Quinone compounds for targeted inhibition of GPX4 and preparation method and application thereof

Abstract

The present invention discloses a quinone compound for targeted inhibition of GPX4 and a preparation method and application thereof, belonging to the field of biomedical technology. Based on the drug design principle of drug chemistry splicing and the use of chemical means, the present invention synthesizes a series of quinone compounds that can have a significant killing effect on GPX4 high-expressing cell lines, which is of great significance for tumor targeted therapy.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a class of quinone compounds with GPX4 inhibitory activity, a preparation method thereof, and an application thereof in anti-tumor. Background Art

[0002] There are many ways of programmed cell death, including apoptosis, necrosis and pyroptosis. Ferroptosis, also known as iron-dependent programmed necrosis, is caused by membrane damage mediated by a large amount of lipid peroxidation. It is a new type of non-apoptotic programmed cell death. It is characterized by iron accumulation and lipid peroxidation. Among them, the activity of glutathione peroxidase 4 (GPX4) is reduced, which reduces the antioxidant capacity of cells, thereby increasing the level of lipid peroxidation and reactive oxygen, and ultimately leading to ferroptosis. Therefore, GPX4 is the core regulatory protein of ferroptosis. The catalytic active center of GPX4 is selenocysteine. With GSH as a cofactor, GPX4 can reduce lipid hydroperoxides in cells to non-toxic lipid alcohol compounds. It can also catalyze the reduction of other organic peroxides such as hydrogen peroxide, thus protecting cells from oxidative stress and inhibiting the occurrence of ferroptosis. Therefore, inhibiting the activity of GPX4 will affect the ability of GPX4 to remove lipid peroxides, ultimately leading to the occurrence of cell ferroptosis. In addition, inhibiting the function of GPX4 will trigger sustained ferroptosis of cells and prevent tumor recurrence, and is therefore one of the strategies to address drug resistance.

[0003] At present, there are still certain challenges for small molecule inhibitors targeting GPX4, and there are no reports of GPX4 inhibitors entering the clinical research stage. The main reason is that since the GPX4 protein lacks a traditional druggable pocket, it is difficult to design compounds targeting GPX4. However, small molecule covalent inhibitors can still destroy its enzyme activity by irreversibly binding to the selenocysteine ​​residues in the active center of GPX4. Therefore, the current GPX4 inhibitors are mainly covalent inhibitors, which work by binding to the selenocysteine ​​in the active site of GPX4, but have problems such as low selectivity. Therefore, it is of great significance to find new active small molecules targeting GPX4. Summary of the invention

[0004] In view of the problems existing in the prior art,

[0005] The purpose of the present invention is to provide a quinone compound for targeted inhibition of GPX4 and a preparation method and application thereof. The quinone compound of the present invention can effectively induce the death of tumor cells with high expression of GPX4 protein, thereby exerting an anti-tumor effect.

[0006] In order to achieve the above invention objectives, the present invention adopts the following technical solutions:

[0007] The present invention provides a quinone compound or a pharmacologically or physiologically acceptable salt thereof for inhibiting GPX4 protein as shown in formula (I) and (II):

[0008]

[0009] in:

[0010] R is any one of a hydrogen atom, a methyl group, a methoxy group, a halogen group, or a fused benzene ring;

[0011] R 1 is any one of a hydrogen atom, a methyl group, a methoxy group, or a halogen group;

[0012] Linker is a connecting group, which represents an -alkylene group or an -alkoxy group or a -piperazinyl group or a piperidinyl group, wherein the -alkylene group or the -alkoxy group or the -piperazinyl group or the piperidinyl group is selected from any one of the following groups or any combination thereof, wherein m and n represent a natural number from 1 to 20:

[0013] -(CH2CH2O) m -(CH2) n NH- or -(CH2CH2O) m -(CH2) n NH- or -(CH2) n NH- or -(CH2) n S-or

[0014] Furthermore, the quinone compounds provided by the present invention are the compounds shown below or their stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically or physiologically acceptable salts or prodrugs;

[0015]

[0016]

[0017] The pharmacologically or physiologically acceptable salts of the present invention refer to salts formed by the quinone compounds of the present invention and pharmacologically or physiologically acceptable acids or bases.

