Artemisinin PROTACs and its preparation method and application

Abstract

The present invention discloses an artemisinin PROTACs, a preparation method and application thereof, wherein the structural formula is: The present invention uses artemisinin modified product artesunate as POI ligand, lenalidomide and pomalidomide as E3 ligands, and linkers selected from amino acid chains, PEG chains and fatty chains. The anticancer activity thereof is superior to that of artesunate, and the present invention has good potential application prospects in anticancer drugs and treatment of critical malaria.

Description

Technical Field

[0001] The present invention belongs to the technical field of organic synthesis, and specifically relates to artemisinin PROTACs and a preparation method and application thereof. Background Art

[0002] Protein degradation targeting chimeras (PROTACs) are a new type of drug with good prospects. Its structure is similar to a dumbbell and is mainly composed of three parts, including E3 ligase ligand (E3 ligand), target protein ligand (POI ligand) and a linker connecting the two. One end of the PROTACs molecule binds to the target protein, and the other end binds to the E3 ubiquitin ligase. The E3 ubiquitin ligase can "stick" a small protein called ubiquitin to the target protein, marking the target protein as a defective or damaged protein, and then use the cell's intrinsic proteolytic enzyme (i.e., 26S proteasome) to specifically recognize and degrade the marked target protein. E3 ligands can recruit E3 ligases, and POI ligands can recognize and bind to target proteins. Therefore, PROTACs molecules form a POI:PROTAC:E3 ligase ternary complex in the cell. In the patient's body, the POI ligand of PROTACs binds to the target protein, and the E3 ligand binds to the substrate binding region of the E3 ubiquitin ligase in the cell. The target protein can be "pulled" to the side of the E3 ubiquitin ligase through the Linker, enabling the ubiquitin-proteasome (UPS) system to degrade the target protein. PROTACs molecules do not need to be highly embedded in the highly active region of the target. They only need to have a short-term adhesion effect on its surface, that is, only through some special intermolecular forces and other low-energy weak binding interactions to achieve the degradation of the target molecule. Currently, because the unique driving model of protein degradation mediated by PROTACs has huge therapeutic potential, this technology is currently mainly used in drug development, and some PROTACs have been clinically tested.

[0003] PROTAC drugs have many advantages over traditional small molecule drugs, which are roughly as follows:

[0004] (1) PROTAC drug design is not limited by Lipinski's five-fold rule;

[0005] (2) The pharmacological effects of traditional small molecules rely on the occupation of key sites on the target protein (i.e., occupancy-driven). The principle is to bind to the active site of the enzyme or receptor to exert its effect, and the project efficacy is poor. Traditional drugs need to maintain a certain drug concentration in the body, and the requirements for small molecule binding sites are high. However, PROTACs can capture the target protein through any corner position. Its pharmacological mechanism is to activate potential targets, thereby degrading the target protein and achieving pharmacodynamic effects. Data show that in the process of drug efficacy, very high drug concentrations are not required to achieve it. The requirements for binding sites are relatively low, and the drug efficacy is better.

[0006] (3) About 50% of proteins in the human body are found to be related to human diseases, such as cancer. However, only about 20% of these proteins can be bound by traditional small molecules, while PROTACs can theoretically bind to any part of the protein.

[0007] (4) PROTACs are more resistant to drug resistance than traditional small molecules. In theory, PROTACs can better tolerate mutations in target proteins.

[0008] (5) PROTACs are more durable and have stronger inhibitory effects than traditional drug molecules; Summary of the Invention

[0009] The present invention aims to provide artemisinin PROTACs.

[0010] Another object of the present invention is to provide a method for preparing the above-mentioned artemisinin PROTACs.

[0011] Another object of the present invention is to provide applications of the above-mentioned artemisinin PROTACs.

[0012] The technical solutions of the present invention are as follows:

[0013] An artemisinin PROTACs, the structural formula of which is in,

[0014] E3 Ligase is lenalidomide or pomalidomide,

[0015] Linker is an amino acid chain, a fatty chain or a PEG chain, the amino acid chain is -NH-(CH2)n1-NH-(CH2)3-CO-, the fatty chain is -NH-(CH2)n2-CO-, the PEG chain is -(O-CH2-CH2)n3-CH2 CH2-CO-, n1 is a natural number from 2 to 10, n2 is a natural number from 2 to 7, and n3 is 2 or 3.

[0016] In a preferred embodiment of the present invention, its structural formula is selected from at least one of the following:

[0017]

[0018]

[0019]

[0020]

[0021]

[0022] The preparation method of the above-mentioned artemisinin PROTACs has the following reaction scheme:

[0023]

[0024] or

[0025]

[0026] or

[0027]

[0028] Use of the above-mentioned artemisinin PROTACs or pharmacologically or physiologically acceptable salts thereof in preparing a cancer treatment composition.

[0029] In a preferred embodiment of the present invention, the cancer includes brain cancer, breast cancer, cervical cancer, stomach cancer, liver cancer and lung cancer.

[0030] A cancer treatment composition, the active ingredient of which includes the above-mentioned artemisinin PROTACs or a pharmacologically or physiologically acceptable salt thereof.

[0031] In a preferred embodiment of the present invention, the cancer includes brain cancer, breast cancer, cervical cancer, stomach cancer, liver cancer and lung cancer.

[0032] The beneficial effects of the present invention are: the present invention uses artemisinin-modified product artesunate as POI ligand, lenalidomide and pomalidomide as E3 ligands, and linkers selected from amino acid chains, PEG chains and fatty chains. Its anti-cancer activity is better than that of artesunate, and it has good potential application prospects in anti-cancer drugs and the treatment of critical malaria. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 This is a diagram of the experimental results of Example 7 of the present invention.

[0034] Figure 2 1 is a diagram showing the experimental results of Example 8 of the present invention.

[0035] Figure 3This is one of the experimental result diagrams of Example 9 of the present invention.

[0036] Figure 4 This is the second diagram of the experimental results of Example 9 of the present invention.

[0037] Figure 5 This is the third figure of the experimental results of Example 9 of the present invention. DETAILED DESCRIPTION

[0038] The technical solution of the present invention is further illustrated and described below through specific implementation methods in conjunction with the accompanying drawings.

[0039] The artemisinin PROTACs prepared in Examples 2 to 5 are listed below:

[0040]

[0041]

[0042]

[0043]

[0044]

[0045]

[0046]

[0047] Example 1: Preparation of artemisinin derivative - artesunate

[0048] Weigh 500 mg of artemisinin and 500 μL of 1,4-dioxane into a reaction flask. Dissolve the mixture in methanol at room temperature with stirring. Maintain the reaction temperature at 0°C. Add 250 mg of lithium borohydride in batches over 60 minutes. Keep the temperature below 5°C for 30 minutes. Monitor the reaction using thin-film chromatography (TLC) until the artemisinin spots disappear. Keep the reaction temperature below 10°C and adjust the pH to approximately 7 by adding phosphoric acid or concentrated sulfuric acid to terminate the reduction reaction. Add 1 g of succinic anhydride and 6 g of DCC. Keep the temperature at 45°C for 3 hours. Monitor the reaction using TLC until the dihydroartemisinin spots disappear. Maintain the temperature at 45°C and concentrate under reduced pressure until no solvent is present. Add ethyl acetate and 0.1% sulfuric acid solution, adjust the pH to 4, and extract with stirring for 15 minutes. Separate the organic phase and extract once more with ethyl acetate. Combine the organic phases, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain crude artesunate. The crude product was dissolved in ethyl acetate and concentrated appropriately, and then recrystallized to obtain 635 mg of fine artesunate (white solid).

[0049] Example 2:

[0050] (1) Preparation of intermediates s1-s12 (preparation of E3 ligand)

[0051] A. The structural formula of intermediate Q1 (n = 1) / Q3 (n = 2) / Q5 (n = 3) / Q7 (n = 4) / Q9 (n = 5) / Q11 (n = 6) is The specific synthesis method is as follows:

[0052] Boc-3-aminopropionic acid (1 g, 4.9 mmol) and DIPEA (1.71 mL, 9.8 mmol) were dissolved in 1 mL of dry DMF and stirred at room temperature for 5 min. HATU (1.49 g, 3.92 mmol) was added and the reaction was monitored by TLC for approximately 0.5 h. Lenalidomide (826 mg, 3.2 mmol) was added and stirred at room temperature. TLC followed the reaction, indicating completion in approximately 12 h. Crushed ice was added to the reaction solution to quench the reaction. The mixture was extracted three times with ethyl acetate. The organic layers were combined, washed sequentially with saturated ammonium chloride solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The reaction was analyzed by PTLC (dichloromethane:methanol = 20:1) to afford Intermediate Q1 (white solid, 1.28 g, 90%).

[0053] Boc-4-aminobutyric acid (1 g, 5.2 mmol) and DIPEA (1.81 mL, 10.4 mmol) were dissolved in 1 mL of dry DMF and stirred at room temperature for 5 min. HATU (1.6 g, 4.16 mmol) was added and the reaction was monitored by TLC for approximately 0.5 h. Lenalidomide (876 mg, 3.38 mmol) was added and stirred at room temperature. TLC followed the reaction, indicating completion in approximately 12 h. The reaction solution was quenched by the addition of crushed ice and extracted three times with ethyl acetate. The organic layers were combined, washed sequentially with saturated ammonium chloride solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The reaction was analyzed by PTLC (dichloromethane:methanol = 20:1) to afford Intermediate Q3 (white solid, 1.3 g, 90%).

[0054] Boc-5-aminovaleric acid (1 g, 4.6 mmol) and DIPEA (1.6 mL, 9.2 mmol) were dissolved in 1 mL of dry DMF and stirred at room temperature for 5 min. HATU (1.4 g, 3.68 mmol) was added and the reaction was monitored by TLC for approximately 0.5 h. Lenalidomide (775 mg, 2.99 mmol) was added and stirred at room temperature. TLC followed the reaction, indicating completion in approximately 12 h. The reaction solution was quenched by the addition of crushed ice and extracted three times with ethyl acetate. The organic layers were combined, washed sequentially with saturated ammonium chloride solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The reaction was analyzed by PTLC (dichloromethane:methanol = 20:1) to afford Intermediate Q5 (white solid, 1.25 g, 91%).

[0055] Boc-6-aminohexanoic acid (1 g, 4.3 mmol) and DIPEA (1.5 mL, 8.6 mmol) were dissolved in 1 mL of dry DMF solution and stirred at room temperature for 5 min. HATU (1.31 g, 3.44 mmol) was added. After approximately 0.5 h of reaction, lenalidomide (725 mg, 2.8 mmol) was added and stirred at room temperature. TLC monitoring of the reaction indicated completion of the reaction after approximately 12 h. The reaction solution was quenched by the addition of crushed ice and extracted three times with ethyl acetate. The organic layers were combined, washed sequentially with saturated ammonium chloride solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The reaction was analyzed by PTLC (dichloromethane:methanol = 20:1) to afford Intermediate Q7 (white solid, 1.2 g, 92%).

[0056] Boc-7-aminoheptanoic acid (1 g, 4.1 mmol) and DIPEA (1.43 mL, 8.2 mmol) were dissolved in 1 mL of dry DMF and stirred at room temperature for 5 min. HATU (1.25 g, 3.28 mmol) was added and the reaction was monitored by TLC for approximately 0.5 h. Lenalidomide (691 mg, 3.38 mmol) was added and stirred at room temperature. TLC followed the reaction, indicating completion in approximately 12 h. The reaction solution was quenched by the addition of crushed ice and extracted three times with ethyl acetate. The organic layers were combined, washed sequentially with saturated ammonium chloride solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The reaction was then analyzed by PTLC (dichloromethane:methanol = 20:1) to afford Intermediate Q9 (white solid, 1.53 g, 93%).

[0057] Boc-8-aminooctanoic acid (1 g, 3.8 mmol) and DIPEA (1.3 mL, 7.6 mmol) were dissolved sequentially in 1 mL of dry DMF and stirred at room temperature for 5 min. HATU (1.16 g, 3.04 mmol) was added and the reaction was monitored by TLC for approximately 0.5 h. Lenalidomide (640 mg, 2.47 mmol) was added and stirred at room temperature. TLC followed the reaction, indicating completion in approximately 12 h. The reaction solution was quenched by the addition of crushed ice and extracted three times with ethyl acetate. The organic layers were combined, washed sequentially with saturated ammonium chloride solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The reaction was analyzed by PTLC (dichloromethane:methanol = 20:1) to afford Intermediate Q11 (white solid, 1.11 g, 90%).

[0058] Intermediate Q1 (1 g, 2.25 mmol) was dissolved in a DCM / MeOH (v:v = 3:1) mixture (10 mL). TFA (3 mL) was added dropwise under an ice bath. After addition, the mixture was slowly brought to room temperature and monitored by TLC. The reaction was complete in approximately 12 hours. After cessation of the reaction, the solvent was removed by concentration under reduced pressure to afford Intermediate Q2 (white solid, 713 mg, 96.0%), which was used directly in the next step without purification.

