SERPINE1 protein degradation agent and application thereof

Abstract

The invention belongs to the field of medicines, and discloses an SERPINE1 protein degradation agent and application thereof. The general formula of the SERPINE1 protein degradation agent is # imgabs0 # or # imgabs1 #. The research finds that a structural fragment for inhibiting SERPINE1 protein and a structural sheet or a hydrophobic label of an E3 protein ligand are coupled together through a linker, the length of the linker is adjusted, the obtained SERPINE1 protein degradation agent has an unexpected anti-tumor effect, the anti-tumor effect of the SERPINE1 protein degradation agent is superior to that of a single SERPINE1 inhibitor, and the SERPINE1 protein degradation agent has a good application prospect. The drug resistance of tumor cells can be overcome, and the drug-forming target property is good.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and specifically relates to a SERPINE1 protein degradation agent and application thereof. Background Art

[0002] In recent years, protein degradation technology has developed rapidly, and the development of degradation agents for pathogenic target proteins has become a powerful tool in the development of new drugs. Protein degradation agents use the ubiquitin-proteasome system in the body to reduce protein expression rather than inhibit protein function by connecting the target protein to the ubiquitination tag, thereby achieving the purpose of treating diseases. At present, some degradation agents have been successfully developed for difficult-to-drug protein targets such as K-ras, and certain therapeutic effects have been achieved. In theory, degradation agents can be recycled and reused. Compared with inhibitors and small molecule antibody drugs, degradation agents have the advantage that catalytic amounts can work. Therefore, protein degradation agent technology can provide a wider range of application prospects for various targeted treatments of cancer.

[0003] Serpin Family E Member 1 (SERPINE1) is a secreted protein with a molecular weight of 52 kDa. Under physiological conditions, it is mainly involved in regulating fibrinolysis and thus affects the body's coagulation function. The secreted active SERPINE1 protein is a 45 kDa single-chain glycoprotein composed of 379-381 amino acids, with 9 α helices and 3 β folded fragments. SERPINE1 prevents the formation of plasmin and inhibits fibrinolysis and thrombolysis by binding to and inhibiting tissue plasminogen activator (tPA) and urokinase plasminogen activator (uPA). Its reduced activity after mutation can easily cause hemorrhagic diseases in the body.

[0004] In recent years, studies have reported that SERPINE1 is one of the important proteins in many cancer-promoting signaling pathways. Its complex dynamic structure gives it pleiotropic biological functions, which are closely related to tumor growth, metastasis and treatment prognosis. Many clinical and preclinical studies have reported that SERPINE1 expression is significantly increased in various types of tumor biopsy tissues and plasma, with multiple inducers such as growth factors, hormones, inflammatory responses and oxidative stress. High expression of SERPINE1 in tumors can help cancer cells degrade the extracellular matrix, thereby invading and spreading, promoting tumor angiogenesis, and ultimately leading to tumor metastasis. In addition, elevated SERPINE1 can inhibit caspase3 and FasL to help cancer cells resist chemotherapy-induced apoptosis and promote the increase in the production of anti-apoptotic proteins such as Bcl-2. It is worth noting that the pleiotropic cancer-promoting phenomenon of SERPINE1 cannot be explained only by the classical function of inhibiting the activity of fibrinolytic-related enzymes. The mechanism of its non-classical function in cancer is not yet fully understood and needs further study.

[0005] Currently, some small molecule inhibitors targeting SERPINE1 have been reported. The vast majority of inhibitors bind to the classical active center of the SERPINE1 protein, induce conformational changes to inhibit tPA and uPA, thereby irreversibly converting SERPINE1 into an inactive form. These inhibitors have achieved certain results mainly in the research of diseases such as cardiovascular diseases (thrombolysis), pulmonary fibrosis, and Alzheimer's disease. The small molecule inhibitors TM5441 and Tiplaxtinin have been reported to have certain anti-tumor activities in colorectal cancer, pancreatic cancer, bladder cancer, and cervical cancer, and the combination of chemotherapy drugs and PD-1 monoclonal antibodies can improve the curative effect. These research results preliminarily confirm that inhibiting SERPINE1 can lead to poor tumor uptake and slow tumor growth. However, the inhibition mechanism of SERPINE1 protein inhibitors requires conformational changes, which also leads to a decrease in their affinity for non-protease ligands, weak binding to them, and difficulty in becoming targets. Currently, none of these inhibitors have been tested in therapeutic clinical trials for cancer. SERPINE1 inhibitors still cannot solve the disease problems caused by the non-classical protein functions of SERPINE1. Summary of the Invention

[0006] An object of the present invention is to overcome at least one deficiency of the prior art and provide a SERPINE1 protein degrader and its application.

[0007] The technical solution adopted by the present invention is as follows:

[0008] In the first aspect of the present invention, there is provided: a SERPINE1 protein degrader, comprising:

[0009] 1) a structural fragment that inhibits the SERPINE1 protein, whose structural formula is and 2) an E3 ligase VHL ligand structural fragment or a hydrophobic tag structural fragment, and the structural fragment 1) and the structural fragment 2) are connected by a linker whose main chain contains 1 to 14 atoms. The general structural formula of the degrader is shown in Formula I or II:

[0010]

[0011] In Formula I and Formula II, R is selected from H, C1-C4 alkyl.

[0012] In some examples of the SERPINE1 protein degrader, the main chain of the linker is an alkyl chain, a polyethylene glycol chain, an ether-containing alkyl chain, an alkyl chain containing a triple bond, or an alkyl chain containing a triple bond and an ether group.