[0018] The present invention also provides a pharmaceutical composition, which includes the quinone compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs.

[0019] The pharmaceutical composition further includes a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

[0020] The pharmaceutical composition is an injection, an oral agent, or a mucosal administration agent.

[0021] The pharmaceutical composition further includes other drugs having the effect of treating or preventing tumors.

[0022] The present invention also provides the use of quinone compounds or pharmaceutical compositions containing the compounds for targeted inhibition of GPX4. The specific application is as follows:

[0023] The use of the quinone compound for targeted inhibition of GPX4 or a pharmaceutical composition comprising the compound in the preparation of a drug for degrading GPX4.

[0024] The use of the quinone compound for targeted inhibition of GPX4 or a pharmaceutical composition containing the compound in the preparation of a drug for treating GPX4-related diseases. The GPX4-related diseases are tumors, neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease.

[0025] The use of the quinone compound for targeted inhibition of GPX4 or the pharmaceutical composition containing the compound in anti-tumor drugs. The tumor is gastric cancer, breast cancer, lung cancer, ovarian cancer, colon adenocarcinoma, renal chromophobe cell carcinoma, renal clear cell carcinoma, lung adenocarcinoma, prostate cancer, rectal adenocarcinoma, thyroid cancer and endometrial cancer. Further, the tumor is a tumor with high expression of GPX4.

[0026] The present invention also proposes a synthetic route for quinone compounds for targeted inhibition of GPX4 as shown in general formula I or general formula II, which specifically comprises the following steps:

[0027] The compound represented by general formula I or general formula II is mainly formed by connecting two chemical functional groups through click reaction or amide condensation reaction. One end is an analogue of ML-162, which is prepared by Ugi Reaction; the other end is prepared by substitution reaction to prepare quinone unit; the amide condensation reaction requires the use of common condensation agents HATU and TCFH.

[0028] Compared with the prior art, the beneficial effects of the present invention are mainly reflected in:

[0029] The present invention synthesizes a novel and diverse GPX4 inhibitor by chemically modifying the nucleus structure of the reported GPX4 inhibitor ML-162. The inventors have confirmed through CCK-8 experiments that the quinone compounds of the present invention can effectively kill the gastric cancer cell line MGC-803 and the fibrosarcoma HT1080 cell line with high GPX4 expression. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is a graph showing the results of lipid peroxide determination;

[0031] Figure 2 Molecular docking simulation of representative compound GI-C-11 and GPX4;

[0032] Figure 3 The synthetic route of quinone compounds GI-C-1 to GI-C-20 with GPX4 inhibitory activity;

[0033] Figure 4 This is a synthetic route for quinone compounds GI-C-21 to GI-C-22 with GPX4 inhibitory activity. DETAILED DESCRIPTION

[0034] The present invention is described in detail below in conjunction with the accompanying drawings and specific examples of the specification. The technical and scientific terms used in the following examples have the same meanings as those generally understood by technicians in the field to which the present invention belongs. Basic raw materials and reagents are obtained from commercial sources with a purity of 97% or more. The room temperature described in the present invention is 25-30°C. The present invention provides a general and specific description of the materials and experimental methods used in the experiments. Although many materials and operating methods used to achieve the purpose of the present invention are well known in the art, the present invention is still described as much as possible here.

[0035] Example 1 Synthesis and structure confirmation of GPX4 quinone inhibitors

[0036] The synthetic route of the final products GI-C-1 to GI-C-20 is shown in Figure 3 shown.

[0037] Synthesis of intermediate 1

[0038] Dissolve p-phenylenediamine (2.97 g, 27.44 mmol) and 2-thiophenecarboxaldehyde (3.08 g, 27.44 mmol) in methanol (25 mL), activate at 25°C for 1 h, add (2-isocyanoethyl)benzene (3 g, 22.87 mmol) and chloroacetic acid (2.16 g, 22.87 mmol), and stir at room temperature overnight. After the reaction is completed, the reaction solution is concentrated to dryness under reduced pressure, and the concentrate is purified by silica gel column chromatography (ethyl acetate: petroleum ether in a volume ratio of 1:1), and dried at 25°C to obtain intermediate 1 (white solid, 2 g, yield 20%).

