethyl)benzyl)amino)butyl)phenyl)carbamate (Compound 10)

[0056] Compound 9 (0.07 g, 0.165 mmol), (4-(trifluoromethyl)phenyl)methanamine (0.045 g, 0.25 mmol) and K2CO3(0.035 g, 0.25 mmol) were added to a reaction vial with 1 mL DMF as solvent, and reacted at 60 °C overnight. After the reaction was completed, the reaction solution was poured into 15 mL water, and the water layer was extracted with DCM (50 mL x 3), and the combined organic layers were concentrated. The mixture was separated by column chromatography (PE:EA = 4:1) to obtain Compound 10 (0.06 g, 70%) as a light yellow oil. 1 H NMR (400 MHz, Chloroform-d) δ 7.55 (d, J = 8.1 Hz, 2H), 7.41 (d, J = 8.0 Hz, 2H), 7.29 (s, 1H), 6.80 (d, J = 7.1 Hz, 1H), 6.66 (d, J = 8.2 Hz, 1H), 3.89 - 3.75 (m, 2H), 3.16 - 3.05 (m, 1H), 2.69 (p, J = 6.2 Hz, 1H), 2.55 (q, J = 8.5 Hz, 2H), 2.05 - 1.94 (m, 2H), 1.81 - 1.70 (m, 3H), 1.68 - 1.60 (m, 3H), 1.50 (s, 9H), 1.38 - 1.12 (m, 7H), 1.11 (d, J = 6.2 Hz, 3H).

[0057] 10. N 1 - cyclohexyl-4-(3-((4-(trifluoromethyl)benzyl)amino)butyl)benzene-1,2-diamine (Compound I-1)

[0058] Compound 10 obtained in the previous step was added to a reaction flask, and 2N hydrochloric acid was added. The reaction was carried out for 5 hours. The solvent was evaporated, the pH was adjusted to 10 with K2CO3, the aqueous layer was extracted with EA, and then the organic layer was dried and concentrated to obtain a yellow oily substance I-1 (0.04 g, 83.3%). 1 H NMR (400MHz, Methanol-d4) δ7.65(q,J=8.4Hz,4H),7.55(d,J=8.5Hz,2H),7.21(d,J=8.4Hz,1H),4.26(d,J=6.1Hz,2H),2.95(dt,J=9.3,5.0Hz,1H) ,2.81–2.74(m,1H),2.30–2.21(m,1H),2.15(d,J=12.7Hz,2H),1.89(dq, J=32.3,16.2Hz,8H),1.59(q,J=14.4,12.7Hz,3H),1.46(d,J=6.5Hz,3H). 13 C NMR (101MHz, Methanol-d4) δ136.57,134.77,130.21,125.54,123.46,118. 11,110.59,55.45,53.47,35.57,35.12,32.78,31.17,25.31,25.03,15.26.

[0059] 11. 4-(3-(benzylamino)butyl)-N 1 -Cyclohexylphenyl-1,2-diamine (compound I-2)

[0060] The synthesis method is the same as I-1, producing a yellow oily substance with a yield of 79%. 1 H NMR(400MHz,Chloroform-d)δ7.35–7.26(m,5H),6.60(s,2H),6.53(s,1H),3 .85–3.72(m,2H),3.18(td,J=10.0,5.0Hz,1H),2.76–2.71(m,1H),2.52(tt,J =10.5,5.2Hz,2H),2.09–2.03(m,2H),1.81–1.74(m,3H),1.64(ddd,J=14.1,1 0.3,7.2Hz,2H),1.41–1.33(m,2H),1.24–1.16(m,3H),1.14(d,J=6.3Hz,3H). 13C NMR (101 MHz, Chloroform-d) δ 140.72, 134.99, 134.39, 132.74, 128.44, 126.89, 120.00, 116.83, 113.63, 52.16, 51.27, 38.87, 33.78, 31.60, 26.12, 25.10, 20.35.

[0061] 12、N 1 - Cyclohexyl-4-(3-((4-methylbenzyl)amino)butyl)benzene-1,2-diamine (Compound I-3)

[0062] Synthesis as for 1-1, yellow oil, yield 81%. 1 H NMR (400 MHz, Chloroform-d) δ 7.20 - 7.17 (m, 2H), 7.12 (d, J = 7.0 Hz, 2H), 6.58 (s, 2H), 6.52 (s, 1 H), 3.80 - 3.66 (m, 2H), 3.17 (dq, J = 10.1, 4.8, 4.2 Hz, 1 H), 2.74 - 2.69 (m, 1 H), 2.53 - 2.47 (m, 2H), 2.33 (s, 3H), 2.05 (d, J = 11.2 Hz, 2H), 1.76 (dt, J = 13.6, 4.4 Hz, 3H), 1.68 - 1.59 (m, 2H), 1.42 - 1.30 (m, 3H), 1.18 (d, J = 12.1 Hz, 2H), 1.12 (dd, J = 6.3, 1.8 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 137.65, 136.50, 134.99, 134.44, 132.48, 129.18, 128.30, 120.06, 116.88, 113.59, 52.20, 52.04, 51.02, 38.92, 33.82, 31.64, 29.82, 26.16, 25.16, 21.22, 20.36.

[0063] 13、N 1 - Cyclohexyl-4-(3-((4-methylbenzyl)amino)butyl)benzene-1,2-diamine (Compound I-3)

[0064] Synthesis as for 1-1, yellow oil, yield 81%. 1H NMR (400 MHz, Chloroform-d) δ 7.26 - 7.21 (m, 2H), 6.98 (ddd, J = 8.7, 6.8, 3.5 Hz, 2H), 6.58 (d, J = 3.5 Hz, 2H), 6.52 (d, J = 2.9 Hz, 1H), 3.81 - 3.64 (m, 2H), 3.19 - 3.12 (m, 1H), 2.70 (dt, J = 11.8, 5.7 Hz, 1H), 2.58 - 2.44 (m, 2H), 2.04 (d, J = 12.1 Hz, 2H), 1.82 - 1.71 (m, 3H), 1.69 - 1.57 (m, 2H), 1.42 - 1.33 (m, 3H), 1.14 - 1.09 (m, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 160.72, 135.00, 134.48, 132.63, 129.83, 120.05, 116.84, 115.33, 115.12, 113.58, 52.20, 52.11, 50.53, 38.81, 33.82, 31.66, 29.82, 26.16, 25.15, 20.38.

[0065] 14、N 1 - Cyclohexyl-4-(3-((4-chlorobenzyl)amino)butyl)benzene-1,2-diamine (Compound I-5)

[0066] Synthesis as for I-1, yellow oil, yield 78%. 1 H NMR (400 MHz, Chloroform-d) δ 7.27 - 7.18 (m, 4H), 6.57 (d, J = 2.1 Hz, 2H), 6.50 (s, 1H), 3.80 - 3.64 (m, 2H), 3.16 (tt, J = 10.0, 5.6 Hz, 1H), 2.67 (p, J = 6.2 Hz, 1H), 2.57 - 2.43 (m, 2H), 2.08 - 1.99 (m, 2H), 1.75 (ddt, J = 9.7, 6.7, 3.9 Hz, 3H), 1.67 - 1.56 (m, 2H), 1.44 - 1.26 (m, 5H), 1.11 (d, J = 6.3 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 139.28, 134.99, 134.48, 132.61, 129.69, 128.56, 120.05, 116.83, 113.55, 52.19, 52.02, 50.55, 38.86, 33.82, 31.64, 26.16, 25.16, 20.42.