[0013] In some examples of SERPINE1 protein degraders, the backbone of the linker contains 4 to 8 atoms, more preferably 6 atoms. Structure-activity analysis shows that a linker of this length can yield SERPINE1 protein degraders with better activity. In particular, the linker is

[0014] In some examples of SERPINE1 protein degraders, the structural formula I is selected from one of the formulas X1 to X7 shown below:

[0015]

[0016] In some examples of SERPINE1 protein degraders, the structural formula II is selected from one of the formulas X8 to X18 shown below:

[0017]

[0018]

[0019] In some examples of SERPINE1 protein degraders, its structural formula is one of the formulas D1 to D39:

[0020]

[0021]

[0022] In formulas D1 to D39, R1 = R2 = R3 = R4 =

[0025] In a second aspect of the present invention, there is provided: a pharmaceutical composition comprising an active ingredient and an excipient, wherein the active ingredient comprises the SERPINE1 protein degrader described in the first aspect of the present invention, or one of its pharmaceutically acceptable salts, crystals, or solvates.

[0026] The pharmaceutically acceptable salts are those commonly used in the art, and depending on the compound, they can be acid addition salts or base addition salts.

[0027] In some examples of the pharmaceutical composition, its dosage form is an injection, an oral formulation, or a transdermal absorbent.

[0028] In a third aspect of the present invention, there is provided: the use of the SERPINE1 protein degrader described in the first aspect of the present invention in the preparation of an anti-tumor drug.

[0029] In some application examples, the tumor is a solid tumor or a hematological tumor.

[0030] In some application examples, the solid tumor is selected from colorectal cancer, pancreatic cancer, bladder cancer or cervical cancer; the hematological tumor is selected from leukemia or lymphoma.

[0031] The beneficial effects of the present invention are as follows:

[0032] The inventors found through research that by coupling a structural fragment that inhibits the SERPINE1 protein and a structural fragment of an E3 protein ligand or a hydrophobic tag together through a linker and adjusting the length of the linker, the obtained SERPINE1 protein degrader has an unexpected anti-tumor effect, and its anti-tumor effect is superior to that of a single SERPINE1 inhibitor. The obtained SERPINE1 protein degrader unexpectedly can overcome the drug resistance of tumor cells and has good druggable target properties. Description of the Drawings

[0033] Figure 1 To screen the SERPINE1 protein degrader of the compound of the present invention.

[0034] Figure 2 This shows the degradation effect of compound D10 of the present invention on SERPINE1 protein at different times.

[0035] Figure 3 This shows the killing effect of compound D10 of the present invention and the control compound SERPINE1 inhibitor Tiplaxtinin on the proliferation of tumor cells.

[0036] Figure 4 This shows the efficacy evaluation of compound D10 on the anti-growth and metastasis of AsPC-1 tumor cells. Detailed Embodiments

[0037] The present invention will be further described below with reference to the drawings. It should be noted that the following examples are based on the present technical solution and give detailed implementation manners and specific operation processes, but the protection scope of the present invention is not limited to this embodiment.

[0038] In the following examples, the intermediates have the same numbers, that is, the intermediates with the same numbers have the same structures.

[0039] Example 1: Synthesis of Compounds D1 and D2

[0040] Synthesis of Intermediates 7-9

[0041]

[0042] Reagents and reaction conditions: (a) Pd(PPh3)4, K2CO3, Toluene / EtOH / H2O, 100 °C; (b) Oxalyl chloride, EtOH, Et2O, 0 °C - rt, (c) NaH, DMF, 0 °C - rt.

[0043] As shown in the above reaction process, the synthesis was carried out using commercially purchased 5-bromoindole 1 and 4-(trifluoromethoxy)phenylboronic acid 2 as starting materials. In a 250 mL round-bottom flask, 5-bromoindole (6 g, 30.61 mmol, 1 equiv), 4-(trifluoromethoxy)phenylboronic acid (9.45 g, 45.91 mmol, 1.5 equiv), tetrakis(triphenylphosphine)palladium (1.77 g, 1.53 mmol, 0.05 equiv), potassium phosphate (19.49 g, 91.81 mmol, 3 equiv), and a mixed solvent of toluene / water / ethanol (72 mL, toluene / water / ethanol = 36 mL / 24 mL / 12 mL) were successively added under an argon atmosphere. The reaction system was placed in an environment of c and stirred for 12 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted three times with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. Then, it was separated and purified by column chromatography (PE / EA = 50 / 1). The separated product obtained was dried and weighed to obtain the intermediate 3 (4.2 g, 50%).

[0044] Compound 3 (4 g, 14.43 mmol, 1 equiv) was dissolved in ether under the protection of argon. Oxalyl chloride 4 (65.86 g, 46.17 mmol, 3.2 equiv) was slowly added dropwise in an ice bath. After stirring for 15 min, the system was warmed to room temperature and stirred for an additional 6 h. Ethanol (2.31 g, 72.14 mmol, 5 equiv) was added and stirred for 30 min. The system was placed in a -20 °C refrigerator for 15 min, washed and filtered with ice-cold ether to obtain a yellow solid 5 (4.8 g, 92%).

[0045] Compound 5 (500 mg, 1 equiv) was dissolved in DMF. Sodium hydride (49.54 mg, 2.06 mmol, 1.5 equiv) was added in portions at 0 °C. After stirring for 15 min, benzyl bromide compound (2.06 mmol, 1.5 equiv) was added. The reaction system was stirred for 15 min and then warmed to room temperature and stirred for an additional 12 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted three times with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. Then, it was separated and purified by column chromatography (PE / EA = 5 / 1). The separated product obtained was dried and weighed to obtain the target products 7 (1.078, 67%), 8 (554 mg, 72.7%), and 9 (425 mg, 54%).

[0046]

[0047] Reagents and reaction conditions: (a) Boc2O, NaOH, MeOH, 0 °C - rt; (b) Pd(OAc)2, KOAc, DMA, 150 °C; (c) HCl / MeOH, rt; (d) HATU, DIPEA, THF, 0 °C - rt; (e) HCl / MeOH, 0 °C - rt; (f) HATU, DIPEA, DMF, rt; (g) HCl / MeOH, rt.