[0039] Synthesis of intermediate 2a

[0040] 3-aminopropionic acid (890 mg, 10 mmol) was dissolved in an aqueous solution (20 mL), 1,4-naphthoquinone (3.84 g, 20 mmol) was dissolved in a hot solution of ethanol (150 mL), the solution was mixed and stirred at room temperature for 24 hours. Sodium hydroxide aqueous solution was added, extracted twice with ethyl acetate, 3N HCl solution was added to the aqueous layer, the pH value was adjusted to acidic (pH 1-2) and a red solid appeared. Intermediate 2a (red solid, 0.92 g, yield 38%) was obtained by suction filtration.

[0041] The synthesis method of intermediates 2b to 2t is the same as above. The starting reactants are naphthoquinone or anthraquinone compounds. According to the properties of the reactants, R 2 The substituent is a hydrogen atom or a chlorine atom.

[0042] Synthesis of the final product GI-C-1

[0043] 1 (100 mg, 0.24 mmol) was dissolved in acetonitrile (3 mL) and compound 2a (60 mg, 0.24 mmol), tetramethyl chlorouronium hexafluorophosphate (82.38 mg, 0.29 mmol), N-methylimidazole (70.37 mg, 0.85 mmol), and stirred at room temperature overnight. The reaction solution was diluted with ethyl acetate (200 mL), washed with water and saturated sodium chloride solution once each, dried with sodium sulfate, filtered, and the filtrate was rotary evaporated to dryness to obtain a concentrate, which was purified by silica gel column chromatography (ethyl acetate: petroleum ether volume ratio 1:3) to obtain GI-C-1 (red solid, 60 mg, yield 37%). 1 H NMR(400MHz,MeOD)δ7.97–7.89(m,2H),7.67(td,J=7.6,1.4Hz,1H),7.58(td ,J=7.6,1.4Hz,1H),7.38(d,J=8.2Hz,2H),7.21–7.00(m,7H),6.74–6.66(m,2 H),6.10(s,1H),5.70(s,1H),3.81(d,J=2.0Hz,2H),3.51(t,J=6.6Hz,2H),3 .42(dt,J=13.3,7.2Hz,1H),3.33–3.24(m,1H),2.65(dt,J=13.7,6.9Hz,4H).

[0044] The synthesis method of the final products GI-C-2 to GI-C-20 is the same as above.

[0045] Final product GI-C-2 (red solid, 30 mg, yield 30%). 1H NMR (400MHz, CDCl3) δ8.24(s,1H),7.96(ddd,J=22.0,7.8,1.3Hz,2H),7.65(td,J=7.6,1.4Hz ,1H),7.54(td,J=7.6,1.3Hz,1H),7.47–7.39(m,2H),7.19–7.00(m,6H),6.83–6.71(m,2H),6 .50(s,1H),6.17(t,J=5.9Hz,1H),6.06(s,1H),5.70(s,1H),3.77–3.66(m,2H),3.54–3.42(m ,2H),3.24(q,J=6.4Hz,2H),2.75–2.66(m,2H),2.45(t,J=6.9Hz,2H),2.03(p,J=6.8Hz,2H).

[0046] Final product GI-C-3 (red solid, 34 mg, yield 31%). 1 H NMR(400MHz, CDCl3)δ8.00(ddd,J=15.6,7.8,1.3Hz,2H),7.75–7.32(m,5H ),7.18–7.00(m,5H),6.91–6.69(m,3H),6.20–6.00(m,4H),5.70(s,1H),3. 73(dd,J=5.3,2.9Hz,2H),3.48(qd,J=9.7,8.9,4.4Hz,3H),3.19(q,J=6.3H z, 2H), 2.72 (q, J = 7.0Hz, 1H), 2.37 (t, J = 6.8Hz, 2H), 1.74 (q, J = 7.5Hz, 5H).

[0047] Final product GI-C-4 (red solid, 28 mg, yield 27%). 1 H NMR (400MHz, CDCl3) δ8.14–8.01(m,2H),7.88(s,1H),7.78–7.69(m,1H),7.63(t,J =7.5Hz,1H),7.50(d,J=8.2Hz,2H),7.26–7.07(m,6H),6.91–6.78(m,2H),6.21(dd, J=13.2,7.4Hz,2H),6.11(s,1H),5.77(s,1H),3.85–3.73(m,2H),3.63–3.48(m,2H) ,3.24(s,2H),2.82(dp,J=20.1,6.7Hz,2H),2.39(s,2H),1.75(s,4H),1.48(s,2H).