[0059] B. The structural formula of intermediate Q2 (n=1) / Q4 (n=2) / Q6 (n=3) / Q8 (n=4) / Q10 (n=5) / Q12 (n=6) is The specific synthesis method is as follows:

[0060] Intermediate Q3 (1 g, 2.25 mmol) was dissolved in a DCM / MeOH (v:v = 3:1) mixture (10 mL). TFA (3 mL) was added dropwise under an ice bath. After addition, the mixture was slowly brought to room temperature and monitored by TLC. The reaction was complete in approximately 12 hours. After cessation of the reaction, the solvent was removed by concentration under reduced pressure to afford Intermediate Q4 (white solid, 675 mg, 90%), which was used directly in the next step without purification.

[0061] Intermediate Q5 (1 g, 2.25 mmol) was dissolved in a DCM / MeOH (v:v = 3:1) mixture (10 mL). TFA (3 mL) was added dropwise under an ice bath. After addition, the mixture was slowly brought to room temperature and monitored by TLC. The reaction was complete in approximately 12 hours. After cessation of the reaction, the solvent was removed by concentration under reduced pressure to afford Intermediate Q6 (white solid, 705 mg, 93.0%), which was used directly in the next step without purification.

[0062] Intermediate Q7 (1 g, 2.25 mmol) was dissolved in a DCM / MeOH (v:v = 3:1) mixture (10 mL). TFA (3 mL) was added dropwise under an ice bath. After addition, the mixture was slowly brought to room temperature and monitored by TLC. The reaction was complete in approximately 12 hours. After cessation of the reaction, the solvent was removed by concentration under reduced pressure to afford Intermediate Q8 (white solid, 690 mg, 92.0%), which was used directly in the next step without purification.

[0063] Intermediate Q9 (1 g, 2.25 mmol) was dissolved in a DCM / MeOH (v:v = 3:1) mixture (10 mL). TFA (3 mL) was added dropwise under an ice bath. After addition, the mixture was slowly brought to room temperature and monitored by TLC. The reaction was complete in approximately 12 hours. After cessation of the reaction, the solvent was removed by concentration under reduced pressure to afford Intermediate Q10 (white solid, 668 mg, 93.0%), which was used directly in the next step without purification.

[0064] Intermediate Q11 (1 g, 2.25 mmol) was dissolved in a DCM / MeOH (v:v = 3:1) mixture (10 mL). TFA (3 mL) was added dropwise under an ice bath. After addition, the mixture was slowly brought to room temperature and monitored by TLC. The reaction was complete in approximately 12 hours. After cessation of the reaction, the solvent was removed by concentration under reduced pressure to afford Intermediate Q12 (white solid, 683 mg, 91.0%), which was used directly in the next step without purification.

[0065] (2) Preparation of final product Q1-6

[0066] A 25 mL round-bottom flask was charged with artesunate (100 mg, 0.26 mmol), linker and E3 ligase ligands Q2 / Q4 / Q6 / Q8 / Q10 / Q12 (360.37 g) (0.51 mol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCl) (0.51 mol), 4-dimethylaminopyridine (DMAP) (0.26 mol), and DCM (5 mL). The reaction was stirred at room temperature for 5-8 h, extracted with DCM, and the organic phase was washed with 5% NaHCO₃ solution and saturated sodium chloride, then dried over anhydrous Na₂SO₄. The solid was removed by filtration, and the solvent was evaporated under reduced pressure to obtain the crude product, which was then purified by column chromatography (PE-EA = 10:1 to 1:1, v / v). Its NMR data are as follows:

[0067] Final product Q1 (white solid, yield 33.5%): 1H NMR(500MHz,DMSO-d6)δ11.03(s,1H),9.86(s,1H),8.08(d,J=9.9Hz,1H),7.81(d d,J=7.3,1.7Hz,1H),7.52(d,J=6.0Hz,1H),7.51–7.47(m,1H),5.62(dd,J=9.8,2 .7Hz,1H),5.52(d,J=7.9Hz,1H),5.15(dd,J=13.3,5.1Hz,1H),4.48–4.24(m,2H) ,2.92(d,J=1.7Hz,1H),2.63(dd,J=4.5,2.4Hz,1H),2.60(p,J=4.1,3.5Hz,2H),2 .43–2.37(m,2H),2.37–2.32(m,1H),2.27(dd,J=4.4,2.6Hz,1H),2.22–2.13(m,1 H),2.04–1.94(m,2H),1.80(s,1H),1.60(tt,J=8.4,3.5Hz,2H),1.56–1.50(m,1H ),1.45–1.39(m,2H),1.34–1.29(m,2H),1.28(s,3H),1.23(d,J=4.1Hz,2H),1.18 –1.12(m,1H),0.99–0.92(m,1H),0.88(d,J=6.4Hz,3H),0.75(d,J=7.1Hz,3H).13C NMR (126MHz, DMSO-d6) δ172.95,171.30,171.11,170.71,169.65,167.89,133.68,132.70,129.54,128.63,125.46,119.15,103.63,91.67,90.6 2,79.91,51.53,51.15,46.48,44.60,36.02,35.93,35.25,33.73,31.66 ,31.26,29.53,29.05,28.94,25.55,24.23,22.67,21.05,20.10,11.77.

[0068] Final product Q2 (white solid, yield 39.6%): 1H NMR(500MHz,Chloroform-d)δ9.41(d,J=14.3Hz,1H),9.36(d,J=16.1Hz,1H),7 .89(t,J=7.5Hz,1H),7.56(dd,J=7.4,2.7Hz,1H),7.37(t,J=7.5Hz,1H),6.93– 6.78(m,1H),5.67(d,J=9.8Hz,1H),5.38(d,J=16.9Hz,1H),5.09(dd,J=13.2,5 .2Hz,1H),4.40(d,J=7.0Hz,2H),3.25(td,J=15.9,14.6,7.8Hz,2H),2.75(s,2H ),2.65(t,J=6.6Hz,2H),2.46(d,J=6.7Hz,2H),2.35(t,J=6.7Hz,2H),2.33–2. 25(m,2H),2.09(dd,J=18.9,6.8Hz,2H),1.99–1.93(m,1H),1.90–1.81(m,2H),1 .80(d,J=7.1Hz,2H),1.70–1.62(m,2H),1.55(dt,J=13.9,4.4Hz,1H),1.32(s, 2H),1.29(s,2H),1.22(s,3H),0.90(d,J=5.6Hz,3H),0.76(t,J=6.4Hz,3H).13C NMR (125 MHz, Common NMR Solvents)δ172.74,172.22,172.06,171.73,169.90,168.45,140.03,129.08,126.93,126.10,125.77,117.83,104.72,96.92,93.51,81.11 ,55.39,48.18,46.28,42.18,39.95,37.83,36.38,35.45,35.42,33.77 ,31.85,30.27,29.74,27.29,26.44,25.24,24.20,24.09,19.44,13.36.

[0069] Final product Q3 (white solid, yield 38.8%): 1H NMR (500 MHz, Chloroform-d) δ 9.13 (d, J = 14.9 Hz, 1H), 8.70 (d, J = 22.2 Hz, 1H), 7.79 (dd, J = 16.1, 7.9 Hz, 1H), 7.63 (d, J = 7.4 Hz, 1H), 7.43 (t, J = 7.8 Hz, 1H), 6.36 (d, J = 6.6 Hz, 1H), 5.67 (dd, J = 13.5, 9.8 Hz, 1H), 5.30 (d, J = 7.5 Hz, 1H), 5.19–5.01 (m, 1H), 4.39 (d, J = 6.4 Hz, 2H), 3.36–3.11 (m, 2H), 2.75 (d, J = 23.3 Hz, 2H), 2.71–2.60 (m, 2H), 2.53–2.45 (m, 2H), 2.45–2.41 (m, 2H), 2.38–2.31 (m, 2H), 2.13 (s, 2H), 2.01 (dt, J = 15.6, 3.9 Hz, 2H), 1.90–1.85 (m, 1H), 1.70 (d, J = 8.2 Hz, 2H), 1.66 (d, J = 15.6 Hz, 2H), 1.58 (q, J = 4.9 Hz, 1H), 1.53 (q, J = 6.8, 5.8 Hz, 2H), 1.41 (q, J = 4.5, , 3.8 Hz, 2H), 1.37 (d, J = 4.9 Hz, 2H), 1.10 (s, 2H), 0.97–0.93 (m, 3H), 0.86 (q, J = 6.3 Hz, 2H), 0.79 (dd, J = 7.1, 2.8 Hz, 3H). 13C NMR (126 MHz, Chloroform-d) δ 171.78, 133.13, 132.43, 104.47, 104.43, 92.30, 91.39, 80.04, 59.39, 51.80, 51.33, 44.96, 38.02, 37.04, 36.03, 33.86, 31. , 80, 31.54, 31.38, 31.11, 30.69, 29.67, 29.57, 29.24, 28.60, 25.71, 24.40, 23.05, 22.62, 21.72, 20.05, 14.00, 11.89.

[0070] Final product Q4 (white solid, yield 39.9%): 1H NMR (500 MHz, Chloroform-d) δ 9.21 (d, J = 12.9 Hz, 1H), 8.79 (d, J = 20.2 Hz, 1H), 7.77 (dd, J = 8.1, 3.6 Hz, 1H), 7.63 (d, J = 7.5 Hz, 1H), 7.44 (td, J = 7.8, 2.5 Hz, 1H), 6.31 (t, J=5.8Hz,1H),5.71(dd,J=9.8,2.5Hz,1H),5.42(d,J=9.7Hz,1H),5.07(td,J=12.5, 5.0Hz,1H),4.38(qd,J=16.8,16.1,8.1Hz,2H),3.14(q,J=6.7Hz,2H),2.85–2.69(m ,2H),2.69–2.55(m,2H),2.55–2.47(m,1H),2.43(q,J=5.9,5.3Hz,2H),2.40–2.35( m,2H),2.35–2.20(m,2H),2.20–2.06(m,2H),2.05–1.94(m,2H),1.87(dq,J=10.5,3 .5Hz,1H),1.77–1.69(m,2H),1.69–1.63(m,2H),1.59(dt,J=13.9,4.5Hz,1H),1.47 –1.42(m,2H),1.36(d,J=11.2Hz,3H),0.98–0.93(m,3H),0.82(t,J=9.9Hz,3H).13C NMR (126MHz, Chloroform-d) δ133.15,128.93,120.40,104.46,104.43,92.23,91.42,80.05,53.31,51.75,51.35,44.99,39.07,37. 08,36.37,36.05,33.88,31.62,31.35,30.51,29.57,29.53,28.79,25.93,25.71,25.68,24.91,24.40,23.02,21.79,20.06,11.92.

[0071] Final product Q5 (white solid, yield 40.2%): 1H NMR (500 MHz, DMSO-d6) δ 11.03 (s, 1H), 9.78 (s, 1H), 7.86 (t, J = 5.6 Hz, 1H), 7.81 (dd, J = 7.2, 1.8 Hz, 1H), 7.50 (d, J = 5.7 Hz, 1H), 7.48 (d, J = 7.5 Hz, 1H), 5.65–5.62 (m, 1H), 5.5 3(d,J=2.3Hz,1H),5.14(dd,J=13.3,5.2Hz,1H),4.40–4.29(m,2H),3.02(q,J=6.6Hz ,2H),2.95–2.87(m,1H),2.65–2.59(m,2H),2.59–2.56(m,2H),2.41–2.36(m,2H),2.3 3(s,1H),2.26(ddd,J=9.7,7.1,4.4Hz,2H),2.19–2.13(m,1H),2.04–1.97(m,2H),1. 91–1.68(m,2H),1.65–1.60(m,2H),1.58(d,J=6.0Hz,2H),1.53(dt,J=8.2,3.8Hz,2H) ,1.43–1.39(m,2H),1.36(d,J=5.5Hz,2H),1.31(d,J=4.6Hz,2H),1.28(s,2H),1.27( s,3H),1.15(dd,J=11.3,6.6Hz,2H),0.87(d,J=3.9Hz,3H),0.74(d,J=7.3Hz,3H).13C NMR(126MHz,DMSO-d6)δ172.93,171.42,171.27,171.12,170.28,167.88 ,133.84,132.69,128.66,103.60,91.63,90.61,79.90,51.56,51.14,46 .52,44.59,38.50,35.95(d,J=8.3Hz),35.81,33.71,31.67,31.24,29.5 7,29.04,28.42,26.17,25.55,25.09,24.22,22.66,21.03,20.09,11.76.