[0048] To a solution of 4-bromophenylbenzylamine 10 (10 g, 53.75 mmol) in methanol (80 mL) at 0 °C was added Boc2O (18.52 mL, 80.62 mmol) and NaOH (107.5 mmol, 1 M). The reaction mixture was warmed to room temperature and stirred for 5 h. Methanol was removed under reduced pressure, and the aqueous solution was adjusted to pH = 5 with 1 M aqueous HCl. The aqueous solution was extracted with dichloromethane (150 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered. Concentration under reduced pressure gave a colorless liquid 11 (14.4 g, 93.6%) which was used in the next step without further purification. Under argon, to a solution of the above-obtained liquid (14.4 g, 50.42 mmol) in DMA (50 mL) were added palladium acetate (226.45 mg, 1.01 mmol), potassium acetate (9.9 g, 100.86 mmol), and 4-methylthiazole 12 (4.59 mL, 50.43 mmol). The reaction mixture was stirred at 150 °C for 10 h. Then, dichloromethane (150 mL) and water (100 mL) were added, and the aqueous phase was extracted with dichloromethane (150 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a yellow liquid 13 (14.35 g, 93.7%) which was used directly in the next step. After removing the Boc group in 13 (6.5 g, 21.35 mmol) under acidic conditions, the solution was adjusted to weakly basic with ammonia water to obtain the free amine residue (5.14 g, 20.77 mmol). A solution of Boc-L-hydroxyproline 14 (5.92 g, 25.62 mmol) and diisopropylethylamine (10.59 mL, 64.05 mmol) in anhydrous tetrahydrofuran (75 mL) was stirred at 0 °C and the above-obtained amine (5.14 g, 20.77 mmol) and HATU (9.74 g, 25.62 mmol) were added. The resulting mixture was warmed to room temperature and stirred for 2 h. After removing THF by rotary evaporation under reduced pressure, dichloromethane (120 mL) and water (80 mL) were added, and the aqueous phase was extracted with dichloromethane (100 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a yellow liquid 15 (8.67 g, 82.5%) which was used in the next step without further purification. After removing the Boc group in 15 (5 g, 11.98 mmol) under acidic conditions, N-Boc-L-tert-leucine 16 (2.76 g, 11.93 mmol), diisopropylethylamine (5.91 mL, 35.78 mmol), and HATU (6.8 g, 17.89 mmol) in DMF (45 mL) were added. The reaction mixture was stirred at room temperature for 5 h. The reaction was monitored by TLC. After the reaction was complete, dichloromethane (120 mL) and water (60 mL) were added, and the aqueous phase was extracted with dichloromethane (100 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure.The residue was purified by silica gel column chromatography (DCM / MeOH = 50 / 1) to obtain a yellow solid 17 (4.5 g, 70.8%). The Boc group of the yellow solid was removed under acidic conditions to obtain compound 18.

[0049]

[0050] Reagents and reaction conditions: (a) Pd(PPh3)2Cl2, CuI, DIPEA, THF, rt; (b) TFA / DCM, rt; (c) HATU, DIPEA, DMF, rt; (d) Pd / C, H2, MeOH, rt; (e) LiOH, THF / H2O, rt.

[0051] In a 25 mL Schlenk tube, compound 7 (100 mg, 10.173 mmol, 1 equiv), tert-butyl propiolate 19 (32.66 mg, 0.259 mmol, 1.5 equiv), bis(triphenylphosphine)palladium(II) dichloride (6.06 mg, 8.63 μmol, 0.1 equiv), copper(I) iodide (3.29 mg, 17.26 μmol, 0.1 equiv), diisopropylethylamine (90.2 μL, 0.518 mmol, 3 equiv), and 2 mL of tetrahydrofuran were successively added under an argon atmosphere. The reaction system was stirred at room temperature for 12 h. After the reaction was completed, the solvent was evaporated under reduced pressure, extracted with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. Then, it was separated and purified by column chromatography (PE / EA = 5 / 1), and the separated product was dried and weighed. The target product 20 (90 mg, 90.3%) was obtained.

[0052] Compound 20 (90 mg, 155.83 mmol) was dissolved in 2 mL of dichloromethane solution, and 0.4 mL of trifluoroacetic acid was slowly added dropwise. The mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure. Then, 18 (49.54 mg, 115.07 μmol), HATU (87.50 mg, 230.13 μmol), and diisopropylethylamine (0.12 ml, 0.69 mmol) were successively added under an argon atmosphere. After reacting for 2 h, the reaction was monitored by TLC. After the reaction was complete, the mixture was extracted three times with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. Then, it was separated and purified by column chromatography (PE / EA = 5 / 1). The separated product obtained was dried and weighed to obtain the target product 21 (40 mg, 27%). Compound 21 (20 mg) was dissolved in methanol (4 mL), and palladium on carbon (2 mg) was added and reacted under a hydrogen atmosphere. The reaction was monitored by TLC. After the reaction was complete, it was concentrated under vacuum, and the crude product was purified by silica gel column (DCM / MeOH = 20 / 1) to obtain compound D2 (20 mg, 98%). Compound D2 (20 mg) was dissolved in a mixed solvent of tetrahydrofuran and water (2 mL, tetrahydrofuran / water = 1 mL / 1 mL), and lithium hydroxide (1.5 mg, 3 equiv) was added. After reacting at room temperature for 2 h, the reaction was monitored by TLC. After the reaction was complete, it was concentrated under vacuum, and compound D1 (19 mg, 96%) was obtained by extracting three times with ethyl acetate. 1 HNMR (500 MHz, CDCl3) δ 8.60–8.52 (m, 2H), 8.37 (d, J = 5.0 Hz, 1H), 7.58–7.51 (m, 2H), 7.40–7.29 (m, 3H), 7.27–7.16 (m, 8H), 7.16–7.09 (m, 1H), 7.08–6.99 (m, 2H), 6.65–6.53 (m, 1H), 6.30 (dd, J = 17.3, 8.9 Hz, 1H), 5.88–5.78 (m, 1H), 5.25 (s, 2H), 4.44 (d, J = 8.8 Hz, 1H), 4.41–4.29 (m, 3H), 4.16 (dd, J = 15.1, 5.4 Hz, 1H), 3.91–3.82 (m, 1H), 3.48 (dd, J = 11.2, 3.5 Hz, 1H), 2.42–2.38 (m, 3H), 2.26–2.20 (m, 1H), 1.96–1.87 (m, 1H), 0.89 (s, 1H), 0.75 (s, 9H).