[0048] Final product GI-C-5 (red solid, 50 mg, yield 45%). 1H NMR (400 MHz, CDCl3) δ 8.10–7.81 (m, 3H), 7.63–7.30 (m, 4H), 7.20–7.01 (m, 5H), 6.78 (tt, J = 4.9, 2.5 Hz, 2H), 6.06 (d, J = 11.4 Hz, 2H), 3.87 (t, J = 6.0 Hz, 2H), 3.79–3.66 (m, 2H), 3.49 (q, J = 6.2, 5.7 Hz, 2H), 2.87–2.69 (m, 2H), 2.64 (t, J = 6.1 Hz, 2H), 2.14 (s, 3H).

[0049] Final product GI-C-6 (red solid, 18 mg, yield 15%). 1 H NMR (400MHz, DMSO) δ10.09(s,1H),8.32(t,J=5.6Hz,1H),7.98(ddd,J=7.1,5.3,1.3Hz,2H),7.79(dtd,J=34.1,7.5,1.3Hz,2H),7.54–7.35(m,4 H),7.32–7.08(m,5H),6.87–6.75(m,2H),6.23(s,1H),4.10–3.99(m,2H ),3.45–3.37(m,1H),3.26(qd,J=7.1,4.1Hz,1H),2.70(q,J=6.7Hz,4H).

[0050] Final product GI-C-7 (red solid, 22 mg, yield 20%). 1H NMR (400MHz, CDCl3) δ8.09 (ddd, J = 20.1, 5.7, 3.4 Hz, 2H), 7.77–7.69 (m, 2H), 7.51–7.35 (m, 6H), 7.32–7.17 (m, 6H), 7.11 (dd, J = 6.6, 5.3 Hz, 1H), 3.92 (d, J = 1.5 Hz, 2H), 3.45 (tdd, J = 5.7, 4.9, 1.6 Hz, 2H), 3.30–3.18 (m, 2H), 2.81 (tdt, J = 5.6, 2.0, 1.0 Hz, 2H), 2.65 (td, J = 5.9, 3.0 Hz, 2H).

[0051] Final product GI-C-8 (red solid, 30 mg, yield 30%). 1H NMR(400MHz,MeOD)δ8.17–7.94(m,3H),7.71(dtd,J=34.1,7.5,1.4Hz,2H),7 .51–7.38(m,2H),7.32–7.25(m,1H),7.21(tt,J=6.9,1.2Hz,2H),7.14(tt,J= 8.1,1.5Hz,3H),6.84–6.77(m,2H),6.19(s,1H),3.95–3.86(m,4H),3.56–3.3 4(m,2H),2.77(t,J=7.2Hz,2H),2.47(t,J=7.1Hz,2H),2.06(p,J=7.0Hz,2H).

[0052] Final product GI-C-9 (red solid, 43 mg, yield 40%). 1 H NMR(400MHz, CDCl3)δ7.85(dt,J=14.4,8.6Hz,1H),7.65–7.30(m,3H),7.18–6.52(m,11H),6.02(d,J=6.7Hz,1H),5.60(d,J =22.3Hz,1H),5.06(s,1H),3.84(s,4H),3.66(s,2H),3.55–2.96(m,5H),2.80–2.56(m,J=6.0Hz,3H),1.99(t,J=6.4Hz,2H).

[0053] Final product GI-C-10 (red solid, 7 mg, yield 5%). 1 H NMR(400MHz, CDCl3)δ8.34(s,1H),8.04–7.91(m,2H),7.70–7.40(m,4H),7 .19–7.02(m,6H),6.83–6.73(m,2H),6.12(dt,J=16.9,5.7Hz,2H),6.00(s ,1H),3.78(t,J=5.7Hz,2H),3.74–3.64(m,5H),3.48(ddt,J=15.5,13.5,6 .8Hz,2H),3.33(q,J=5.3Hz,2H),2.79–2.68(m,2H),2.58(t,J=5.7Hz,2H).