[0072] Final product Q6 (white solid, yield 38.9%): 1H NMR (500 MHz, Chloroform-d) δ 9.11 (d, J = 3.8 Hz, 1H), 8.73 (d, J = 17.6 Hz, 1H), 7.78 (t, J = 8.4 Hz, 1H), 7.68 (d, J = 7.5 Hz, 1H), 7.48 (t, J = 7.7 Hz, 1H), 6.18 (d, J = 5.2 Hz, 1H), 5.75 (d, J = 9.8 Hz, 1H), 5.44 (d, J =5.4Hz,1H),5.20–5.04(m,1H),4.51–4.31(m,2H),3.17(q,J=5.7,5.3Hz,2H),2.79(s,1H),2.73(d t,J=18.2,6.2Hz,2H),2.68(d,J=6.7Hz,1H),2.59–2.52(m,1H),2.47(p,J=6.8Hz,2H),2.42(dd,J=9 .0,5.8Hz,2H),2.38–2.24(m,2H),2.14(s,1H),2.03(dq,J=14.3,4.4,3.6Hz,2H),1.93–1.87(m,2H ),1.80–1.73(m,2H),1.72–1.68(m,2H),1.62(dt,J=13.8,4.4Hz,1H),1.40(d,J=4.3Hz,3H),1.37(d ,J=3.2Hz,1H),1.35–1.32(m,2H),1.32–1.30(m,2H),1.30–1.28(m,1H),1.27(d,J=3.0Hz,2H),1.2 6–1.21(m,2H),1.01(d,J=12.3Hz,1H),0.97(dd,J=5.9,1.5Hz,3H),0.84(dd,J=7.1,2.0Hz,3H).13C NMR(126MHz,Chloroform-d)δ172.14,172.08,171.70,170.32,169.32,133.3 8,132.74,129.17,104.68,104.66,92.47,91.65,80.29,53.57,51.93,51.63, 45.28,39.47,37.36,36.72,36.31,34.16,31.87,31.62,30.89,29.84,29.36,29.32,28.77,28.40,26.36,25.97,25.46,24.67,23.36,22.06,20.32,12.18.

[0073] Example 3:

[0074] (1) Preparation of intermediates N7-N16 and NJ7-NJ16

[0075] A. Preparation of Intermediate N1

[0076] Potassium phthalimide (2 g, 0.011 mol, 1 eq) was weighed into a 50 mL round-bottom flask, and 20 mL of acetone was added. TBAB (0.32 g, 0.001 mol, 0.1 eq) and 1,2-dibromoethane (3.9 mL, 0.033 mol, 3 eq) were then added sequentially with stirring. The mixture was refluxed at 70°C for 2 h. TIC analysis (developing solvent: PE:EtOAc = 6:1) indicated complete reaction of the starting material. Post-treatment: Cool, quench with ice water, and repeatedly extract with DCM. Distill under reduced pressure to obtain a yellow liquid, which was purified by column chromatography (PE:EtOAc = 10:1) to afford N1-NN2 (white solid, 86.8% yield). Its NMR data: 1H NMR (500 MHz, Chloroform-d) δ 7.87 (dd, J = 5.4, 3.1 Hz, 2H), 7.74 (dd, J = 5.5, 3.0 Hz, 2H), 4.10 (t, J = 6.7 Hz, 2H), 3.61 (t, J = 6.7 Hz, 2H). 13C NMR (125 MHz, Chloroform-d) δ 167.83, 134.24, 131.85, 123.54, 39.31, 28.15.

[0077] The synthesis method of intermediates N1-NN3 to N1-NN6 is similar to that of N1-NN2. The structural formula of N1-NN2 to N1-NN6 is Its NMR data are as follows:

[0078] N1-NN3 (white solid, yield 80.9%): 1H NMR (500 MHz, Chloroform-d) δ 7.86–7.84 (m, 2H), 7.73–7.71 (m, 2H), 3.84 (t, J = 6.9 Hz, 2H), 3.41 (s, 2H), 2.25 (d, J = 6.8 Hz, 2H). 13C NMR (125 MHz, Chloroform) δ 168.26, 134.07, 132.03, 123.35, 36.75, 31.65, 29.78.

[0079] N1-NN4 (white solid, yield 80.5%): 1H NMR (500 MHz, Chloroform-d) δ 7.84 (dd, J = 5.4, 3.0 Hz, 2H), 7.71 (dd, J = 5.4, 3.0 Hz, 2H), 3.72 (t, J = 6.7 Hz, 2H), 3.44 (t, J = 6.4 Hz, 2H), 1.92–1.88 (m, 2H), 1.86–1.82 (m, 2H). 13C NMR (125 MHz, Chloroform) δ 168.38, 134.00, 132.06, 123.28, 36.97, 32.78, 29.85, 27.25.

[0080] N1-NN5 (white solid, yield 82.7%): 1H NMR (500 MHz, Chloroform-d) δ 7.87–7.76 (m, 2H), 7.75–7.63 (m, 2H), 3.67 (t, J = 7.3 Hz, 2H), 3.37 (t, J = 6.8 Hz, 2H), 1.88 (dt, J = 15.0, 6.9 Hz, 2H), 1.73–1.64 (m, 2H), 1.54–1.41 (m, 2H). 13C NMR (125 MHz, Chloroform) δ 167.94, 133.61, 131.83, 123.33, 40.94, 40.25, 28.53, 27.77, 26.03.

[0081] N1-NN6 (white solid, yield 78.7%): 1H NMR (500 MHz, Chloroform-d) δ 7.90–7.78 (m, 2H), 7.75–7.65 (m, 2H), 3.68 (t, J = 7.2 Hz, 2H), 3.38 (t, J = 6.8 Hz, 2H), 1.91–1.79 (m, 2H), 1.74–1.64 (m, 2H), 1.54–1.42 (m, 2H), 1.41–1.31 (m, 2H). 13C NMR (125 MHz, Chloroform) δ 168.19, 133.51, 131.83, 123.21, 41.71, 39.46, 32.46, 27.88, 27.17, 26.72.

[0082] B. Preparation of intermediate N2

[0083] At room temperature, N1-NN2 (1 g, 0.004 mol, 1 eq) was weighed into a 25 mL round-bottom flask. DMF (15 mL) was added and stirred thoroughly. K2CO3 (1.7 g, 0.012 mol, 3 eq) and KI (0.2 g, 0.0012 mmol, 0.3 eq) were then added. The temperature was raised to 75°C, and γ-aminobutyric acid (0.82 g, 0.008 mol, 2 eq) was slowly added. TIC (developing solvent: PE:EtOAc = 7:1) confirmed the absence of N1-NN2 starting material after 24 h of reaction. Workup: The reaction was quenched with ice water, resulting in the precipitation of a yellow solid. The solid was stirred at room temperature for 30 min and thoroughly extracted with EtOAc. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford N2-J7 as a colorless, transparent liquid, which was used directly in the next reaction (a yellow oily liquid, 85.1% yield) without purification. The synthesis methods of intermediates N2-J8 to N2-J9 ​​are similar to those of N2-J7. N2-J8: yellow oily liquid, yield 85.6%. N2-J9: yellow oily liquid, yield 85.3%.

[0084] At room temperature, N1-NN3 (1 g, 0.0037 mol, 1 eq) was weighed into a 25 mL round-bottom flask. DMF (5 mL) was added and stirred thoroughly. K2CO3 (1.53 g, 0.011 mol, 3 eq) and KI (0.015 g, 0.0001 mmol, 0.3 eq) were then added. The temperature was raised to 75°C, and 6-aminohexanoic acid (970 mg, 0.0074 mol, 2 eq) was slowly added. TIC (developing solvent: PE:EtOAc = 7:1) was used to detect the presence of N1-NN3 starting material after 24 h of reaction. Post-treatment: Add ice water to quench the reaction, and a yellow solid precipitates. Stir at room temperature for 30 minutes, then fully extract with EtOAc. The organic layer is washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated acid under reduced pressure to obtain a colorless, transparent liquid N2-J10, which can be used directly in the next step without purification (yellow oily liquid, yield 83.2%). The synthesis method of intermediates N2-J11 to N2-J13 is similar to that of N2-J10. N2-J11: yellow oily liquid, yield 82.3%. N2-J12: yellow oily liquid, yield 81.3%. N2-J13: yellow oily liquid, yield 80.7%.

[0085] At room temperature, N1-NN2 (1 g, 0.0033 mol, 1 eq) was weighed into a 25 mL round-bottom flask. DMF (5 mL) was added and stirred thoroughly. K2CO3 (1.37 g, 0.0099 mol, 3 eq) and KI (0.15 g, 0.0009 mol, 0.3 eq) were then added. The temperature was raised to 75°C, and 11-aminoundecanoic acid (1.3 g, 0.0099 mol, 3 eq) was slowly added. TIC (developing solvent: PE:EtOAc = 7:1) was used to detect the presence of N1-NN2 starting material after 24 h of reaction. Post-treatment: Add ice water to quench the reaction, and a yellow solid precipitates. Stir at room temperature for 30 minutes, adjust to neutral with 1M HCl, and thoroughly extract with EtOAc. The organic layer is washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated acid under reduced pressure to obtain a colorless, transparent liquid N2-J14, which can be used directly in the next step without purification (yellow oily liquid, yield 80.4%). The synthesis method of intermediates N2-J15 to N2-J16 is similar to that of N2-J14. N2-J15: yellow oily liquid, yield 81.6%. N2-J16: yellow oily liquid, yield 80.5%.

[0086] C. Preparation of intermediate N3

[0087] N2-J7 (0.8 g, 0.0028 mol, 2 eq) was weighed into a 50 mL flask and DMF (10 mL) was added. After stirring for several minutes, DIPEA (890 μl, 0.0051 mol, 3 eq) and HATU (780 mg, 0.002 mol, 1.2 eq) were added sequentially. The solution immediately turned deep red upon addition. Stir for approximately 10 minutes. TIC analysis (developing solvent: DCM:MeOH = 20:1) was performed until the active ester was formed. Lenalidomide (500 mg, 0.0019 mol, 1 eq) was added. The reaction was allowed to proceed at room temperature for 12 hours. TIC analysis (developing solvent: DCM:MeOH = 20:1) confirmed complete reaction of the lenalidomide. The reaction mixture was quenched by adding crushed ice and thoroughly extracted with DCM. The organic layer was washed with saturated ammonium chloride solution and saturated brine in sequence, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a yellow liquid. After standing, a yellow solid precipitated and was purified by column chromatography to obtain N3-J7 (yellow solid, yield 59.6%). N3-J7: 1H NMR(500MHz,DMSO-d6)δ7.18(t,J=7.6Hz,3H),6.91(dd,J=7.4,0.9Hz,3H),6.80 (dd,J=8.0,0.9Hz,3H),5.10(dd,J=13.3,5.1Hz,3H),4.21(d,J=16.9Hz,3H),4.1 0(d,J=16.9Hz,3H),2.92(ddd,J=17.0,13.4,5.3Hz,3H),2.61(ddt,J=17.4,4.0, 2.1Hz,3H),2.30(qd,J=13.3,4.4Hz,3H),2.02(dtd,J=12.8,5.2,2.2Hz,3H).13C NMR (125MHz, DMSO) δ173.40,171.72,169.39,168.55,159.71,144.13,134.80,132.72,129.30,127.57,126.07,116 .90,114.05,110.87,110.54,55.40,51.99,51.99,49.06,46.06,46.06,40.56,40.47,31.71,31.60,23.32,23.25.

[0088] The synthesis of intermediates N3-J8 to N3-J18 is similar to that of N3-J7, except that the number of carbon atoms in the intermediate connecting alkane chain is different. Intermediate N3 is obtained by reacting the corresponding N2 of the same chain length with lenalidomide. The distance between the two amino groups on both sides is the same as the serial number, and the chain length gradually increases. Its NMR data are as follows: N3-J8 (yellow solid, yield 58.2%): 1H NMR(500MHz,DMSO-d6)δ7.18(t,J=7.7Hz,3H),6.95–6.88(m,3H),6.84–6.76(m,3 H),5.10(dd,J=13.3,5.1Hz,3H),4.21(d,J=16.9Hz,3H),4.11(d,J=17.0Hz,2H),3 .17(d,J=5.2Hz,2H),2.91(ddd,J=17.5,13.7,5.5Hz,3H),2.61(ddd,J=17.6,4.2 ,1.9Hz,3H),2.30(qd,J=13.3,4.5Hz,3H),2.02(dtd,J=12.8,5.3,2.2Hz,3H).13C NMR (125MHz, DMSO) δ173.40,171.72,169.40,162.81,144.12,140.38,139.84,137.95,132.71,129.31,126.08,116.91,1 10.89,66.57,62.53,60.00,58.24,55.40,52.00,49.06,46.06,40.99,39.45,36.26,31.71,31.25,30.77,23.24,21.28.