[0053] Example 2: Synthesis of Compounds D5 - D11

[0054]

[0055] Reagents and reaction conditions: (a) Pd(PPh3)4, CuI, DIPEA, THF, 100 °C; (b) HATU, DIPEA, DMF; (c) Pd / C, H2, MeOH; (d) LiOH, THF / H2O.

[0056] Take a 25 mL Schlenk tube and successively add compound 7 (200 mg, 0.345 mmol, 1 equiv), alkynoic acid 22 (0.517 mmol, 1.5 equiv), palladium(II) bis(triphenylphosphine) dichloride (24.23 mg, 34.52 μmol, 0.1 equiv), copper(I) iodide (6.58 mg, 34.52 μmol, 0.1 equiv), diisopropylethylamine (0.18 mL, 1.04 mmol, 3 equiv), and 3 mL of tetrahydrofuran under an argon atmosphere. Stir the reaction system at room temperature for 12 h. After the reaction is completed, rotary evaporate the solvent under reduced pressure, extract with ethyl acetate, dry over anhydrous magnesium sulfate, filter, rotary evaporate the solvent from the filtrate under reduced pressure, and then separate and purify by column chromatography (DCM / MeOH = 40 / 1). Weigh the separated product after drying. Obtain the target product 23 (n = 1, 199 mg, 78%; n = 2, 120 mg, 61.7%). Add 18 (0.12 mmol, 1.2 equiv), HATU (76.92 mg, 0.2 mmol, 2 equiv), diisopropylethylamine (0.05 mL, 0.3 mmol), and 2 mL of DMF successively to a round-bottom flask containing compound 23 (0.1 mmol) under an argon atmosphere. Stir the reaction system at room temperature for 2 h. Monitor the reaction by TLC. After the reaction is complete, extract three times with ethyl acetate, dry over anhydrous magnesium sulfate, filter, rotary evaporate the solvent from the filtrate under reduced pressure, and then separate and purify by column chromatography (DCM / MeOH = 40 / 1). Weigh the separated product after drying to obtain the target product D10 (68 mg, 69%). 11H NMR (500 MHz, CDCl3) δ 8.68–8.62 (m, 2H), 8.46 (s, 1H), 7.65 (d, J = 8.7 Hz, 2H), 7.46 (dd, J = 8.5, 1.7 Hz, 1H), 7.37–7.27 (m, 11H), 7.10 (d, J = 8.2 Hz, 2H), 6.19 (d, J = 8.6 Hz, 1H), 5.36 (s, 2H), 4.69 (t, J = 7.9 Hz, 1H), 4.56–4.47 (m, 3H), 4.31 (dd, J = 14.9, 5.2 Hz, 1H), 4.05 (d, J = 11.4 Hz, 1H), 3.94 (s, 3H), 3.60 (dd, J = 11.3, 3.6 Hz, 1H), 2.49 (s, 3H), 2.43 (d, J = 6.1 Hz, 2H), 2.38–2.33 (m, 2H), 2.11 (dd, J = 13.4, 8.1 Hz, 1H), 1.98 (s, 1H), 1.87 (dd, J = 15.5, 7.3 Hz, 2H), 0.91 (d, J = 10.0 Hz, 9H); D5 (73 mg, 73%), 1 1H NMR (500 MHz, CDCl3) δ 8.60 (s, 1H), 8.57 (d, J = 1.4 Hz, 1H), 8.38 (s, 1H), 7.61–7.55 (m, 2H), 7.39 (dd, J = 8.5, 1.7 Hz, 1H), 7.30–7.18 (m, 10H), 7.13 (d, J = 5.8 Hz, 1H), 7.00 (d, J = 8.2 Hz, 2H), 6.43 (d, J = 8.7 Hz, 1H), 5.28 (s, 2H), 4.57 (t, J = 7.9 Hz, 1H), 4.47 (dd, J = 15.2, 7.6 Hz, 3H), 4.22 (dd, J = 15.0, 5.2 Hz, 1H), 3.97 (d, J = 11.4 Hz, 1H), 3.87 (s, 3H), 3.54 (dd, J = 11.3, 3.6 Hz, 1H), 2.69–2.57 (m, 2H), 2.44–2.36 (m, 6H), 2.01 (dd, J = 13.4, 8.1 Hz, 1H), 1.97 (d, J = 2.3 Hz, 1H), 0.84 (s, 9H); D5 (50 mg) was dissolved in methanol (2 mL), palladium on carbon (5 mg) was added, and the reaction was carried out in a hydrogen atmosphere. The reaction was monitored by TLC. After the reaction was complete, it was concentrated in vacuo, and the crude product was purified by silica gel column (DCM / MeOH = 40 / 1) to obtain the target product D6, 11H NMR (500 MHz, CDCl3) δ 8.63–8.56 (m, 2H), 8.39 (s, 1H), 7.59 (d, J = 8.7 Hz, 2H), 7.42 (dd, J = 8.5, 1.8 Hz, 1H), 7.27 (dd, J = 16.8, 8.5 Hz, 5H), 7.23 (s, 2H), 7.19 (s, 2H), 7.05 (q, J = 8.2 Hz, 4H), 5.29 (s, 2H), 4.70 (d, J = 2.8 Hz, 5H), 4.61 (t, J = 8.0 Hz, 1H), 4.50 (dd, J = 14.9, 6.7 Hz, 1H), 4.46–4.40 (m, 2H), 4.25 (dd, J = 14.9, 5.1 Hz, 1H), 4.02 (d, J = 11.5 Hz, 1H), 3.87 (s, 3H), 3.51 (dd, J = 11.4, 3.4 Hz, 1H), 3.14 (s, 1H), 2.52 (t, J = 6.8 Hz, 2H), 2.44 (s, 3H), 2.13 (t, J = 7.0 Hz, 2H), 2.07–2.01 (m, 1H), 0.83 (s, 9H). Weigh 20 mg of each of compounds D10, D5, and D6 respectively, add lithium hydroxide (3 equiv), and stir in a mixed solution of tetrahydrofuran and water (1 mL / 1 mL) for 0.5 h. Monitor the reaction by TLC. After the reaction is complete, extract with ethyl acetate three times, dry with anhydrous magnesium sulfate, and rotary evaporate the filtrate under reduced pressure to remove the solvent to obtain the target products D7-9 and D11 in quantitative yield.