[0054] Final product GI-C-11 (red solid, 15 mg, yield 14%). 1H NMR(400MHz, CDCl3)δ8.07(dt,J=7.8,1.6Hz,1H),7.96–7.83(m,2H),7.61(dt d,J=37.3,7.5,1.3Hz,2H),7.39(d,J=8.4Hz,2H),7.18–7.04(m,5H),6.83–6. 74(m,2H),6.25(s,1H),6.02(d,J=7.7Hz,2H),4.08–3.99(m,2H),3.84–3.67( m,6H),3.50(dq,J=11.3,6.8Hz,2H),2.83–2.68(m,2H),2.58(t,J=5.6Hz,2H).

[0055] Final product GI-C-12 (red solid, 13 mg, yield 10%). 1 H NMR(400MHz, CDCl3)δ8.78(s,1H),8.10–8.03(m,2H),7.78–7.69(m,2H),7.51 –7.46(m,2H),7.45(d,J=1.3Hz,1H),7.42–7.35(m,3H),7.32–7.17(m,6H),7.1 1(dd,J=6.6,5.3Hz,1H),6.11(s,1H),3.92(d,J=1.5Hz,2H),3.61–3.40(m,6H ),2.81(tdt,J=5.6,2.0,1.0Hz,2H),2.68(p,J=5.1Hz,1H),1.96–1.83(m,4H).

[0056] Final product GI-C-13 (red solid, 30 mg, yield 27%). 1 H NMR (400MHz, CDCl3) δ8.04 (dd, J=7.3, 1.8Hz, 1H), 7.94 (dd, J=7.2, 1.7Hz, 1H), 7.75 (s ,1H),7.68–7.56(m,2H),7.49–7.37(m,2H),7.19–7.01(m,6H),6.84–6.71(m,2H),6.23 –5.95(m,2H),3.85–3.68(m,4H),3.54–3.47(m,2H),3.27(ddd,J=13.7,10.8,3.3Hz,2H ),2.82–2.65(m,2H),2.50(tt,J=10.4,4.5Hz,1H),1.98(qd,J=12.7,11.6,3.7Hz,4H).

[0057] Final product GI-C-14 (red solid, 22 mg, yield 21%). 1 H NMR (400MHz, CDCl3) δ7.99 (dd, J=7.6, 1.3Hz, 1H), 7.89 (dd, J=7.6, 1.4Hz, 1H), 7.71 (s, 1H), 7.60 (td,J=7.6,1.4Hz,1H),7.50(td,J=7.5,1.3Hz,1H),7.38(d,J=8.4Hz,2H),7.19–7.03(m,5H),6.7 8(tt,J=5.0,2.5Hz,2H),6.13–6.03(m,2H),3.80–3.67(m,2H),3.58(t,J=6.7Hz,2H),3.49(ddd, J=8.9,7.0,4.3Hz,2H),2.82–2.67(m,2H),2.41(t,J=7.0Hz,2H),2.13(s,3H),2.03–1.95(m,2H).

[0058] Final product GI-C-15 (red solid, 23 mg, yield 22%). 1 H NMR (400MHz, CDCl3) δ7.74–7.57(m,1H),7.53–7.38(m,5H),7.18–7.00(m,8H),6.86–6.75(m,4H),5.98(d,J=7.9Hz,2H),3.94(s, 3H),3.77(t,J=5.6Hz,2H),3.73–3.67(m,4H),3.53–3.44(m,2H),3.33(q,J=5.3Hz,2H),2.79–2.69(m,2H),2.56(t,J=5.7Hz,2H).

[0059] Final product GI-C-16 (red solid, 30 mg, yield 29%). 1 H NMR(400MHz, CDCl3)δ8.15(d,J=9.9Hz,1H),7.92(dd,J=19.8,8.6Hz,1H),7.52–7.36(m,3H),7.18–6.72(m,8H),6.25–5.95(m,3H),5.70(d,J=5.2 Hz,1H),3.86(d,J=11.4Hz,3H),3.82–3.65(m,6H),3.48(dh,J=13.3,6.7 Hz,2H),3.33(q,J=5.3Hz,2H),2.83–2.66(m,2H),2.57(t,J=5.6Hz,2H).