[0089] N3-J9 (yellow solid, yield 57.8%): 1H NMR(500MHz,DMSO-d6)δ7.18(t,J=7.6Hz,3H),6.90(dd,J=7.4,0.9Hz,3H),6.80( dd,J=7.9,0.9Hz,3H),5.10(dd,J=13.3,5.1Hz,3H),4.21(d,J=17.0Hz,3H),4.10( d,J=16.9Hz,3H),3.16(d,J=5.0Hz,2H),2.99–2.87(m,3H),2.61(ddt,J=17.5,4.1 ,2.0Hz,3H),2.30(qd,J=13.2,4.4Hz,3H),2.02(dtd,J=12.7,5.2,2.2Hz,1H).13C NMR (125MHz, DMSO) δ173.39,171.72,169.38,168.45,165.21,144.14,134.87,132.71,132.15,129.29,126.07,123.46,1 16.88,110.85,51.98,50.47,49.04,46.06,42.38,40.56,39.47,37.21,33.03,31.71,27.06,25.74,23.24,23.20,20.83.

[0090] N3-J10 (yellow solid, yield 57.7%): 1H NMR(500MHz,DMSO-d6)δ7.18(t,J=7.6Hz,3H),6.91(dd,J=7.4,0.9Hz,3H),6.80 (dd,J=8.0,0.9Hz,3H),5.10(dd,J=13.3,5.1Hz,3H),4.21(d,J=16.9Hz,3H),4.1 0(d,J=16.9Hz,3H),2.92(ddd,J=17.0,13.4,5.3Hz,3H),2.61(ddt,J=17.4,4.0, 2.1Hz,3H),2.30(qd,J=13.3,4.4Hz,3H),2.02(dtd,J=12.8,5.2,2.2Hz,3H).13C NMR (125MHz, DMSO) δ173.40,171.72,169.39,168.55,159.71,144.13,134.80,132.72,129.30,127.57,126.07,116 .90,114.05,110.87,110.54,55.40,51.99,51.99,49.06,46.06,46.06,40.56,40.47,31.71,31.60,23.32,23.25.

[0091] N3-J11 (yellow solid, yield 58.6%): 1H NMR(500MHz,DMSO-d6)δ7.18(t,J=7.7Hz,3H),6.95–6.88(m,3H),6.84–6.76(m,3 H),5.10(dd,J=13.3,5.1Hz,3H),4.21(d,J=16.9Hz,3H),4.11(d,J=17.0Hz,2H),3 .17(d,J=5.2Hz,2H),2.91(ddd,J=17.5,13.7,5.5Hz,3H),2.61(ddd,J=17.6,4.2 ,1.9Hz,3H),2.30(qd,J=13.3,4.5Hz,3H),2.02(dtd,J=12.8,5.3,2.2Hz,3H).13C NMR (125MHz, DMSO) δ173.40,171.72,169.40,162.81,144.12,140.38,139.84,137.95,132.71,129.31,126.08,116.91,1 10.89,66.57,62.53,60.00,58.24,55.40,52.00,49.06,46.06,40.99,39.45,36.26,31.71,31.25,30.77,23.24,21.28.

[0092] N3-J12 (yellow solid, yield 59.7%): 1H NMR(500MHz,DMSO-d6)δ8.09(s,1H),7.20(s,1H),7.19(s,1H),7.17(s,1H),6.9 1(d,J=7.4Hz,2H),6.80(d,J=7.9Hz,4H),5.10(dd,J=13.3,5.1Hz,3H),4.18(s, 2H),4.12(s,2H),4.08(s,2H),3.65–3.56(m,2H),3.14(t,J=7.6Hz,2H),2.91(d ,J=4.5Hz,2H),2.88(d,J=2.1Hz,2H),2.73(s,1H),2.63(t,J=3.2Hz,2H),2.60(s ,1H),2.35–2.25(m,4H),2.05–1.99(m,4H),0.88(dt,J=19.1,7.3Hz,4H).13CNM R(125MHz,DMSO)δ173.40,173.30,171.73,169.37,165.53,144.11,144.05,134. 22,132.72,129.97,129.31,126.05,116.87,110.88,54.02,51.97,46.01,42.28,31.71,30.38,28.85,23.87,23.24,22.86,18.53,17.18,14.38,12.95,11.38.

[0093] N3-J13 (yellow solid, yield 55.6%): 1H NMR (500 MHz, DMSO-d6) δ 7.19 (t, J = 7.6 Hz, 3H), 6.92 (d, J = 7.3 Hz, 3H), 6.81 (d, J = 7.8 Hz, 3H), 5.11 (dd, J = 13.3, 5.1 Hz, 3H), 4.22 (d, J = 16.9 Hz, 3H), 4.11 (d, J = 17.0 Hz, 3H) ),3.17(d,J=3.1Hz,1H),2.92(ddd,J=16.9,13.4,5.2Hz,3H),2.62(ddd,J=17.4,4. 0,1.8Hz,3H),2.31(qd,J=13.3,4.4Hz,3H),2.03(dtd,J=12.8,5.4,2.3Hz,3H).13C NMR (125MHz, DMSO) δ173.39,171.72,169.38,169.37,168.78,144.32,144.12,132.72,132.46,129.30,126.08,116.90,1 16.61,110.88,55.40,51.99,51.93,49.06,46.37,46.12,46.05,40.47,39.46,31.77,31.71,31.64,24.06,23.34,23.25.

[0094] N3-J14 (yellow solid, yield 57.3%): 1H NMR (500 MHz, DMSO-d6) δ 7.19 (t, J = 7.6 Hz, 3H), 6.91 (d, J = 7.3 Hz, 3H), 6.79 (d, J = 7.9 Hz, 3H), 5.75 (s, 1H), 5.10 (dd, J = 13.3, 5.1 Hz, 3H), 4.18 (s, 3H), 4.10 (d, J = 16.7 Hz, 4H), 2.90 (d, J = 15.2 Hz, 4H), 2.65–2.58 (m, 3H), 2.33–2.27 (m, 3H), 2.05–2.00 (m, 3H). 13C NMR (125MHz, DMSO) δ173.39,171.73,169.37,165.53,162.78,144.11,134. 22,132.72,130.62,129.96,129.31,126.06,123.49,123.44,116.87,110.8 8,67.60,55.38,54.06,51.97,49.07,46.00,42.30,36.25,31.71,31.23,30.38,28.85,23.87,23.24,22.87,18.61,18.54,17.18,14.38,12.96,11.38.

[0095] N3-J15 (yellow solid, yield 57.1%): 1H NMR (500 MHz, DMSO-d6) δ 8.08 (s, 1H), 7.18 (t, J = 7.6 Hz, 3H), 6.91 (d, J = 7.3 Hz, 3H), 6.80 (d, J = 7.8 Hz, 3H), 5.10 (dd, J = 13.3, 5.1 Hz, 3H), 4.25–4.16 (m, 3H), 4.10 (d, J = 17.0 Hz, 3H) ,3.54–3.47(m,1H),2.95–2.85(m,3H),2.72(s,1H),2.63–2.56(m,3H),2.33–2.26(m,3H ),2.03(tt,J=4.4,2.2Hz,3H),1.29(s,3H),1.26(d,J=2.0Hz,3H),1.24–1.22(m,4H).13C NMR (125MHz, DMSO) δ173.39,171.72,169.38,165.52,162.80,144.12,134.83,134.22,132.71,129.96,129.30,126.07,116.89,110.87,70.25 ,67.61,53.73,51.99,46.05,42.01,38.71,37.53,36.26,31.71,30.38 ,28.85,24.97,23.87,23.25,22.86,18.42,17.16,14.37,12.63,11.38.

[0096] N3-J16 (yellow solid, yield 54.7%): 1H NMR (500 MHz, DMSO-d6) δ 7.95 (s, 1H), 7.18 (t, J = 7.6 Hz, 3H), 6.91 (d, J = 7.2 Hz, 3H), 6.80 (d, J = 7.8 Hz, 3H), 5.10 (dd, J = 13.3, 5.1 Hz, 3H), 4.21 (d, J = 16.9 Hz, 3H), 2.88 (s, 4H), 2.72 (s, 3H), 2.61 (ddd, J = 17.7, 4.7, 2.4 Hz, 3H), 2.30 (qd, J = 13.3, 4.5 Hz, 3H), 2.02 (dtd, J = 12.6, 5.2, 2.2 Hz, 3H). 13C NMR (125MHz, DMSO) δ174.65,173.39,171.72,170.46,169.38,169.35,162. 80,160.98,144.13,138.90,134.87,132.71,132.41,129.30,126.07,123. 51,116.89,110.86,110.18,54.81,51.99,47.63,46.05,43.38,40.56,39.47,37.83,36.26,33.47,31.71,31.25,29.23,25.37,23.24,21.85,20.58.

[0097] D. Preparation of intermediates NJ7 to NJ16

[0098]

[0099] N3-NJ7 (50 mg, 0.0094 mmol, 1 equivalent) was weighed and placed in a 25 mL round-bottom flask. Anhydrous EtOH (5 mL) was added and stirred thoroughly. 80% hydrazine hydrate (600 μl) was then added. The reaction was heated to 100°C and stirred for 2 h before termination. Post-treatment: 1 M HCl was added to adjust the pH to neutral. The solvent was then evaporated to obtain NJ7 as a white solid (98.5% yield).

[0100] Intermediates NJ8-NJ16 were synthesized using the same deprotection method as NJ7. Intermediates NJ8-NJ18 were obtained by deprotecting the corresponding intermediates N3 of the same chain length using the same method as NJ7. NJ8: White solid, 97.5% yield. NJ9: White solid, 98.6% yield. NJ10: White solid, 97.8% yield. NJ11: White solid, 98.4% yield. NJ12: White solid, 99.7% yield. NJ13: White solid, 99.3% yield. NJ14: White solid, 96.9% yield. NJ15: White solid, 99.5% yield. NJ16: White solid, 98.5% yield.

[0101] (2) Preparation of final products Q7 to Q16

[0102] Q (27 mg, 0.06 mmol, 1.5 equiv) was added to a 25 mL flask with DMF (3 mL). After stirring for several minutes, DIPEA (35 μl, 0.186 mmol, 3 equiv) and HATU (28 mg, 0.014 mol, 1.2 equiv) were added sequentially. The mixture was stirred for approximately 20 minutes and analyzed by TIC (developing solvent: DCM:MeOH = 20:1). After the active ester was formed, NJ7 (40 mg, 0.09 mmol, 1.5 equiv) was added. The reaction was allowed to proceed at room temperature for 12 hours. TIC analysis (developing solvent: dichloromethane:methanol = 20:1) confirmed complete reaction. The reaction mixture was quenched by adding crushed ice and thoroughly extracted with DCM. The organic layer was washed sequentially with saturated ammonium chloride solution and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield a yellow liquid. Purification by PTLC gave the final product, Q7, as a white solid in a 51.7% yield.

[0103] Final product Q7 (white solid, yield 50.7%): 1H NMR (500 MHz, Chloroform-d) δ 9.36 (s, 1H), 8.97 (s, 1H), 7.89 (dd, J = 8.0, 1.1 Hz, 1H), 7.59 (t, J = 7.1 Hz, 1H), 7.56 (dd, J = 8.5, 1.2 Hz, 1H), 7.22 (t, J = 8.2 Hz, 1H), 5.49 (ddd, J = 8.1, 3.8, 2.6 Hz, 1H), 4.94(dd,J=3.7,2.5Hz,1H),4.84(tt,J=6.8,6.1Hz,1H),4.49(t,J=8.4Hz,1H),4.24(d,J=24.5H z,2H),3.34(qd,J=6.8,4.7Hz,2H),2.89(qd,J=6.8,5.2Hz,2H),2.75(qd,J=6.4,0.9Hz,2H),2.6 9–2.63(m,2H),2.63–2.52(m,2H),2.52–2.43(m,2H),2.30(t,J=10.1Hz,2H),2.16–2.10(m,1H), 2.10–2.05(m,2H),2.05–2.03(m,1H),2.03–2.00(m,1H),2.00–1.95(m,1H),1.94–1.86(m,1H),1 .85–1.80(m,1H),1.80–1.74(m,2H),1.74–1.66(m,1H),1.65–1.62(m,1H),1.61–1.51(m,2H),1. 51–1.44(m,1H),1.36(s,3H),1.35–1.30(m,1H),0.95(d,J=8.3Hz,3H),0.90(d,J=7.3Hz,3H).13C NMR(125MHz,Common NMR Solvents)δ172.74,172.06,171.78,171.60,169.90,168.45,139.63,129 .08,127.07,126.10,125.08,117.83,104.72,96.48,93.06,81.11,54.53, 48.18,46.83,46.28,45.55,42.03,41.97,38.98,36.48,35.76,35.61,33. 63,31.94,30.38,29.74,27.95,26.48,24.67,24.24,24.20,19.44,13.31.

