A GPX4 protein degrader, its preparation method and application, and an anti-tumor cell drug

Abstract

The present invention provides a GPX4 protein degrader, a preparation method and application thereof, and an anti-tumor cell drug, belonging to the technical field of drug applications. The degrader provided by the present invention has a molecular structure of proteolysis targeting chimeras (PROTACs), with the ML210 unit as the targeting unit, which can effectively bind to the target protein. The A2 substituent serves as a small molecule ligand that binds to the E3 ubiquitin ligase complex, and the A1 substituent serves as a linker that connects the two ligands. Since PROTAC needs to form a conformationally stable ternary complex with the target protein and the E3 ligase in space to exert its degradation activity, the PROTAC strategy has the advantage of enhancing the target selectivity of drug molecules. Therefore, the GPX4 protein degrader can specifically recognize and target the GPX4 protein, and effectively ubiquitinate the GPX4 protein, thereby inducing ferroptosis in tumor cells.

Description

Technical Field

[0001] The present invention relates to the technical field of drug application, and in particular to a GPX4 protein degrading agent, a preparation method and application thereof, and an anti-tumor cell drug. Background Art

[0002] Ferroptosis is a type of programmed cell death that can be traced back to the beginning of this century. In 2012, Dixon's research group discovered that this non-apoptotic form of cell death is iron-dependent, so they called it "ferroptosis."

[0003] Glutathione peroxidase 4 (GPX4) is one of the peroxidases most associated with ferroptosis. GPX4 utilizes glutathione (GSH) and converts it into an oxidized form (GSSG), while reducing lipid peroxides (L-OOH) to an alcohol form (L-OH). GPX4 is highly expressed in cancer cells to counteract oxidative stress. Decreased GPX4 activity causes a large accumulation of lipid peroxides, driving ferroptosis. Therefore, targeted inhibition of GPX4 to cause ferroptosis has become a new anti-tumor strategy. Currently, GPX4 inhibitors are mainly covalent inhibitors. Common GPX4 protein inhibitors include: (1S, 3R)-RSL3, ML162, ML210, and JKE-1674, but all have the problem of low targeting and easy off-target.

[0004] Summary of the invention

[0005] In view of this, the purpose of the present invention is to provide a GPX4 protein degrader and a preparation method and application thereof, and an anti-tumor cell drug. The GPX4 protein degrader provided by the present invention has a protein degradation targeting chimera (PROTAC) molecular structure, which can effectively target and degrade GPX4 protein, thereby inducing ferroptosis of tumor cells.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a GPX4 protein degradation agent having a structure shown in Formula I:

[0008]

[0009] In Formula I, A1 is

[0010] A2 is

[0011] Among them, m=1~20, n=1~20.

[0012] Preferably, it has the structure shown in any one of Formula I-1 to I-15:

[0013]

[0014] In Formula I-1 to I-4, m = 1 to 20;

[0015]

[0016]

[0017] In Formula I-5 to I-8, n = 1 to 20;

[0018]

[0019]

[0020] The present invention provides a method for preparing the above-mentioned GPX4 protein degrader, comprising the following steps:

[0021] ① When A1 is The method for preparing the GPX4 protein degrader comprises the following steps:

[0022] A compound having the structure shown in Formula a reacts with a compound having the structure of NH2-A2 in a substitution reaction to obtain a compound having the structure shown in Formula b;

[0023] The compound having the structure shown in Formula b undergoes a deprotection reaction to obtain a compound having the structure shown in Formula c;

[0024] The compound having the structure shown in Formula c reacts with a compound having the structure shown in Formula d in a condensation reaction to obtain the GPX4 protein degrader;

[0025]

[0026] ② When A1 is The method for preparing the GPX4 protein degrader comprises the following steps:

[0027] A compound having the structure shown in Formula e reacts with a compound having the structure of F-A2 in a substitution reaction to obtain a compound having the structure shown in Formula f;

[0028] The compound having the structure shown in Formula f undergoes a deprotection reaction to obtain a compound having the structure shown in Formula g;

[0029] The compound having the structure shown in Formula g reacts with a compound having the structure shown in Formula d in a condensation reaction to obtain the GPX4 protein degrader;

[0030]

[0031] ③ When A1 is the preparation method of the GPX4 protein degrader includes the following steps:

[0032] The compound with the structure shown in formula h reacts with the compound with the F-A2 structure through a substitution reaction to obtain a compound with the structure shown in formula i;

[0033] The compound with the structure shown in formula i undergoes a deprotection reaction to obtain a compound with the structure shown in formula j;

[0034] The compound with the structure shown in formula j reacts with the compound with the structure shown in formula k through a substitution reaction to obtain a compound with the structure shown in formula l;

[0035] The compound with the structure shown in formula l undergoes a condensation reaction with the compound with the structure shown in formula d to obtain the GPX4 protein degrader;

[0036]

[0037] ④ When A1 is the preparation method of the GPX4 protein degrader includes the following steps:

[0038] The compound with the structure shown in formula m reacts with the compound with the F-A2 structure through a substitution reaction to obtain a compound with the structure shown in formula n;

[0039]

[0040] The compound with the structure shown in formula n undergoes a condensation reaction and a deprotection reaction with the compound with the structure shown in formula d to obtain the GPX4 protein degrader.

[0041] Preferably, the preparation method of the compound with the structure shown in formula d includes the following steps:

[0042] 4-chloro-4'-hydroxybenzophenone reacts with tert-butyl bromoacetate through a substitution reaction to obtain a compound with the structure shown in formula d-1;

[0043] Under the action of a reducing agent, the compound with the structure shown in formula d-1 undergoes a reduction reaction to obtain a compound with the structure shown in formula d-2;

[0044] The compound with the structure shown in formula d-2 undergoes a chlorination reaction with oxalyl chloride to obtain a compound with the structure shown in formula d-3;

[0045] The compound with the structure shown in formula d-3 reacts with piperazine through a substitution reaction to obtain a compound with the structure shown in formula d-4;

[0046] 6-Methylisoxazole-3-carboxylic acid undergoes a nitration reaction with potassium nitrate to obtain a compound with the structure shown in Formula d-5;

[0047] A compound with the structure shown in Formula d-4, a compound with the structure shown in Formula d-5, and oxalyl chloride undergo a coupling reaction to obtain a compound with the structure shown in Formula d-6;

[0048] The compound with the structure shown in Formula d-6 undergoes a deprotection reaction to obtain a compound with the structure shown in Formula d.

[0049]

[0050] Preferably, when A2 is The preparation method of the compound with the structure of NH2-A2 includes the following steps:

[0051] The compound with the structure shown in Formula v1 undergoes a substitution reaction with 4-methylthiazole to obtain a compound with the structure shown in Formula v2;

[0052] The compound with the structure shown in Formula v2 undergoes a deprotection reaction to obtain a compound with the structure shown in Formula v3;

[0053] The compound with the structure shown in Formula v3 undergoes a condensation reaction with the compound with the structure shown in Formula w1 to obtain a compound with the structure shown in Formula v4;

[0054] The compound with the structure shown in Formula v4 undergoes a deprotection reaction to obtain a compound with the structure shown in Formula v5;

[0055] The compound with the structure shown in Formula v5 undergoes a condensation reaction with the compound with the structure shown in Formula w2 to obtain a compound with the structure shown in Formula v6;

[0056] The compound with the structure shown in Formula v6 undergoes a deprotection reaction to obtain a compound with the structure shown in Formula v7;

[0057]

[0058] Preferably, the preparation method of the compound with the structure shown in Formula a includes the following steps:

[0059] The compound with the structure shown in Formula a-1 undergoes a substitution reaction with 1-Boc-piperazine to obtain a compound with the structure shown in Formula a-2;

[0060]

[0061] The compound with the structure shown in Formula a-2 undergoes a substitution reaction with p-toluenesulfonyl chloride to obtain a compound with the structure shown in Formula a.

[0062] Preferably, the preparation method of the compound having the structure shown in Formula e includes the following steps:

[0063] Phthalimide undergoes a substitution reaction with the compound having the structure shown in Formula e-1 to obtain a compound having the structure shown in Formula e-2;

[0064] The compound having the structure shown in Formula e-2 undergoes a substitution reaction with 1-Boc piperazine to obtain a compound having the structure shown in Formula e-3;

[0065] The compound having the structure shown in Formula e-3 undergoes a deprotection reaction to obtain a compound having the structure shown in Formula e-4;

[0066] The compound having the structure shown in Formula e-4 undergoes a substitution reaction with 1-Boc-4-bromomethylpiperidine to obtain a compound having the structure shown in Formula e-5;

[0067] Under the action of hydrazine hydrate, the compound having the structure shown in Formula e-5 undergoes a deimination reaction to obtain a compound having the structure shown in Formula e;

[0068]

[0069] Preferably, the preparation method of the compound having the structure shown in Formula h includes the following steps:

[0070] The compound having the structure shown in Formula h-1 undergoes a substitution reaction with the compound having the structure shown in Formula h-2 to obtain a compound having the structure shown in Formula h-3;

[0071]

[0072] The compound having the structure shown in Formula h-3 undergoes a de-Cbz protection reaction to obtain a compound having the structure shown in Formula h;

[0073] The preparation method of the compound having the structure shown in Formula m includes the following steps:

[0074] The compound having the structure shown in Formula m-1 undergoes a substitution reaction and a deprotection reaction with the compound having the structure shown in Formula m-2 to obtain a compound having the structure shown in Formula m-3;

[0075]

[0076] The compound having the structure shown in Formula m-3 undergoes a substitution reaction with the compound having the structure shown in Formula m-4 to obtain a compound having the structure shown in Formula m-5;

[0077]

[0078] The compound with the structure shown in Formula m-5 is subjected to a de-Cbz protecting group reaction to obtain a compound with the structure shown in Formula m.

[0079] The present invention provides the use of the above-mentioned GPX4 protein degrader in the preparation of anti-tumor drugs and anti-drug-resistant tumor drugs.

[0080] The present invention provides an anti-tumor drug, which includes a drug active component and a drug excipient; the drug active component is the above-mentioned GPX4 protein degrader.

[0081] The present invention provides a GPX4 protein degrader with the structure shown in Formula I. The proteolysis targeting chimera (PROTACs) strategy directly induces the degradation of target proteins. PROTACs have a unique "event-driven" mode of action, the advantage of targeting undruggable proteins, and high-efficiency and long-lasting activity. The degrader provided by the present invention has a proteolysis targeting chimera (PROTACs) molecular structure, with the ML210 unit as the targeting unit, which can effectively bind to the target protein. The A2 substituent serves as a small molecule ligand that binds to the E3 ubiquitin ligase complex, and the A1 substituent serves as a linker that connects the two ligands. Since PROTAC needs to form a conformationally stable ternary complex with the target protein and the E3 ligase in space to exert its degradation activity, the PROTAC strategy has the advantage of enhancing the target selectivity of drug molecules. Therefore, the GPX4 protein degrader can specifically recognize and target the GPX4 protein, and effectively ubiquitinate the GPX4 protein, thereby inducing ferroptosis in tumor cells. The results of the examples show that the GPX4 protein degrader provided by the present invention exhibits obvious degradation activity and anti-tumor activity in vivo and in vitro. By degrading GPX4, a large amount of intracellular reactive oxygen species (ROS) accumulates, ultimately leading to ferroptosis of cells. Description of the Drawings

[0082] Figure 1 It shows the degradation effect of the GPX4 protein degrader on the GPX4 protein after acting on HT1080 cells;

[0083] Figure 2 It shows the relationship between the degradation activity of DC2 and the concentration and time;

[0084] Figure 3 It shows the evaluation results of the inhibitory effect of DC2 on cell proliferation;

[0085] Figure 4 It shows the relationship between the degradation effect of DC2 on GPX4 in xenograft human fibrosarcoma (HT1080) tissue and the administration time;

[0086] Figure 5 It shows the changes in the tumor volume and body weight of mice after multiple administrations. Detailed Embodiments

[0087] The present invention provides a GPX4 protein degrader having the structure shown in Formula I:

[0088]

[0089] In Formula I, A1 is

[0090] A2 is

[0091] Wherein, m = 1 to 20, preferably 7 to 14, more preferably 8 to 12, and further preferably 10 to 11;

[0092] n = 1 to 20, preferably 2 to 15, more preferably 2 to 10, and further preferably 4 to 8.

[0093] In the present invention, "*" represents the connection site.

[0094] In the present invention, the GPX4 protein degrader has the structure shown in any one of Formulas I-1 to I-15:

[0095]

[0096] In Formulas I-1 to I-4, m = 1 to 20;

[0097]

[0098]

[0099] In Formulas I-5 to I-8, n = 1 to 20;

[0100]

[0101]

[0102] As a specific embodiment of the present invention, the structural formula of the GPX4 protein degrader is shown in Table 1.

[0103] Table 1 Structural formula of the GPX4 protein degrader

[0104]

[0105]

[0106]

[0107] The present invention provides a preparation method of the above GPX4 protein degrader, comprising the following steps:

[0108] ①When A1 is the preparation method of the GPX4 protein degrader includes the following steps:

[0109] The compound with the structure shown in formula a reacts with the compound with the structure of NH2-A2 through a substitution reaction to obtain a compound with the structure shown in formula b;

[0110] The compound with the structure shown in formula b undergoes a deprotection reaction to obtain a compound with the structure shown in formula c;

[0111] The compound with the structure shown in formula c reacts with the compound with the structure shown in formula d through a condensation reaction to obtain the GPX4 protein degrader;

[0112]

[0113] In the present invention, the compound with the structure shown in formula a reacts with the compound with the structure of NH2-A2 through a substitution reaction to obtain a compound with the structure shown in formula b. In the present invention, the preparation method of the compound with the structure shown in formula a includes the following steps:

[0114] The compound with the structure shown in formula a-1 reacts with 1-Boc-piperazine through a substitution reaction to obtain a compound with the structure shown in formula a-2;

[0115]

[0116] In the present invention, the molar ratio of the compound with the structure shown in formula a-1 to 1-Boc-piperazine is preferably 1:1. In the present invention, the catalyst used in the substitution reaction is preferably K2CO3 and KI. In the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably the reflux temperature, and the time is preferably 2 h.

[0117] The compound with the structure shown in formula a-2 reacts with p-toluenesulfonyl chloride through a substitution reaction to obtain a compound with the structure shown in formula a.

[0118] In the present invention, the molar ratio of the compound with the structure shown in formula a-2 to p-toluenesulfonyl chloride is preferably 1:1. In the present invention, the catalyst of the substitution reaction is preferably triethylamine. In the present invention, the organic solvent used in the substitution reaction is preferably dichloromethane; the temperature of the substitution reaction is preferably 0 °C to room temperature, and the time is preferably overnight.

[0119] In the present invention, a compound having the structure shown in formula a undergoes a substitution reaction with a compound having the structure NH2-A2. In the present invention, the molar ratio of the compound having the structure shown in formula a to the compound having the structure NH2-A2 is preferably 1:1. In the present invention, the catalyst for the substitution reaction is preferably K2CO3 and KI. In the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably the reflux temperature, and the time is preferably 2 h.