[0060] Final product GI-C-17 (red solid, 22 mg, yield 19%). 1 H NMR (400MHz, CDCl3) δ8.43 (d, J = 20.1Hz, 2H), 8.09 (s, 1H), 7.90 (ddd, J = 17.9, 8.2, 1. 3Hz,2H),7.71–7.36(m,4H),7.18–6.99(m,5H),6.83–6.73(m,2H),6.27(t,J=5.8Hz,1 H),6.15–6.01(m,2H),5.77(s,1H),3.78–3.66(m,2H),3.49(qd,J=6.7,3.2Hz,2H),3 .24(q,J=6.5Hz,2H),2.82–2.66(m,2H),2.47(t,J=7.0Hz,2H),2.07(p,J=6.6Hz,2H).

[0061] Final product GI-C-18 (red solid, 20 mg, yield 18%). 1 H NMR (400MHz, CDCl3) δ8.78 (s, 1H), 8.28 (d, J = 28.0Hz, 2H), 7.82 (t, J = 8.2Hz, 2H), 7.67–7.38 (m, 4H), 7. 18–6.64(m,12H),6.08(d,J=19.6Hz,3H),3.90–3.61(m,4H),3.48(q,J=6.6Hz,2H),2.86–2.69(m,4H).

[0062] Final product GI-C-19 (red solid, 11 mg, yield 8%). 1 H NMR(400MHz, CDCl3)δ8.15(d,J=9.9Hz,1H),7.92(dd,J=19.8,8.6Hz,1H),7.52–7.36(m,3H),7.18–6.72(m,8H),6.25–5.95(m,3H),5.70(d,J=5.2 Hz,1H),3.86(d,J=11.4Hz,3H),3.82–3.65(m,6H),3.48(dh,J=13.3,6.7 Hz,2H),3.33(q,J=5.3Hz,2H),2.83–2.66(m,2H),2.57(t,J=5.6Hz,2H).

[0063] Final product GI-C-20 (red solid, 10 mg, yield 8%). 1H NMR(500MHz,Chloroform-d)δ8.14(d,J=8.3Hz,1H),7.57(d,J=2.7Hz,1H),7.51–7.43(m,3H),7.42–7.35(m,3H),7.32–7.17(m,6H),7.17–7. 08(m,2H),3.92(d,J=1.4Hz,2H),3.79(s,2H),3.61–3.40(m,6H),2.81 (tdt,J=5.6,2.0,1.0Hz,2H),2.68(p,J=5.1Hz,1H),1.95–1.83(m,4H).

[0064] The synthetic route of the final product GI-C-21 to GI-C-22 is shown in Figure 4 shown.

[0065] Synthesis of intermediate 4

[0066] 3-Chloro-4-(propan-2-hexyloxy)aniline (4.96 g, 27.44 mmol) and 2-thiophenecarboxaldehyde (3.08 g, 27.44 mmol) were dissolved in methanol (25 mL), activated at 25°C for 1 h, and then (2-isocyanoethyl)benzene (3 g, 22.87 mmol) and chloroacetic acid (2.16 g, 22.87 mmol) were added and stirred at room temperature overnight. The reaction solution was concentrated to dryness under reduced pressure, and the concentrate was purified by silica gel column chromatography (ethyl acetate: petroleum ether, volume ratio 1:1), and dried at 25°C to obtain intermediate 4 (white solid, 2 g, yield 20%).

[0067] Synthesis of intermediate 5a

[0068] 3-Aminoazetidine-1-carboxylic acid tert-butyl ester (1.72 g, 10 mmol) and 1,4-naphthoquinone (1.12 g, 5 mmol) were dissolved in ethanol (20 mL), the solution was mixed and stirred at 80 ° C for 12 hours. The reaction solution was concentrated to dryness under reduced pressure, and the concentrate was purified by silica gel column chromatography (ethyl acetate: petroleum ether with a volume ratio of 3:1) and dried at 25 ° C to obtain a red solid. The red solid was dissolved in dichloromethane (20 ml), and trifluoroacetic acid (4 ml) was added. After reacting for 2 hours, the solution was rotary evaporated to dryness and then dried in an oven at 40 ° C to obtain intermediate 5a (red solid, 830 mg, yield 64%).

[0069] The synthesis method of intermediate 5b is the same as above.