[0067] 15. N 1 -cyclohexyl-4-(3-((4-isopropylbenzyl)amino)butyl)phenyl-1,2-diamine (compound I-6)

[0068] The synthesis method is the same as I-1, producing a yellow oily substance with a yield of 83%. 1 H NMR(400MHz,Chloroform-d)δ7.26–7.24(m,2H),7.21–7.17(m,2H),6.59(s,2H),6. 54(s,1H),3.83–3.68(m,2H),3.17(tt,J=10.1,3.6Hz,1H),2.90(p,J=6.9Hz,1H),2. 75(p,J=6.2Hz,1H),2.60–2.45(m,2H),2.09–2.02(m,2H),1.77(dt,J=12.0,3.8Hz, 3H),1.70–1.59(m,2H),1.42–1.31(m,3H),1.27–1.23(m,9H),1.15(d,J=6.2Hz,3H). 13 C NMR(101MHz,Chloroform-d)δ147.69,135.01,134.47,132.68,128.45,126.58,120. 08,116.88,113.56,52.20,50.91,38.72,33.91,31.61,26.18,25.17,24.18,20.22.

[0069] 16. 4-(3-((4-(tert-butyl)benzyl)amino)butyl)-N 1 -Cyclohexylphenyl-1,2-diamine (compound I-7)

[0070] The synthesis method is the same as I-1, producing a yellow oily substance with a yield of 73%. 1H NMR (400 MHz, Chloroform-d) δ 7.36 - 7.32 (m, 2H), 7.25 (s, 2H), 6.58 (d, J = 1.1 Hz, 2H), 6.54 (s, 1H), 3.83 - 3.65 (m, 2H), 3.17 (tt, J = 10.1, 3.7 Hz, 1H), 2.73 (p, J = 6.2 Hz, 1H), 2.60 - 2.44 (m, 2H), 2.05 (dd, J = 12.6, 2.9 Hz, 2H), 1.77 (ddd, J = 9.5, 7.7, 3.8 Hz, 3H), 1.63 (dtt, J = 13.6, 6.8, 3.5 Hz, 2H), 1.40 - 1.33 (m, 2H), 1.31 (d, J = 2.9 Hz, 9H), 1.20 (ddd, J = 17.1, 9.2, 3.0 Hz, 3H), 1.13 (d, J = 6.3 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 149.85, 134.99, 134.45, 132.78, 128.07, 125.42, 120.08, 116.88, 113.56, 52.25, 50.94, 38.89, 34.56, 33.83, 31.63, 31.50, 26.17, 25.16, 20.37.

[0071] 17、N 1 - Cyclohexyl-4-(3-((2-(trifluoromethyl)benzyl)amino)butyl)benzene-1,2-diamine (Compound I-8)

[0072] Synthesis as for I-1, yellow oil, yield 71%. 1 H NMR (400 MHz, Chloroform-d) δ 7.61 (t, J = 8.0 Hz, 2H), 7.49 (t, J = 7.5 Hz, 1H), 7.32 (t, J = 7.6 Hz, 1H), 6.58 (s, 2H), 6.55 (s, 1H), 3.92 (q, J = 13.9 Hz, 2H), 3.16 (ddd, J = 13.7, 8.3, 3.7 Hz, 1H), 2.75 (h, J = 6.2 Hz, 1H), 2.52 (dt, J = 9.2, 6.1 Hz, 2H), 2.08 - 2.00 (m, 2H), 1.76 (dt, J = 12.9, 3.5 Hz, 3H), 1.63 (ddd, J = 13.6, 9.3, 6.7 Hz, 2H), 1.40 - 1.30 (m, 3H), 1.24 - 1.17 (m, 2H), 1.13 (d, J = 6.3 Hz, 3H). 13C NMR (101 MHz, Chloroform-d) δ 135.01, 134.46, 132.70, 132.01, 130.82, 126.89, 120.04, 116.84, 113.61, 52.65, 52.08, 47.34, 38.98, 33.81, 31.65, 26.16, 25.15, 20.47.

[0073] 18、N 1 - Cyclohexyl-4-(3-((pyridin-4-ylmethyl)amino)butyl)benzene-1,2-diamine (Compound I-9)

[0074] Synthesis as for I-1, yellow oil, yield 82%. 1 H NMR (400 MHz, Chloroform-d) δ 8.53 (s, 1H), 8.47 (d, J = 4.8 Hz, 1H), 7.65 - 7.59 (m, 1H), 7.24 - 7.19 (m, 1H), 6.57 (s, 2H), 6.51 (s, 1H), 3.83 - 3.68 (m, 2H), 3.19 - 3.11 (m, 1H), 2.69 (h, J = 6.0 Hz, 1H), 2.49 (dp, J = 14.0, 6.9 Hz, 2H), 2.03 (d, J = 11.6 Hz, 2H), 1.80 - 1.68 (m, 3H), 1.68 - 1.57 (m, 2H), 1.35 (q, J = 12.1 Hz, 2H), 1.23 - 1.15 (m, 3H), 1.11 (dd, J = 6.3, 1.8 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 149.79, 148.42, 136.30, 135.99, 135.03, 134.48, 132.58, 123.48, 120.00, 116.80, 113.57, 52.27, 52.18, 48.65, 38.94, 33.80, 31.66, 26.15, 25.15, 20.50.

[0075] 19、N 1 - Cyclohexyl-4-(3-(phenylamino)butyl)benzene-1,2-diamine (Compound I-10)

[0076] Synthesis as for I-1, yellow oil, yield 84%. 1H NMR (400 MHz, Chloroform-d) δ 7.55 (d, J = 8.1 Hz, 2H), 7.41 (d, J = 8.0 Hz, 2H), 7.29 (s, 1H), 6.80 (d, J = 7.1 Hz, 1H), 6.66 (d, J = 8.2 Hz, 1H), 3.89 - 3.75 (m, 2H), 3.16 - 3.05 (m, 1H), 2.69 (p, J = 6.2 Hz, 1H), 2.55 (q, J = 8.5 Hz, 2H), 2.05 - 1.94 (m, 2H), 1.81 - 1.70 (m, 3H), 1.68 - 1.60 (m, 3H), 1.50 (s, 9H), 1.38 - 1.12 (m, 7H), 1.11 (d, J = 6.2 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 147.65, 129.24, 120.01, 116.99, 116.78, 113.15, 47.87, 39.01, 33.53, 31.69, 29.70, 26.01, 25.03, 20.85.

[0077] 20、N 1 - Cyclohexyl-4-(3-(cyclohexylamino)butyl)benzene-1,2-diamine (Compound I-11)

[0078] Synthesis as for I-1, yellow oil, yield 74%. 1 H NMR (400 MHz, Chloroform-d) δ 7.64 (ddd, J = 12.0, 8.2, 1.3 Hz, 1H), 7.56 - 7.41 (m, 2H), 3.17 - 3.09 (m, 1H), 3.08 - 2.97 (m, 1H), 2.80 - 2.71 (m, 1H), 2.57 (ddd, J = 14.6, 9.5, 5.4 Hz, 1H), 2.43 - 2.33 (m, 1H), 2.01 (d, J = 10.9 Hz, 4H), 1.69 (tdt, J = 34.4, 17.8, 9.6 Hz, 9H), 1.36 - 1.28 (m, 4H), 1.25 (d, J = 6.4 Hz, 3H), 1.18 - 1.12 (m, 5H). 13 C NMR (101 MHz, Chloroform-d) δ 134.64, 132.21, 128.68, 120.02, 116.76, 113.48, 53.79, 52.12, 36.04, 33.74, 31.37, 31.09, 30.64, 26.13, 25.41, 25.12, 24.99.

[0079] 21、N1 Cyclohexyl-4-(3-(piperidin-l-yl)butyl)benzene-l,2-diamine (Compound I-12)

[0080] Synthetic procedure same as I-1, yellow oil, yield 75%. 1 H NMR (400 MHz, Methanol-d4) δ 6.64 - 6.62 (m, 1H), 6.60 - 6.53 (m, 2H), 3.30 (dt, J = 3.2, 1.5 Hz, 1H), 2.72 - 2.59 (m, 1H), 2.49 - 2.39 (m, 1H), 2.15 - 1.99 (m, 4H), 1.89 - 1.73 (m, 9H), 1.70 - 1.58 (m, 4H), 1.37 (dd, J = 15.8, 4.6 Hz, 5H), 1.25 - 1.15 (m, 3H). 13 C NMR (101 MHz, Methanol-d4) δ 135.42, 134.07, 129.99, 119.14, 116.27, 113.64, 61.85, 52.31, 49.08, 33.07, 32.57, 31.18, 25.81, 24.95, 23.21, 21.72, 12.43.