[0057] Example 3: Synthesis of Compounds D12-D14, D17-18

[0058]

[0059] Reagents and reaction conditions: (a) NaH, DMF; (b) Pd(PPh3)4, CuI, DIPEA, THF, 100 °C; (c) TFA / DCM, rt; (d) HATU, DIPEA, DMF; (e) Pd / C, H2, MeOH; (f) LiOH, THF / H2O.

[0060] Compound 24 (10.25 mmol) was dissolved in DMF. Sodium hydride (615.15 mg, 15.38 mmol, 1.5 equiv) was added portionwise at 0 °C. After stirring for 15 min, tert-butyl bromoacetate 25 (2.16 g, 30.76 mmol, 3 equiv) was added. The reaction mixture was stirred for 15 min and then warmed to room temperature and stirred for an additional 12 h. The reaction was monitored by TLC. After completion of the reaction, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (PE / EA = 20 / 1). The isolated product was dried and weighed to obtain the target product 26. A 25-mL Schlenk tube was charged successively with compound 7 (200 mg, 0.345 mmol, 1 equiv), 26 (0.517 mmol, 1.5 equiv), bis(triphenylphosphine)palladium(II) dichloride (24.23 mg, 34.52 μmol, 0.1 equiv), copper(I) iodide (6.58 mg, 34.52 μmol, 0.1 equiv), diisopropylethylamine (0.18 mL, 1.04 mmol, 3 equiv), and 3 mL of tetrahydrofuran under an argon atmosphere. The reaction mixture was stirred at room temperature for 12 h. After completion of the reaction, the solvent was removed under reduced pressure, and the residue was extracted with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (PE / EA = 20 / 1). The isolated product was dried and weighed to obtain the target product 27 n = 2 (141 mg, 63%), n = 1 (65 mg, 30%). Compound 27 (152.61 mmol) was dissolved in 2 mL of dichloromethane solution. 0.4 mL of trifluoroacetic acid was added dropwise slowly, and the mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After completion of the reaction, the solvent was removed under reduced pressure from the filtrate. Then, 18 (74.97 mg, 174.11 μmol), HATU (110.34 mg, 290.18 μmol), and diisopropylethylamine (0.126 mL, 0.72 mmol) were added successively under an argon atmosphere. After reacting for 2 h, the reaction was monitored by TLC. After completion of the reaction, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (DCM / MeOH = 20 / 1). The isolated product was dried and weighed to obtain the target product D12 (84 mg, 56%). 11H NMR (500 MHz, CDCl3) δ 8.58–8.54 (m, 2H), 8.38 (d, J = 0.8 Hz, 1H), 7.57 (dd, J = 8.6, 0.8 Hz, 2H), 7.37 (dd, J = 8.5, 1.6 Hz, 1H), 7.29 (d, J = 7.4 Hz, 2H), 7.27–7.19 (m, 8H), 7.15 (d, J = 8.8 Hz, 1H), 7.01 (d, J = 8.0 Hz, 2H), 5.27 (s, 2H), 4.60 (d, J = 1.0 Hz, 1H), 4.45–4.39 (m, 3H), 4.25 (dd, J = 15.1, 5.5 Hz, 1H), 3.94 (dd, J = 17.8, 3.9 Hz, 3H), 3.86 (d, J = 0.9 Hz, 3H), 3.65–3.53 (m, 4H), 2.63 (t, J = 6.3 Hz, 2H), 2.40 (d, J = 1.3 Hz, 3H), 2.35 (dd, J = 11.5, 6.4 Hz, 1H), 2.06–2.00 (m, 1H), 0.85 (s, 9H) and PH1553 (70 mg, 46%). Compound D12 (50.4 μmol) was dissolved in methanol solution (2 mL), palladium on carbon (5 mg) was added, and the reaction was carried out under a hydrogen atmosphere. The reaction was monitored by TLC. After the reaction was complete, it was concentrated in vacuo, and the crude product was purified by silica gel column (DCM / MeOH = 40 / 1) to obtain the target product 28. 11H NMR (400 MHz, CDCl3) δ 8.57 (t, J = 6.8 Hz, 2H), 8.37 (dd, J = 13.6, 7.2 Hz, 1H), 7.56 (t, J = 5.7 Hz, 2H), 7.38 (t, J = 8.5 Hz, 1H), 7.33–7.16 (m, 10H), 7.10–7.00 (m, 3H), 5.26 (dd, J = 12.8, 8.8 Hz, 2H), 4.60 (t, J = 7.9 Hz, 1H), 4.47–4.36 (m, 3H), 4.25 (dd, J = 15.0, 5.4 Hz, 1H), 4.00–3.89 (m, 1H), 3.88–3.76 (m, 5H), 3.68–3.45 (m, 4H), 3.43–3.36 (m, 1H), 3.07 (dd, J = 7.3, 3.4 Hz, 1H), 2.51 (td, J = 12.4, 6.4 Hz, 2H), 2.43–2.31 (m, 4H), 2.08–1.98 (m, 1H), 1.61–1.48 (m, 1H), 0.84 (t, J = 7.1 Hz, 9H). 20 mg each of compounds D12 and D28 were dissolved in a mixed solution of tetrahydrofuran and water (tetrahydrofuran / water = 1 mL / 1 mL), lithium hydroxide (3 equiv) was added, and the mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation under reduced pressure to obtain the target products D13, D14, D17, and D18 in quantitative yield.