[0120] In the present invention, a compound having the structure shown in formula b undergoes a deprotection reaction to obtain a compound having the structure shown in formula c. In the present invention, the catalyst for the deprotection reaction is preferably trifluoroacetic acid. In the present invention, the organic solvent for the deprotection reaction is preferably dichloromethane; the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 30 min.

[0121] In the present invention, the compound having the structure shown in formula c undergoes a condensation reaction with the compound having the structure shown in formula d to obtain a GPX4 protein degrader. In the present invention, the preparation method of the compound having the structure shown in formula d preferably includes the following steps:

[0122] 4-Chloro-4'-hydroxybenzophenone undergoes a substitution reaction with tert-butyl bromoacetate to obtain a compound having the structure shown in formula d-1;

[0123] Under the action of a reducing agent, the compound having the structure shown in formula d-1 undergoes a reduction reaction to obtain a compound having the structure shown in formula d-2;

[0124] The compound having the structure shown in formula d-2 undergoes a chlorination reaction with oxalyl chloride to obtain a compound having the structure shown in formula d-3;

[0125] The compound having the structure shown in formula d-3 undergoes a substitution reaction with piperazine to obtain a compound having the structure shown in formula d-4;

[0126] 6-Methylisoxazole-3-carboxylic acid undergoes a nitration reaction with potassium nitrate to obtain a compound having the structure shown in formula d-5;

[0127] The compound having the structure shown in formula d-4, the compound having the structure shown in formula d-5 and oxalyl chloride undergo a coupling reaction to obtain a compound having the structure shown in formula d-6;

[0128] The compound having the structure shown in formula d-6 undergoes a deprotection reaction to obtain a compound having the structure shown in formula d;

[0129]

[0130] In the present invention, 4-chloro-4'-hydroxybenzophenone undergoes a substitution reaction with tert-butyl bromoacetate to obtain a compound having the structure shown in Formula d-1. In the present invention, the molar ratio of 4-chloro-4'-hydroxybenzophenone to tert-butyl bromoacetate is preferably 1:1.5. In the present invention, the catalyst for the substitution reaction is preferably K2CO3. In the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably 85 °C, and the time is preferably 2 h.

[0131] In the present invention, under the action of a reducing agent, the compound having the structure shown in Formula d-1 undergoes a reduction reaction to obtain a compound having the structure shown in Formula d-2. In the present invention, the reducing agent is preferably sodium borohydride. In the present invention, the molar ratio of the reducing agent to the compound having the structure shown in Formula d-1 is preferably 1:1. In the present invention, the organic solvents used in the reduction reaction are preferably tetrahydrofuran and methanol. In the present invention, the temperature of the reduction reaction is preferably 0 °C, and the time is preferably 30 min.

[0132] In the present invention, the compound having the structure shown in Formula d-2 undergoes a chlorination reaction with oxalyl chloride to obtain a compound having the structure shown in Formula d-3. In the present invention, the molar ratio of the compound having the structure shown in Formula d-2 to oxalyl chloride is preferably 1:2; in the present invention, the organic solvents for the chlorination reaction are preferably dichloromethane and dimethylformamide. In the present invention, the temperature of the chlorination reaction is preferably 0 °C, and the time is preferably 8 h.

[0133] In the present invention, the compound having the structure shown in Formula d-3 undergoes a substitution reaction with piperazine to obtain a compound having the structure shown in Formula d-4. In the present invention, the molar ratio of the compound having the structure shown in Formula d-3 to piperazine is preferably 1:10; in the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably 85 °C, more preferably 90 °C.

[0134] In the present invention, 6-methylisoxazole-3-carboxylic acid undergoes a nitration reaction with potassium nitrate to obtain a compound having the structure shown in Formula d-5. In the present invention, the molar ratio of 6-methylisoxazole-3-carboxylic acid to potassium nitrate is preferably 1:1.5.

[0135] In the present invention, the nitration reaction is preferably carried out in a sulfuric acid environment, and the temperature of the nitration reaction is preferably 50 °C, and the time is preferably 4 h.

[0136] In the present invention, a compound having the structure shown in Formula d-4 and a compound having the structure shown in Formula d-5 are subjected to a coupling reaction with oxalyl chloride to obtain a compound having the structure shown in Formula d-6. In the present invention, the molar ratio of the compound having the structure shown in Formula d-4, the compound having the structure shown in Formula d-5, and oxalyl chloride is preferably 1:1.5.

[0137] In the present invention, the catalyst for the coupling reaction is preferably triethylamine; the organic solvent used in the coupling reaction is preferably acetonitrile; the temperature of the coupling reaction is preferably room temperature, and the time is preferably 5 h.

[0138] In the present invention, the compound having the structure shown in Formula d-6 is subjected to a deprotection reaction to obtain a compound having the structure shown in Formula d. In the present invention, the deprotection reagent used in the deprotection reaction is preferably trifluoroacetic acid. In the present invention, the organic solvent used in the deprotection reaction is preferably dichloromethane. In the present invention, the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 5 h.

[0139] In the present invention, the compound having the structure shown in Formula c and the compound having the structure shown in Formula d are subjected to a condensation reaction to obtain a GPX4 protein degrader. In the present invention, the molar ratio of the compound having the structure shown in Formula c to the compound having the structure shown in Formula d is preferably 1:1.

[0140] In the present invention, the condensation reagent used in the condensation reaction is preferably HATU and DIPEA. In the present invention, the organic solvent used in the condensation reaction is preferably dimethylformamide. In the present invention, the temperature of the condensation reaction is preferably room temperature, and the time is preferably 2 h.

[0141] In the present invention, the compound having the NH2-A2 structure is preferably

[0142] In the present invention, the preparation method of the compound having the structure shown in Formula V7 preferably includes the following steps:

[0143] A compound having the structure shown in Formula v1 is subjected to a substitution reaction with 4-methylthiazole to obtain a compound having the structure shown in Formula v2;

[0144]

[0145] In the present invention, the molar ratio of the compound having the structure shown by formula v1 to 4-methylthiazole is preferably 1:2. In the present invention, the catalyst for the substitution reaction is preferably palladium acetate and potassium acetate. In the present invention, the solvent for the substitution reaction is preferably dry dimethylformamide; the substitution reaction is preferably carried out under the protection of Ar gas, the temperature of the substitution reaction is preferably 90 °C, and the time is preferably 2 h.

[0146] In the present invention, the compound having the structure shown by formula v2 undergoes a deprotection reaction to obtain a compound having the structure shown by formula v3;

[0147]

[0148] In the present invention, the deprotection reagent used for the deprotection reaction is preferably trifluoroacetic acid; in the present invention, the organic solvent used for the deprotection reaction is preferably dichloromethane; the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 30 min.

[0149] In the present invention, the compound having the structure shown by formula v3 and the compound having the structure shown by formula w1 undergo a condensation reaction to obtain a compound having the structure shown by formula v4;

[0150]

[0151] In the present invention, the molar ratio of the compound having the structure shown by formula v3 to the compound having the structure shown by formula w1 is preferably 1:1. In the present invention, the condensation reagent used for the condensation reaction is preferably HATU and DIPEA. In the present invention, the organic solvent used for the condensation reaction is preferably dimethylformamide; the temperature of the condensation reaction is preferably room temperature, and the time is preferably overnight.

[0152] In the present invention, the compound having the structure shown by formula v4 undergoes a deprotection reaction to obtain a compound having the structure shown by formula v5;

[0153]

[0154] In the present invention, the deprotection reagent used for the deprotection reaction is preferably trifluoroacetic acid; the organic solvent used for the deprotection reaction is preferably dichloromethane; the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 30 min.

[0155] In the present invention, the compound having the structure shown by formula v5 and the compound having the structure shown by formula w2 undergo a condensation reaction to obtain a compound having the structure shown by formula v6;

[0156]

[0157] In the present invention, the molar ratio of the compound having the structure shown by formula v5 to the compound having the structure shown by formula w2 is preferably 1:1. In the present invention, the condensation reagent used in the condensation reaction is preferably HATU and DIPEA. In the present invention, the organic solvent used in the condensation reaction is preferably dimethylformamide; the temperature of the condensation reaction is preferably room temperature, and the time is preferably overnight.

[0158] The compound having the structure shown by formula v6 is subjected to a deprotection reaction to obtain a compound having the structure shown by formula v7.

[0159] In the present invention, the deprotection reagent used in the deprotection reaction is preferably trifluoroacetic acid; the organic solvent used in the deprotection reaction is preferably dichloromethane; the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 30 min.

[0160] ② When A1 is The preparation method of the GPX4 protein degrader includes the following steps:

[0161] The compound having the structure shown by formula e and the compound having the F-A2 structure are subjected to a substitution reaction to obtain a compound having the structure shown by formula f;

[0162]

[0163] The compound having the structure shown by formula f is subjected to a deprotection reaction to obtain a compound having the structure shown by formula g;

[0164]

[0165] The compound having the structure shown by formula g and the compound having the structure shown by formula d are subjected to a condensation reaction to obtain the GPX4 protein degrader.

[0166] In the present invention, the compound having the structure shown by formula e and the compound having the NH2-A2 structure are subjected to a substitution reaction to obtain a compound having the structure shown by formula f. In the present invention, the preparation method of the compound having the structure shown by formula e preferably includes the following steps:

[0167] Phthalimide and the compound having the structure shown by formula e-1 are subjected to a substitution reaction to obtain a compound having the structure shown by formula e-2;

[0168]

[0169] In the present invention, the molar ratio of phthalimide to the compound having the structure shown in Formula e-1 is preferably 1:1.2. In the present invention, the catalyst for the substitution reaction is preferably tetrabutylammonium bromide and K2CO3. In the present invention, the organic solvent used in the substitution reaction is preferably dimethylformamide; the temperature of the substitution reaction is preferably room temperature, and the time is preferably 3 h.

[0170] In the present invention, the compound having the structure shown in Formula e-2 undergoes a substitution reaction with 1-Boc piperazine to obtain a compound having the structure shown in Formula e-3;

[0171]

[0172] In the present invention, the catalyst for the substitution reaction is preferably triethanolamine and K2CO3. In the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably the reflux temperature, and the time is preferably overnight.

[0173] In the present invention, the compound having the structure shown in Formula e-3 undergoes a deprotection reaction to obtain a compound having the structure shown in Formula e-4;

[0174]

[0175] In the present invention, the deprotection reagent used in the deprotection reaction is preferably trifluoroacetic acid; the organic solvent for the deprotection reaction is preferably dichloroethane; the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 3 h.

[0176] In the present invention, the compound having the structure shown in Formula e-4 undergoes a substitution reaction with 1-Boc-4-bromomethylpiperidine to obtain a compound having the structure shown in Formula e-5;

[0177]

[0178] In the present invention, the molar ratio of the compound having the structure shown in Formula e-4 to 1-Boc-4-bromomethylpiperidine is preferably 1:1.1. In the present invention, the catalyst for the substitution reaction is preferably triethanolamine and K2CO3. In the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably the reflux temperature, and the time is preferably overnight.

[0179] Under the action of hydrazine hydrate, the compound having the structure shown in Formula e-5 undergoes a deacylimidation reaction to obtain a compound having the structure shown in Formula e. In the present invention, the molar ratio of the compound having the structure shown in Formula e-5 to hydrazine hydrate is preferably 1:2. In the present invention, the solvent for the deacylimidation reaction is preferably ethanol; the temperature of the deacylimidation reaction is preferably the reflux temperature, and the time is preferably overnight.

[0180] In the present invention, a compound having the structure shown in formula e undergoes a substitution reaction with a compound having the F-A2 structure to obtain a compound having the structure shown in formula f. In the present invention, the compound having the F-A2 structure is preferably fluorinated thalidomide or N-methyl fluorinated thalidomide.

[0181]

[0182] In the present invention, the molar ratio of the compound having the structure shown in formula e to the compound having the F-A2 structure is preferably 1:1. In the present invention, the condensing agent used in the substitution reaction is preferably DIPEA. In the present invention, the organic solvent used in the substitution reaction is preferably dimethyl sulfoxide. In the present invention, the temperature of the substitution reaction is preferably 130 °C, and the time is preferably 0.5 h.

[0183] In the present invention, the compound having the structure shown in formula f undergoes a deprotection reaction to obtain a compound having the structure shown in formula g. In the present invention, the deprotecting agent used in the deprotection reaction is preferably trifluoroacetic acid. In the present invention, the organic solvent used in the deprotection reaction is preferably dichloromethane; the temperature of the deprotection reaction is preferably room temperature, and the time is preferably overnight.

[0184] In the present invention, the compound having the structure shown in formula g undergoes a condensation reaction with the compound having the structure shown in formula d to obtain a GPX4 protein degrader. In the present invention, the molar ratio of the compound having the structure shown in formula g to the compound having the structure shown in formula d is preferably 1:1; the condensing agent used in the condensation reaction is preferably HATU and DIPEA. In the present invention, the organic solvent used in the condensation reaction is preferably dimethylformamide; the temperature of the condensation reaction is preferably room temperature, and the time is preferably overnight.

[0185] ③ When A1 is the preparation method of the GPX4 protein degrader includes the following steps:

[0186] A compound having the structure shown in formula h undergoes a substitution reaction with a compound having the F-A2 structure to obtain a compound having the structure shown in formula i;

[0187] The compound having the structure shown in formula i undergoes a deprotection reaction to obtain a compound having the structure shown in formula j;

[0188] The compound having the structure shown in formula j undergoes a substitution reaction with the compound having the structure shown in formula k to obtain a compound having the structure shown in formula l;

[0189] The compound with the structure shown in Formula l undergoes a condensation reaction with the compound with the structure shown in Formula d to obtain a GPX4 protein degrader;

[0190]

[0191] In the present invention, the compound with the structure shown in Formula h undergoes a substitution reaction with the compound with the F-A2 structure to obtain the compound with the structure shown in Formula i. In the present invention, the preparation method of the compound with the structure shown in Formula h preferably includes the following steps:

[0192] The compound with the structure shown in Formula h-1 undergoes a substitution reaction with the compound with the structure shown in Formula h-2 to obtain the compound with the structure shown in Formula h-3;

[0193]

[0194] In the present invention, the molar ratio of the compound with the structure shown in Formula h-1 to the compound with the structure shown in Formula h-2 is preferably 1:1. In the present invention, the catalyst for the substitution reaction is preferably K2CO3 and KI. In the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably the reflux temperature, and the time is preferably overnight.

[0195] In the present invention, the compound with the structure shown in Formula h-3 undergoes a de-Cbz protecting group reaction to obtain the compound with the structure shown in Formula h. In the present invention, the deprotecting reagent used in the de-Cbz protecting group reaction is preferably trifluoroacetic acid; the organic solvent used in the de-Cbz protecting group reaction is preferably dichloromethane; the temperature of the de-Cbz protecting group reaction is preferably room temperature, and the time is preferably 3 h.