[0081] 22、N 1 Cyclohexyl-4-(3-((naphthalen-l-ylmethyl)amino)butyl)benzene-l,2-diamine (Compound I-13)

[0082] Synthetic procedure same as I-1, yellow oil, yield 85%. 1 H NMR (400 MHz, Chloroform-d) δ 8.09 (d, J = 8.4 Hz, 1H), 7.81 (d, J = 8.2 Hz, 1H), 7.76 (d, J = 8.3 Hz, 1H), 7.68 - 7.60 (m, 1H), 7.57 - 7.51 (m, 1H), 7.51 - 7.37 (m, 3H), 6.50 - 6.41 (m, 2H), 4.36 (q, J = 13.3 Hz, 2H), 2.97 - 2.87 (m, 1H), 2.46 (ddd, J = 59.1, 15.1, 8.2 Hz, 2H), 2.14 - 1.96 (m, 3H), 1.85 (dd, J = 12.7, 8.8 Hz, 1H), 1.73 (d, J = 12.4 Hz, 2H), 1.63 (d, J = 12.8 Hz, 1H), 1.29 - 1.08 (m, 9H). 13C NMR (101 MHz, Chloroform-d) δ 132.74, 131.11, 131.01, 130.58, 127.96, 127.81, 127.56, 127.44, 125.78, 124.97, 124.45, 122.28, 118.93, 115.71, 51.71, 51.51, 32.54, 30.12, 28.68, 25.00, 24.01.

[0083] 23、N 1 - Cyclohexyl-4-(3-(6,7-dimethoxy-3,4-dihydroquinolin-l(2H)-yl)butyl)-l,2- diamine (Compound I-14)

[0084] Synthesis as for I-1, yellow oil, 84% yield. 1 H NMR (400 MHz, Chloroform-d) δ 6.59 (dd, J = 9.6, 3.9 Hz, 4H), 6.52 (s, 1H), 3.86 - 3.77 (m, 7H), 3.72 - 3.57 (m, 2H), 3.19 - 3.13 (m, 1H), 2.78 (d, J = 9.7 Hz, 4H), 2.68 (q, J = 7.8, 6.4 Hz, 1H), 2.54 (tq, J = 13.9, 8.3, 7.9 Hz, 2H), 2.04 (d, J = 11.8 Hz, 2H), 1.96 - 1.86 (m, 1H), 1.80 - 1.70 (m, 2H), 1.68 - 1.57 (m, 2H), 1.39 - 1.30 (m, 2H), 1.23 - 1.14 (m, 3H), 1.08 (d, J = 5.7 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 147.10, 135.01, 134.38, 133.12, 127.72, 126.80, 120.09, 116.98, 113.61, 111.50, 109.65, 58.18, 56.01, 52.22, 50.92, 45.95, 35.87, 33.83, 32.44, 29.71, 26.17, 25.17, 14.01.

[0085] 24、N 1 - Cyclohexyl-4-(3-(6,7-dimethoxy-3,4-dihydroquinolin-l(2H)-yl)butyl)-l,2- diamine (Compound I-14)

[0086] Synthesis as for I-1, yellow oil, 84% yield. 1H NMR (400 MHz, Chloroform-d) δ 7.00 (d, J = 6.1 Hz, 2H), 6.69 (t, J = 8.8 Hz, 1H), 6.54 (dd, J = 12.2, 7.4 Hz, 3H), 3.87 (d, J = 26.9 Hz, 2H), 3.12 (dq, J = 21.9, 6.9 Hz, 2H), 2.81 (dt, J = 12.3, 6.3 Hz, 1H), 2.59 (ddd, J = 14.0, 9.0, 6.0 Hz, 1H), 2.42 (dt, J = 14.3, 7.9 Hz, 1H), 1.98 (dd, J = 20.8, 8.7 Hz, 3H), 1.76 (td, J = 11.7, 9.2, 5.0 Hz, 3H), 1.69 - 1.60 (m, 1H), 1.39 - 1.31 (m, 5H), 1.26 (d, J = 4.5 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 164.37, 161.76, 135.20, 131.32, 120.02, 116.75, 113.64, 112.28, 112.03, 52.31, 52.06, 48.50, 45.88, 36.54, 33.64, 31.13, 29.76, 26.10, 25.12.

[0087] 25、N 1 - Cyclohexyl-4-(3-((3,5-dichlorobenzyl)amino)butyl)benzene-1,2-diamine (Compound I-16)

[0088] Synthesis as for I-1, yellow oil, yield 83%. 1 H NMR (400 MHz, Chloroform-d) δ 7.22 (s, 3H), 6.58 (d, J = 1.6 Hz, 2H), 6.53 (s, 1H), 3.80 - 3.64 (m, 2H), 3.15 (ddt, J = 10.1, 7.5, 3.7 Hz, 1H), 2.69 (q, J = 6.3 Hz, 1H), 2.59 - 2.45 (m, 2H), 2.03 (dd, J = 9.1, 3.4 Hz, 2H), 1.79 - 1.72 (m, 3H), 1.68 - 1.60 (m, 2H), 1.34 (ddd, J = 15.1, 12.5, 3.2 Hz, 3H), 1.21 - 1.15 (m, 3H), 1.12 (d, J = 6.3 Hz, 3H). 13C NMR (101 MHz, Chloroform-d) δ 135.04, 134.89, 134.50, 127.18, 126.78, 120.02, 116.79, 113.59, 52.19, 50.09, 38.60, 33.79, 31.58, 29.82, 26.16, 25.16, 20.23.

[0089] 26. 4-(3-((3,5-bis(trifluoromethyl)benzyl)amino)butyl)-N 1 - Cyclohexylbenzene-1,2-diamine (Compound 1-17)

[0090] Synthesis as for 1-1, yellow oil, yield 81 %. 1 H NMR (400 MHz, Chloroform-d) δ 7.78 (d, J = 29.7 Hz, 2H), 6.62 - 6.51 (m, 4H), 3.97 - 3.75 (m, 2H), 3.16 (tq, J = 10.2, 3.6 Hz, 1 H), 2.75 - 2.68 (m, 1 H), 2.60 - 2.48 (m, 3H), 2.06 - 2.00 (m, 2H), 1.80 - 1.69 (m, 5H), 1.69 - 1.59 (m, 3H), 1.14 (d, J = 6.3 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 135.05, 134.33, 131.66, 131.33, 128.30, 119.99, 119.87, 116.84, 116.67, 113.64, 67.66, 52.60, 52.19, 50.08, 41.08, 33.68, 31.52, 29.71, 26.06, 25.04, 23.57.

[0091] 27. N 1 - Cyclohexyl-4-(3-((3,5-dimethoxybenzyl)amino)butyl)benzene-1,2-diamine (Compound 1-18)

[0092] Synthesis as for 1-1, yellow oil, yield 80%. 1H NMR (400 MHz, Chloroform-d) δ 6.56 (s, 2H), 6.53 - 6.46 (m, 3H), 6.35 (t, J = 2.2 Hz, 1H), 3.77 (s, 8H), 3.19 - 3.11 (m, 1H), 2.75 (q, J = 6.2 Hz, 1H), 2.49 (dddd, J = 23.8, 17.5, 11.9, 6.5 Hz, 2H), 2.07 - 1.97 (m, 2H), 1.80 - 1.71 (m, 3H), 1.68 - 1.60 (m, 2H), 1.40 - 1.31 (m, 2H), 1.13 (d, J = 6.3 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 160.93, 135.00, 134.49, 132.37, 128.68, 120.03, 116.84, 113.54, 106.36, 99.30, 55.44, 52.17, 52.00, 50.86, 38.32, 33.80, 31.53, 29.81, 26.15, 25.15, 19.73.