[0104] The synthesis of Q8 to Q16 is similar to that of compound Q7, with only the intermediate connecting chains being different, corresponding to NJ8 to NJ16, respectively. Their specific characterization data are as follows:

[0105] Final product Q8 (white solid, yield 51.5%): 1H NMR (500 MHz, Chloroform-d) δ 9.36 (s, 1H), 8.97 (s, 1H), 7.89 (dd, J = 8.1, 1.1 Hz, 1H), 7.65 (t, J = 6.8 Hz, 1H), 7.56 (dd, J = 8.5, 1.2 Hz, 1H), 7.22 (t, J = 8.2 Hz, 1H), 5.49 (ddd, J = 8.1, 3.8, 2.6 Hz, 1H), 4 .94(dd,J=3.7,2.5Hz,1H),4.90(q,J=6.4Hz,1H),4.49(t,J=8.4Hz,1H),4.24(d,J=24.5Hz,2H),3 .22–3.02(m,2H),2.75–2.70(m,2H),2.70–2.66(m,2H),2.66–2.62(m,2H),2.62–2.53(m,2H),2.5 3–2.43(m,2H),2.30(t,J=10.1Hz,2H),2.15–2.10(m,1H),2.10–2.05(m,2H),2.05–2.03(m,1H),2 .03–2.00(m,1H),2.00–1.94(m,1H),1.93–1.87(m,1H),1.86–1.81(m,1H),1.80(d,J=3.2Hz,1H), 1.79–1.77(m,2H),1.77–1.72(m,1H),1.72–1.65(m,1H),1.65–1.62(m,1H),1.62–1.50(m,2H),1. 50–1.45(m,1H),1.36(s,3H),1.36–1.30(m,1H),0.95(d,J=8.3Hz,3H),0.90(d,J=7.3Hz,3H).13C NMR(125MHz,Common NMR Solvents)δ172.88,172.74,172.06,171.60,169.90,168.45,140.03,129.0 8,127.07,126.10,125.08,118.38,104.72,97.27,93.06,81.11,54.53,47. 88,47.28,46.28,45.66,41.79,38.98,38.46,36.48,35.45,35.42,33.63,31.94,30.59,29.74,27.36,27.29,26.48,25.09,24.24,24.20,19.21,13.31.

[0106] Final product Q9 (white solid, yield 54.5%): 1H NMR (500 MHz, Chloroform-d) δ 9.36 (s, 1H), 8.97 (s, 1H), 7.89 (dd, J = 8.1, 1.1 Hz, 1H), 7.63 (t, J = 6.7 Hz, 1H), 7.56 (dd, J = 8.5, 1.2 Hz, 1H), 7.22 (t, J = 8.2 Hz, 1H), 5.49 (ddd, J = 8.1, 3.8, 2.6 Hz, 1H), 4.94 (dd, J = 3.8, 2.5 Hz, 1H), 4.55 (p, J = 6.2Hz, 1H), 4.49 (t, J = 8.4Hz, 1H), 4.24 (d, J = 24.5Hz, 2H), 3.11 (q, J = 6.5Hz, 2H), 2.71 (q, J = 6 .5Hz,2H),2.66(dd,J=10.5,1.3Hz,1H),2.65–2.64(m,1H),2.64–2.63(m,1H),2.63–2.62(m,1H),2.62–2.60( m,1H),2.59–2.54(m,1H),2.53–2.45(m,2H),2.30(t,J=10.1Hz,2H),2.16–2.11(m,1H),2.10–2.05(m,2H),2 .05–2.03(m,1H),2.03–2.00(m,1H),2.00–1.96(m,1H),1.94–1.87(m,1H),1.86–1.81(m,1H),1.81–1.74(m,2 H),1.73–1.67(m,1H),1.66–1.62(m,1H),1.62–1.58(m,1H),1.58–1.53(m,2H),1.53–1.51(m,1H),1.51–1.4 8(m,1H),1.48–1.42(m,2H),1.37(s,3H),1.36–1.30(m,1H),0.95(d,J=8.3Hz,3H),0.90(d,J=7.3Hz,3H).13C NMR(125MHz,Common NMR Solvents)δ172.88,172.85,172.06,171.60,169.58,168.45,139.63,129.08,127.07,126.10,125.08,118.38,104.72,96.48,93.06 ,81.11,54.53,47.88,47.39,47.28,46.28,41.97,39.36,38.98,36.48,35.45,35.42,33.63,31.94,30.59,29.60,28.20,27.29,27.11,26.48,25.09,24.24,24.20,19.03,13.31.

[0107] Final product Q10 (white solid, yield 52.2%): 1H NMR (500 MHz, Chloroform-d) δ 9.36 (s, 1H), 8.97 (s, 1H), 7.89 (dd, J = 8.1, 1.1 Hz, 1H), 7.56 (dd, J = 8.5, 1.2 Hz, 1H), 7.53 (t, J = 6.7 Hz, 1H), 7.22 (t, J = 8.2 Hz, 1H), 5.43 (ddd, J = 8.2, 3.8, 2.6 Hz, 1H), 4.94 (dd, J = 3.8, 2.5 Hz, 1H), 4.72 (p, J = 6.3 Hz,1H),4.49(t,J=8.4Hz,1H),4.24(d,J=24.5Hz,2H),3.06(q,J=6.7Hz,2H),2.71(q,J=6.5Hz,2H),2.69–2.66(m,1H) ,2.66–2.65(m,1H),2.65–2.64(m,1H),2.63(d,J=3.2Hz,1H),2.62–2.59(m,1H),2.56(dd,J=10.4,7.9Hz,1H),2.53–2. 45(m,2H),2.30(t,J=10.1Hz,2H),2.16–2.11(m,1H),2.11–2.05(m,2H),2.05–2.02(m,1H),2.01(ddd,J=6.1,2.9,1.0 Hz,1H),2.00–1.96(m,1H),1.94–1.87(m,1H),1.85–1.80(m,1H),1.80–1.74(m,2H),1.73–1.66(m,1H),1.65–1.62(m,1 H),1.62–1.58(m,1H),1.58–1.54(m,1H),1.54–1.53(m,1H),1.53–1.51(m,1H),1.51–1.49(m,2H),1.49–1.44(m,1H), 1.37(s,3H),1.37–1.35(m,1H),1.35–1.33(m,1H),1.33–1.26(m,1H),0.95(d,J=8.4Hz,3H),0.89(d,J=7.3Hz,3H).13C NMR(125MHz,Common NMR Solvents)δ172.88,172.85,172.06,171.60,169.58,168.16,139.54,129.08,127.07,126.10 ,125.08,118.38,104.72,96.48,93.06,81.11,54.53,47.88,47.29,47.28,46.28,41.97,39.29,38.98,36.48,35.45,35.42,33.63,31.94,30.59,29.60,29.22,28.98,27.29,26.48,26.18,25.09,24.24,24.20,19.03,13.31.

[0108] Final product Q11 (white solid, yield 51.3%): 1H NMR (500 MHz, Chloroform-d) δ 9.36 (s, 1H), 8.97 (s, 1H), 7.89 (dd, J = 8.1, 1.1 Hz, 1H), 7.60–7.57 (m, 1H), 7.57–7.54 (m, 1H), 7.22 (t, J = 8.2 Hz, 1H), 5.43 (ddd, J = 8.2, 3.8, 2.6 Hz, 1H), 4.94 (dd, J = 3.8, 2.5 Hz, 1H), 4.72 (p, J = 6.3 Hz, 1H), 4.49 (t, J = 8. 4Hz,1H),4.24(d,J=24.5Hz,2H),3.06(q,J=6.7Hz,2H),2.71(q,J=6.5Hz,2H),2.68–2.65(m,1H),2.65(d,J=2.3Hz,1H),2. 64–2.63(m,1H),2.62(d,J=1.5Hz,1H),2.62–2.59(m,1H),2.56(dd,J=10.4,7.9Hz,1H),2.48(td,J=11.8,1.5Hz,2H),2.30 (t,J=10.1Hz,2H),2.16–2.10(m,1H),2.10–2.05(m,2H),2.05–2.02(m,1H),2.01(ddd,J=6.1,2.9,1.0Hz,1H),2.00–1.94( m,1H),1.94–1.87(m,1H),1.85–1.80(m,1H),1.80–1.74(m,2H),1.74–1.66(m,1H),1.66–1.62(m,1H),1.62–1.58(m,1H),1 .58–1.54(m,1H),1.54–1.52(m,1H),1.52(q,J=1.7Hz,1H),1.51–1.49(m,2H),1.49–1.45(m,1H),1.42–1.38(m,1H),1.37( s,3H),1.36–1.35(m,1H),1.35(tt,J=2.4,1.3Hz,1H),1.34–1.27(m,1H),0.95(d,J=8.4Hz,3H),0.89(d,J=7.3Hz,3H).13C NMR(125MHz,Common NMR Solvents)δ172.96,172.85,172.06,171.60,169.58,168.16,139.54,129 .08,127.07,126.10,125.08,117.64,104.72,96.48,93.06,81.11,54.53,47.88,47.28,47.17,45.07,42.05,40.10,39.72,36.48,35.45,35.01,33.63,31.94,30.79,29.60,29.18,28.47,28.19,28.05,27.95,26.48,25.09,24.24,24.20,19.03,13.36.

[0109] Final product Q12 (white solid, yield 52.2%): 1H NMR (500 MHz, Chloroform-d) δ 9.36 (s, 1H), 8.97 (s, 1H), 7.89 (dd, J = 8.1, 1.2 Hz, 1H), 7.62 (dd, J = 8.5, 1.2 Hz, 1H), 7.58 (t, J = 6.7 Hz, 1H), 7.24 (t, J = 8.3 Hz, 1H), 5.43 (ddd, J = 8.2, 3.8, 2.6 Hz, 1H), 4.94 (dd, J = 3.8, 2.5 Hz, 1H), 4.72 (p, J = 6.3 Hz, 1H), 4.47 (t, J = 8.4 Hz, 1H), 4.24 (d, J = 24.5 Hz, 2H), 3.07 (q, J = 6.7 Hz, 2H), 2.71 (q, J = 6.5 Hz, 2H), 2.68–2.64 (m, 2H), 2.63 (d, J = 0.9 Hz, 1H), 2.62 (d, J = 1.7 Hz, 1H), 2.62–2.59 (m, 1H), 2.56 (dd, J = 10.4, 7.9 Hz, 1H), 2.48 (td, J = 11.8, 1.5 Hz, 2H), 2.30 (t, J = 10.1 Hz, 2H), 2.18–2.12 (m, 1H), 2.12–2.05 (m, 2H), 2.05–2.02 (m, 1H), 2.02–2.00 (m, 1H), 2.00–1.95 (m, 1H), 1.94–1.87 (m, 1H), 1.85–1.80 (m, 1H), 1.80–1.74 (m, 2H), 1.74–1.66 (m, 1H), 1.66–1.62 (m, 1H), 1.62–1.58 (m, 1H), 1.58–1.54 (m, 2H), 1.52 (dd, J = 3.5, 2.6 Hz, 1H), 1.52–1.49 (m, 2H), 1.49–1.45 (m, 1H), 1.42–1.37 (m, 1H), 1.37 (s, 3H), 1.36–1.35 (m, 1H), 1.35 (dd, J = 2.7, 1.7 Hz, 1H), 1.34–1.32 (m, 2H), 1.32–1.25 (m, 2H), 0.91 (d, J = 8.4 Hz, 3H), 0.89 (d, J = 7.3 Hz, 3H). 13C NMR (125 MHz, Common NMR Solvents) δ 172.96, 172.85, 172.06, 171.60, 169.58, 168.16,17,45.07,42.05,40.10,39.73,36.48,35.45,35.01,33.63,31.94,30.79,29.60,29.18,29.08,28.47,27.95,27.76,27.53,26.48,25.09,24.24,24.20,19.03,13.31.

[0110] Final product Q13 (white solid, yield 51.2%): 1H NMR (500 MHz, Chloroform-d) δ 9.36 (s, 1H), 8.97 (s, 1H), 7.89 (dd, J = 8.1, 1.2 Hz, 1H), 7.62 (dd, J = 8.4, 1.1 Hz, 1H), 7.59 (d, J = 6.6 Hz, 1H), 7.24 (t, J = 8.3 Hz, 1H), 5.43 (ddd, J = 8.2, 3.8, 2.6 Hz, 1H), 4.94 (dd, J = 3.8, 2.5 Hz, 1H), 4.72 (p, J = 6.3 Hz, 1H ),4.47(t,J=8.4Hz,1H),4.24(d,J=24.5Hz,2H),3.07(q,J=6.7Hz,2H),2.71(q,J=6.5Hz,2H),2.68–2.63(m,2H),2.63–2 .62(m,2H),2.62–2.59(m,1H),2.56(dd,J=10.4,7.9Hz,1H),2.48(td,J=11.8,1.5Hz,2H),2.30(t,J=10.1Hz,2H),2.18–2 .12(m,1H),2.12–2.05(m,2H),2.05–2.02(m,1H),2.02–2.00(m,1H),2.00–1.96(m,1H),1.94–1.87(m,1H),1.85–1.80(m ,1H),1.80–1.74(m,2H),1.74–1.66(m,1H),1.65–1.62(m,1H),1.62–1.58(m,1H),1.58–1.53(m,2H),1.53–1.52(m,1H),1 .52–1.49(m,2H),1.49–1.45(m,1H),1.37(s,3H),1.37–1.36(m,1H),1.36–1.34(m,1H),1.34–1.32(m,1H),1.32–1.28(m, 2H),1.28(s,1H),1.27(d,J=1.0Hz,1H),1.27(s,1H),1.26–1.24(m,1H),0.89(d,J=7.3Hz,3H),0.85(d,J=8.4Hz,3H).13C NMR(125MHz,Common NMR Solvents)δ172.96,172.85,172.06,171.60,169.58,168.16,139.54,129.08,127. 07,126.10,125.08,117.64,104.72,96.48,93.06,81.11,53.88,47.88,47.28,47.17,45.07,42.88,40.10,39.75,36.44,35.45,35.01,33.63,31.94,30.79,29.60,29.18,28.52,27.95,27.60,27.53,26.48,25.09,24.24,24.20,19.03,13.05.