[0061] Example 4: Synthesis of Compounds D20 - D21, D26 - D27

[0062]

[0063] Reagents and reaction conditions: (a) NaH, DMF; (b) Pd(PPh3)4, CuI, DIPEA, THF, 100 °C; (c) TFA / DCM, rt; (d) HATU, DIPEA, DMF; (e) Pd / C, H2, MeOH; (f) LiOH, THF / H2O.

[0064] Compound 29 (4 mmol) was dissolved in THF. Sodium hydride (271.98 mg, 6.4 mmol, 1.7 equiv) was added portionwise at 0 °C. After stirring for 15 min, tert-butyl bromoacetate 30 (936.27, 4.8 mmol, 1.2 equiv) was added. The reaction mixture was stirred for 15 min and then warmed to room temperature and reacted for another 12 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. Then it was separated and purified by column chromatography (PE / EA = 20 / 1). The separated product was dried and weighed to obtain the target product 31 (n = 1, 172 mg, 20%; n = 2, 310 mg, 30%). A 25-mL Schlenk tube was taken, and compound 31 (200 mg, 0.345 mmol, 1 equiv), 7 (0.517 mmol, 1.5 equiv), bis(triphenylphosphine)palladium(II) dichloride (24.23 mg, 34.52 μmol, 0.1 equiv), copper(I) iodide (6.58 mg, 34.52 μmol, 0.1 equiv), diisopropylethylamine (0.18 mL, 1.04 mmol, 3 equiv), and 3 mL of tetrahydrofuran were added successively under an argon atmosphere. The reaction mixture was stirred at room temperature for 12 h. After the reaction was completed, the solvent was removed by rotary evaporation under reduced pressure. The mixture was extracted with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. Then it was separated and purified by column chromatography (PE / EA = 20 / 1). The separated product was dried and weighed to obtain the target product n = 1 (194 mg, 79%), n = 2 (118 mg, 50%). Compound 32 (138.2 μmol) was dissolved in 2 mL of dichloromethane solution. 0.4 mL of trifluoroacetic acid was slowly added dropwise, and the mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure. Then, 18 (61.44 mg, 138.21 μmol), HATU (105.1 mg, 276.4 μmol), and diisopropylethylamine (0.123 mL, 0.69 mmol) were added successively under an argon atmosphere. After reacting for 2 h, the reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. Then it was separated and purified by column chromatography (DCM / MeOH = 40 / 1). The separated product was dried and weighed to obtain the target product 33 (84 mg, 64%). 11H NMR (400 MHz, CDCl3) δ 8.70–8.59 (m, 2H), 8.46 (d, J = 17.8 Hz, 1H), 7.70–7.63 (m, 1H), 7.53–7.26 (m, 11H), 7.16–7.05 (m, 2H), 5.37 (d, J = 22.6 Hz, 2H), 4.72 (t, J = 8.0 Hz, 1H), 4.59–4.46 (m, 3H), 4.46–4.29 (m, 3H), 4.06 (d, J = 12.4 Hz, 1H), 4.00 (d, J = 5.0 Hz, 2H), 3.95 (d, J = 5.3 Hz, 3H), 3.74 (ddd, J = 10.9, 7.3, 3.9 Hz, 4H), 3.63 (dd, J = 11.3, 3.2 Hz, 1H), 2.49 (s, 3H), 2.33–2.06 (m, 4H), 0.97 (s, 9H). Compound 33 (60 mg) was dissolved in a methanol solution (2 mL), palladium on carbon (5 mg) was added, and the reaction was carried out under a hydrogen atmosphere. The reaction was monitored by TLC. After the reaction was complete, it was concentrated in vacuo, and the crude product was purified by silica gel column (DCM / MeOH = 40 / 1) to quantitatively obtain the target product 34. 1 1H NMR (400 MHz, CDCl3) δ 8.76–8.59 (m, 2H), 8.46 (d, J = 17.0 Hz, 1H), 7.75–7.62 (m, 2H), 7.60–7.20 (m, 12H), 7.18–7.05 (m, 2H), 5.38 (d, J = 21.9 Hz, 2H), 4.71 (d, J = 7.9 Hz, 1H), 4.58–4.50 (m, 2H), 4.46–4.27 (m, 3H), 3.98 (dd, J = 17.2, 7.0 Hz, 5H), 3.78–3.60 (m, 8H), 2.96 (s, 1H), 2.89 (s, 1H), 2.81 (s, 2H), 2.51 (s, 3H), 2.25–1.94 (m, 6H), 0.96 (s, 9H). 20 mg each of compounds 33 and 34 were dissolved in a mixed solution of tetrahydrofuran and water (tetrahydrofuran / water = 1 mL / 1 mL), lithium hydroxide (3 equiv) was added, and the mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, it was extracted three times with ethyl acetate, dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation under reduced pressure to obtain the target products D20, D21, D26, and D27 in quantitative yield.

[0065] Example 5: Synthesis of Compounds D3 and D4

[0066]

[0067] Reagents and reaction conditions: (a) TFA, DCM; (b) HATU, DIPEA, DMF; (c) TFA / DCM, rt; (d) HATU, DIPEA, DMF; (e) LiOH, THF, H2O.