[0093] 28、N 1 - Cyclohexyl-4-(3-(phenethylamino)butyl)benzene-1,2-diamine (Compound I-19)

[0094] Synthesis as for I-1, yellow oil, yield 86%. 1 H NMR (400 MHz, Chloroform-d) δ 7.30 - 7.27 (m, 1H), 7.20 (ddd, J = 10.0, 6.7, 3.9 Hz, 4H), 6.56 - 6.47 (m, 3H), 3.14 (ddd, J = 9.7, 7.7, 3.3 Hz, 2H), 2.97 - 2.90 (m, 5H), 2.90 - 2.82 (m, 3H), 2.53 (tt, J = 9.3, 4.6 Hz, 1H), 2.46 - 2.34 (m, 1H), 2.02 (dd, J = 13.7, 4.1 Hz, 2H), 1.96 - 1.89 (m, 1H), 1.78 - 1.68 (m, 3H), 1.64 (dd, J = 10.3, 5.8 Hz, 1H), 1.22 (dd, J = 9.8, 5.2 Hz, 5H). 13C NMR (101 MHz, Chloroform-d) δ 138.71, 135.02, 134.58, 131.35, 128.76, 128.61, 126.52, 119.95, 116.67, 113.51, 53.04, 51.94, 47.30, 36.72, 33.70, 31.22, 26.06, 25.04, 18.36.

[0095] 29、N 1 - Cyclohexyl-4-(3-morpholinopropyl)benzene-1,2-diamine (Compound I-20)

[0096] Synthesis as for I-1, yellow oil, 83% yield. 1 H NMR (400 MHz, Chloroform-d) δ 6.58 (s, 2H), 6.55 (s, 1H), 3.73 (s, 4H), 3.19 - 3.12 (m, 1H), 2.61 - 2.44 (m, 8H), 2.03 (dd, J = 8.5, 3.8 Hz, 2H), 1.79 - 1.70 (m, 2H), 1.68 - 1.60 (m, 1H), 1.57 - 1.47 (m, 1H), 1.39 - 1.29 (m, 2H), 1.23 - 1.12 (m, 3H), 1.03 (d, J = 5.9 Hz, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 135.03, 134.33, 132.57, 119.98, 116.81, 113.66, 67.03, 58.91, 52.18, 48.62, 34.98, 33.71, 32.01, 26.07, 25.05, 13.94.

[0097] 30、N 1 - Cyclohexyl-4-(3-morpholinopropyl)benzene-1,2-diamine (Compound I-20)

[0098] Synthesis as for I-1, yellow oil, 83% yield. 1H NMR (400 MHz, Chloroform-d) δ 7.60 (s, 1H), 7.55 - 7.45 (m, 2H), 7.45 - 7.38 (m, 1H), 6.55 (dd, J = 16.0, 3.6 Hz, 3H), 3.91 - 3.72 (m, 2H), 3.21 - 3.10 (m, 1H), 2.78 - 2.65 (m, 1H), 2.59 - 2.42 (m, 2H), 2.07 - 1.97 (m, 2H), 1.84 - 1.68 (m, 3H), 1.70 - 1.56 (m, 2H), 1.42 - 1.24 (m, 5H), 1.16 - 1.12 (m, 3H).

[0099] 31、N 1 - Cyclohexyl-4-(3-((1,2,3,4-tetrahydronaphthalen-1-yl)amino)butyl)benzene-1,2-diamine (Compound I-22)

[0100] Synthetic procedure same as I-1, yellow oil, yield 71%. 1 H NMR (400 MHz, Chloroform-d) δ 7.32 - 7.28 (m, 1H), 7.12 (dd, J = 5.4, 2.8 Hz, 2H), 7.08 - 7.04 (m, 1H), 6.57 (d, J = 5.3 Hz, 3H), 3.95 - 3.81 (m, 1H), 3.21 - 3.11 (m, 1H), 2.96 - 2.76 (m, 1H), 2.74 - 2.64 (m, 1H), 2.59 - 2.49 (m, 2H), 2.08 - 1.91 (m, 4H), 1.87 - 1.59 (m, 10H), 1.22 - 1.16 (m, 6H).

[0101] Example 2

[0102] 1, 1, 2-Bis(bromomethyl)benzene (Compound 12)

[0103] O-Xylene (0.5 g, 4.7 mmol), NBS (1.84 g, 10.36 mmol) and AIBN (0.08 g, 0.47 mmol) were added to a flask containing 40 mL of chloroform and the reaction was refluxed under N2until the reaction was complete by TLC. The mixture was separated by column chromatography to give a white solid (1.02 g, 82%). 1 H NMR (400 MHz, Chloroform-d) δ 7.38 - 7.34 (m, 2H), 7.32 - 7.29 (m, 2H), 4.66 (s, 4H).

[0104] 2-Methyl-3-butyn-2-yl isatin (Compound 11)

[0105] Methyl 2,3-bis(bromomethyl)benzoate (0.25 g, 0.76 mmol), 2-methylbut-3-yn-2- amine (0.06 g, 0.72 mmol) and K2CO3(0.29 g, 2.2 mmol) were added into a reaction vial, THF (25 mL) was added as solvent, and the mixture was stirred at 60 °C for 18 h. The reaction was poured into 50 mL water, the aqueous layer was extracted with EA (50 mL x 3), and the combined organic layers were concentrated. The mixture was separated by column chromatography (PE:EA = 10:1) to give compound 13 (0.04 g, 21%) as a white solid. 1 H NMR (400 MHz, Chloroform-d) δ 8.17 (d, J = 8.4 Hz, 1H), 7.98 (d, J = 7.5 Hz, 1H), 7.54 (t, J = 7.6 Hz, 1H), 4.92 (s, 2H), 3.96 (d, J = 8.1 Hz, 5H), 2.52 (s, 1H), 1.89 (s, 6H).

[0106] 3, N-cyclohexyl-4-(3-(isoindolin-2-yl)-3-methylbut-1-yn-1-yl)-2-nitroaniline (Compound 14)

[0107] 2-(2-methylbut-3-yn-2-yl)isoindoline (0.03 g, 0.16 mmol), 4-bromo-N-cyclohexyl-2- nitroaniline (0.06 g, 0.19 mmol), Pd(PPh3)2Cl2(0.034 g, 0.0046 mmol) and Cul (0.015 g, 0.008 mmol) were added into a reaction vial, triethylamine (5 mL) was added as solvent, N2 protection, and the mixture was stirred at 40 °C overnight to give a yellow solid (0.04 g. 61%). 1 H NMR (400 MHz, Chloroform-d) δ 8.10 (d, J = 8.8 Hz, 1H), 7.81 (d, J = 7.4 Hz, 1H), 7.52 (dd, J = 7.6, 1.0 Hz, 1H), 7.45 (t, J = 8.0 Hz, 2H), 6.89 (d, J = 1.5 Hz, 1H), 6.62 (dd, J = 8.9, 1.6 Hz, 1H), 4.65 (s, 2H), 3.52 (m, 1H), 2.06 - 2.00 (m, 2H), 1.97 (s, 6H), 1.83 - 1.76 (m, 2H), 1.47 - 1.31 (m, 6H).

[0108] 4, N 1 -cyclohexyl-4-(3-(isoindolin-2-yl)-3-methylbutyl)benzene-1,2-diamine (Compound II-1)

[0109] N-cyclohexyl-4-(3-(isoindoline-2-yl)-3-methylbut-1-yn-1-yl)-2-nitroaniline (0.04 g) was added to a reaction flask containing 20 mL of solvent, and palladium on carbon (0.012 g) was added. The reaction was carried out under H2 conditions. After the reaction was completed, the palladium on carbon was filtered off, and the mixture was separated by column chromatography to give a pale yellow solid (0.024 g, 64%). 1 H NMR (400MHz, Methanol-d4) δ7.72(d,J=7.6Hz,1H),7.57(d,J=7.3Hz,1H),7.53(d,J=7.6 Hz,1H),7.47(t,J=7.5Hz,1H),6.60(d,J=7.7Hz,1H),6.46(s,1H),6.39(d,J=7.8Hz,1H), 4.59(s,2H),3.18(s,1H),2.46–2.40(m,2H),2.31(dd,J=11.1,5.6Hz,2H),2.02(d,J=12. 9Hz,2H),1.77(d,J=13.8Hz,3H),1.67(d,J=12.1Hz,1H),1.57(s,6H),1.44–1.32(m,4H). 13 C NMR (101MHz, Methanol-d4) δ141.63,135.90,133.88,133.43,131.15,127.55,122.51,122. 22,117.78,116.80,113.45,57.23,52.02,49.44,41.38,33.04,30.35,25.80,25.48,24.88.