[0196] In the present invention, the compound with the structure shown in Formula h undergoes a substitution reaction with the compound with the F-A2 structure to obtain the compound with the structure shown in Formula i. In the present invention, the compound with the F-A2 structure is preferably fluorinated thalidomide. In the present invention, the structural formula of the fluorinated thalidomide is

[0197] In the present invention, the molar ratio of the compound with the structure shown in Formula h to the compound with the F-A2 structure is preferably 1:1. In the present invention, the condensing agent used in the substitution reaction is preferably DIPEA; the organic solvent used in the substitution reaction is preferably dimethyl sulfoxide. In the present invention, the temperature of the substitution reaction is preferably 130 °C, and the time is preferably 1 h.

[0198] In the present invention, a compound having the structure shown in Formula I undergoes a deprotection reaction to obtain a compound having the structure shown in Formula J. In the present invention, the deprotection reagent used in the deprotection reaction is preferably trifluoroethylamine; the organic solvent used in the deprotection reaction is preferably dichloromethane; the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 3 h.

[0199] In the present invention, a compound having the structure shown in Formula J reacts with a compound having the structure shown in Formula K through a substitution reaction to obtain a compound having the structure shown in Formula L. In the present invention, the molar ratio of the compound having the structure shown in Formula J to the compound having the structure shown in Formula K is preferably 1:1.5. In the present invention, the catalyst used in the substitution reaction is preferably K2CO3 and KI. In the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably the reflux temperature, and the time is preferably overnight.

[0200] In the present invention, a compound having the structure shown in Formula L reacts with a compound having the structure shown in Formula D through a condensation reaction to obtain a GPX4 protein degrader. In the present invention, the compound having the structure shown in Formula L is preferably first subjected to a deprotection reaction, and the deprotection reagent used in the deprotection reaction is preferably trifluoroacetic acid. In the present invention, the organic solvent used in the deprotection reaction is preferably dichloromethane; the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 3 h.

[0201] In the present invention, the condensation reagent used in the condensation reaction is preferably HATU and DIPEA. In the present invention, the organic solvent used in the condensation reaction is preferably dimethylformamide; the temperature of the condensation reaction is preferably room temperature, and the time is preferably overnight.

[0202] ④ When A1 is The preparation method of the GPX4 protein degrader includes the following steps:

[0203] A compound having the structure shown in Formula M reacts with a compound having the F-A2 structure through a substitution reaction to obtain a compound having the structure shown in Formula N;

[0204]

[0205] The compound having the structure shown in Formula N reacts with the compound having the structure shown in Formula D through a condensation reaction and a deprotection reaction to obtain a GPX4 protein degrader.

[0206] In the present invention, a compound having the structure shown in Formula M reacts with a compound having the F-A2 structure through a substitution reaction to obtain a compound having the structure shown in Formula N. In the present invention, the preparation method of the compound having the structure shown in Formula M includes the following steps:

[0207] The compound having the structure shown in formula m-1 and the compound having the structure shown in formula m-2 undergo a substitution reaction and a deprotection reaction to obtain a compound having the structure shown in formula m-3;

[0208]

[0209] In the present invention, the molar ratio of the compound having the structure shown in formula m-1 to the compound having the structure shown in formula m-2 is preferably 1:1. In the present invention, the catalyst for the substitution reaction is preferably K2CO3 and KI. In the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably the reflux temperature, and the time is preferably overnight.

[0210] In the present invention, the compound having the structure shown in formula m-3 and the compound having the structure shown in formula m-4 undergo a substitution reaction to obtain a compound having the structure shown in formula m-5;

[0211]

[0212] In the present invention, the molar ratio of the compound having the structure shown in formula m-3 to the compound having the structure shown in formula m-4 is preferably 1:1, and the catalyst for the substitution reaction is preferably K2CO3 and KI. In the present invention, the organic solvent used in the substitution reaction is preferably acetonitrile; the temperature of the substitution reaction is preferably the reflux temperature, and the time is preferably overnight.

[0213] In the present invention, the compound having the structure shown in formula m-5 undergoes a de-Cbz protection group reaction to obtain a compound having the structure shown in formula m. In the present invention, the deprotection reagent used in the de-Cbz protection group reaction is preferably trifluoroacetic acid; the organic solvent used in the de-Cbz protection group reaction is preferably dichloromethane; the temperature of the de-Cbz protection group reaction is preferably room temperature, and the time is preferably 3 h.

[0214] In the present invention, the compound having the structure shown in formula m and the compound having the F-A2 structure undergo a substitution reaction to obtain a compound having the structure shown in formula n. In the present invention, the compound having the F-A2 structure is preferably thalidomide fluoride. In the present invention, the condensing agent used in the substitution reaction is preferably DIPEA. In the present invention, the organic solvent used in the substitution reaction is preferably dimethyl sulfoxide; the temperature of the substitution reaction is preferably 130 °C, and the time is preferably 1 h.

[0215] In the present invention, a compound represented by formula n and a compound having the structure represented by formula d are subjected to a condensation reaction and a deprotection reaction to obtain a GPX4 protein degrader. In the present invention, the molar ratio of the compound represented by formula n to the compound having the structure represented by formula d is preferably 1:1; the condensation reagent used in the condensation reaction is preferably HATU and DIPEA. In the present invention, the organic solvent used in the condensation reaction is preferably dimethylformamide; the temperature of the condensation reaction is preferably room temperature, and the time is preferably overnight.

[0216] In the present invention, the deprotection reagent used in the deprotection reaction is preferably trifluoroacetic acid; the organic solvent used in the deprotection reaction is preferably dichloromethane; the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 3 h.

[0217] The present invention provides the use of the above GPX4 protein degrader in the preparation of anti-tumor drugs. In the present invention, the anti-tumor drug is preferably a drug against fibrosarcoma, oral squamous cell carcinoma, glioma, diffuse large B-cell lymphoma, liver cancer, pancreatic cancer, prostate cancer, breast cancer, neuroblastoma, ovarian cancer, melanoma, kidney cancer, gastric cancer, colorectal cancer, thyroid cancer, lung cancer, head and neck cancer, urothelial cancer, cervical cancer or endometrial cancer.

[0218] The present invention provides an anti-tumor cell drug, comprising a drug active component and a drug excipient; the drug active component is the above GPX4 protein degrader; the present invention has no special requirements for the drug excipient, and the drug excipients well-known to those skilled in the art can be used. In the present invention, the effective content of the drug active ingredient in the drug is preferably 1 to 50 wt%, more preferably 5 to 30 wt%.

[0219] The following is a detailed description of a GPX4 protein degrader provided by the present invention, its preparation method and application, and an anti-tumor cell drug in conjunction with examples, but they should not be construed as limiting the protection scope of the present invention.

[0220] The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified. The structural formulas of some intermediates in the following examples are shown in Table 2.

[0221] Table 2 Structural formulas of some intermediates

[0222]

[0223]

[0224] Example 1 Synthesis of GPX4 protein degrader MC1

[0225] (1) Synthesis of intermediate SM3

[0226] 5-Methylisoxazole-3-carboxylic acid (10 g, 78.68 mmol) was added portionwise to a mixture of KNO3 (11.93 g, 118.02 mmol) and concentrated sulfuric acid (30 ml). After complete dissolution, the reaction mixture was heated at 50 °C for 4 h. After the reaction was completed, it was cooled to room temperature. Cold water was slowly added to the system, and then the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed successively with saturated NaCl solution and dried over anhydrous Na2SO4. The solvent was removed by distillation under reduced pressure to obtain a yellow solid (3.23 g), which was used in the next step without purification.

[0227] (2) Synthesis of intermediate M1

[0228] 4-Chloro-4'-hydroxybenzophenone (10 g, 42.98 mmol) was dissolved in 150 ml of acetonitrile. K2CO3 (8.95 g, 64.77 mmol) and tert-butyl bromoacetate (10.06 g, 51.58 mmol) were added successively, and the reaction mixture was refluxed at 85 °C for 2 h. After the reaction was completed, cold water was added to the system to quench the reaction mixture, and then the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed successively with saturated NaCl solution and dried over anhydrous Na2SO4. Column chromatography (P / E = 30 / 1 - 10 / 1) gave a white solid (13.9 g, 94%).

[0229] 1 H NMR (400 MHz, DMSO-d 6 ) δ 7.72 (q, J = 8.84, 16.40 Hz, 4H, Ar-H), 7.58 (d, J = 8.48 Hz, 2H, Ar-H a ), 7.06 (d, J = 8.84 Hz, 2H, Ar-H b ), 4.78 (s, 2H, CH2), 1.43 (s, 3H, CH3×3); 13 C NMR (100 MHz, DMSO-d 6 ) δ 193.59, 167.73, 161.94, 137.50, 136.75, 132.48, 131.56, 130.03, 128.99, 114.89, 82.10, 81.90, 65.50, 29.00, 28.11, 27.87.

[0230] (3) Synthesis of intermediate M2

[0231] M1 (0.5 g, 1.44 mmol) was dissolved in a mixed solvent of THF / MeOH (5 / 5 ml), stirred for 10 min under an ice bath condition, and then NaBH4 (49 mg, 1.44 mmol) was added thereto in portions, followed by continued stirring for 30 min. After the reaction was completed, the reaction was quenched with saturated NH4Cl solution, extracted three times with ethyl acetate, the organic phases were combined, washed successively with saturated NaCl solution and dried over anhydrous Na2SO4. Column chromatography (P / E = 10 / 1 - 6 / 1) gave a colorless transparent liquid (0.4 g, 80%).

[0232] 1 H NMR (400 MHz, CDCl3) δ 7.29 (brs, 4H, Ar-H), 7.23 (d, J = 8.60 Hz, 2H, Ar-H a ), 6.84 (d, J = 8.76 Hz, 2H, Ar-H b ); 5.75 (s, 1H, OH), 4.48 (s, 2H, CH2), 1.48 (s, 3H, CH3×3); 13 C NMR (100 MHz, CDCl3) δ 167.96, 157.55, 142.35, 136.65, 133.14, 128.53, 127.96, 127.82, 114.69, 82.47, 75.07, 65.70, 28.05.

[0233] (4) Synthesis of intermediate M3

[0234] M2 (385 mg, 1.10 mmol) was dissolved in 5 ml of dry dichloromethane, stirred for 10 min under an ice bath condition, and then oxalyl chloride (280 mg, 2.21 mmol) and 3 drops of DMF were added, and the mixture was stirred at room temperature overnight. After the reaction was completed, the solvent and excess oxalyl chloride were removed under reduced pressure to obtain a residue, which was subjected to the next reaction without purification.

[0235] (5) Synthesis of intermediate M4

[0236] The M3 obtained from the above reaction was dissolved in acetonitrile, and then anhydrous piperazine was added thereto, and the mixture was refluxed at 90 °C overnight. After the reaction was completed, the reaction was quenched with cold water, extracted three times with ethyl acetate, the organic phases were combined, washed successively with saturated NaCl solution and dried over anhydrous Na2SO4. Column chromatography (D / M = 30 / 1) gave a light yellow solid M4 (2.3 g).

[0237] 1 H NMR (400 MHz, DMSO-d 6)δ 7.40 (d, J = 8.52 Hz, 2H, Ar-H), 7.32 (d, J = 8.48 Hz, 2H, Ar-H), 7.27 (d, J = 8.72 Hz, 2H, Ar-H), 6.81 (d, J = 8.72 Hz, 2H, Ar-H), 4.58 (s, 2H, CH2), 4.21 (s, 1H, NH), 2.68 (t, J = 4.76 Hz, 2H, CH2×2), 2.18 (brs, 4H, CH2×2); 1.40 (s, 3H, CH3×3); 13 13C NMR (100 MHz, DMSO-d 6 )δ 168.29, 157.07, 142.68, 135.24, 131.56, 129.78, 129.15, 128.84, 114.95, 81.78, 74.47, 65.44, 53.24, 46.18, 28.13.

[0238] (6) Synthesis of Intermediate M5

[0239] Dissolve SM3 (0.50 g, 2.90 mmol) in dry dichloromethane (20 mL), stir for 5 min under ice bath conditions, add oxalyl chloride (0.76 g, 5.80 mmol) and DMF (1 drop), and stir overnight at room temperature. Remove the solvent and excess oxalyl chloride by distillation under reduced pressure. Dissolve the residue in dry dichloromethane, stir for 10 min under ice bath conditions, and then dropwise add a dichloromethane (20 ml) solution of M4 (1.2 g, 2.90 mmol) and triethylamine (0.61 g, 5.8 mmol). Stir at room temperature for 5 h. After the reaction is completed, quench the reaction with cold water, extract three times with dichloromethane, combine the organic phases, wash successively with saturated NaCl solution and dry over anhydrous Na2SO4. Column chromatography (P / E = 30 / 1 - 3 / 1) gives a white solid (1.42 g, 88%).

[0240] 1 1H NMR (400 MHz, CDCl3) δ 7.32 (d, J = 8.24 Hz, 2H, Ar-H), 7.23 - 7.26 (m, 4H, Ar-H), 6.80 (d, J = 8.20 Hz, 2H, Ar-H), 4.48 (s, 2H, CH2), 4.21 (s, 1H, CH), 3.81 (brs, 2H, CH2), 3.34 (t, J = 5.12 Hz, 2H, CH2), 2.84 (s, 3H, CH3), 2.50 (t, J = 5.12 Hz, 2H, CH2), 2.35 (brs, 2H, CH2), 1.46 (s, 3H, CH3×3); 1313C NMR (100 MHz, CDCl3) δ 171.71, 167.92, 157.37, 156.50, 153.05, 128.99, 128.88, 114.95, 82.41, 74.30, 65.73, 53.45, 51.56, 51.00, 46.99, 42.36, 28.04, 13.46.

[0241] (7) Synthesis of Intermediate M6

[0242] Dissolve M5 (0.5 g, 0.88 mmol) in dichloromethane (30 mL), add trifluoroacetic acid (1.00 g, 8.80 mmol), and stir at room temperature for 5 h. After completion of the reaction, remove the solvent and excess trifluoroacetic acid by distillation under reduced pressure, dissolve in dichloromethane, wash with saturated NaCl solution, and dry over anhydrous Na2SO4. Remove the solvent under reduced pressure to obtain a white solid (0.38 g, 84%). Without purification, directly proceed to the next step of the reaction.

[0243] (8) Synthesis of Intermediate C7-1

[0244] Dissolve 7-bromo-1-heptanol (4.00 g, 20.50 mmol) in acetonitrile (80 mL), sequentially add K2CO3 (5.70 g, 41.00 mmol), KI (1.02 g, 6.15 mmol), and 1-Boc-piperazine (2.22 g, 20.50 mmol), and reflux at 85 °C for 3 h. After completion of the reaction, quench the reaction with cold water, extract three times with ethyl acetate, combine the organic phases, wash with saturated NaCl solution, and dry over anhydrous Na2SO4. Separate by column chromatography (D / M = 60 / 1) to obtain a pale yellow solid (5.04 g, 82%).