[0068] Compound 8 (100 mg, 180.66 μmol) was dissolved in 2 mL of dichloromethane solution, and 0.4 mL of trifluoroacetic acid was slowly added dropwise. The mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure. Then, tert-butyl glycinate 34 (35.6 mg, 271.4 μmol), HATU (137.59 mg, 361.86 μmol), and diisopropylethylamine (0.09 mL, 542.79 μmol) were successively added under an argon atmosphere. After reacting for 2 h, the reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. Then, it was separated and purified by column chromatography (PE / EA = 1.5 / 1). The separated product obtained was dried and weighed to obtain the target product 35 (75 mg, 68%). Compound 33 (75 mg, 180.66 μmol) was dissolved in 2 mL of dichloromethane solution, and 0.4 mL of trifluoroacetic acid was slowly added dropwise. The mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure. Then, 18 (52.89 mg, 122.84 μmol), HATU (93.41 mg, 245.67 μmol), and diisopropylethylamine (0.128 mL, 737.02 μmol) were added. After reacting for 2 h, the reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. Then, it was separated and purified by column chromatography (DCM / CH3OH = 40 / 1). The separated product obtained was dried and weighed to obtain the target product D4 (80 mg, 67%). D4 (70 mg) was dissolved in a mixed solution of tetrahydrofuran and water (1 mL / 1 mL), lithium hydroxide (3 equiv) was added, and the mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation under reduced pressure to obtain the target product D3 in quantitative yield. 11H NMR (500 MHz, MeOD) δ 9.58 (s, 1H), 8.63 (s, 1H), 8.44 (s, 1H), 7.74 (d, J = 7.9 Hz, 3H), 7.59 (d, J = 8.4 Hz, 2H), 7.39 (d, J = 7.5 Hz, 4H), 7.31 (d, J = 8.0 Hz, 2H), 7.24 (dd, J = 15.3, 8.1 Hz, 5H), 5.51 (s, 2H), 4.54 (d, J = 6.5 Hz, 1H), 4.47–4.36 (m, 3H), 4.23 (d, J = 15.7 Hz, 1H), 3.96 (s, 2H), 3.77 (d, J = 11.0 Hz, 1H), 3.67 (dd, J = 11.0, 3.5 Hz, 1H), 2.41 (s, 3H), 2.12–2.08 (m, 1H), 1.95 (ddd, J = 13.2, 9.3, 4.2 Hz, 1H), 1.30 (s, 2H), 0.90 (s, 9H).

[0069] Example 6: Synthesis of Compounds D23 and D27

[0070]

[0071] Reagents and reaction conditions: (a) HATU, DIPEA, DMF; (b) TFA, DCM; (c) TFA, DCM; (d) HATU, DIPEA, DMF; (e) LiOH, THF, H2O

[0072] In an argon atmosphere, compound 36 (1 mmol), 18 (430.56 mg, 1 mmol), HATU (760.45 mg, 2 mmol), and diisopropyl ethylamine (0.5 mL, 3 mmol) were added respectively. After reacting for 2 h, the reaction was monitored by TLC. After the reaction was complete, it was extracted three times with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, the filtrate was rotary evaporated under reduced pressure to remove the solvent, and then separated and purified by column chromatography (DCM / MeOH = 40 / 1). The separated product obtained was dried and weighed to obtain the target product 37 (n = 5, 577.86 mg, 86%; n = 8, 332.71 mg, 46.6%). Compound 37 (n = 5, 120 mg, 0.178 mmol; n = 8, 130 mg, 0.182 mmol) was dissolved in 2 mL of dichloromethane solution, and 0.4 mL of trifluoroacetic acid was slowly added dropwise, and stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the filtrate was rotary evaporated under reduced pressure to remove the solvent and directly used for the next step. Compound 8 (100 mg, 180.66 μmol) was dissolved in 2 mL of dichloromethane solution, and 0.4 mL of trifluoroacetic acid was slowly added dropwise, and stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the filtrate was rotary evaporated under reduced pressure to remove the solvent, and then 38 (0.178 mmol), HATU (137.59 mg, 361.86 μmol), and diisopropyl ethylamine (0.09 mL, 542.79 μmol) were successively added in an argon atmosphere. After reacting for 2 h, the reaction was monitored by TLC. After the reaction was complete, it was extracted three times with ethyl acetate, dried over anhydrous magnesium sulfate, filtered, the filtrate was rotary evaporated under reduced pressure to remove the solvent, and then separated and purified by column chromatography (PE / EA = 1.5 / 1). The separated product obtained was dried and weighed to obtain the target product. 50 mg of each of the above target products was dissolved in a mixed solution of tetrahydrofuran and water, lithium hydroxide (3 equiv) was added, and stirred at room temperature for 0.5 h, and then the reaction was monitored by TLC. After the reaction was complete, it was extracted three times with ethyl acetate, dried over anhydrous magnesium sulfate, the filtrate was rotary evaporated under reduced pressure to remove the solvent, and the target products D23 (115 mg, 60%) and D25 (100 mg, 73%) were obtained in quantitative yield

[0073] Example 7: Synthesis of Compounds D29 and D33

[0074]

[0075] Reagents and reaction conditions: (a) K2CO3, 80 °C, DMF; (b) LiOH, THF, H2O; (c) HATU, DIPEA, DMF; (d) SOCl2; (e) NaH, DMF; (f) LiOH, THF, H2O