[0070] Synthesis of intermediate 6a

[0071] The intermediate 5a (307 mg, 1.17 mmol) was dissolved in acetonitrile (5 ml), and azidoacetic acid (118 mg, 1.17 mmol), tetramethyl chlorouronium hexafluorophosphate (357.62 mg, 1.40 mmol), and N-methylimidazole (259.53 mg, 4.1 mmol) were added, and stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to a concentrate, and ethyl acetate: petroleum ether with a volume ratio of 1:1 was used as a developing solvent, and thin layer chromatography was used for tracking and monitoring. The filtrate was collected, distilled under reduced pressure, and then vacuum dried at 25°C to obtain the intermediate 6a (white solid, 300 mg, yield 74%).

[0072] The synthesis method of intermediate 6b is the same as above.

[0073] Synthesis of the final product GI-C-21

[0074] Intermediate 4 (145 mg, 0.289 mmol) and compound 6a (100 mg, 0.289 mmol) were dissolved in N,N-dimethylformamide (3 ml), copper sulfate (22.97 mg, 0.145 mmol) and sodium ascorbate (143 mg, 0.723 mmol) were dissolved in water (3 ml), the two solutions were mixed under nitrogen protection, and stirred at room temperature for 12 h. After the reaction was completed, the reaction solution was diluted with water (100 ml), extracted with ethyl acetate solution 3 times, dried with sodium sulfate, filtered, and the filtrate was rotary evaporated to dryness to obtain a concentrate, and a mixture of ethyl acetate / petroleum ether with a volume ratio of 1:1 was used as the eluent, and thin layer chromatography was used for tracking detection. The filtrate was collected, distilled under reduced pressure, and then dried in vacuo at 25°C to obtain the final product GI-C-21 (red solid, 50 mg, yield 30%).

[0075] The synthesis method of the final product GI-C-22 is the same as above.

[0076] Final product GI-C-21 (red solid, yield 63%). 1H NMR(400MHz, CDCl3)δ8.15–7.78(m,3H),7.65(dtd,J=34.3,7.5,1.4Hz,2H),7.19(s,2H),7.19–6.98(m,4 H),6.79(dt,J=6.6,3.7Hz,2H),6.11(d,J=5.5Hz,1H),6.00(d,J=12.3Hz,2H),5.19(s,2H),5.13–5.04(m ,1H),4.99(d,J=3.5Hz,2H),4.58(t,J=8.3Hz,1H),4.44(dd,J=10.9,7.6Hz,1H),4.16(dd,J=9.4,4.8Hz, 1H), 3.97 (dd, J=11.0, 5.0Hz, 1H), 3.75 (d, J=1.7Hz, 2H), 3.56–3.41 (m, J=6.8Hz, 2H), 2.84–2.64 (m, 2H).

[0077] Final product GI-C-22 (red solid, yield 56%). 1 H NMR(400MHz, CDCl3)δ8.03(dd,J=7.7,1.3Hz,1H),7.98–7.83(m,2H),7.63(td,J=7.6 ,1.4Hz,1H),7.53(td,J=7.5,1.3Hz,1H),7.26–7.01(m,8H),6.99–6.75(m,3H),6.51 (s,1H),6.02(d,J=3.8Hz,2H),5.17(s,2H),5.08(s,2H),3.81–3.73(m,4H),3.47(dq ,J=15.7,6.8Hz,2H),3.35(q,J=6.2Hz,2H),2.79–2.68(m,2H),1.82(p,J=6.5Hz,2H).

[0078] Example 2 Verification of the killing effect of the synthesized compound on GPX4 high-expressing tumor cell lines

[0079] Cell viability assay: CCK8 assay was used to evaluate the cytotoxicity and IC of all target compounds in HT1080 cells (DMEM medium) and MGC803 cells (1640 medium). 50 ML162 was used as a positive control. The cells were cultured at 5×10 3Cells were inoculated in 96-well plates at a density of 10 cells / well for 24 hours. Then, the cells were treated with different concentrations of compounds (0.1μM, 1μM, 5μM, 10μM, 30μM) for 48h. Subsequently, 10μL of CCK8 solution was added to each well, and after culturing for 1.5h, the absorbance at 450nm was detected using a microplate reader (TECAN). After the absorbance value was converted into inhibition rate, Graphpad Prism 5 was used to calculate IC 50 value.