[0110] 5. The synthesis method of methyl 2-(4-(3-amino-4-(cyclohexylamino)phenyl)-2-methylbut-2-yl)isoindoline-4-carboxylic acid (compound II-2) is the same as that of II-1. 1H NMR (400 MHz, Chloroform-d) δ 8.15 (dd, J = 7.7, 1.1 Hz, 1H), 7.98 (dd, J = 7.6, 1.0 Hz, 1H), 7.69 - 7.59 (m, 1H), 7.53 (t, J = 7.7 Hz, 1H), 7.24 - 7.15 (m, 1H), 7.09 (d, J = 4.7 Hz, 1H), 4.79 (s, 2H), 4.20 - 4.10 (m, 1H), 3.95 (s, 3H), 2.76 - 2.68 (m, 2H), 2.48 - 2.39 (m, 2H), 2.21 - 2.15 (m, 2H), 1.95 (d, J = 13.5 Hz, 2H), 1.83 - 1.70 (m, 4H), 1.64 (s, 6H), 1.55 - 1.47 (m, 2H). 13 C NMR (101 MHz, Chloroform-d) δ 166.63, 141.66, 135.84, 134.52, 131.43, 127.25, 126.72, 123.72, 122.06, 56.23, 51.21, 49.61, 40.99, 32.36, 30.36, 28.68, 25.71, 24.68, 24.40.

[0111] 6、N 1 - Cyclohexyl-4-(3-methyl-3-(5-(methylsulfonyl)isoindolin-2-yl)butyl)benzene-1,2-diamine (Compound II-3) was synthesized by the same method as II-1. 1 H NMR (400 MHz, Chloroform-d) δ 7.80 (d, J = 8.5 Hz, 2H), 7.39 (d, J = 7.8 Hz, 1H), 6.66 (d, J = 7.6 Hz, 1H), 6.48 (d, J = 7.2 Hz, 2H), 4.17 (s, 4H), 3.23 (m, 1H), 3.03 (s, 3H), 2.63 - 2.56 (m, 2H), 2.09 - 2.01 (m, 2H), 1.83 - 1.72 (m, 5H), 1.70 - 1.58 (m, 2H), 1.39 (q, J = 12.8, 12.0 Hz, 3H), 1.22 (s, 6H). 13 C NMR (101 MHz, Chloroform-d) δ 139.26, 137.12, 131.83, 126.33, 122.93, 121.70, 117.24, 51.69, 51.46, 33.61, 30.20, 29.70, 26.02, 25.00.

[0112] 7. 3-Amino-4-(cyclohexylamino)-N-(2-(isoindolin-2-yl)ethyl)benzamide (Compound II-4)

[0113] Synthetic procedure similar to II-1. 1 H NMR (400 MHz, Chloroform-d) δ 7.82 (d, J = 8.0 Hz, 1H), 7.56 - 7.51 (m, 1H), 7.48 - 7.42 (m, 2H), 7.18 (d, J = 1.8 Hz, 1H), 7.09 (dd, J = 7.8, 1.7 Hz, 2H), 6.67 (dd, J = 8.0, 0.6 Hz, 1H), 4.50 (s, 2H), 3.92 - 3.87 (m, 2H), 3.73 (q, J = 5.4 Hz, 2H), 3.35 - 3.25 (m, 1H), 2.10 - 2.01 (m, 2H), 1.76 (m, 2H), 1.66 (m, 1H), 1.47 - 1.35 (m, 3H), 1.21 - 1.12 (m, 2H). 13 C NMR (101 MHz, Chloroform-d) δ 167.98, 141.41, 140.57, 132.49, 132.29, 131.54, 128.07, 123.59, 122.87, 122.19, 120.66, 116.30, 110.02, 51.41, 50.86, 42.28, 39.73, 33.30, 25.88, 24.94.

[0114] 8. Methyl 2-(2-(3-amino-4-(cyclohexylamino)benzamido)ethyl)isoindoline-4- carboxylate (Compound II-5) Synthetic procedure similar to II-1. 1 H NMR (400 MHz, Chloroform-d) δ 8.57 (d, J = 2.1 Hz, 1H), 8.30 (d, J = 7.4 Hz, 1H), 7.93 - 7.85 (m, 2H), 7.39 (d, J = 7.4 Hz, 1H), 7.29 (t, J = 7.6 Hz, 1H), 6.87 (d, J = 9.1 Hz, 1H), 6.77 (t, J = 4.3 Hz, 1H), 4.33 (s, 2H), 4.02 (s, 2H), 3.89 (d, J = 0.9 Hz, 3H), 3.64 (m, 2H), 3.53 (m, 1H), 3.01 (t, J = 5.8 Hz, 2H), 2.03 (d, J = 9.8 Hz, 2H), 1.79 (m, 2H), 1.69 - 1.62 (m, 1H), 1.47 - 1.32 (m, 5H). 13C NMR (101 MHz, Chloroform-d) δ 167.80, 166.72, 140.57, 132.57, 128.77, 127.33, 126.70, 125.15, 122.77, 120.55, 116.42, 110.03, 60.02, 58.34, 54.79, 53.43, 52.04, 51.43, 38.07, 33.28, 29.71, 25.89, 24.93.

[0115] 9. 3-Amino-4-(cyclohexylamino)-N-(2-(5-(methylsulfonyl)isoindolin-2- yl)ethyl)benzamide (Compound II-6)

[0116] Synthetic procedure same as II-1. 1 H NMR (400 MHz, Chloroform-d) δ 7.84 - 7.75 (m, 2H), 7.39 (d, J = 7.9 Hz, 1H), 7.21 (s, 1H), 6.72 - 6.66 (m, 1H), 6.55 (d, J = 8.0 Hz, 1H), 4.10 (s, 4H), 3.62 (t, J = 5.3 Hz, 2H), 3.31 - 3.21 (m, 1H), 3.04 (d, J = 5.7 Hz, 5H), 2.03 (m, 2H), 1.80 - 1.71 (m, 2H), 1.65 (m, 1H), 1.41 - 1.26 (m, 3H), 1.19 (d, J = 12.4 Hz, 2H). 13 C NMR (101 MHz, Methanol-d4) δ 169.71, 144.77, 140.21, 140.06, 132.90, 126.68, 123.17, 121.33, 119.82, 114.93, 109.18, 58.38, 58.20, 54.80, 51.32, 43.07, 37.65, 32.82, 25.69, 25.30, 24.85.

[0117] 10. 3-Amino-4-(cyclohexylamino)-N-(2-(5-fluoroisoindolin-2- yl)ethyl)benzamide (Compound II-7)

[0118] Compound 4-(cyclohexylamino)-N-(2-(5-fluoroisoindolin-2-yl)ethyl)-3- nitrobenzamide (0.1 g, 0.23 mmol), zinc dust (0.045 g, 0.69 mmol) and catalytic amount of hydrochloric acid were taken in a 20 mL methanol vial and allowed to react at room temperature till completion. The mixture was filtered through celite and the solution concentrated. The resulting mixture was separated by column chromatography to get the compound as a light yellow color (0.73 g, 79%).1 H NMR (400 MHz, Chloroform-d) δ 8.64 (s, 1H), 8.26 (d, J = 6.8 Hz, 1H), 7.82 (d, J = 9.1 Hz, 1H), 7.54 (s, 1H), 7.44 - 7.36 (m, 1H), 7.22 - 7.09 (m, 1H), 6.83 (d, J = 9.0 Hz, 1H), 4.49 (d, J = 4.1 Hz, 2H), 3.88 (d, J = 4.6 Hz, 2H), 3.74 (s, 2H), 3.54 - 3.48 (m, 1H), 2.06 - 1.96 (m, 2H), 1.83 - 1.74 (m, 2H), 1.69 - 1.59 (m, 1H), 1.37 (m, 5H). 13 C NMR (101 MHz, Methanol-d4) δ 169.78, 163.90, 139.98, 132.92, 123.70, 121.02, 119.86, 114.92, 114.08, 109.55, 109.32, 109.14, 58.57, 58.06, 54.89, 51.31, 37.39, 32.82, 25.69, 24.8.