[0245] 1 1H NMR (400 MHz, CDCl3) δ 3.65 (q, J = 11.96, 6.20 Hz, 2H, CH2), 3.50 (t, J = 5.32 Hz, 4H, CH2×2), 2.46 (brs, 4H, CH2×2), 2.40 (t, J = 7.52 Hz, 2H, CH2), 1.53 - 1.61 (m, 4H, CH2×2), 1.48 (s, 9H, CH3×3), 1.32 - 1.43 (m, 7H, OH&CH2×3); 13 13C NMR (100 MHz, CDCl3) δ 154.67, 79.74, 62.92, 58.65, 58.64, 52.93, 32.68, 29.23, 28.42, 27.39, 26.36, 25.64. HRMS (ESI + ): m / z calculated for C16 H 33 N2O3(M + H) + : 301.2491; found 301.2491.

[0246] (9) Synthesis of Intermediate C7-2

[0247] Dissolve C7-1 (6.02 g, 20.50 mmol) in dichloromethane (40 ml), add triethylamine (5.7 ml, 41.00 mmol), and stir for 10 min under an ice bath. Dropwise add a solution of p-toluenesulfonyl chloride (7.82 g, 41.00 mmol) in dichloromethane (20 ml) thereto, and react overnight at room temperature. After completion of the reaction, quench the reaction with water, extract three times with ethyl acetate, combine the organic phases, wash successively with saturated NaCl solution and dry over anhydrous Na2SO4. Column chromatography (D / M = 80 / 1) gives a light yellow transparent liquid (8.03 g, 86%).

[0248] 1 H NMR (400 MHz, CDCl3) δ 7.80 (d, J = 8.32 Hz, 2H, Ar-H×2), 7.36 (d, J = 8.00 Hz, 2H, Ar-H×2), 4.03 (t, J = 6.48 Hz, 2H, CH2), 3.48 (br, 4H, CH2×2), 2.47 (s, 3H, CH3), 2.35 - 2.43 (m, 6H, CH2×2, CH2), 1.54 - 1.69 (m, 2H, CH2), 1.45 - 1.54 (m, 11H, CH3×3, CH2), 1.27 - 1.36 (m, 6H, CH2×3); 13 C NMR (100 MHz, CDCl3) δ 154.67, 144.63, 133.22, 129.79, 127.86, 79.72, 70.57, 58.55, 58.55, 52.93, 28.78, 28.76, 28.42, 27.19, 36.36, 25.27, 21.64. HRMS (ESI + ): m / z calculated for C 23 H 39 N2O5S(M + H) + : 455.2580; found 455.2576.

[0249] (10) Synthesis of Intermediate C7-3

[0250] In the future, lenalidomide (570 mg, 2.20 mmol) was dissolved in acetonitrile (40 ml), and then C7-2 (1000 mg, 0.98 mmol), K2CO3 (365 mg, 2.64 mmol), and KI (109 mg, 0.66 mmol) were added successively. The mixture was refluxed at 90 °C for 2 h. After the reaction was completed, the reaction was quenched by adding water, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed successively with saturated NaCl solution and dried over anhydrous Na2SO4. Column chromatography (D / M = 60 / 1) gave white solid C7-3 (515 mg, 42%).

[0251] 1 H NMR (400 MHz, CDCl3) δ 7.29 - 7.36 (m, 2H, Ar-H×2), 6.87 (dd, J = 7.40, 1.28 Hz, 1H, Ar-H), 5.20 (dd, J = 13.44, 5.16 Hz, 1H, CH), 4.26 (q, J = 52.32, 15.40 Hz, 2H, CH2), 3.73 - 3.83 (m, 4H, CH2×2), 3.48 (t, J = 4.52 Hz, 4H, CH2×2), 2.96 - 3.02 (m, 1H, CH 2-a ), 2.81 - 2.91 (m, 1H, CH 2-b ), 2.35 - 2.45 (m, 6H, CH2×2, CH2), 2.25 - 2.32 (m, 1H, CH 2-b ), 2.15 - 2.21 (m, 1H, CH 2-a ), 1.51 - 1.54 (m, 2H, CH2), 1.47 (s, 9H, CH3×3), 1.29 - 1.35 (m, 6H, CH2×3); 13 C NMR (100 MHz, CDCl3) δ 170.98, 169.91, 169.81, 154.64, 141.17, 132.40, 129.49, 126.28, 118.09, 114.47, 79.74, 58.73, 58.57, 52.90, 52.49, 45.02, 40.56, 32.23, 29.47, 28.93, 28.41, 27.82, 27.23, 26.74, 22.90. HRMS (ESI + ): m / z calculated for C 29 H 44 N5O5 (M + H) + : 542.3342; found 542.3343.

[0252] (11) Synthesis of Intermediate C7-4

[0253] Dissolve C7-3 (515 mg, 0.95 mmol) in dichloromethane (10 ml), add trifluoroacetic acid (1083 mg, 9.50 mmol), and stir at room temperature for 2 h. After the reaction is completed, remove the solvent and part of the remaining trifluoroacetic acid under reduced pressure to obtain 370 mg of a yellow oil, which is used for the next reaction without separation.

[0254] (12) Synthesis of GPX4 protein degrader

[0255] Dissolve M6 (541 mg, 1.05 mmol) in DMF (10 ml), and successively add HATU (479 mg, 1.26 mmol), DIPEA (271 mg, 2.10 mmol), and compound C7-4 (465 mg, 1.05 mmol), and react at room temperature for 2 h. After the reaction is completed, add cold water to the system to quench the reaction solution, extract it three times with ethyl acetate, combine the organic phases, wash them successively with saturated NaCl solution and dry over anhydrous Na2SO4. Column separation (D / M = 50 / 1 - 30 / 1) gives a yellow solid (120 mg, 15%).

[0256] 1 H NMR (400 MHz, CDCl3) δ 7.29 - 7.31 (m, 2H, Ar-H), 7.20 - 7.26 (m, 6H, Ar-H), 6.79 - 6.82 (m, 3H, Ar-H), 5.11 (dd, J = 5.23, 13.13 Hz, 1H, CH), 4.59 (s, 2H, CH2), 4.11 - 4.25 (m, 3H, CH2&CH), 3.83 (s, 2H, CH2), 3.76 - 3.79 (m, 2H, CH2), 3.68 - 3.73 (m, 2H, CH2), 3.56 - 3.78 (m, 2H, CH2), 3.48 - 3.50 (m, 2H, CH2), 3.29 - 3.32 (m, 2H, CH2), 2.87 - 2.94 (m, 1H, CH 2a ), 2.74 - 3.83 (m, 1H, CH 2b ), 2.79 (s, 3H, CH3), 2.46 (t, J = 5.13 Hz, 2H, CH2), 2.31 - 2.38 (m, 4H, CH2×2), 2.25 - 2.28 (m, 2H, CH2), 2.06 - 2.13 (m, 1H, CH 2a ), 1.38 - 1.50 (m, 4H, CH2×2), 1.23 - 1.26 (m, 7H, CH 2b &CH2×3); 1313C NMR (100 MHz, CDCl3) δ 171.78, 171.04, 169.94, 169.83, 166.13, 157.25, 156.51, 153.04, 141.43, 140.78, 134.39, 132.80, 132.36, 129.44, 128.95, 128.90, 128.85, 128.74, 126.25, 118.01, 114.96, 114.09, 74.20, 67.47, 58.33, 53.49, 53.21, 52.71, 52.52, 51.50, 50.93, 46.99, 45.11, 42.37, 41.96, 40.53, 32.18, 28.94, 27.77, 27.15, 26.76, 26.50, 22.78, 13.44. M.P. 133 - 134 °C. HRMS (ESI + ): m / z calculated for C 48 H 56 ClN9O9 (M + H) + : 938.3890; found 938.3988.

[0257] Synthesis of GPX4 protein degraders MC2 - MC8 in Examples 2 - 8

[0258] The differences between Examples 2 - 8 and Example 1 lie in the different m values of the starting materials when preparing intermediate C7 - 1.

[0259] Spectral data of GPX4 protein degrader MC2: 1 1H NMR (400 MHz, CDCl3) δ 7.24 - 7.34 (m, 8H, Ar - H), 6.83 - 6.87 (m, 3H, Ar - H), 6.24 (s, 1H, NH), 5.16 (dd, J = 5.09, 13.32 Hz, 1H, CH), 4.63 (s, 2H, CH2), 4.15 - 4.30 (m, 3H, CH2 & CH), 3.72 - 3.83 (m, 6H, CH2×3), 3.61 - 3.64 (m, 2H, CH2), 3.53 - 3.56 (m, 2H, CH2), 3.33 - 3.36 (m, 1H, CH 2a ), 2.92 - 2.99 (m, 1H, CH 2b ), 2.79 - 2.88 (m, 2H, CH2), 2.45 - 2.51 (m, 1H, CH 2a ), 2.26 - 2.51 (m, 12H, CH3 & CH2×4 & CH 2b ), 2.12 - 2.18 (m, 1H, CH2a ), 1.44 - 1.54 (m, 4H, CH2×2), 1.26 - 1.28 (m, 9H, CH 2b &CH2×4); 13 C NMR (100 MHz, CDCl3) δ 171.72, 171.02, 169.92, 169.88, 169.83, 166.18, 159.65, 158.61, 157.25, 157.18, 156.51, 153.06, 141.31, 140.82, 140.76, 134.50, 134.41, 132.85, 132.76, 132.39, 129.48, 129.04, 129.00, 128.94, 128.87, 128.79, 126.27, 118.07, 114.96, 114.88, 114.29, 102.71, 74.33, 74.24, 67.58, 58.40, 53.24, 52.69, 52.51, 52.14, 51.52, 51.35, 50.95, 47.01, 45.04, 42.59, 42.38, 41.92, 40.59, 32.20, 29.21, 28.99, 27.83, 27.23, 26.78, 26.51, 22.84, 13.45, 12.11. M.P. 131 - 132 °C. HRMS (ESI + ): m / z calculated for C 49 H 58 ClN9O9 (M + H) + : 952.4046; found 952.4146.

[0260] Spectral data of the GPX4 protein degrader MC3: 1 H NMR (400 MHz, CDCl3) δ 7.21 - 7.34 (m, 7H, Ar - H×6 & NH), 7.05 - 7.17 (m, 2H, Ar - H), 6.83 - 6.88 (m, 3H, Ar - H), 5.16 (dd, J = 5.15, 13.71 Hz, 1H, CH), 4.63 - 4.64 (m, 2H, CH2), 4.14 - 4.30 (m, 3H, CH2 & CH), 3.80 - 3.83 (m, 2H, CH2), 3.70 - 3.77 (m, 2H, CH2), 3.61 - 3.64 (m, 2H, CH2), 3.52 - 3.56 (m, 2H, CH2), 3.33 - 3.36 (m, 1H, CH 2a ), 2.92 - 2.98 (m, 1H, CH 2b), 2.82 - 2.87 (m, 2H, CH2), 2.49 (t, J = 4.96 Hz, 2H, CH2), 2.38 - 2.43 (m, 5H, CH3 & CH2), 2.28 - 2.37 (m, 4H, CH2×2), 2.11 - 2.17 (m, 1H, CH 2a ), 1.42 - 1.54 (m, 6H, CH2×3), 1.23 - 1.32 (m, 11H, CH 2b & CH2×5); 13 C NMR (100 MHz, CDCl3) δ 171.74, 171.02, 169.92, 169.84, 166.16, 157.25, 156.52, 153.05, 141.33, 140.76, 134.42, 132.84, 132.38, 129.54, 129.46, 129.23, 128.95, 128.92, 128.86, 128.77, 128.58, 128.42, 126.27, 118.05, 114.96, 114.63, 114.26, 88.78, 74.23, 67.65, 67.51, 58.45, 53.27, 52.71, 52.51, 51.52, 50.94, 47.00, 45.16, 45.04, 42.38, 41.97, 40.63, 32.19, 29.29, 29.05, 27.87, 27.27, 26.86, 26.57, 26.36, 25.88, 22.83, 13.44. M.P. 128 - 129 °C. HRMS (ESI + ): m / z calculated for C 50 H 60 ClN9O9 (M + H) + : 966.4203; found 966.4311.

[0261] Spectral data of the GPX4 protein degrader MC4: 1 H NMR (400 MHz, CDCl3) δ 7.22 - 7.32 (m, 8H, Ar - H×7 & NH), 7.04 - 7.16 (m, 1H, Ar - H), 6.82 - 6.84 (m, 3H, Ar - H), 5.15 (dd, J = 4.87, 13.39 Hz, 1H, CH), 4.61 (s, 2H, CH2), 4.13 - 4.28 (m, 3H, CH2 & CH), 3.77 - 3.81 (m, 3H, CH2 & CH 2a), 3.70 - 3.76 (m, 2H, CH2), 3.51 - 3.63 (m, 4H, CH2×2), 3.33 (t, J = 4.95 Hz, 2H, CH2), 2.91 - 2.97 (m, 1H, CH 2b ), 2.82 (s, 3H, CH3), 2.48 (t, J = 4.70 Hz, 2H, CH2), 2.38 - 2.43 (m, 4H, CH2×2), 2.30 - 2.35 (m, 4H, CH2×2), 2.10 - 2.16 (m, 1H, CH 2a ), 1.42 - 1.50 (m, 4H, CH2×2), 1.22 - 1.28 (m, 13H, CH 2b &CH2×6); 13 C NMR (100 MHz, CDCl3) δ 171.73, 171.02, 169.92, 169.83, 166.18, 157.24, 156.51, 153.06, 141.30, 140.76, 134.44, 132.86, 132.39, 129.48, 128.94, 128.92, 128.87, 128.77, 126.27, 118.06, 114.97, 114.29, 74.24, 67.52, 58.46, 53.22, 52.69, 52.51, 51.52, 50.95, 47.00, 45.02, 42.38, 41.90, 40.65, 32.20, 29.35, 29.32, 29.10, 27.89, 27.30, 26.88, 26.54, 22.85, 13.45. M.P. 118 - 119 °C. HRMS (ESI + ): m / z calculated for C 51 H 62 ClN9O9 (M + H) + : 980.4359; found 980.4466.

[0262] Spectral data of the GPX4 protein degrader MC5: 1 H NMR (400 MHz, CDCl3) δ 7.22 - 7.35 (m, 8H, Ar - H×7&NH), 7.05 - 7.18 (m, 1H, Ar - H), 6.83 - 6.86 (m, 3H, Ar - H), 5.17 (dd, J = 5.02, 8.11 Hz, 1H, CH), 4.63 (s, 2H, CH2), 4.11 - 4.30 (m, 3H, CH2&CH), 3.79 - 3.83 (m, 3H, CH2&CH 2a), 3.71 - 3.77 (m, 2H, CH2), 3.54 - 3.64 (m, 4H, CH2×2), 3.34 (t, J = 4.51 Hz, 2H, CH2), 2.92 - 2.99 (m, 1H, CH 2b ), 2.83 (s, 3H, CH3), 2.50 (t, J = 4.81 Hz, 2H, CH2), 2.42 (brs, 4H, CH2×2), 2.28 - 2.37 (m, 4H, CH2×2), 2.11 - 2.18 (m, 1H, CH 2a ), 1.43 - 1.53 (m, 4H, CH2×2), 1.24 - 1.29 (m, 15H, CH 2b & CH2×7); 13 C NMR (100 MHz, CDCl3) δ 171.72, 171.01, 169.91, 169.81, 166.14, 157.25, 156.58, 156.50, 153.06, 141.30, 140.76, 134.42, 132.85, 132.39, 129.46, 128.94, 128.91, 128.87, 128.77, 128.59, 126.27, 118.04, 114.96, 114.28, 74.24, 67.69, 67.54, 60.37, 58.48, 53.26, 52.70, 52.50, 51.52, 50.95, 47.00, 45.13, 45.01, 42.38, 41.94, 40.66, 32.20, 29.41, 29.14, 27.92, 27.35, 26.92, 26.58, 22.86, 13.45. M.P. 112 - 113 °C. HRMS (ESI + ): m / z calculated for C 52 H 64 ClN9O9 (M + H) + : 994.4516; found 994.4922.