[0080] The killing effects of the synthesized target compounds on GPX4 high-expressing cell lines are shown in Table 1:

[0081] Table 1

[0082]

[0083]

[0084] From the data in the above table, we can see that most of the synthesized quinone compounds have a good killing effect on HT1080 and MGC803 cell lines with high expression of GPX4. 50 The values ​​were all below 10 μM. Representative compounds included GI-C-11 and GD-C-16, which had the ability to kill gastric cancer cell line MGC803 comparable to the positive control ML162.

[0085] Example 3 Lipid peroxide determination

[0086] HT1080 cells were cultured in 6-well plates, with 6×10 cells per well. 5 After drug (5 μM) treatment, cells were collected and suspended in 500 μL PBS containing 2 μM C11-BODIPY (581 / 591), and incubated at 37 degrees for 10 min, followed by flow cytometry analysis. In this experiment, compound GI-C-16 was randomly selected as a representative to test its effect on intracellular lipid peroxides. The results are shown in Figure 1 .

[0087] Figure 1 Lipid peroxide assay

[0088] from Figure 1 It is not difficult to see that at the same concentration, GI-C-16 has a stronger ability to induce lipid peroxides than the positive control ML162, and the accumulation of peroxides will trigger ferroptosis.

[0089] Example 4 Molecular docking

[0090] The molecular docking software Autodock Vina was used to simulate the molecular structure of the representative compound GI-C-11 (e.g. Figure 2The experimental results showed that, unlike the ML162 covalent inhibitor, this class of compounds may interact with proteins in a non-covalent manner. GI-C-11 firmly occupied the active catalytic pocket of GPX4 and formed four hydrogen bonds with the four amino acid residues W136, N137, G47 and K48, which may affect the catalytic activity of GPX4 and induce ferroptosis in cells.

Claims

1. A quinone compound or a pharmacologically or physiologically acceptable salt thereof for targeted inhibition of GPX4, characterized in that: The general formula of the quinone compound or its pharmacologically or physiologically acceptable salt is shown in Formula I: In the formula I: R is any one of a hydrogen atom, a methyl group, a methoxy group, a halogen group, or a fused benzene ring substituent; R 1 is any one of a hydrogen atom, a methyl group, a methoxy group or a halogen substituent; Linker is a connecting group selected from any one of the following groups or any combination thereof, wherein m and n represent a natural number from 1 to 20: -(CH2CH2O) m -(CH2) n NH- or -(CH2) n NH- or -(CH2) n S-or 2. A quinone compound or a pharmacologically or physiologically acceptable salt thereof for targeted inhibition of GPX4, characterized in that: The quinone compound is any one of the following compounds GI-C-1 to GI-C-22:

3. A method for preparing the quinone compound or the pharmacologically or physiologically acceptable salt thereof according to claim 2, characterized in that Prepared by synthetic route 1: Synthetic route 1: Among them, R 1 , R 2 , the linker group corresponds to the substituent of the specific compound of claim 2.

4. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the quinone compound or a pharmacologically or physiologically acceptable salt thereof according to any one of claims 1 to 2, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

5. Use of the quinone compound according to any one of claims 1 to 2 or its pharmacologically or physiologically acceptable salt or the pharmaceutical composition according to claim 4 in the preparation of a quinone GPX4 inhibitor.

6. Use of the quinone compound according to any one of claims 1 to 2 or its pharmacologically or physiologically acceptable salt or the pharmaceutical composition according to claim 4 in the preparation of a drug for treating GPX4-related diseases.

7. The use according to claim 6, characterized in that The GPX4-related diseases are tumors and neurodegenerative diseases.

8. Use of a quinone compound according to any one of claims 1 to 2 or a pharmacologically or physiologically acceptable salt thereof or a pharmaceutical composition according to claim 4 in an anti-tumor drug, wherein the tumor is gastric cancer, breast cancer, lung cancer, ovarian cancer, colon adenocarcinoma, renal clear cell carcinoma, lung adenocarcinoma, prostate cancer, rectal adenocarcinoma, thyroid cancer and endometrial cancer.

9. The use according to claim 8, characterized in that The tumor is a tumor with high expression of GPX4.

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