[0111] Final product Q14 (white solid, yield 49.6%): 1H NMR (500 MHz, Chloroform-d) δ 9.36 (s, 1H), 8.97 (s, 1H), 7.89 (dd, J = 8.1, 1.2 Hz, 1H), 7.64–7.61 (m, 1H), 7.59 (d, J = 6.6 Hz, 1H), 7.33–7.10 (m, 1H), 5.43 (ddd, J = 8.2, 3.8, 2.6 Hz, 1H), 4.94 (dd, J = 3.8, 2.5 Hz, 1H), 4.72 (p, J = 6.3 Hz, 1H), 4.47 (t, J = 8.4 Hz, 1H), 4.24 (d, J = 24.5 Hz, 2H), 3.07 (q, J = 6.6 Hz, 2H), 2.71 (q, J = 6.5 Hz, 2H), 2.68–2.63 (m, 2H), 2.63–2.62 (m, 2H), 2.62–2.59 (m, 1H), 2.56 (dd, J = 10.5, 7.9 Hz, 1H), 2.48 (td, J = 11.9, 1.6 Hz, 2H), 2.30 (t, J = 10.1 Hz, 2H), 2.18–2.12 (m, 1H), 2.12–2.05 (m, 2H), 2.05–2.02 (m, 1H), 2.02–2.00 (m, 1H), 2.00–1.95 (m, 1H), 1.94–1.86 (m, 1H), 1.85–1.80 (m, 1H), 1.80–1.74 (m, 2H), 1.73–1.67 (m, 1H), 1.66–1.63 (m, 1H), 1.62–1.57 (m, 1H), 1.57–1.54 (m, 2H), 1.53 (dd, J = 4.0, 1.0 Hz, 1H), 1.52–1.51 (m, 1H), 1.51–1.46 (m, 2H), 1.41–1.37 (m, 1H), 1.37 (s, 3H), 1.36–1.35 (m, 1H), 1.35–1.33 (m, 1H), 1.33–1.29 (m, 2H), 1.29–1.26 (m, 2H), 1.26 (dt, J = 1.7, 1.0 Hz, 2H), 1.25–1.21 (m, 2H), 0.89 (d, J = 7.3 Hz, 3H), 0.85 (d, J = 8.4 Hz, 3H). 13C NMR (125 MHz, Common NMR Solvents) δ 172.96, 172.85, 172.06, 171.73, 169.58, 168.16, 139.6, 129.08, 127.07, 126.1, 125.08, 117.64, 104.72, 96.48, 93.06, 81.11, 53.88, 47.%, 47.28, 47.17,45.07,43.29,40.10,39.73,36.44,35.45,34.78,33.63,31.94,30.79,29.75,29.60,29.48,29.19,29.19,28.34,27.95,27.60,27.53,26.45,25.09,24.24,24.20,19.03,13.05.

[0112] Final product Q15 (white solid, yield 50.6%): 1H NMR (500 MHz, Chloroform-d) δ 9.58 (s, 1H), 8.95 (s, 1H), 7.75 (dd, J = 8.1, 1.2 Hz, 1H), 7.56 (dd, J = 8.2, 1.1 Hz, 1H), 7.42 (t, J = 8.2 Hz, 1H), 6.78 (t, J = 4.9 Hz, 1H), 5.77 (ddd, J = 7.7, 3.8, 2.5 Hz, 1H), 5.19 (dd, J = 3.7, 2.5 Hz, 1H), 4.62 (t, J = 5.8 Hz, 1H), 4.44 (d, J=12.3Hz,1H),4.31(d,J=12.4Hz,1H),3.12(tdd,J=5.7,4.8,1.1Hz,2H),2.80–2.74(m,2H),2.74–2.70(m,2H),2.62–2.57(m, 1H),2.57–2.54(m,2H),2.54–2.52(m,2H),2.52–2.47(m,1H),2.45(p,J=5.0Hz,1H),2.31(t,J=8.2Hz,2H),2.28–2.22(m,1H), 2.22–2.18(m,1H),2.18–2.15(m,1H),2.15–2.09(m,1H),2.09–2.00(m,1H),1.89–1.86(m,1H),1.86–1.85(m,1H),1.84(d,J=1 .1Hz,1H),1.84(d,J=2.5Hz,1H),1.83(d,J=1.2Hz,1H),1.83–1.78(m,1H),1.74–1.69(m,1H),1.69–1.66(m,1H),1.66–1.58(m ,1H),1.54–1.48(m,2H),1.48–1.44(m,2H),1.44–1.41(m,1H),1.40(s,3H),1.34–1.29(m,2H),1.29(d,J=0.7Hz,2H),1.28(d, J=2.2Hz,2H),1.28(d,J=0.9Hz,2H),1.27(t,J=1.1Hz,4H),1.26–1.22(m,1H),0.94(d,J=7.8Hz,3H), 0.90(d,J=7.1Hz,3H).13C NMR (125MHz, Common NMR Solvents) δ172.84,172.50,172.09,170.34,169.63,167.54,138.89,132.38,131.99,126.20,122.95,117.29,104.27,98.90,89.60,81.72,56.42,50.66,49.23,48.51,45.96,45.78,43.07,37.14,36.72,36.04,34.40,33.44,31.58,31.40,31.06,30.73,29.47,29.40,29.26,29.16,28.24,27.27,26.01,25.39,24.64,24.48,24.27,21.73,20.38,14.32.

[0113] Final product Q16 (white solid, yield 51.6%): 1H NMR (500 MHz, Chloroform-d) δ 9.36 (s, 1H), 8.97 (s, 1H), 7.89 (dd, J = 8.0, 1.1 Hz, 1H), 7.64–7.61 (m, 1H), 7.59 (d, J = 6.6 Hz, 1H), 7.38–7.09 (m, 1H), 5.44 (ddd, J = 8.2, 3.8, 2.5 Hz, 1H), 4.94 (dd, J = 3.8, 2.5 Hz, 1H), 4.72 (p, J = 6.3 Hz, 1H), 4.47 (t, J = 8.4 Hz, 1H), 4.24 (d, J = 24.5 Hz, 2H), 3.07 (q, J = 6.6 Hz, 2H), 2.71 (q, J = 6.5 Hz, 2H), 2.68–2.63 (m, 2H), 2.63–2.62 (m, 2H), 2.62–2.59 (m, 1H), 2.56 (dd, J = 10.5, 7.9 Hz, 1H), 2.48 (td, J = 11.9, 1.6 Hz, 2H), 2.30 (t, J = 10.1 Hz, 2H), 2.17–2.11 (m, 1H), 2.11–2.05 (m, 2H), 2.05–2.02 (m, 1H), 2.02–2.00 (m, 1H), 2.00–1.95 (m, 1H), 1.94–1.86 (m, 1H), 1.85–1.80 (m, 1H), 1.80–1.74 (m, 2H), 1.74–1.67 (m, 1H), 1.67–1.63 (m, 1H), 1.63–1.59 (m, 1H), 1.59–1.54 (m, 2H), 1.53 (d, J = 2.8 Hz, 1H), 1.52–1.51 (m, 1H), 1.51–1.47 (m, 2H), 1.37 (s, 3H), 1.37–1.35 (m, 2H), 1.34 (dd, J = 5.6, 2.5 Hz, 1H), 1.33–1.29 (m, 2H), 1.29–1.26 (m, 2H), 1.26–1.25 (m, 1H), 1.25 (d, J = 1.5 Hz, 1H), 1.25 (s, 1H), 1.24 (d, J = 0.9 Hz, 1H), 1.24 (s, 1H), 1.24–1.20 (m, 2H), 0.90 (d, J = 6.4 Hz, 3H), 0.89 (d, J = 5.4 Hz, 3H). 13C NMR (125 MHz, Common NMR Solvents) δ 172.96, 172.85, 172.06, 171.73, 169.58, 168.16, 140.03, 128.30, 127.07, 126.10, 125.08, 117.64, 104.72, 96.48,93.06,81.11,53.88,47.28,47.17,47.16,45.07,44.58,39.75,39.52,36.44,35.45,34.78,33.63,31.94,30.79,29.75,29.57,29.33,29.25,29.19,28.52,27.60,27.53,27.29,26.32,25.24,24.24,24.20,19.03,13.05.

[0114] Example 4:

[0115] (1) Preparation of intermediates p1 to p2

[0116] The structural formulas of intermediates p1 (n=2) and p2 (n=3) are The specific synthesis method is as follows: Fluorothalidamide (2g, 7.241mmol, 1eq) is dissolved in 20mL of DMF and stirred to dissolve. DIEA (1.872mL, 14.481mmol, 2eq) and N-Boc-2,2'-(ethylenedioxy)diethylamine (2mL, 8.689mmol, 1.2eq) are then added and the mixture is heated to 90°C and refluxed for 12h. After completion, the reaction is quenched with ice water and extracted with ethyl acetate. The combined organic phases are washed with saturated NaHCO3 and saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography (PE:EA = 1:1) to obtain F-polyethylene glycol (intermediate p1, a yellow transparent oil, 228.6mg, 14% yield).

[0117] The synthesis of intermediate p2 is similar to that of p1, except that the starting material is tert-butyl (2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)carbamate. Intermediate P2 is a yellow transparent oil with a yield of 18%).

[0118] (2) Preparation of final products Q17-Q18:

[0119] Preparation of the final product Q17: To a 25 mL round-bottom flask, artesunate (100 mg, 0.26 mmol), linker and E3 ligase ligand p1 (118 mg, 0.51 mol), HOBT (0.51 mol), 4-dimethylaminopyridine (DMAP) (0.26 mol), and DCM (5 mL) were added. The reaction was stirred at room temperature for 5-8 h, extracted with DCM, and the organic phase was washed with 5% NaHCO₃ solution and saturated sodium chloride, then dried over anhydrous Na₂SO₄. The solid was removed by filtration, and the solvent was evaporated under reduced pressure to obtain the crude product, which was then purified by column chromatography (PE-EA = 10:1 to 1:1, v / v).

[0120] Final product Q17 (fluorescent yellow powder, yield 61.4%): 1H NMR (500 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.96 (t, J = 5.6 Hz, 1H), 7.58 (dd, J = 8.6, 7.1 Hz, 1H), 7.14 (d, J = 8.6 Hz, 1H), 7.04 (d, J = 7.0 Hz, 1H), 6.61 (t, J = 5.8 Hz, 1H), 5.70 (d, J = 55.1 Hz, 1H), 5.5 3(s,1H),5.06(dd,J=12.7,5.4Hz,1H),3.62(t,J=5.4Hz,2H),3.57(dd,J=6.2,3.6Hz,2H),3. 53–3.49(m,2H),3.47(q,J=5.6Hz,2H),3.39(t,J=5.9Hz,2H),3.18(q,J=5.9Hz,2H),2.88(dd d,J=16.9,13.7,5.4Hz,1H),2.65–2.57(m,2H),2.57–2.51(m,2H),2.38(dt,J=6.9,3.3Hz,2H ),2.30–2.12(m,2H),2.00(dddd,J=17.7,14.5,5.9,3.7Hz,2H),1.79(dddd,J=11.7,5.7,2.5H z,1H),1.66–1.56(m,2H),1.52(dt,J=13.7,4.2Hz,1H),1.47–1.38(m,2H),1.28(s,3H),1.26 –1.19(m,2H),1.15(td,J=11.5,6.6Hz,1H),0.87(d,J=6.4Hz,3H),0.75(d,J=7.1Hz,3H).13C NMR (126MHz, DMSO-d6) δ172.85,171.26,170.61,170.12,168.97,167.32,146. 42,136.27,132.12,117.47,110.71,109.27,103.60,91.63,90.60,79.88,69.7 1,69.62,69.17,68.90,54.94,51.13,48.58,44.59,41.72,38.62,36.00,35.91,33.72,31.65,31.01,29.46,28.95,25.54,24.21,22.17,21.04,20.08,11.74.