[0263] Spectral data of the GPX4 protein degrader MC6: 11H NMR (400 MHz, CDCl3) δ 7.25 - 7.35 (m, 9H, Ar - H × 8 & NH), 7.05 - 7.18 (m, 1H, Ar - H), 6.85 - 6.88 (m, 3H, Ar - H), 5.19 (dd, J = 5.37, 13.50 Hz, 1H, CH), 4.64 (s, 2H, CH2), 4.14 - 4.33 (m, 3H, CH2 & CH), 3.82 - 3.84 (m, 2H, CH2), 3.76 - 3.78 (m, 3H, CH2 & CH 2a ), 3.58 - 3.65 (m, 4H, CH2 × 2), 3.36 (t, J = 5.01 Hz, 2H, CH2), 2.95 - 2.99 (m, 1H, CH 2b ), 2.86 (s, 3H, CH3), 2.51 (t, J = 5.11 Hz, 2H, CH2), 2.44 (brs, 4H, CH2 × 2), 2.34 - 2.37 (m, 4H, CH2 × 2), 2.14 - 2.20 (m, 1H, CH 2a ), 1.48 - 1.56 (m, 4H, CH2 × 2), 1.26 - 1.30 (m, 17H, CH 2b & CH2 × 8); 13 13C NMR (100 MHz, CDCl3) δ 171.70, 171.00, 169.90, 169.80, 166.15, 157.24, 156.50, 153.06, 141.26, 140.75, 134.45, 132.87, 132.41, 129.47, 128.94, 128.92, 128.87, 128.77, 126.27, 118.06, 114.95, 114.35, 74.25, 67.57, 58.46, 53.22, 52.66, 52.50, 51.52, 50.96, 47.00, 44.99, 42.38, 40.67, 32.21, 29.46, 29.42, 29.17, 27.93, 27.34, 26.93, 22.87, 13.45, -0.01. M.P. 122 - 123 °C. HRMS (ESI + ): m / z calculated for C 53 H 66 ClN9O9 (M + H) + : 1008.4672; found 1008.4780.

[0264] Spectral data of the GPX4 protein degrader MC7: 11H NMR (400 MHz, CDCl3) δ 7.24 - 7.28 (m, 9H, Ar - H × 8 & NH), 6.33 - 6.81 (m, 1H, Ar - H), 6.85 - 6.88 (m, 3H, Ar - H), 5.17 - 5.21 (m, 1H, CH), 4.58 - 4.64 (m, 2H, CH2), 4.19 - 4.32 (m, 3H, CH2 & CH), 3.72 - 3.79 (m, 7H, CH2 × 3 & CH 2a ), 3.62 - 3.63 (m, 2H, CH2), 3.35 (brs, 2H, CH2), 2.95 - 3.00 (m, 1H, CH 2b ), 2.85 - 2.88 (m, 2H, CH2), 2.36 - 2.49 (m, 10H, CH3 & CH2 × 3 & CH 2a ), 2.12 - 2.22 (m, 2H, CH 2b ), 1.51 - 1.56 (m, 4H, CH2 × 2), 1.24 - 1.28 (m, 19H, )2.86 (s, 3H, CH3), 2.51 (t, J = 5.11 Hz, 2H, CH2), 2.44 (brs, 4H, CH2 × 2), 2.34 - 2.37 (m, 4H, CH2 × 2), 2.14 - 2.20 (m, 1H, CH 2a ), 1.48 - 1.56 (m, 4H, CH2 × 2), 1.26 - 1.30 (m, 19H, CH 2b & CH2 × 9); 13 13C NMR (100 MHz, CDCl3) δ 172.50, 172.09, 169.63, 167.52, 166.48, 158.60, 155.96, 148.87, 145.93, 139.98, 135.11, 134.71, 131.40, 129.51, 129.33, 129.06, 127.31, 126.25, 123.47, 117.07, 116.56, 115.80, 74.06, 66.01, 56.30, 55.80, 52.57, 51.92, 46.82, 45.85, 45.43, 44.08, 30.81, 29.43, 29.32, 29.23, 29.20, 29.17, 29.13, 29.09, 29.02, 27.56, 27.48, 27.25, 25.99, 13.78. M.P. 111 - 112 °C. HRMS (ESI + ): m / z calculated for C 54 H 68 ClN9O9 (M + H) +: 1022.4829; found 1022.5059.

[0265] Spectral data of GPX4 protein degrader MC8: 1 H NMR (400 MHz, CDCl3) δ 7.22 - 7.36 (m, 8H, Ar - H×7 & NH), 7.07 - 7.18 (m, 2H, Ar - H), 6.85 - 6.90 (m, 3H, Ar - H), 5.20 (dd, J=5.17, 13.27 Hz, 1H, CH), 4.64 - 4.66 (m, 2H, CH2), 4.20 - 4.33 (m, 3H, CH2 & CH 2a ), 3.73 - 3.85 (m, 5H, CH2×2 & CH 2b ), 3.60 - 3.69 (m, 6H, CH2×3), 3.55 - 3.37 (m, 1H, CH 2a ), 2.95 - 3.01 (m, 1H, CH 2b ), 2.86 (s, 3H, CH3), 2.47 - 2.53 (m, 5H, CH2×2 & CH 2a ), 2.34 - 2.44 (brs, 4H, CH2×2), 2.27 - 2.32 (m, 1H, CH 2b ), 2.14 - 2.21 (m, 1H, CH 2b ), 1.48 - 1.56 (m, 4H, CH2×2), 1.26 - 1.31 (m, 21H, CH 13 CNMR (100 MHz, CDCl3) δ 172.50, 172.09, 169.63, 167.52, 166.48, 158.60, 155.96, 148.87, 145.93, 139.98, 135.11, 134.71, 131.40, 129.51, 129.33, 129.06, 127.31, 126.25, 123.47, 117.07, 116.56, 115.80, 74.06, 66.01, 56.30, 55.80, 52.57, 51.92, 46.82, 45.85, 45.43, 44.08, 30.81, 29.43, 29.32, 29.23, 29.21, 29.20, 29.17, 29.13, 29.09, 29.02, 27.56, 27.48, 27.25, 25.99, 13.78. M.P. 102 - 103℃. HRMS (ESI + ): m / z calculated for C 55 H 70 ClN9O9 (M + H)+ : 1036.4985; found 1036.5074.

[0266] Example 9

[0267] Synthesis of GPX4 protein degrader MC9

[0268] (1) Synthesis of intermediate C9-9

[0269] Dissolve azidotrimethylsilane (0.92 g, 7.98 mmol) in 15 mL of dry DMF, add KF (0.47 g, 7.98 mmol), stir at room temperature for 30 min, then add C9-2 (1.28 g, 2.66 mmol), and react at 60 °C for 2 h. After the reaction is completed, quench the reaction with cold NaOH solution, extract with ethyl acetate three times, combine the organic phases, wash successively with saturated NaCl solution and dry over anhydrous Na2SO4. Concentrate under reduced pressure to obtain a pale yellow liquid C9-9 (1.14 g, 89%), and carry out the next reaction without separation.

[0270] (2) Synthesis of intermediate C9-10

[0271] Dissolve C9-9 (1.14 g, 2.36 mmol) in 15 mL of methanol, add 5% Pd / C (180 mg), and carry out a hydrogenation reaction for 2 h. After the reaction is completed, filter off the palladium carbon, and concentrate the filtrate to obtain 0.73 g of a gray solid, and carry out the next reaction without separation.

[0272] (3) Synthesis of intermediate C9-11

[0273] Dissolve fluorothalidomide (0.20 g, 0.72 mmol) in 5 mL of dry DMF, successively add C9-10 (0.84 g, 1.44 mmol) and DIPEA (0.19 g, 1.44 mmol), and react at 90 °C overnight. After the reaction is completed, quench the reaction with water, extract with ethyl acetate three times, combine the organic phases, wash successively with saturated NaCl solution and dry over anhydrous Na2SO4. Separate by column chromatography (D / M = 60 / 1) to obtain a yellow solid C9-11 (210 mg, 25%). 11H NMR (400 MHz, CDCl3) δ 8.70 (s, 1H, NH), 7.48 - 7.52 (m, 1H, Ar - H), 7.10 (d, J = 7.04 Hz, 1H, Ar - H), 6.89 (d, J = 8.52 Hz, 1H, Ar - H), 6.24 (t, J = 5.40 Hz, NH), 4.91 - 4.95 (m, 1H, CH), 3.49 (br, 4H, CH2×2), 3.27 (q, J = 12.64, 6.84 Hz, 2H, CH2), 2.70 - 2.92 (m, 3H, CH 2-a ×2 & CH 2-b ), 2.35 - 2.51 (m, 6H, CH2×2 & CH2), 2.11 - 2.17 (m, 1H, CH 2-b ), 1.63 - 1.71 (m, 2H, CH2), 1.50 - 1.58 (m, 2H, CH2), 1.47 (s, 9H, CH3×3), 1.30 - 1.45 (m, 10H, CH2×5); 13 13C NMR (100 MHz, CDCl3) δ 171.13, 169.50, 168.46, 167.64, 154.69, 147.03, 136.07, 132.49, 116.62, 111.32, 109.83, 79.73, 58.63, 52.85, 52.85, 48.89, 42.61, 31.48, 29.47, 29.44, 29.36, 29.17, 28.42, 27.46, 26.81, 26.45, 22.84. HRMS (ESI+): m / z calculated for C31H46N5O6 (M + H)+: 583.3370; found 583.3752.

[0274] (4) Synthesis of GPX4 protein degrader

[0275] Dissolve C9 - 11 (0.10 g, 0.16 mmol) in 10 mL of DCM, add 1 mL of TFA, and stir at room temperature for 30 min. After the reaction is completed, remove the solvent under reduced pressure to obtain a light yellow liquid mixture of C9 - 11, which is used for the next reaction without separation. Dissolve M6 (82 mg, 0.16 mmol) in DMF (10 ml), and successively add HATU (91 mg, 0.24 mmol), DIPEA (70 mg, 0.48 mmol) and the crude product from the previous step, and react at room temperature for 2 h. After the reaction is completed, add cold water to the system to quench the reaction solution, extract it three times with ethyl acetate, combine the organic phases, wash them successively with saturated NaCl solution and dry over anhydrous Na2SO4. Column separation (D / M = 50 / 1 - 30 / 1) gives a yellow solid (47 mg, 30%).1 1H NMR (400 MHz, CDCl3) δ 9.02 (s, 1H, NH), 7.47 (dd, J = 7.39, 8.34 Hz, 1H, Ar-H), 7.33 - 7.30 (m, 2H, Ar-H×2), 7.27 - 7.22 (m, 4H, Ar-H×4), 7.07 (d, J = 7.13 Hz, 1H, Ar-H×1), 6.87 - 6.82 (m, 3H, Ar-H×3), 6.23 (t, J = 5.52 Hz, 1H, NH), 5.29 (s, 1H, CH), 4.62 (s, 2H, CH2), 4.21 (s, 1H, CH), 3.82 - 2.80 (m, 2H, CH2), 3.60 (d, J = 24.50 Hz, 4H, CH2×2), 3.35 - 3.32 (m, 2H, CH2), 2.88 - 2.66 (s, 5H, CH3&CH2), 2.51 - 2.44 (m, 6H, CH2×3), 2.36 - 2.31 (m, 2H, CH2), 2.25 (q, J = 6.57 Hz, 2H, CH2), 2.14 - 2.08 (m, 1H, CH 2a ) 1.42 - 1.25 (m, 17H, CH2×8&CH 2b ); 13 13C NMR (100 MHz, CDCl3) δ 171.65, 171.30, 169.50, 168.64, 167.58, 166.13, 157.14, 156.44, 153.00, 146.97, 140.71, 136.04, 134.41, 132.80, 132.44, 128.87, 128.82, 128.72, 116.58, 114.88, 111.28, 109.81, 74.20, 67.52, 58.33, 55.40, 53.40, 52.50, 51.48, 50.91, 48.80, 48.95, 42.43, 42.31, 31.37, 29.64, 29.17, 29.08, 28.92, 28.90, 28.60, 28.49, 27.31, 27.16, 26.54, 25.24, 22.81, 22.64, 17.07, 14.08, 13.43. HRMS (ESI + ): m / z calculated for C 50 H 58 ClN9O 10 S (M + H)+: 980.3995; found 980.4091.

[0276] Example 10

[0277] Synthesis of GPX4 Protein Degrader MC10

[0278] (1) Synthesis of Intermediate C9-12

[0279] Dissolve fluorinated thalidomide (200 mg, 0.72 mmol) in anhydrous DMF (5 mL), and successively add compound C9-2 (347 mg, 0.72 mmol) and DIPEA (186 mg, 1.44 mmol). React at 90 °C for 1 h. After the reaction is completed, cool to room temperature, add cold water to the system to quench the reaction, extract three times with ethyl acetate, combine the organic phases, wash successively with saturated NaCl solution and dry over anhydrous Na2SO4. Column chromatography (P / E = 10 / 1 - 6 / 1) gives a white solid (163 mg, 30%).