[0119] 11. 3-Amino-N-(2-(4-bromoisoindolin-2-yl)ethyl)-4- (cyclohexylamino)benzamide (Compound II-8)

[0120] Synthesis method is the same as II-7. 1 H NMR (400 MHz, Chloroform-d) δ 7.31 (d, J = 7.6 Hz, 1H), 7.24 (dd, J = 4.4, 1.1 Hz, 1H), 7.07 (dt, J = 15.1, 7.4 Hz, 2H), 6.74 (t, J = 4.5 Hz, 1H), 6.54 (d, J = 8.1 Hz, 1H), 4.02 (m, 4H), 3.57 (q, J = 5.6 Hz, 2H), 3.25 (m, 1H), 2.93 (t, J = 5.9 Hz, 2H), 2.05 - 1.98 (m, 2H), 1.74 (m, 2H), 1.67 - 1.60 (m, 1H), 1.40 - 1.30 (m, 2H), 1.18 (m, 3H). 13C NMR (101 MHz, Chloroform-d) δ 167.85, 140.51, 140.36, 132.69, 129.96, 128.80, 122.70, 121.19, 120.44, 117.28, 116.29, 109.96, 59.96, 59.75, 54.55, 51.43, 38.23, 33.31, 25.91, 24.96.

[0121] 12. 3-Amino-4-(cyclohexylamino)-N-(2-(4-fluoroisoindolin-2-yl)ethyl)benzamide (Compound II-9)

[0122] Synthetic procedure same as II-7. 1 H NMR (400 MHz, Chloroform-d) δ 7.24 - 7.21 (m, 2H), 7.18 (dt, J = 8.0, 3.7 Hz, 1H), 6.98 (d, J = 7.4 Hz, 1H), 6.89 (t, J = 8.6 Hz, 1H), 6.73 (s, 1H), 6.58 - 6.54 (m, 1H), 4.05 (d, J = 16.1 Hz, 4H), 3.60 (q, J = 5.5 Hz, 2H), 3.26 (dm, 1H), 2.98 (t, J = 5.8 Hz, 2H), 2.07 - 2.00 (m, 2H), 1.75 (m, 2H), 1.65 (m, 1H), 1.41 - 1.31 (m, 2H), 1.19 (m, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 167.81, 158.90, 140.58, 132.60, 129.06, 122.70, 120.46, 118.06, 116.38, 113.82, 113.62, 109.99, 58.93, 55.45, 54.61, 51.43, 38.14, 33.31, 29.70, 25.89, 24.93.

[0123] 13. 3-Amino-N-(2-(5-chloroisoindolin-2-yl)ethyl)-4-(cyclohexylamino)benzamide (Compound II-10) Synthetic procedure same as II-7. 1H NMR (400 MHz, Chloroform-d) δ 7.23 (d, J = 5.4 Hz, 2H), 7.16 (d, J = 6.7 Hz, 2H), 7.08 (dd, J = 6.4, 1.8 Hz, 1H), 6.72 (d, J = 5.2 Hz, 1H), 6.56 (d, J = 8.8 Hz, 1H), 4.06 (s, 4H), 3.60 (q, J = 5.7 Hz, 2H), 3.27 (m, 1H), 2.98 (t, J = 5.9 Hz, 2H), 2.10 - 1.96 (m, 2H), 1.75 (m, 2H), 1.64 (m, 1H), 1.43 - 1.23 (m, 4H). 13 C NMR (101 MHz, Chloroform-d) δ 167.79, 141.57, 140.56, 138.03, 132.61, 128.77, 128.65, 127.07, 122.74, 120.62, 120.47, 116.37, 110.00, 59.49, 58.14, 54.61, 51.42, 38.12, 33.31, 25.88, 24.93.

[0124] 14. 3-Amino-4-(cyclohexylamino)-N-(3-(isoindolin-2-yl)propyl)benzamide (Compound II-11)

[0125] Synthetic procedure same as II-7. 1 H NMR (400 MHz, Chloroform-d) δ 7.85 (d, J = 7.5 Hz, 1H), 7.54 (t, J = 7.4 Hz, 2H), 7.45 (q, J = 8.2 Hz, 3H), 7.35 (s, 1H), 6.67 (d, J = 8.3 Hz, 1H), 4.40 (s, 2H), 3.77 - 3.70 (m, 2H), 3.37 (q, J = 5.9 Hz, 2H), 3.30 (dd, J = 8.5, 5.1 Hz, 1H), 2.10 - 2.01 (m, 2H), 1.88 (p, J = 6.4 Hz, 2H), 1.77 (m, 2H), 1.66 (m, 1H), 1.44 - 1.30 (m, 2H), 1.20 (m, 3H). 13 C NMR (101 MHz, Chloroform-d) δ 169.67, 141.22, 132.36, 131.56, 128.21, 123.71, 122.87, 120.59, 116.63, 51.93, 50.15, 39.48, 35.64, 33.13, 29.70, 27.74, 25.87, 24.96.

[0126] 15. 3-Amino-4-(cyclohexylamino)-N-(4-(isoindolin-2-yl)butyl)benzamide (Compound II-12)

[0127] Synthesized as for II-7. 1 H NMR (400 MHz, Chloroform-d) δ 7.85 - 7.80 (m, 1H), 7.52 (t, J = 7.4 Hz, 1H), 7.44 (t, J = 8.7 Hz, 2H), 7.23 (d, J = 8.7 Hz, 1H), 6.61 (d, J = 8.4 Hz, 1H), 6.41 (d, J = 5.5 Hz, 1H), 4.37 (s, 2H), 3.66 (t, J = 7.1 Hz, 2H), 3.47 (q, J = 6.4 Hz, 2H), 3.34 - 3.24 (m, 1H), 2.04 (d, J = 12.5 Hz, 2H), 1.76 (q, J = 6.1 Hz, 4H), 1.65 (q, J = 6.7 Hz, 3H), 1.44 - 1.24 (m, 5H). 13 C NMR (101 MHz, Chloroform-d) δ 168.82, 141.16, 132.81, 131.28, 128.04, 123.61, 122.75, 50.01, 42.06, 39.52, 32.88, 26.67, 26.09, 25.77, 24.91.

[0128] 16. 3-Amino-4-(cyclopentylamino)-N-(2-(isoindolin-2-yl)ethyl)benzamide (Compound II-13)

[0129] Synthesized as for II-7. 1 H NMR (400 MHz, Chloroform-d) δ 7.22 (d, J = 4.6 Hz, 5H), 6.84 (s, 1H), 6.58 (d, J = 8.2 Hz, 1H), 4.05 (s, 4H), 3.82 - 3.76 (m, 1H), 3.65 - 3.59 (m, 2H), 3.02 (t, J = 5.8 Hz, 2H), 2.03 (m, 2H), 1.77 - 1.69 (m, 2H), 1.66 - 1.57 (m, 2H), 1.54 - 1.44 (m, 2H). 13 C NMR (101 MHz, Chloroform-d) δ 141.12, 132.88, 126.99, 123.06, 122.35, 120.35, 115.89, 58.89, 54.70, 54.39, 38.16, 33.58, 24.24.