[0121] Preparation of the final product Q18: To a 25 mL round-bottom flask were added artesunate (100 mg, 0.26 mmol), linker and E3 ligase ligand p2 (141 mg, 0.51 mol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCl) (0.51 mol), 4-dimethylaminopyridine (DMAP) (0.26 mol), and DCM (5 mL). The reaction was stirred at room temperature for 5-8 h, extracted with DCM, and the organic phase was washed with 5% NaHCO₃ solution and saturated sodium chloride, then dried over anhydrous Na₂SO₄. The solid was removed by filtration, and the solvent was evaporated under reduced pressure to obtain the crude product, which was then purified by column chromatography (PE-EA = 10:1 to 1:1, v / v).

[0122] Final product Q18 (fluorescent yellow powder, yield: 61.2%): 1H NMR (500 MHz, DMSO-d6) δ11.10 (s, 1H), 8.04–7.83 (m, 1H), 7.59 (dd, J = 8.6, 7.1 Hz, 1H), 7.15 (d, J = 8.6 Hz, 1H), 7.04 (d, J = 7.1 Hz, 1H), 6.61 (t, J = 5.8 Hz, 1H), 5.70 (d, J = 53.6 Hz, 1H), 5.63 (s, 1H), 5.06 (dd, J = 12.8, 5.4 Hz,1H),4.24(dd,J=5.6,3.7Hz,1H),3.62(t,J=5.4Hz,2H),3.57(dd,J=5.7,2.9Hz,2H),3.55–3.53(m,2H) ,3.52(t,J=2.9Hz,2H),3.50–3.48(m,2H),3.47(dd,J=7.0,4.2Hz,2H),3.18(q,J=5.8Hz,2H),2.96–2.82( m,1H),2.63–2.59(m,1H),2.57(dd,J=6.9,4.7Hz,2H),2.43–2.35(m,2H),2.27(ddd,J=9.7,7.1,4.3Hz,1H ),2.17(td,J=14.0,3.9Hz,1H),2.06–1.97(m,2H),1.80(dq,J=10.1,3.3Hz,1H),1.60(td,J=14.0,13.5,3 .4Hz,2H),1.53(dt,J=13.6,4.2Hz,1H),1.48–1.40(m,2H),1.40–1.36(m,1H),1.28(s,3H),1.24(d,J=12. 1Hz,2H),1.17(dd,J=11.4,6.5Hz,1H),0.96–0.91(m,1H),0.88(d,J=6.5Hz,3H),0.75(d,J=7.1Hz,3H).13C NMR(126MHz,DMSO-d6)δ172.87,171.26,170.62,170.12,146.44,132.12,12 9.53,117.50,110.72,109.26,103.60,91.64,90.60,79.89,69.82,69.80,6 9.62,69.11,68.91,54.95,51.14,48.58,44.60,41.72,38.63,36.00,35.92,33.72,31.66,31.00,29.47,28.96,25.54,24.21,22.43,22.17,21.04,20.09,11.74,10.94.

[0123] Example 5:

[0124] (1) Preparation of intermediates w2-8

[0125] The structural formula of intermediate w2(n=1) / w3(n=2) / w4(n=3) / w5(n=4) / w6(n=5) / w7(n=6) / w8(n=7) is The specific synthesis method is as follows:

[0126] Fluorothalidamide (2 g, 7.241 mmol) was dissolved in 20 mL of DMF with stirring. DIEA (1.872 mL, 14.481 mmol) and N-Boc-1,2-ethylenediamine (2 mL, 8.689 mmol) were then added and the mixture was heated to 90°C and refluxed for 12 h. After completion, the reaction was quenched with ice water and extracted with ethyl acetate. The combined organic phases were washed sequentially with saturated NaHCO₃ and saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography (PE:EA = 1:1) to afford w2 (a yellow, transparent oil, 1.3533 g, 60% yield).

[0127] The synthesis of w3-w8 was similar to that of w2, except that N-Boc-1,2-ethylenediamine was replaced by N-Boc-1,3-propylenediamine (2 eq), N-Boc-1,4-butylenediamine (2 eq), N-Boc-1,6-hexanediamine (2 eq), N-Boc-1,7-heptanediamine (2 eq), and N-Boc-1,8-octanediamine (2 eq).

[0128] (2) Preparation of final product Q19-25

[0129] A 25 mL round-bottom flask was charged with artesunate (100 mg, 0.26 mmol), linker and E3 ligase ligands w2 / w3 / w4 / w5 / w6 / w7 / w8 (2 eqq) (360.37 g) (0.51 mol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCl) (0.51 mol), 4-dimethylaminopyridine (DMAP) (0.26 mol), and DCM (5 mL). The reaction was stirred at room temperature for 5-8 h, extracted with DCM, and the organic phase was washed with 5% NaHCO₃ solution and saturated sodium chloride, then dried over anhydrous Na₂SO₄. The solid was removed by filtration, and the solvent was evaporated under reduced pressure to obtain the crude product, which was then purified by column chromatography (PE-EA = 10:1 to 1:1, v / v) to obtain the final products Q19 / 20 / 21 / 22 / 23 / 24 / 25. Their NMR data are as follows:

[0130] Final product Q19 (fluorescent yellow powder, yield 63.6%): 1H NMR (500 MHz, Chloroform-d) δ 8.65 (d, J = 12.9 Hz, 1H), 7.50 (t, J = 7.8 Hz, 1H), 7.09 (d, J = 7.1 Hz, 1H), 6.98 (d, J = 8.7 Hz, 1H), 6.52 (d, J = 5.8 Hz, 1H), 6.42 (d, J = 5.5 Hz, 1H), 5.75 (d, J = 9.7 Hz, 1H), 5.44 (s, 1H), 4 .93(dd,J=12.0,5.4Hz,1H),3.60–3.44(m,2H),3.43(d,J=5.7Hz,2H),2.91–2.78(m,2H),2.76(dd,J= 8.2,4.0Hz,2H),2.72(dt,J=9.2,2.7Hz,2H),2.59–2.53(m,1H),2.52–2.49(m,1H),2.48–2.44(m,1H), 2.44–2.36(m,1H),2.34(dd,J=14.0,4.0Hz,1H),2.14–2.10(m,1H),2.06–2.02(m,1H),2.00(dd,J=4. 9,2.9Hz,1H),1.89(dd,J=6.8,3.5Hz,1H),1.86(dt,J=6.6,3.6Hz,1H),1.75(dd,J=9.0,4.8Hz,1H),1. 73–1.70(m,1H),1.68(d,J=3.4Hz,1H),1.61(dd,J=9.0,4.6Hz,1H),1.59–1.52(m,1H),1.52–1.45(m, 1H),1.40(s,3H),1.28(d,J=6.4Hz,1H),1.24(s,2H),0.94(d,J=6.0Hz,3H),0.82(d,J=7.1Hz,3H).13C NMR (126MHz, CDCl3) δ172.22,171.84,171.54,169.48,168.82,167.62,146.79,136.33,133.47,117.52,111.87,110.25,104.53,92.84,91.49 ,80.18,51.51,50.20,48.45,45.18,42.02,38.97,37.24,35.62,33.58 ,31.74,31.45,30.79,29.79,27.12,24.57,22.74,21.95,20.21,12.05.

[0131] Final product Q20 (fluorescent yellow powder, yield 61.6%): 1H NMR (500 MHz, Chloroform-d) δ 8.58 (d, J = 28.8 Hz, 1H), 7.48 (dd, J = 8.6, 7.1 Hz, 1H), 7.07 (d, J = 7.1 Hz, 1H), 6.89 (dd, J = 8.6, 2.3 Hz, 1H), 6.40 (s, 1H), 6.22 (d, J = 5.9 Hz, 1H), 5.78 (d, J=9.8Hz,1H),5.74(dd,J=9.9,1.9Hz,1H),5.43(s,1H),5.41(d,J=12.3Hz,1H),4.97–4. 86(m,1H),3.50–3.34(m,2H),3.32(d,J=6.6Hz,2H),2.91–2.79(m,2H),2.79–2.75(m,2H) ,2.74–2.71(m,2H),2.58–2.54(m,1H),2.54–2.48(m,2H),2.46(dd,J=7.5,2.2Hz,1H),2 .35(ddt,J=13.9,9.2,4.6Hz,2H),2.16–2.08(m,1H),2.02(ddt,J=14.4,5.1,2.6Hz,2H), 1.84(dt,J=9.4,3.3Hz,2H),1.75(ddt,J=13.1,9.6,3.4Hz,2H),1.71–1.65(m,2H),1.60( dt,J=14.7,5.1Hz,2H),1.40(s,3H),1.24(s,2H),0.94(s,3H),0.83(d,J=2.2Hz,3H).13C NMR (126MHz, CDCl3) δ172.09,172.02,171.61,169.56,168.77,167.77,148.15,138.18,132.03,120.97,113.17,110.56,104.63,93.58,91 .56,81.22,51.58,50.87,49.00,45.26,40.38,37.36,36.27,34.14,3 1.80,31.15,30.03,29.22,25.98,24.65,22.87,22.04,20.29,12.16.

[0132] Final product Q21 (fluorescent yellow powder, yield 60.6%): 1H NMR (500MHz, DMSO-d6) δ11.11(s,1H),7.97(t,J=5.7Hz,1H),7.58(dd,J=8.6,7.0H z,1H),7.11(d,J=8.6Hz,1H),7.02(d,J=7.0Hz,1H),6.55(t,J=6.0Hz,1H),5.64(d ,J=9.8Hz,1H),5.56(d,J=21.7Hz,1H),5.05(dd,J=12.9,5.5Hz,1H),4.16(q,J=5. 2Hz,2H),3.30(q,J=6.7Hz,2H),3.17(s,2H),3.16(s,2H),3.07(q,J=6.5Hz,2H),2 .95–2.86(m,1H),2.61–2.57(m,2H),2.38(t,J=7.2Hz,2H),2.27(ddd,J=11.0,7.1 ,4.3Hz,1H),2.18(td,J=14.0,3.9Hz,1H),2.02(ddt,J=16.1,13.3,7.7Hz,2H),1. 86–1.75(m,1H),1.65–1.58(m,2H),1.57–1.53(m,2H),1.53–1.46(m,2H),1.42(dd ,J=15.7,5.7Hz,2H),1.28(s,3H),0.88(d,J=6.3Hz,3H),0.75(d,J=7.1Hz,3H).13C NMR(126MHz,DMSO-d6)δ172.83,171.23,170.34,170.10,168.94,167.32,1 46.41,136.29,117.25,110.40,109.00,103.58,91.60,90.58,79.87,54.95 ,51.12,48.59,48.54,44.57,41.55,38.18,35.98,35.90,33.71,31.64,31.00,29.58,29.05,26.50,26.17,25.53,24.20,22.18,21.02,20.07,11.73.

[0133] Final product Q22 (fluorescent yellow powder, yield 59.6%): 1H NMR(500MHz,Chloroform-d)δ8.46(s,1H),7.51–7.45(m,1H),7.08(t,J=6.6Hz,1H ),6.87(dd,J=8.6,2.8Hz,1H),6.23(s,1H),5.91(q,J=5.0Hz,1H),5.74(dd,J=9.8 ,3.6Hz,1H),5.47–5.29(m,1H),4.92(dd,J=12.1,5.4Hz,1H),3.41–3.26(m,2H),3 .25(dd,J=6.4,3.1Hz,2H),2.96–2.78(m,2H),2.78–2.75(m,2H),2.75–2.58(m,2H) ,2.58–2.48(m,2H),2.46(dt,J=7.1,3.5Hz,1H),2.35(td,J=13.9,3.9Hz,1H),2.1 2(dt,J=14.6,4.7Hz,1H),2.01(dt,J=14.6,4.2Hz,2H),1.88(ddq,J=13.4,6.5,3.7 Hz,2H),1.79–1.73(m,1H),1.71–1.67(m,2H),1.53(q,J=7.3Hz,2H),1.43–1.41(m ,3H),1.25(d,J=4.6Hz,2H),0.94(dd,J=6.0,3.5Hz,3H),0.84(t,J=6.1Hz,3H).13C NMR (126MHz, CDCl3) δ172.12,171.52,171.42,169.72,168.67,167.77,147 .06,136.29,132.60,116.84,111.58,110.06,104.64,92.36,91.56,80.26 ,51.63,49.02,45.29,42.40,39.34,37.35,36.31,34.18,31.83,31.55,31.05,29.98,29.18,28.64,26.02,24.68,24.07,22.91,22.07,20.72,11.04.