[0280] (2) Synthesis of GPX4 Protein Degrader

[0281] Dissolve C9-12 (0.12 g, 0.16 mmol) in 10 mL of DCM, add 1 mL of TFA, and stir at room temperature for 30 min. After the reaction is completed, remove the solvent under reduced pressure to obtain a light yellow liquid mixture C9-11, which is used in the next step without separation. Dissolve M6 (82 mg, 0.16 mmol) in DMF (10 ml), and successively add HATU (91 mg, 0.24 mmol), DIPEA (70 mg, 0.48 mmol) and the crude product from the previous step, and react at room temperature for 2 h. After the reaction is completed, add cold water to the system to quench the reaction solution, extract three times with ethyl acetate, combine the organic phases, wash successively with saturated NaCl solution and dry over anhydrous Na2SO4. Column chromatography (D / M = 50 / 1 - 30 / 1) gives a pale yellow solid (36 mg, 30%). 11H NMR (400 MHz, CDCl3) δ 8.64 (s, 1H, Ar-H), 7.81 (d, J = 7.73 Hz, 1H, NH), 7.38 - 7.33 (m, 4H, Ar-H×4), 7.31 - 7.28 (m, 2H, Ar-H×2), 7.25 - 7.20 (m, 4H, Ar-H×4), 6.84 - 6.80 (m, 2H, Ar-H×2), 5.05 - 4.98 (m, 1H, CH), 4.80 (dd, J = 5.31, 8.31 Hz, 1H, CH), 4.50 (s, 2H, CH2), 4.57 - 4.53 (m, 1H, CH), 4.19 (s, 1H, CH), 3.80 - 3.77 (m, 2H, CH2), 3.51 (t, J = 4.98 Hz, 2H, CH2), 3.42 (s, 10H, CH2×5), 3.32 (t, J = 5.01 Hz, 2H, CH2), 3.09 (s, 1H, NH), 2.81 (s, 3H, CH3), 2.62 - 2.56 (m, 1H, CH 2a ), 2.49 - 2.44 (m, 6H, CH3 & CH2 & CH 2a ), 2.39 - 2.32 (m, 7H, CH2×3 & CH 2b ), 1.93 - 1.86 (m, 1H, CH 2b ), 2.27 (t, J = 7.62 Hz, 2H, CH2), 1.44 - 1.41 (m, 7H, CH2×2 & CH2), 1.23 (s, 10H, CH2×5), 0.98 (s, 9H, CH3×3); 13 13C NMR (100 MHz, CDCl3) δ 175.90, 171.66, 169.71, 166.16, 157.09, 156.47, 153.91, 150.31, 148.26, 143.17, 140.62, 134.35, 132.75, 131.59, 130.64, 129.45, 128.82, 128.77, 128.65, 126.34, 114.84, 74.14, 69.75, 67.39, 58.41, 58.15, 55.54, 53.18, 52.95, 52.58, 51.41, 50.83, 50.43, 48.86, 48.81, 46.91, 45.04, 43.17, 42.29, 41.86, 35.13, 35.06, 30.13, 29.29, 29.27, 29.25, 27.23, 27.08, 26.68, 26.48, 22.19, 15.90, 13.35. HRMS (ESI +): m / z calculated for C 60 H 79 ClN 10 O9S(M+H)+: 1151.5441; found 1151.5447.

[0282] Synthesis of GPX4 protein degrader DC1 in Example 11

[0283] (1) Synthesis of intermediate T6-2

[0284] Dissolve the compound (114 mg, 0.30 mmol) with the structure shown in T5-2 purchased in DMSO (10 mL), add thalidomide fluoride (83 mg, 0.30 mmol) and TEA (63 μL, 0.45) in sequence, and react at 130 °C for 0.5 h. After the reaction is completed, add cold water to the system to quench the reaction, extract three times with ethyl acetate, combine the organic phases, wash with saturated NaCl solution and dry with anhydrous Na2SO4 in sequence. Separate by column chromatography (D / M = 50 / 1 - 20 / 1) to obtain a bright yellow solid (77 mg, 44%).

[0285] 1 H NMR (400 MHz, CDCl3) δ 8.44 (s, 1H, NH), 7.48 (t, J = 7.80 Hz, 1H, NH), 7.08 (d, J = 7.12 Hz, 1H, Ar-H), 6.87 (d, J = 8.52 Hz, 1H, Ar-H), 6.64 (t, J = 5.03 Hz, 1H, Ar-H), 4.90 - 4.94 (m, 1H, CH), 4.07 (brs, 2H, CH2), 3.33 (q, J = 6.38, 11.02 Hz, 2H, CH2), 2.48 - 2.90 (m, 16H, CH2×8), 2.19 (d, J = 6.32 Hz, 2H, CH2), 2.08 - 2.13 (m, 1H, CH), 1.17 (d, J = 13.77 Hz, 2H, CH2), 1.44 (s, 9H, CH3×3), 1.02 - 1.11 (m, 2H, CH2); 13 C NMR (100 MHz, CDCl3) δ 171.13, 171.11, 169.10, 168.41, 167.67, 154.89, 146.67, 136.03, 132.54, 116.78, 111.38, 110.16, 79.20, 64.38, 56.22, 53.51, 52.90, 48.82, 39.37, 33.53, 31.43, 30.72, 28.44, 22.74.

[0286] (2) Synthesis of intermediate T7-2

[0287] Dissolve compound T6-2 (72 mg, 0.13 mmol) in DCM, add an excess of TFA (0.5 mL), and stir at room temperature for 2 h. After completion of the reaction, remove the solvent and excess TFA under reduced pressure to obtain a yellow oil (72 mg), which is directly used in the next reaction without purification.

[0288] (3) Synthesis of Degradant DC1

[0289] Dissolve compound M6 (67 mg, 0.13 mmol) in DMF (10 ml), and successively add HATU (59 mg, 0.15 mmol), DIPEA (25 mg, 0.19 mmol), and compound T7-2 (63 mg, 0.13 mmol), and react at room temperature for 2 h. After completion of the reaction, add water to the system to quench the reaction solution, extract it three times with ethyl acetate, combine the organic phases, wash successively with saturated NaCl solution and dry over anhydrous Na2SO4. Column chromatography (D / M = 50 / 1 - 30 / 1) gives a yellow solid (25 mg, 19%).

[0290] 1 H NMR (400 MHz, CDCl3) δ 8.63 (s, 1H, NH), 7.47 (t, J = 8.30 Hz, 1H, Ar-H), 7.32 (d, J = 7.75 Hz, 2H, Ar-H), 7.20 - 7.26 (m, 4H, Ar-H), 7.06 (d, J = 7.75 Hz, 2H, Ar-H), 6.84 - 6.87 (m, 3H, Ar-H), 6.61 - 6.64 (m, 1H, NH), 4.89 - 4.93 (m, 1H, CH), 4.62 (s, 2H, CH2), 4.21 (s, 1H, CH), 3.80 - 3.82 (m, 2H, CH2), 2.98 - 3.04 (m, 1H, CH 2a ), 2.78 - 2.88 (m, 4H, CH 2b &CH3), 2.64 - 2.67 (m, 4H, CH2×2), 2.48 - 2.53 (m, 9H, CH2×4&CH 2a ), 2.34 - 2.36 (brs, 3H, CH 2b &CH2), 2.16 - 2.18 (m, 2H, CH2), 2.08 - 2.12 (m, 1H, CH 2a ), 1.68 - 1.82 (m, 4H, CH2×2), 1.42 (s, 2H, CH2), 1.23 - 1.27 (m, 2H, CH2), 1.07 - 1.13 (m, 2H, CH2), 0.82 - 0.93 (m, 1H, CH); 1313C NMR (100 MHz, CDCl3) δ 171.69, 171.26, 169.13, 168.53, 167.67, 166.10, 157.36, 156.50, 153.04, 146.66, 140.79, 136.06, 134.29, 132.82, 132.54, 128.93, 128.87, 128.85, 128.77, 116.80, 114.95, 111.40, 110.18, 74.26, 67.62, 64.03, 56.21, 53.45, 52.83, 51.53, 50.95, 48.85, 47.01, 45.44, 42.39, 39.36, 33.49, 31.46, 31.30, 30.39, 29.69, 28.43, 28.40, 22.77, 13.47. M.P. 132 - 133 °C. HRMS (ESI + ): m / z calculated for C 49 H 55 ClN 10 O 10 (M + H) + : 979.3791; found 979.3864.

[0291] Example 12 Synthesis of GPX4 protein degrader DC2

[0292] (1) Synthesis of intermediate T1-3

[0293] Phthalimide (3000 mg, 20.39 mmol), anhydrous K2CO3 (8454 mg, 61.17 mmol), and tetrabutylammonium bromide (197 mg, 0.61 mmol) were successively added to DMF (40 mL) and mixed well. 1,3-Dibromopropane (4940 mg, 24.47 mmol) was added, and the reaction was carried out at room temperature for 3 h. After the reaction was completed, water was added to the system to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed successively with saturated NaCl solution and dried over anhydrous Na2SO4. Column chromatography separation (P / E = 10 / 1 - 3 / 1) gave a white solid (3827 mg, 70%).

[0294] 1 1H NMR (400 MHz, CDCl3) δ 7.76 - 7.80 (m, 2H, Ar-H), 7.65 - 7.69 (m, 2H, Ar-H), 3.77 (t, J = 6.96 Hz, 2H, CH2), 3.36 (t, J = 6.72 Hz, 2H, CH2), 2.20 (quint, J = 6.80 Hz, 6.76 Hz, 2H, CH2); 1313C NMR (100 MHz, CDCl3) δ 168.15, 134.03, 131.94, 123.26, 36.68, 31.62, 29.93.

[0295] (2) Synthesis of Intermediate T2-3

[0296] T1-3 (1000 mg, 3.73 mmol), 1-Boc piperazine (834 mg, 4.48 mmol) and K2CO3 (1289 mg, 9.32 mmol) were successively added to acetonitrile (30 mL), mixed well, and triethylamine (16 μL, 0.11 mmol) was added dropwise. The mixture was refluxed at 85 °C for 12 h. After the reaction was completed, water was added to the system to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed successively with saturated NaCl solution and dried over anhydrous Na2SO4. Column chromatography separation (P / E = 10 / 1 - 3 / 1) gave a white solid (1300 mg, 93%).

[0297] 1 1H NMR (400 MHz, CDCl3) δ 7.79 - 7.83 (m, 2H, Ar-H), 7.67 - 7.71 (m, 2H, Ar-H), 3.75 (t, J = 6.60 Hz, CH2), 3.23 (t, J = 5.20 Hz, CH2), 2.39 (t, J = 7.00 Hz, CH2), 2.27 - 2.29 (m, 4H, CH2×2), 1.84 (quint, J = 6.80, 6.76 Hz, 2H, CH2), 1.41 (s, 9H, CH3×3); 13 13C NMR (100 MHz, CDCl3) δ 177.57, 157.34, 143.56, 131.41, 118.45, 83.95, 56.07, 51.66, 34.53, 29.89, 22.14.

[0298] (3) Synthesis of Intermediate T3-3

[0299] Compound T2-3 (3.60 g, 9.64 mmol) was dissolved in DCM (30 mL), and excess TFA (3 mL) was added. The reaction was carried out at room temperature for 2 h. After the reaction was completed, the solvent and the remaining TFA were removed under reduced pressure. Water was added thereto and extracted with EA. The organic phase was discarded. 1N NaOH solution was added to the aqueous phase to adjust the pH to alkaline, and the mixture was extracted with EA three times. The organic phases were collected and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure to obtain a pale yellow solid (2.64 g, 99%), which was directly used in the next step without purification.

[0300] (4) Synthesis of Intermediate T4-3

[0301] Compound T3-3 (2.64 g, 9.64 mmol) was dissolved in acetonitrile (30 mL). K2CO3 (3.33 g, 24.1 mmol), 1-Boc-4-bromomethylpiperidine (2.07 g, 10.60 mmol) and KI (0.48 g, 2.89 mmol) were added successively, and the mixture was refluxed at 85 °C overnight. After the reaction was completed, cold water was added to the system to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed successively with saturated NaCl solution and dried over anhydrous Na2SO4. Column chromatography separation (P / E = 10 / 1 - 3 / 1) gave a pale yellow solid (1.90 g, 42%).

[0302] 1 H NMR (400 MHz, CDCl3) δ 7.81 - 7.85 (m, 2H, Ar-H), 7.69 - 7.73 (m, 2H, Ar-H), 4.06 (brs, 2H, CH2), 3.75 (t, J = 6.92 Hz, CH2), 2.65 (t, J = 13.04 Hz, CH2), 2.26 - 2.42 (m, 8H, CH2×4), 2.04 (d, J = 7.08 Hz, 2H, CH2), 1.82 - 1.89 (m, 2H, CH2), 1.67 (d, J = 13.16 Hz, 2H, CH2), 1.54 - 1.59 (m, 1H, CH), 1.43 - 1.46 (m, 11H, CH3×3&CH2), 0.97 - 1.07 (m, 2H, CH2); 13 C NMR (100 MHz, CDCl3) δ 168.46, 154.89, 133.78, 132.33, 123.13, 80.13, 64.49, 56.05, 53.48, 53.11, 35.90, 33.90, 31.39, 27.85, 24.87.

[0303] (5) Synthesis of intermediate T5-3

[0304] Compound T4-3 (750 mg, 1.59 mmol) was dissolved in ethanol, and 80% hydrazine hydrate (161 mg, 3.17 mmol) was added. The mixture was refluxed at 80 °C for 5 h. After the reaction was completed, the solvent was removed under reduced pressure to obtain a yellowish-white solid (250 mg, 46%), which was directly used in the next step without purification.

[0305] (6) Synthesis of intermediate T6-3

[0306] The operation and treatment were the same as those for compound T6-2, and a bright yellow solid (210 mg, 48%) was obtained.

[0307] 11H NMR (400 MHz, CDCl3) δ 8.75 (s, 1H, NH), 7.46 (t, J = 8.08 Hz, 1H, NH), 7.06 (d, J = 7.04 Hz, 1H, Ar-H), 6.91 (d, J = 8.53 Hz, 1H, Ar-H), 6.41 (t, J = 5.82 Hz, 1H, Ar-H), 4.88 - 4.92 (m, 1H, CH), 4.06 (brs, 2H, CH2), 3.32 (q, J = 6.09, 12.70 Hz, 2H, CH2), 2.46 - 2.88 (m, 16H, CH2×8), 2.17 (d, J = 6.76 Hz, 2H, CH2), 1.78 - 1.85 (m, 2H, CH2), 1.70 (d, J = 12.75 Hz, 2H, CH2), 1.59 - 1.65 (m, 1H, CH), 1.44 (s, 9H, CH3×3), 1.00 - 1.10 (m, 2H, CH2); 13 13C NMR (100 MHz, CDCl3) δ 171.27, 169.35, 168.56, 167.64, 154.91, 146.97, 136.03, 132.54, 116.72, 111.37, 109.94, 79.21, 64.47, 55.75, 53.39, 53.18, 48.86, 41.04, 33.53, 31.45, 30.76, 28.47, 26.29, 22.82.

[0308] (7) Synthesis of Intermediate T7-3

[0309] The operation and treatment were the same as those for Compound T7-2, to obtain a yellow oil (158 mg), which was directly subjected to the next reaction without purification.

[0310] (8) Synthesis of GPX4 Protein Degrader DC2

[0311] The operation and treatment were the same as those for Compound DC1, to obtain a yellow solid (92 mg, 29%).