[0130] 17. 3-Amino-4-(cycloheptylamino)-N-(2-(isoindolin-2-yl)ethyl)benzamide (Compound II-14)

[0131] Synthesized as for II-7. 1 H NMR (400 MHz, Chloroform-d) δ 7.23 (d, J = 10.2 Hz, 5H), 6.85 (t, J = 4.1 Hz, 1H), 6.57 (d, J = 8.9 Hz, 1H), 4.04 (s, 4H), 3.99 (m, 2H), 3.65 - 3.59 (m, 2H), 3.51 (m, 3H), 3.01 (t, J = 5.8 Hz, 2H), 2.07 - 1.97 (m, 2H), 1.51 (m, 2H). 13 C NMR (101 MHz, Chloroform-d) δ 167.70, 139.87, 138.67, 132.99, 127.29, 123.46, 122.44, 120.58, 116.55, 110.36, 66.72, 58.96, 54.91, 48.82, 37.88, 33.42.

[0132] 18. 3-Amino-N-(2-(isoindolin-2-yl)ethyl)-4-((tetrahydro-2H-pyran-4-yl)amino)benzamide (Compound II-15)

[0133] Synthesized as for II-7. 1 H NMR (400 MHz, Chloroform-d) δ 7.23 (d, J = 10.2 Hz, 5H), 6.85 (t, J = 4.1 Hz, 1H), 6.57 (d, J = 8.9 Hz, 1H), 4.04 (s, 4H), 3.99 (m, 2H), 3.65 - 3.59 (m, 2H), 3.51 (m, 3H), 3.01 (t, J = 5.8 Hz, 2H), 2.07 - 1.97 (m, 2H), 1.51 (m, 2H). 13 C NMR (101 MHz, Chloroform-d) δ 167.70, 139.87, 138.67, 132.99, 127.29, 123.46, 122.44, 120.58, 116.55, 110.36, 66.72, 58.96, 54.91, 48.82, 37.88, 33.42.

[0134] 19. N 1- 4-(3-(Isoindolin-2-yl)-3-methylbut-1-yn-1-yl)cyclohexan-1-ol (Compound II- 16) was synthesized according to the procedure described for II-7. 1 H NMR (400 MHz, Chloroform-d) δ 7.20 (q, J = 5.0 Hz, 4H), 6.71 (dd, J = 7.9, 1.9 Hz, 1H), 6.65 (s, 1H), 6.56 (d, J = 7.9 Hz, 1H), 4.20 (s, 4H), 3.18 (m, 1H), 2.06 - 1.97 (m, 2H), 1.80 - 1.62 (m, 4H), 1.59 (s, 6H), 1.37 (d, J = 12.5 Hz, 2H), 1.25 (d, J = 11.1 Hz, 2H). 13 C NMR (101 MHz, Chloroform-d) δ 139.76, 136.02, 135.14, 126.64, 122.81, 122.37, 116.08, 114.44, 87.77, 87.32, 54.58, 54.23, 51.66, 33.59, 29.54, 25.97, 24.95.

[0135] Example 3: Test of ferroptosis inhibition activity of compounds

[0136] GPX4 inhibitors such as RSL-3 can induce ferroptosis in cells, and this ferroptosis can also be blocked by other small molecules such as lipophilic antioxidants, such as Ferrostatin-1 (fer-1), Liproxstatin, etc. Therefore, the ability of ferroptosis inhibitors to block ferroptosis can be indicated by the reversal of ferroptosis induced by ferroptosis inducers.

[0137] Cell lines: Human neuroblastoma cell SH-SY5Y, human fibrosarcoma cell HT1080 and mouse hippocampal neuron cell were purchased from Shanghai Cell Bank of Chinese Academy of Sciences.

[0138] Method: MTT method, as follows: Take the logarithmic growth period of human renal cancer cell line SH-SY5Y / HT1080 / HT22, digest and collect and dilute, about 4000-5000 cells per hole in 96 hole plate, 80 μL per hole. Place in 37℃, 5% CO2 incubator overnight. The experiment is divided into DMSO control group and nine different concentrations of compound administration group. Add different concentrations of compounds to the administration group, and set up DMSO control group (the same dilution multiple as the highest concentration of compound), place in 5% CO2, 37℃ incubator for 1h, then add 5 μM / 100 nM / 100 nM RSL3 to induce ferroptosis at each compound concentration, and set up RSL3 control group and DMSO control group, place in 5% CO2, 37℃ incubator for 48h. Add 5mg / mL of 20 μL MTT solution to each hole, continue to incubate the cells in a 37℃ incubator for 2h, add 100 μL DMSO to each hole, and shake for 10min. Place in enzyme-linked immunosorbent assay instrument, detect the optical density value (OD value) of each hole at 570nm wavelength, repeat the experiment 3 times. Calculate according to the following formula: survival rate % = experimental group OD value / DMSO control group OD value x 100%.

[0139] The results are shown in Table 1, which shows that the various compounds of the present application can significantly inhibit ferroptosis and have good ferroptosis inhibition activity.

[0140] Table 1 Inhibition activity of compounds on RSL3-induced SH-SY5Y cell ferroptosis

[0141] In the table: "A" indicates IC 50 ≤0.1 μM, "B" indicates IC 50 >0.1 μM and ≤0.5 μM.

[0142] Table 2 Inhibition activity of compounds on RSL3-induced HT1080 cell ferroptosis

[0143] Table 3 Inhibition activity of compounds on RSL3-induced HT22 cell ferroptosis

[0144] Example 4: Sigma receptor affinity test of compounds

[0145] Sigma receptor inhibitors generally can exhibit strong affinity for Sigma receptors and exert corresponding pharmacological effects. The radioligand competition experiment was tested by Shanghai Pharming Kangde New Drug Development Co., Ltd., and the specific steps are as follows: (1) 1 μl of diluted reference compound and test compound was transferred to the detection plate. 1 μl of non-specific binding compound was added to the detection plate as a control. In addition, 1 μl of dimethyl sulfoxide was transferred to the detection plate to detect the total binding force. (2) 100 μL of membrane stock solution was added to the detection plate. (3) 100 μL of radio ligand was added. (4) The plate was sealed and shaken at 300 rpm under the specified conditions. (5) At room temperature, the Unifilter-96 GF / C filter plate was soaked in 0.3% PEI for at least 0.5 hours. (6) After the completion of the binding assay, the mixture was filtered into a GF / B plate using a Perkin Elmer Filtermate Harvester, and then washed 6 times with cold washing buffer. (7) The filter plate was dried at 50°C for 1 hour. (8) After drying, the bottom of the filter plate hole was sealed with a back sealing tape. 50 μl of Perkin Elmer Microscint 20 cocktail was added, and the top of the filter plate was sealed with a sealing film. (9) The Perkin Elmer MicroBeta2 reader was used to count the 3 H]DTG trapped on the filter. (10) The affinity of the compound was calculated.

[0146] Table 4 Affinity of compounds for Sigma 1 / 2 (σ 1 / 2 )

[0147] In the table: "A" indicates IC 50 ≤0.1 μM, "B" indicates IC 50 >0.1 μM and ≤0.5 μM, "C" indicates IC 50 >0.5 μM and ≤2 μM, "D" indicates IC 50 >2 μM.

[0148] Example 5: Compounds can inhibit the polymerization of Aβ 1-42 monomers into polymers in vitro

[0149] Alzheimer's disease is the most common neurodegenerative disease, one of the main pathological features is the deposition of β-amyloid protein in extracellular senile plaques. In addition, a large number of studies have shown that many metal ions (mainly Cu 2+ , Zn 2+ and Fe 2+The level of metal ions in the brain of AD patients is much higher than that of normal people, and is enriched in senile plaques of AD patients, eventually leading to imbalance of metal ion homeostasis.

[0150] Aβ 1-42 Preparation of monomers: Aβ 1-42 powder was taken out from -70℃ refrigerator and thawed at room temperature for about 30 min. A certain amount of Aβ 1-42 was weighed and dissolved in hexafluoroisopropanol to a final concentration of 1 mg / mL. The solution was then left to stand at room temperature for 2 h. After the solution was clear, it was ultrasonicated under ice bath condition for 30 min to remove possible Aβ 1-42 aggregates. Then the sample was frozen in -70℃ refrigerator for more than 4 h. After that, the sample was freeze-dried in a freeze dryer. After 24 h, the sample was taken out and Aβ 1-42 monomers in fluffy floc were obtained, which were stored in -20℃ refrigerator for later use.