[0134] Final product Q23 (fluorescent yellow powder, yield 63.2%): 1H NMR (500 MHz, DMSO-d6) δ 7.91 (q, J = 5.6, 4.8 Hz, 1H), 7.57 (t, J = 7.8 Hz, 1H), 7.07 (d, J = 8.7 Hz, 1H), 7.00 (d, J = 7.0 Hz, 1H), 6.50 (t, J = 5.9 Hz, 1H), 5.72 (s, 1H), 5.61 (d, J = 9. 7Hz,1H),5.47(s,1H),5.02(dd,J=12.9,5.5Hz,1H),3.28–3.23(m,2H),3.01(q,J=6 .5Hz,2H),2.94–2.64(m,2H),2.63–2.57(m,2H),2.56(d,J=5.9Hz,2H),2.35(t,J=7. 1Hz,2H),2.31–2.15(m,2H),2.13(dd,J=14.0,4.1Hz,1H),1.99(tt,J=10.7,6.0Hz, 2H),1.77(td,J=6.4,3.2Hz,1H),1.60–1.57(m,1H),1.54(t,J=7.5Hz,3H),1.52–1.4 5(m,2H),1.39(d,J=5.7Hz,1H),1.36(d,J=6.2Hz,2H),1.34(s,2H),1.32–1.30(m,2 H),1.29(s,1H),1.22–1.08(m,2H),0.84(d,J=6.4Hz,3H),0.72(d,J=7.2Hz,3H).13C NMR(126MHz,DMSO-d6)δ173.14,171.51,170.70,170.32,169.17,167.57,146 .64,136.59,132.34,117.43,110.66,109.12,103.84,91.83,90.80,80.07,5 5.06,51.25,48.74,44.72,41.94,38.67,36.24,36.06,33.83,31.80,31.15,29.75,29.21,28.80,26.23,26.20,25.69,24.36,22.37,21.23,20.23,11.90.

[0135] Final product Q24 (fluorescent yellow powder, yield 65.2%): 1H NMR (500 MHz, Chloroform-d) δ9.38 (s, 1H), 8.51 (t, J = 5.9 Hz, 1H), 7.90 (dd, J = 7.7, 1.1 Hz, 1H), 7.58 (t, J = 6.7 Hz, 1H), 7.41 (t, J = 7.8 Hz, 1H), 7.18 (dd, J = 8.1, 1.3 Hz, 1H), 5.51–5.49 (m, 1H), 5.49–5.46 (m ,1H),4.94(dd,J=3.7,2.5Hz,1H),3.30(q,J=6.2Hz,2H),3.06(q,J=6.6Hz,2H),2.71–2.63(m,2H),2 .63–2.59(m,2H),2.55–2.46(m,2H),2.20(ddd,J=9.2,7.9,6.4Hz,1H),2.15–2.10(m,1H),2.10(d,J =2.0Hz,1H),2.09–2.07(m,1H),2.07(d,J=7.7Hz,1H),2.05–2.01(m,1H),2.01–1.95(m,1H),1.81(d ddd,J=13.0,9.5,8.2,7.0Hz,1H),1.75–1.70(m,1H),1.70–1.65(m,2H),1.65–1.62(m,1H),1.62–1. 53(m,2H),1.53–1.49(m,2H),1.49–1.45(m,1H),1.41–1.36(m,2H),1.36(s,3H),1.35(d,J=2.3Hz,1 H),1.34(dd,J=3.8,2.1Hz,2H),1.33–1.26(m,2H),0.95(d,J=8.3Hz,3H),0.90(d,J=7.3Hz,3H).13C NMR(125MHz,Common NMR Solvents)δ172.49,172.22,172.06,170.14,167.28,162.24,141.32,1 28.11,125.77,124.01,120.35,117.95,104.72,96.48,93.06,81.11,52.77 ,48.18,43.53,41.97,39.15,38.98,35.76,35.61,33.63,32.03,30.38,29.91,29.74,29.08,28.34,27.41,26.85,24.67,24.24,24.20,19.44,13.31.

[0136] Final product Q25 (fluorescent yellow powder, yield 62.3%): 1H NMR (500 MHz, Chloroform-d) δ 8.61 (s, 1H), 7.47 (dd, J = 8.6, 7.1 Hz, 1H), 7.05 (d, J = 7.1 Hz, 1H), 6.86 (d, J = 8.5 Hz, 1H), 6.22 (d, J = 5.7 Hz, 1H), 5.88 (t, J = 5.7 Hz, 1H), 5.74 (d, J = 9.9 Hz, 1H) ,5.42(d,J=2.2Hz,1H),4.91(dd,J=12.1,5.5Hz,1H),3.24(t,J=6.0Hz,2H),3.21–3.16(m,2 H),2.96–2.80(m,2H),2.81–2.75(m,2H),2.75–2.68(m,2H),2.58–2.47(m,2H),2.46–2.32( m,2H),2.13–2.08(m,1H),2.03–1.98(m,1H),1.86(dq,J=10.0,3.1Hz,1H),1.78–1.67(m,2H ),1.64(dd,J=16.0,8.6Hz,2H),1.60–1.55(m,1H),1.47–1.44(m,2H),1.40(s,3H),1.37(dd ,J=9.0,3.6Hz,2H),1.33(dd,J=6.6,2.6Hz,2H),1.31–1.29(m,2H),1.28(d,J=4.3Hz,2H),1 .26(d,J=4.6Hz,1H),1.23(t,J=3.7Hz,2H),0.93(d,J=6.1Hz,3H),0.82(d,J=7.1Hz,3H).13C NMR(126MHz,Chloroform-d)δ172.05,172.04,171.47,171.45,171.34,169.62,168.71, 168.69,167.76,147.07,136.20,132.56,116.76,111.41,109.89,104.56,92.28,91.54, 80.21,51.59,48.94,45.26,42.64,39.69,37.32,36.27,34.14,31.82,31.49,30.97,29.89,29.54,29.53,29.13,29.11,26.80,26.75,26.00,24.64,22.87,22.03,20.29,12.14.

[0137] Example 6: In vitro anti-tumor activity test (IC50) of artemisinin PROTACs of the present invention

[0138] The artemisinin PROTACs prepared in Examples 2 to 5 of the present invention were tested for their ability to inhibit the proliferation of three tumor cells, using the conventional CCK-8 method. When the tumor cells MCF-7 cells (human breast cancer cells), A549 (human lung cancer cells), and Hela cells (cervical cancer cells) grew to 80% to 90% of the culture dish, the cells were digested from the culture dish, centrifuged and resuspended in fresh DMEM / 1640 complete medium, counted under a microscope, and then the cell suspension was diluted to 50 cells / μL. A circle of sterile PBS was spread on the outermost periphery of the 96-well plate, 100 μL per well, and 100 μL of the diluted cell suspension was spread on each well of the remaining wells and cultured in a 37°C, 5% CO2 incubator. After 24 hours, the medium in the 96-well plate was aspirated, and the drug was diluted with fresh complete medium at a certain concentration gradient and then added to the 96-well plate in sequence. After 48 hours, the medium containing the drug was removed, and CCK-8 liquid diluted with medium was added (100 μL medium + 10 μL CCK-8). The cells were incubated in a 37°C, 5% CO2 incubator for 30-40 minutes. The absorbance was measured at OD 450 on a microplate reader, and the inhibition rate was calculated based on the absorbance at OD 450. The data were used to calculate the half-maximal inhibitory concentration (IC50) using SPSS software.

[0139] The calculation formula is as follows:

[0140] Inhibition rate = [(Ac-As) / (Ac-Ab)] × 100%

[0141] As: absorbance of experimental wells (containing cells, culture medium, CCK-8 solution, and drug solution);

[0142] Ac: absorbance of control well (containing cells, culture medium, and CCK-8 solution, but no drug);

[0143] Ab: absorbance of blank wells (containing culture medium and CCK-8 solution, but not cells or drugs).

[0144] Experimental Results: To further evaluate the in vitro antitumor activity of the target compound, MCF-7 cells (human breast cancer cells), A549 cells (human lung cancer cells), and Hela cells (cervical cancer cells) were selected as test tumor lines, and artemisinin Q was used as a control group. The test results are shown in Table 1. Overall, the antitumor activity of artemisinin PROTACs was superior to that of the artemisinin derivative itself. Based on the IC50 and compound structure, Q4 was selected as the dosing group, and MCF-7 cells (human breast cancer cells) were used as the dosing tumor line for further proteomic analysis.

[0145] Table 1 In vitro antitumor activity (IC50) of Q and Q1-Q25

[0146]

[0147]

[0148] Example 7: Proteomic Analysis of Artemisinin PROTACs of the Present Invention

[0149] In this example, a total of 9 cell samples were set up and divided into 3 groups, namely PROTACs group (drug group), atNP group (control group) and blank group, and the biological replicates corresponding to each group were 3 cases. Three groups of data were generated for comparison, drug group vs. control group, drug group vs. blank group and control group vs. blank group. The original data contains 9 experimental samples. In order to better analyze the data, this example performs a series of preparations and arrangements (Data Management) on the original data. It mainly includes the following steps: filtering for missing values, retaining proteins detected in all samples in the TMT project; screening the number of unique peptides of the protein to which it belongs: retaining the number of unique peptides greater than or equal to 1.

[0150] Experimental results: Based on LC-MS quantitative analysis of proteins, a total of 7738 proteins (groups) were identified in this experiment. Statistical methods were used to screen differentially expressed proteins, where the screening criteria for differentially expressed proteins were: Student's t-test P-VALUE < 0.05 and FOLD CHANGE ≤ 0.67 or FOLD CHANGE ≥ 1.5. The summary data of all differentially expressed proteins are shown in Table 3. According to the screening criteria, the differentially expressed protein analysis in the drug-treated group and the control group obtained a total of 25 differentially expressed proteins, of which 22 were upregulated (fc> = 1.5) and 3 were downregulated (fc < = 0.67). (Table 2). Volcano plot of differential proteins based on the precursor group vs blank group, PROTAC group vs CON group ( Figure 1 Downregulated proteins in the blue region were screened for proteins downregulated between Q2 vs. Con and Q vs. Con. Following the team's previous screening criteria, proteins with downregulated protein levels in the Q2 group compared to the Con group and proteins with no significant downregulation in the Q group compared to the Con group were screened. Potential target proteins with significantly decreased protein levels in the Q2 vs. Con group were identified: HMGB1 and RPL9. Specific differentially expressed protein data are shown in (Tables 3 and 4).

[0151] Table 2 Statistics of differentially expressed proteins

[0152]

[0153] Table 3 Differential protein HMGB1

[0154]

[0155] Table 4 Differential protein RPL9

[0156]

[0157] Example 8: Trends in potential target proteins for the anti-tumor effects of artemisinin PROTACs of the present invention

[0158] In this example, the accuracy of the omics results was verified by immunoblotting. MCF-7 cells were treated with the drug for 48 hours, and the cells were collected and proteins were extracted for immunoblotting analysis. Figure 1 The results showed that Q2 can degrade HMGB1 in a dose-dependent manner. At the same time, this example added Q4 with a different chain length from Q2 to verify that it also showed the same degradation effect. At the same time, this example verified the expression level of the potential target RPL9 protein after drug action through immunoblotting analysis. Figure 2 This was consistent with the omics results.

[0159] Example 9: Verification of potential target proteins for the anti-tumor effects of artemisinin PROTACs of the present invention

[0160] This example further verifies potential targets through CETSA thermal stability experiments. The principle is that the target protein will enhance its thermal stability after binding to the ligand, making it less likely to be degraded by high temperature. Figure 3 As shown in Figure 2, the results of immunoblotting analysis showed that the thermal stability of HMGB1 protein in the artesunate incubation group was significantly higher than that in the solvent group, proving that HMGB1 molecules can directly bind to artesunate. Figure 4 The immunoblot analysis shown above showed that the thermal stability of RPL9 protein in the artesunate incubation group was significantly increased compared with the solvent group, proving that the RPL9 molecule can also directly bind to artesunate.

[0161] This example further validates the target through the Darts experiment. The principle is that after the drug binds to the target protein, the protein structure becomes more stable, thus having the property of resisting protease hydrolysis. Figure 5 As shown, it was demonstrated that artesunate can directly bind to HMGB1 and thus improve the stability of HMGB1 against proteolytic enzymes.

[0162] The above description is merely a preferred embodiment of the present invention and therefore cannot be used to limit the scope of the present invention. In other words, equivalent changes and modifications made within the scope of the present invention and the contents of the specification should still fall within the scope of the present invention.

Claims

1. An artemisinin PROTACs, characterized by: Its structural formula is in, E3 Ligase is lenalidomide or pomalidomide, Linker is an amino acid chain, a fatty chain or a PEG chain, the amino acid chain is -NH-(CH2)n1-NH-(CH2)3-CO-, the fatty chain is -NH-(CH2)n2-CO-, the PEG chain is -(O-CH2-CH2)n3-CH2 CH2-CO-, n1 is a natural number from 2 to 10, n2 is a natural number from 2 to 7, and n3 is 2 or 3.

2. The artemisinin PROTACs according to claim 1, characterized in that: Its structural formula is selected from at least one of the following:

3. Use of the artemisinin PROTACs or pharmacologically or physiologically acceptable salts thereof according to claim 1 or 2 in preparing a composition for treating cancer, wherein the cancer is selected from breast cancer, cervical cancer and lung cancer.

4. A cancer treatment composition, characterized in that: The active ingredient comprises the artemisinin PROTACs or a pharmacologically or physiologically acceptable salt thereof according to claim 1 or 2, and the cancer is selected from breast cancer, cervical cancer and lung cancer.

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