[0312] 11H NMR (400 MHz, CDCl3) δ 8.40 (s, 1H, NH), 7.47 (t, J = 7.90 Hz, 1H, Ar - H), 7.32 (d, J = 8.46 Hz, 2H, Ar - H), 7.23 - 7.25 (m, 4H, Ar - H), 7.07 (d, J = 7.00 Hz, 2H, Ar - H), 6.92 (d, J = 7.31 Hz, 1H, Ar - H), 6.92 (d, J = 8.80 Hz, 2H, Ar - H), 6.61 - 6.64 (m, 1H, NH), 4.88 - 4.92 (m, 1H, CH), 4.62 (s, 2H, CH2), 4.21 (s, 1H, CH), 3.80 - 3.83 (m, 2H, CH2), 2.98 - 3.04 (m, 1H, CH 2a ), 2.80 - 2.89 (m, 4H, CH 2b & CH3), 2.70 - 2.77 (m, 2H, CH2), 2.50 - 2.64 (m, 2H, CH2), 2.44 - 2.51 (m, 11H, CH2×5 & CH 2a ), 2.34 - 2.36 (brs, 3H, CH 2b & CH2), 2.09 - 2.23 (m, 4H, CH2×2), 1.76 - 1.86 (m, 6H, CH2×3), 1.42 (s, 2H, CH2), 1.03 - 1.13 (m, 2H, CH2), 0.86 - 0.89 (m, 1H, CH); 13 13C NMR (100 MHz, CDCl3) δ 171.67, 171.05, 169.34, 168.42, 167.61, 166.08, 157.41, 156.50, 153.08, 146.96, 140.81, 136.07, 134.30, 132.86, 132.54, 128.95, 128.88, 116.74, 114.98, 111.42, 109.95, 74.29, 67.71, 64.11, 55.83, 53.34, 53.14, 51.54, 50.97, 48.85, 47.01, 45.44, 42.38, 41.10, 33.49, 31.42, 31.31, 30.39, 26.19, 22.83, 13.47. M.P. 132 - 133 °C; HRMS (ESI + ): m / z calculated for C 50 H 57 ClN 10 O 10 (M + H) +: 993.3948; found 993.4053.

[0313] Synthesis of GPX4 protein degrading agents DC3 - DC5 in Examples 13 - 15

[0314] The differences between Examples 13 - 15 and Example 12 lie in the different values of the starting material n when preparing intermediate T1 - 3.

[0315] Spectral data of GPX4 protein degrading agent DC3: 1 H NMR(400MHz, CDCl3) δ7.86(s, 1H, NH), 7.58(t, J = 8.04Hz, 1H, Ar - H), 7.42 - 7.45(m, 1H, Ar - H), 7.27 - 7.37(d, J = 8.46Hz, 2H, Ar - H), 7.12(d, J = 8.72Hz, 1H, Ar - H), 7.03(d, J = 7.05Hz, 1H, Ar - H), 6.79 - 6.86(m, 2H, Ar - H), 6.58(s, 1H, NH), 5.03 - 5.07(m, 1H, CH), 4.73(s, 2H, CH2), 4.37(s, 1H, CH), 2.85 - 2.98(m, 3H, CH2&CH 2a ), 2.80 - 2.88(m, 1H, CH 2b ), 2.56 - 2.61(m, 2H, CH2), 2.49 - 2.51(m, 17H, CH2×8&CH 2a ), 2.38 - 2.42(m, 3H, CH 2b &CH2), 1.57 - 1.73(m, 9H, CH2×4&CH 2a ), 1.17 - 1.23(m, 2H, CH2), 0.86 - 1.04(m, 4H, CH 2b &CH2&CH); 13 C NMR(100MHz, DMSO - d 6)δ172.50, 172.08, 169.62, 168.47, 166.70, 166.53, 158.60, 155.96, 148.87, 144.74, 139.98, 135.11, 134.71, 132.37, 132.23, 129.51, 129.33, 129.06, 123.47, 120.58, 120.37, 115.80, 114.07, 74.06, 66.47, 63.73, 56.53, 53.27, 52.42, 51.92, 51.34, 45.85, 43.87, 32.95, 29.89, 28.94, 27.29, 24.96, 24.67, 13.78. M.P. 125 - 126℃. HRMS(ESI + ): m / z calculated for C 51 H 59 ClN 10 O 10 (M + H) + : 1007.4104; found 1007.4211.

[0316] Spectral data of the GPX4 protein degrader DC4: 1 H NMR(400 MHz, CDCl3) δ8.40(s, 1H, NH), 7.47(t, J = 7.90 Hz, 1H, Ar - H), 7.32(d, J = 8.15 Hz, 2H, Ar - H), 7.21 - 7.28(m, 4H, Ar - H), 7.11(d, J = 7.20 Hz, 2H, Ar - H), 6.82(d, J = 7.31 Hz, 1H, Ar - H), 6.77(d, J = 8.80 Hz, 2H, Ar - H), 6.62 - 6.69(m, 1H, NH), 4.63 - 4.92(m, 1H, CH), 4.62(s, 2H, CH2), 4.21(s, 1H, CH), 3.81 - 3.83(m, 2H, CH2), 2.98 - 3.04(m, 1H, CH 2a ), 2.80 - 2.89(m, 4H, CH 2b &CH3), 2.70 - 2.77(m, 2H, CH2), 2.50 - 2.64(m, 2H, CH2), 2.44 - 2.51(m, 15H, CH2×7&CH 2a ), 2.34 - 2.36(brs, 3H, CH 2b&CH2), 2.09 - 2.23 (m, 4H, CH2×2), 1.76 - 1.86 (m, 6H, CH2×3), 1.42 (s, 2H, CH2), 1.03 - 1.15 (m, 2H, CH2), 0.84 - 0.89 (m, 1H, CH); 13 C NMR (100 MHz, CDCl3) δ 172.58, 172.44, 169.63, 168.47, 166.70, 166.56, 158.60, 155.98, 148.87, 144.74, 139.97, 135.11, 134.71, 132.37, 132.29, 129.51, 129.33, 129.06, 123.77, 120.58, 120.37, 115.80, 114.07, 74.06, 66.47, 63.73, 55.82, 53.27, 52.42, 51.92, 51.34, 45.85, 44.05, 43.87, 32.95, 29.88, 29.04, 28.94, 26.68, 25.44, 24.67, 13.78. M.P. 135 - 136 °C. HRMS (ESI + ): m / z calculated for C 52 H 62 ClN 10 O 10 (M + H) + : 1021.4261; found 1021.4364.

[0317] Spectral data of the GPX4 protein degrader DC5: 11H NMR (400 MHz, CDCl3) δ 7.50 (t, J = 7.90 Hz, 1H, Ar-H), 7.35 (d, J = 8.51 Hz, 2H, Ar-H), 7.23 - 7.28 (m, 4H, Ar-H), 7.07 - 7.11 (m, 2H, Ar-H), 6.85 - 6.91 (m, 3H, Ar-H), 6.21 - 6.24 (m, 1H, NH), 4.90 - 4.94 (m, 1H, CH), 4.64 - 4.68 (m, 2H, CH2), 4.51 - 4.54 (m, 1H, CH), 3.82 - 3.91 (m, 2H, CH2), 3.25 - 3.30 (m, 2H, CH2), 2.86 (s, 3H, CH3), 2.64 - 2.78 (m, 4H, CH2×2), 2.51 - 2.67 (m, 8H, CH2×4), 2.12 - 2.25 (m, 2H, CH2), 1.72 - 1.85 (m, 4H, CH2×2), 1.63 - 1.70 (m, 6H, CH2×3), 1.39 - 1.49 (m, 6H, CH2×3), 1.27 (s, 2H, CH2), 1.04 - 1.12 (m, 2H, CH2), 0.88 - 0.91 (m, 1H, CH); 13 13C NMR (100 MHz, CDCl3) δ 171.75, 171.35, 169.62, 168.83, 167.56, 166.18, 157.36, 156.54, 153.06, 146.98, 140.82, 136.23, 134.40, 132.84, 132.47, 129.53, 129.22, 128.95, 128.87, 128.79, 128.43, 116.77, 115.03, 114.70, 111.47, 109.87, 74.25, 63.60, 52.71, 52.09, 51.53, 50.95, 48.84, 48.01, 47.00, 42.39, 31.35, 29.69, 28.73, 26.42, 25.90, 22.89, 18.08, 13.46. M.P. 131 - 132 °C. HRMS (ESI + ): m / z calculated for C 53 H 63 ClN 10 O 10 (M + H) + : 1035.4417; found 1035.4524.

[0318] Synthesis of GPX4 protein degrader TC1 in Example 14

[0319] (1) Synthesis of Intermediate D1

[0320] Dissolve 1 - boc piperazine (200 mg, 1.07 mmol) in CH3CN (15 mL), and successively add N - boc bromomethyl piperidine (300 mg, 1.07 mmol), K2CO3 (220 mg, 1.60 mmol), and reflux at 85 °C overnight. After the reaction is completed, add water to the system to quench the reaction, extract three times with ethyl acetate, combine the organic phases, wash successively with saturated NaCl solution and dry over anhydrous Na2SO4. Separate by column chromatography (P / E = 10 / 1 - 3 / 1) to obtain a white solid (224 mg, 56%).

[0321] 1 H NMR (400 MHz, CDCl3) δ 7.34 - 7.37 (m, 4H, Ar - H), 7.26 - 7.30 (m, 1H, Ar - H), 5.17 (s, 2H, CH2), 3.65 - 3.72 (m, 2H, CH2), 3.24 - 3.41 (m, 6H, CH2×3), 2.36 - 2.60 (m, 6H, CH2×3), 1.72 - 1.85 (m, 5H, CH2×2&CH), 1.42 (s, 9H, CH3×3); 13 C NMR (100 MHz, CDCl3) δ 13 155.74, 155.12, 136.65, 128.47, 128.41, 128.36, 79.41, 67.00, 63.73, 52.99, 44.01, 43.81, 32.96, 28.77, 28.39.

[0322] (2) Synthesis of Intermediate D2

[0323] Dissolve D3 (100 mg, 0.24 mmol) in MeOH (5 mL), add a catalytic amount of Pd / C (0.1 eq), place it in a hydrogenator, introduce high - purity hydrogen, and stir at room temperature for 3 h. After the reaction is completed, filter through diatomaceous earth, collect the filtrate, and remove the solvent by distillation under reduced pressure to obtain a gray - white solid residue, which is directly used in the next step without purification.

[0324] (3) Synthesis of Intermediate D3

[0325] Dissolve D2 (50 mg, 0.18 mmol) in DMSO (10 mL), and successively add the purchased thalidomide fluoride (52 mg, 0.18 mmol) and TEA (30 mg, 0.27 mmol), then reflux at 90 °C for 1 h. After the reaction is completed, add water thereto and extract with EA. Collect the organic phase, wash it three times with saturated sodium chloride solution and dry it with anhydrous Na2SO4. Remove the solvent under reduced pressure, and obtain a yellow solid (68 mg, 45%) by column chromatography (D / M = 50 / 1).

[0326] 1 H NMR (400 MHz, CDCl3) δ 8.44 (s, 1H, NH), 7.48 (t, J = 7.80 Hz, 1H, NH), 7.08 (d, J = 7.12 Hz, 1H, Ar-H), 6.87 (d, J = 8.52 Hz, 1H, Ar-H), 6.64 (t, J = 5.03 Hz, 1H, Ar-H), 4.90 - 4.94 (m, 1H, CH), 4.07 (brs, 2H, CH2), 3.33 (q, J = 6.38, 11.02 Hz, 2H, CH2), 2.48 - 2.94 (m, 12H, CH2×6), 2.19 (d, J = 6.32 Hz, 2H, CH2), 2.08 - 2.13 (m, 1H, CH), 1.17 (d, J = 13.77 Hz, 2H, CH2), 1.44 (s, 9H, CH3×3), 1.02 - 1.11 (m, 2H, CH2); 13 C NMR (100 MHz, CDCl3) δ 172.08, 169.61, 167.63, 166.37, 155.12, 143.07, 133.39, 133.24, 123.22, 122.08, 117.03, 79.41, 63.73, 53.12, 51.03, 50.39, 44.01, 32.96, 29.89, 28.77, 28.39, 24.67.

[0327] (4) Synthesis of intermediate D4

[0328] Dissolve D3 (100 mg, 0.18 mmol) in DCM, add an excess of TFA (0.5 mL), and stir at room temperature for 2 h. After the reaction is completed, remove the solvent and the excess TFA under reduced pressure to obtain a yellow oil (102 mg), which is directly used in the next step without purification.

[0329] (5) Synthesis of intermediate D5

[0330] Dissolve D4 (150 mg, 0.24 mmol) in CH3CN (10 mL), and successively add N-Boc-bromomethyl piperidine (66 mg, 0.24 mmol) and K2CO3 (50 mg, 0.36 mmol), then reflux at 85 °C overnight. After the reaction is completed, add cold water to quench the reaction, extract with ethyl acetate three times, combine the organic phases, wash successively with saturated NaCl solution and dry over anhydrous Na2SO4. Column separation (D / M = 30 / 1) gives a bright yellow solid (84 mg, 42%).

[0331] 1 H NMR (400 MHz, CDCl3) δ 7.85 - 7.87 (m, 1H, Ar-H), 7.52 (t, J = 6.72 Hz, 1H, Ar-H), 7.21 (dd, J = 1.05, 7.48 Hz, 1H, Ar-H), 5.49 (t, J = 3.60 Hz, 1H, CH), 3.65 - 3.72 (m, 2H, CH2), 3.20 - 0.330 (m, 6H, CH2×3), 2.48 - 2.71 (m, 14H, CH2×7), 2.16 - 2.22 (m, 1H, CH 2a ), 1.46 - 1.85 (m, 11H, CH×2&CH 2b &CH2×4), 1.42 (s, 9H, CH3×3); 13 C NMR (100 MHz, CDCl3) δ 172.08, 169.61, 167.63, 166.37, 155.12, 143.07, 133.39, 133.24, 123.22, 122.08, 117.03, 79.41, 63.88, 63.72, 53.53, 53.12, 51.03, 50.39, 44.01, 33.15, 33.00, 29.89, 29.27, 28.77, 28.39, 24.67.

[0332] (6) Synthesis of GPX4 protein degrader TC1

[0333] Dissolve D5 (80 mg, 0.13 mmol) in DCM (5 mL), add an excess of TFA (0.5 mL), and stir at room temperature for 2 h. After the reaction is complete, remove the solvent and excess TFA under reduced pressure to obtain a yellow oil, which is directly used in the next reaction without purification. Dissolve the above oil in DMF, and successively add M6 (67 mg, 0.13 mmol), HATU (74 mg, 0.19 mmol), and DIPEA (25 mg, 0.19 mmol), and stir at room temperature overnight. After the reaction is complete, add cold water to the system to quench the reaction solution, extract it three times with ethyl acetate, combine the organic phases, wash them successively with saturated NaCl solution and dry over anhydrous Na2SO4. Column separation (D / M = 50 / 1 - 30 / 1) gives a yellow solid (54 mg, 40%).