[0151] ThT fluorescence experiment: Thioflavin T (ThT) as a fluorescent dye molecule can specifically bind to the β-sheet structure in Aβ 1-42 fibers, and then under the action of 440 nm excitation light, an absorption peak appears at 480 nm, which can be used to detect the aggregation of Aβ 1-42 . Before the experiment, Aβ 1-42 monomers were mixed with RTH at different ratios, and the final concentration of Aβ 1-42 was always kept at 25 μM. After mixing, the sample was incubated in an air shaker (37℃). After 24 h of incubation, 200 μl of sample was taken and added to 2 mL of ThT solution (25 μM ThT, 20 mM HEPES, pH 7.4). After mixing well, the ThT fluorescence intensity was measured in a fluorescence spectrophotometer. The instrument parameters are as follows: excitation wavelength 440 nm, emission wavelength 480 nm, excitation bandwidth and emission bandwidth both 5 nm. Each experimental point was repeated three times to take the average value, and the standard deviation was calculated.

[0152] The above examples are only used to illustrate the technical solutions of the present application, but not to limit it; although the foregoing embodiments of the present application have been described in detail, those skilled in the art can still modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement for part of the technical features; and such modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions claimed by the present application.

Claims

1. An aromatic alkylamine ferroptosis and / or Sigma receptor bifunctional inhibitor, characterized in that, The aromatic alkylamine ferroptosis and / or Sigma receptor bifunctional inhibitor has a structural formula as shown below: 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; Linker is selected from a direct bond, C1-C6alkyl or C1-C6alkyl containing from 1 to 3 independent substituents, C2-C6alkenyl or C2-C6alkenyl containing from 1 to 3 independent substituents, C0to C6alkyl-C3to C6cycloalkyl-C0to C6alkyl or C0to C6alkyl-C3to C6cycloalkyl-C0to C6alkyl containing from 1 to 3 independent substituents, C0to C6alkyl-Z-C0to C6alkyl or C0to C6alkyl-Z-C0to C6alkyl containing from 1 to 3 independent substituents; wherein Z is selected from N, NR a , -SO2-, OC, OC=O, CO or C=OO; R 3 , R 4 , R 5 and R 6 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 3 , R 4 , R 5 and R 6 are each one of the above substituents, or wherein two, three, or four of R 3 , R 4 , R 5 , R 6 are simultaneously selected from the above substituents; When ring A is absent, X is selected from CH2, O, S, NH, NH (C1-C4 alkyl), N (C1-C4 alkyl)2, NH (C3-C7 cycloalkyl), NHC(O)(C1-C4 alkyl), NC(O)R a NS(O) 1 / 2 R a ; m, n are the number of carbon atoms, and are 0, 1 or 2; R a 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; 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.

2. The aromatic alkylamine ferroptosis and / or Sigma receptor bifunctional inhibitor according to claim 1, characterized in that, The inhibitors are compounds I-1 to I-22, II-1 to II-17, whose structural formulae are specified below:

3. Process for the preparation of the aromatic alkylamine ferroptosis and / or Sigma receptor bifunctional inhibitors according to claim 2, characterized in that, The preparation method comprises the following steps: The synthesis of the compounds I-1~I-22 is as follows: 4-chloro-3-nitrobenzaldehyde is used as a raw material, sodium borohydride reduction gives compound 2, then substitution reaction with the corresponding amine gives compound 3; compound 3 is oxidized by PCC to give compound 4, which undergoes aldol condensation reaction with acetone under alkaline conditions to give compound 5; sodium borohydride reduction of the double bond and carbonyl group of compound 5 gives compound 6, then the nitro group is reduced by hydrogen to give the key intermediate 7; the amino group is protected by (Boc)2O to give the corresponding compound 8; halogenation reaction with CBr4 in the presence of PPh3 gives compound 9; compound 10 is obtained by substitution reaction of compound 9 with the corresponding aromatic amine or aliphatic amine, then the Boc protecting group is removed to give compounds I-1~I-22; Reagents and reaction conditions: (a) sodium borohydride, methanol; (b) cyclohexylamine, potassium carbonate, DMSO; (c) PCC, chloroform; (d) acetone, sodium hydroxide; (e) sodium borohydride, methanol; (f) Pd / C, hydrogen, methanol; (g) (Boc)2O, triethylamine, tetrahydrofuran; (h) carbon tetrabromide, triphenylphosphine, dichloromethane; (i) corresponding amine, potassium carbonate, potassium iodide, DMF; (j) i: HCl / CH3CH2OH; ii: corresponding aldehyde, sodium triacetoxyborohydride, dichloromethane.

4. Process for the preparation of the aromatic alkylamine ferroptosis and / or Sigma receptor bifunctional inhibitors according to claim 2, characterized in that, The preparation method comprises the following steps: The synthesis of the compounds II-1 to II-5, II-16 and II-17 is as follows: Compound 11 undergoes a radical substitution reaction with NBS under the catalysis of AIBN to give compound 12, which then undergoes a nucleophilic substitution reaction with 2-methylbut-3-yn-2-amine to give the key intermediate 13; compound 13 undergoes a coupling reaction with the corresponding aryl halide compound under the catalysis of Pd(PPh3)2Cl2 and Cul to give the key intermediate compound 14; compound II-16 / 17 is obtained by reduction of compound 14 with zinc powder, while compounds II-1 to II-5 need to be further reduced under hydrogen gas conditions; Reagents and reaction conditions: (k) NBS, AIBN, chloroform, reflux; (l) 2-methylbut-3-yn-2-amine, potassium carbonate, tetrahydrofuran, 60°C; (m) 4-bromo-N-cyclohexyl-2-nitroaniline, Pd(PPh3)2Cl2, copper iodide, triethylamine; (n) i: zinc powder, hydrochloric acid, methanol; ii: corresponding aldehyde, sodium triacetoxyborohydride, dichloromethane; (o) Pd / C, hydrogen, methanol.

5. Process for the preparation of the aromatic alkylamine ferroptosis and / or Sigma receptor bifunctional inhibitors according to claim 2, characterized in that, The preparation method comprises the following steps: The synthesis of the compounds II-4~II-15 is as follows: 4-chloro-3-nitrobenzoic acid is used as a starting material, and a condensation reaction with a corresponding amine group fragment to obtain compound 16, followed by a nucleophilic substitution reaction with an amine derivative to obtain compound 17, and after removing the Boc protecting group to obtain intermediate 18, a nucleophilic substitution reaction of compound 18 and a corresponding bromide compound to obtain key intermediate 19, and further reduction with zinc powder to obtain the target compounds II-4~II-15; Reagents and reaction conditions: (p) i: sulfuric acid chloride, dichloromethane, DMF; ii: (2-aminoethyl)carbamic acid tert-butyl ester, triethylamine, dichloromethane; (q) cyclohexylamine, potassium carbonate, DMF; (r) trifluoroacetic acid, dichloromethane; (s) 1,2-bis(bromomethyl)benzene, potassium carbonate, tetrahydrofuran; (t) i: zinc powder, hydrochloric acid, methanol; ii: corresponding aldehyde, sodium triacetoxyborohydride, dichloromethane.

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

7. Use of the inhibitor of claim 1 or 2 in the preparation of a drug for treating a disease related to ferroptosis and / or a disease related to Sigma receptor.

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

9. Use according to claim 7, characterized in that, The disease related to Sigma receptor includes Alzheimer's disease, Parkinson's disease, stroke, drug addiction, neuropathic pain, schizophrenia and depression.

10. A medicament for treating iron death and / or a Sigma receptor related disease, characterized by, The drug contains the compound of claim 1 or 2 and a pharmaceutically acceptable salt, carrier or adjuvant.

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