[0334] 1 H NMR (400 MHz, CDCl3) δ 7.86 (dd, J = 1.28, 7.12 Hz, 1H, Ar-H), 7.52 (t, J = 7.36 Hz, 1H, Ar-H), 7.39 - 7.41 (m, 2H, Ar-H), 7.30 - 7.33 (m, 2H, Ar-H), 7.21 (dd, J = 1.19, 7.37 Hz, 1H, Ar-H), 7.14 - 7.16 (m, 2H, Ar-H), 6.92 - 6.95 (m, 2H, Ar-H), 5.49 (t, J = 3.41 Hz, 1H, CH), 4.79 (s, 2H, CH2), 4.34 - 4.35 (m, 2H, CH2), 3.82 - 3.89 (m, 2H, CH2), 3.69 - 3.75 (m, 2H, CH2), 3.58 - 3.64 (m, 2H, CH2), 3.36 - 3.43 (m, 4H, CH2×2), 3.26 - 3.28 (m, 2H, CH2), 3.20 - 3.22 (m, 4H, CH2×2), 2.72 (s, 3H, CH3), 2.49 - 2.69 (m, 13H, CH&CH2×6), 2.16 - 2.22 (m, 1H, CH 2a ), 1.64 - 1.81 (m, 11H, C H 2b &CH2×5); 1313C NMR (100 MHz, CDCl3) δ 172.50, 172.08, 169.61, 168.47, 167.63, 166.37, 158.60, 155.96, 148.87, 143.07, 139.98, 135.11, 134.71, 133.39, 133.24, 129.51, 129.33, 129.06, 123.47, 123.22, 122.08, 117.03, 115.80, 74.06, 66.47, 63.90, 63.72, 53.53, 53.12, 51.92, 51.03, 50.39, 45.85, 43.87, 33.15, 33.02, 29.89, 29.27, 28.94, 24.67, 13.78.

[0335] Synthesis of GPX4 protein degrader TC2 in Example 15

[0336] Dissolve the compound of the structure shown in L4 (120 mg, 0.23 mmol) in DCM (5 mL), add an excess of TFA (0.5 mL), and stir at room temperature for 2 h. After the reaction is complete, remove the solvent and excess TFA under reduced pressure to obtain a yellow oil, which is directly subjected to the next reaction without purification. Dissolve the above oil in DMF, and successively add M6 (118 mg, 0.23 mmol), HATU (131 mg, 0.34 mmol), and DIPEA (45 mg, 0.34 mmol), and stir at room temperature overnight. After the reaction is complete, add cold water to the system to quench the reaction solution, extract it three times with ethyl acetate, combine the organic phases, wash them successively with saturated NaCl solution and dry them over anhydrous Na2SO4. Column separation (D / M = 50 / 1 - 30 / 1) gives a yellow solid (72 mg, 30%).

[0337] 11H NMR (400 MHz, CDCl3) δ 8.51 (d, J = 7.79 Hz, 1H, NH), 7.86 (dd, J = 1.28, 7.12 Hz, 1H, Ar-H), 7.52 (t, J = 7.36 Hz, 1H, Ar-H), 7.39 - 7.41 (m, 2H, Ar-H), 7.30 - 7.33 (m, 2H, Ar-H), 7.21 (dd, J = 1.19, 7.37 Hz, 1H, Ar-H), 7.14 - 7.16 (m, 2H, Ar-H), 6.92 - 6.95 (m, 2H, Ar-H), 5.49 (t, J = 3.41 Hz, 1H, CH), 4.77 (s, 2H, CH2), 4.34 - 4.36 (m, 2H, CH2), 3.82 - 3.89 (m, 2H, CH2), 3.69 - 3.75 (m, 2H, CH2), 3.58 - 3.64 (m, 2H, CH2), 3.36 - 3.45 (m, 4H, CH2×2), 3.26 - 3.28 (m, 2H, CH2), 3.20 - 3.22 (m, 4H, CH2×2), 2.72 (s, 3H, CH3), 2.49 - 2.69 (m, 13H, CH&CH2×6), 2.16 - 2.22 (m, 1H, CH 2a ), 1.64 - 1.88 (m, 11H, CH 2b &CH2×5); 13 13C NMR (100 MHz, CDCl3) δ 172.50, 172.08, 169.62, 168.47, 166.78, 166.53, 158.60, 155.96, 148.87, 144.29, 139.98, 135.11, 134.71, 132.60, 132.06, 129.51, 129.33, 129.06, 123.47, 121.61, 120.80, 115.80, 114.49, 74.06, 66.47, 63.90, 63.81, 53.53, 51.92, 51.34, 51.27, 50.74, 45.85, 43.87, 33.16, 33.02, 31.31, 29.89, 29.27, 28.94, 24.67, 13.78.

[0338] Test Example Bioactivity Evaluation

[0339] (1) Evaluation of Compound Degradation

[0340] By the WB method, the degradation effect of compounds at different concentrations on GPX4 protein in HT1080 cells after 6 h and 24 h of treatment was detected, as Figure 1 shown. Figure 1Among them, (a) is the degradation activity of the monocyclic GPX4 degrader; (b)-(d) show that MC3 degrades GPX4 in a concentration-dependent and time-dependent manner; (c) is the degradation activity of the bicyclic GPX4 degrader at a concentration of 1 μM; (e) is the degradation activity of the bicyclic GPX4 degrader at a concentration of 0.1 μM; (f) is the degradation activity of the tricyclic GPX4 degrader at a concentration of 1 μM.

[0341] As can be seen from Figure 1 It can be seen that among the monocyclic GPX4 degraders, MC3 has the best degradation effect and can degrade GPX4 in a concentration-dependent and time-dependent manner. The effect of the bicyclic degrader is generally better than that of the monocyclic and tricyclic degraders, and DC2 has the best activity.

[0342] (2) Relationship between the degradation activity of DC2 and concentration and time

[0343] The relationship between the degradation activity of DC2 and concentration and time is as shown in Figure 2 shown. Figure 2 Among them, (a) and (b) show the effects of different concentrations of DC2 on the GPX4 protein level in HT1080 cells after 24 h of treatment; (c) and (d) show the degradation effect of 0.1 μM DC2 on GPX4 in HT1080 cells at different treatment times.

[0344] After selecting DC2, the relationship between its degradation activity and concentration and time was studied. We set 6 dosing concentrations in a gradient, and observed the band changes after 24 h of dosing. The results are as shown in Figure 2 (a) and (c) in. The results show that the degradation effect of DC2 is concentration-dependent, and the DC in HT1080 cells 50 is 0.03 μM, and the degradation of GPX4 is more complete at a concentration of 0.3 μM.

[0345] The relationship between the degradation effect of DC2 and time is as shown in Figure 2 (b) and (d) in. DC2 (0.1 μM) at the same concentration was administered at 8 time points (0.5, 1, 2, 4, 6, 8, 12, 24 h), and then the band changes were observed. The results show that as the time prolongs, the level of GPX4 gradually decreases. When the dosing time is 12 h, DC2 can degrade more than 80% of GPX4. Based on the above experimental results, it can be confirmed that the degradation effect of DC2 has the characteristics of concentration dependence and time dependence.

[0346] (3) Evaluation of the inhibitory effect of DC2 on cell proliferation

[0347] Figure 3 The results of the evaluation of the inhibitory effect of DC2 on cell proliferation are shown in Figure 3(a) shows the inhibitory effects of DC2 and ML210 on the proliferation of HEK293T cells; (b) shows the inhibitory effects of DC2 and ML210 on the proliferation of HT1080 cells.

[0348] HEK293T (human embryonic kidney cells) was selected to evaluate the toxic effect of DC2 on normal cells, and ML210 was selected as a positive control. The inhibitory rates of DC2 and ML210 at different concentrations on the proliferation of HEK293T cells were determined by the MTT method, and the corresponding IC 50 values were calculated using GraphPad Prism 8, and the results are shown in Figure 3 (a). The results showed that the IC 50 value of DC2 was 1.07 μM, while the IC50 value of ML210 was 0.22 μM, indicating that the toxic effect of the degrader on HEK293T was less than that of the inhibitor.

[0349] Subsequently, the anti-proliferative effect of DC2 in tumor cells was evaluated. The cell line used was HT1080 (human fibrosarcoma cells). The inhibitory effects of DC2 and ML210 on the proliferation of HT1080 cells are shown in Figure 3 (b). It can be seen that the IC 50 value of DC2 was 0.3 μM, which was comparable to that of ML210 (0.1 μM).

[0350] (4) Pharmacokinetic study of DC2 in mice

[0351] To evaluate the pharmacokinetic properties of DC2 in vivo and explore the dosing method and dosage for subsequent pharmacodynamic experiments, the present invention selected ICR mice for pharmacokinetic experiments. The mice were randomly divided into two large groups according to the dosing method. One group was administered orally (the dosing dose was 10 mg / kg), and the other group was administered by tail vein injection (the dosing dose was 1 mg / kg). Then, blood was collected at 7 time points (5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h) and the concentration of DC2 was detected. The results are shown in Table 1.

[0352] Table 1 Pharmacokinetic parameters of DC2 in vivo with different dosing methods

[0353]

[0354] C max : Peak concentration; T max : Time to peak concentration; T 1 / 2 : Terminal elimination half-life; AUC: Area under the plasma concentration-time curve; Vz: Apparent volume of distribution; Cl: Clearance rate; MRT: Mean residence time; *: Data not detected;

[0355] Based on the above results, tail vein injection was selected as the administration method for the next pharmacodynamic study.

[0356] (5) Pharmacodynamic study of DC2 in mice with xenograft tumors

[0357] To explore the anti-tumor effect of DC2 at the animal level, the present invention constructed a mouse model of xenograft human fibrosarcoma (HT1080). DC2 was intravenously injected into the tail veins of tumor-bearing mice at a dose of 20 mg / kg. Tumor tissues were obtained at designated time points (3, 6, 12, 24, 48 h), and proteins were extracted for Western Blotting experiments. The relationship between the degradation effect of DC2 on GPX4 in xenograft human fibrosarcoma (HT1080) tissues and the administration time was as Figure 4 shown. The results showed that DC2 exhibited a degradation effect on GPX4 6 h after administration, reached the maximum degradation 12 h after administration, and the degradation activity could be maintained for at least 48 h.

[0358] To explore the safety of DC2, the present invention set three administration concentrations (5 mg / kg, 20 mg / kg, 40 mg / kg), another positive drug group (ML210, 3 mg / kg) and a blank control group (saline), and administered by tail vein injection once every two days, while measuring the body weight of mice. The changes in the body weight of mice after multiple administrations were as Figure 5 shown. It can be seen that although ML210 effectively inhibited tumor growth, it caused a significant decrease in the body weight of mice, while DC2 had almost no effect on the body weight, indicating that DC2 has better safety than ML210.

[0359] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A GPX4 protein degrader having the structure shown in Formula I: In Formula I, A1 is A2 is Among them, m = 1 - 20, n = 1 - 20.

2. The GPX4 protein degrader according to claim 1, wherein Having the structure shown in any one of Formula I-1 to I-8: In Formula I-1 to I-4, m = 1 - 20; In Formula I-5 to I-8, n = 1 - 20.

3. A method for preparing the GPX4 protein degrader according to claim 1 or 2, comprising the following steps: ①When A1 is , the preparation method of the GPX4 protein degrader comprises the following steps: A compound having the structure shown in Formula a reacts with a compound having the structure NH2-A2 in a substitution reaction to obtain a compound having the structure shown in Formula b; The compound having the structure shown in Formula b undergoes a deprotection reaction to obtain a compound having the structure shown in Formula c; The compound having the structure shown in Formula c reacts with a compound having the structure shown in Formula d in a condensation reaction to obtain the GPX4 protein degrader; ②When A1 is , the preparation method of the GPX4 protein degrader comprises the following steps: A compound having the structure shown in Formula e reacts with a compound having the structure F-A2 in a substitution reaction to obtain a compound having the structure shown in Formula f; The compound having the structure shown in Formula f undergoes a deprotection reaction to obtain a compound having the structure shown in Formula g; The compound having the structure shown in Formula g reacts with a compound having the structure shown in Formula d in a condensation reaction to obtain the GPX4 protein degrader.

4. The preparation method according to claim 3, characterized in that, The preparation method of the compound having the structure shown in Formula d comprises the following steps: 4-chloro-4'-hydroxybenzophenone reacts with tert-butyl bromoacetate in a substitution reaction to obtain a compound having the structure shown in Formula d-1; Under the action of a reducing agent, the compound having the structure shown in Formula d-1 undergoes a reduction reaction to obtain a compound having the structure shown in Formula d-2; The compound having the structure shown in Formula d-2 reacts with oxalyl chloride in a chlorination reaction to obtain a compound having the structure shown in Formula d-3; The compound having the structure shown in Formula d-3 reacts with piperazine in a substitution reaction to obtain a compound having the structure shown in Formula d-4; 5-methylisoxazole-3-carboxylic acid reacts with potassium nitrate in a nitration reaction to obtain a compound having the structure shown in Formula d-5; The compound having the structure shown in Formula d-4, the compound having the structure shown in Formula d-5 and oxalyl chloride undergo a coupling reaction to obtain a compound having the structure shown in Formula d-6; The compound having the structure shown in Formula d-6 undergoes a deprotection reaction to obtain a compound having the structure shown in Formula d.

5. The preparation method according to claim 3, characterized in that, When A2 is , a method for preparing a compound having an NH2-A2 structure, comprising the following steps: A compound having the structure shown in Formula v1 reacts with 4-methylthiazole in a substitution reaction to obtain a compound having the structure shown in Formula v2; The compound having the structure shown in Formula v2 undergoes a deprotection reaction to obtain a compound having the structure shown in Formula v3; The compound having the structure shown in Formula v3 reacts with a compound having the structure shown in Formula w1 in a condensation reaction to obtain a compound having the structure shown in Formula v4; The compound having the structure shown in Formula v4 undergoes a deprotection reaction to obtain a compound having the structure shown in Formula v5; The compound having the structure shown in Formula v5 reacts with a compound having the structure shown in Formula w2 in a condensation reaction to obtain a compound having the structure shown in Formula v6; The compound having the structure shown in Formula v6 undergoes a deprotection reaction to obtain a compound having the structure shown in Formula v7; 6. The preparation method according to claim 3, wherein The preparation method of the compound having the structure shown in Formula a comprises the following steps: The compound with the structure shown in Formula a-1 undergoes a substitution reaction with 1-Boc-piperazine to obtain a compound with the structure shown in Formula a-2; The compound with the structure shown in Formula a-2 undergoes a substitution reaction with p-toluenesulfonyl chloride to obtain a compound with the structure shown in Formula a.

7. The preparation method according to claim 3, characterized in that, The preparation method of the compound with the structure shown in Formula e includes the following steps: Phthalimide undergoes a substitution reaction with the compound with the structure shown in Formula e-1 to obtain a compound with the structure shown in Formula e-2; The compound with the structure shown in Formula e-2 undergoes a substitution reaction with 1-Boc piperazine to obtain a compound with the structure shown in Formula e-3; The compound with the structure shown in Formula e-3 undergoes a deprotection reaction to obtain a compound with the structure shown in Formula e-4; The compound with the structure shown in Formula e-4 undergoes a substitution reaction with 1-Boc-4-bromomethylpiperidine to obtain a compound with the structure shown in Formula e-5; Under the action of hydrazine hydrate, the compound with the structure shown in Formula e-5 undergoes a deimination reaction to obtain a compound with the structure shown in Formula e.

8. Use of the GPX4 protein degrader according to claim 1 or 2 or the GPX4 protein degrader prepared by the preparation method according to any one of claims 3 to 7 in the preparation of anti-tumor drugs and anti-drug-resistant tumor drugs.

9. An anti-tumor drug, comprising a drug active component and a drug excipient; the drug active component is the GPX4 protein degrader according to claim 1 or 2 or the GPX4 protein degrader prepared by the preparation method according to claims 3 to 7.

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