[0076] p-Hydroxybenzyl alcohol 39 (500 mg, 4.03 mmol) was dissolved in DMF (5 mL), potassium carbonate (667.98 mg, 4.83 mmol) was added, and the mixture was stirred at 80 °C for 15 min. Then 40 (2.9 mmol) was added, and the reaction was carried out overnight. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. Then it was separated and purified by column chromatography (PE / EA = 10 / 1). The separated product was dried and weighed to obtain the target product (n = 1, 750 mg, 78%; n = 5, 956 mg, 65%). Next, the obtained product was dissolved in a mixed solution of tetrahydrofuran and water, lithium hydroxide (3 equiv) was added, and the reaction was carried out for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure to obtain product 41 quantitatively. Under the protection of argon, 41 (4.01 mmol), 42 (4.01 mmol, 0.66 g), HATU (2.29 g, 6.02 mmol), diisopropylethylamine (3.34 mL, 24.07 mmol), and DMF (5 mL) were added respectively, and the reaction was carried out for 2 h. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The crude product was purified by silica gel column (PE / EA = 3 / 1) to obtain 43 (n = 1, 762 mg, 60%; n = 5, 920 mg, 59%). 43 (0.6 mmol) was dissolved in anhydrous dichloromethane, and thionyl chloride (0.058 mL, 1.3 equiv) was added dropwise at 0 °C. After the addition was complete, the reaction system was warmed to room temperature and the reaction was continued for 1 h. The reaction was monitored by TLC. After the reaction was complete, an appropriate amount of small ice cubes were added and stirred for 10 min to quench the reaction. The mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The crude product was purified by silica gel column (PE / EA = 1.5 / 1) to obtain 44 (n = 1, 107 mg, 44%; n = 5, 117 mg, 50%). Compound 44 was dissolved in DMF (2 mL), and sodium hydride (13.21 mg, 1.2 equiv) was added portionwise at 0 °C. After stirring for 0.5 h, 3 (302.79 mmol) was added to the reaction system and the reaction was carried out overnight. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times and concentrated in vacuo. The crude product was purified by silica gel column (PE / EA = 1.5 / 1) to obtain 45 (n = 1, 72 mg, 40%; n = 5, 100 mg, 78%). 45 (50 mg) was dissolved in a mixed solution of tetrahydrofuran and water, lithium hydroxide (3 equiv) was added, and the mixture was stirred at room temperature for 0.5 h. Then the reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation under reduced pressure to obtain the target product D29 in quantitative yield. 11H NMR (400 MHz, CDCl3) δ 8.96 (s, 1H), 8.60 (s, 1H), 7.67 (d, J = 8.3 Hz, 2H), 7.53 (d, J = 8.4 Hz, 1H), 7.41 (d, J = 8.5 Hz, 1H), 7.34–7.27 (m, 2H), 7.22 (d, J = 8.2 Hz, 2H), 6.93 (d, J = 8.2 Hz, 2H), 6.23 (s, 1H), 5.38 (s, 2H), 4.39 (s, 2H), 2.13–2.01 (m, 10H), 1.70 (s, 5H); PH1421, 1 1H NMR (400 MHz, CDCl3) δ 8.83 (s, 1H), 8.50 (s, 1H), 7.58 (d, J = 8.6 Hz, 3H), 7.43 (d, J = 8.5 Hz, 1H), 7.34 (d, J = 8.5 Hz, 1H), 7.24–7.17 (m, 3H), 7.08 (d, J = 8.3 Hz, 2H), 6.78 (d, J = 8.4 Hz, 2H), 5.25 (s, 2H), 3.85 (t, J = 6.4 Hz, 2H), 2.05 (t, J = 7.1 Hz, 2H), 1.98 (s, 3H), 1.91 (s, 7H), 1.73–1.66 (m, 2H), 1.58 (s, 9H). D33, 1 1H NMR (400 MHz, CDCl3) δ 8.83 (s, 1H), 8.50 (s, 1H), 7.58 (d, J = 8.6 Hz, 3H), 7.43 (d, J = 8.5 Hz, 1H), 7.34 (d, J = 8.5 Hz, 1H), 7.24–7.17 (m, 3H), 7.08 (d, J = 8.3 Hz, 2H), 6.78 (d, J = 8.4 Hz, 2H), 5.25 (s, 2H), 3.85 (t, J = 6.4 Hz, 2H), 2.05 (t, J = 7.1 Hz, 2H), 1.98 (s, 3H), 1.91 (s, 7H), 1.73–1.66 (m, 2H), 1.58 (s, 9H).

[0077] Example 8: Synthesis of Compounds D32 and D36

[0078]

[0079] Reagents and reaction conditions: (a) SOCl2; (b) THF; (c) TsCl, DMAP, Et3N, DCM; K2CO3, 80 °C, DMF; (e) SOCl2; (f) NaH, DMF; (g) LiOH, THF, H2O.

[0080] Adamantane acetic acid 46 (500 mg, 2.57 mmol) was dissolved in thionyl chloride (5 mL) and refluxed for 2 h. The solvent was removed by rotary evaporation under vacuum to obtain adamantane acetyl chloride, which was directly used in the next step. In a solution of THF, adamantane acetyl chloride (2.57 mmol) and compound 47 (6.43 mmol) were successively added, and the mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC. After the reaction was complete, the solvent was removed by rotary evaporation under vacuum. The crude product was purified by silica gel column chromatography (DCM / MeOH = 20 / 1) to obtain 48 quantitatively. In an argon atmosphere, 48 (1.42 mmol), p-toluenesulfonyl chloride (541.99 mg, 2.84 mmol, 2 equiv), dimethylaminopyridine (86.83 mg, 0.71 mmol), and triethylamine (0.395 mL, 2.84 mmol) were successively added to 10 mL of dichloromethane. The mixture was stirred at room temperature for 12 h. After the reaction was complete, it was extracted with ethyl acetate, and the organic phase was concentrated. The crude product was purified by silica gel column chromatography (PE / EA = 3 / 1) to obtain 50 quantitatively (R = O(CH2)2, 250 mg, 40.4%; R = (CH2)2, 384 mg, 45%).

[0081] p-Hydroxybenzyl alcohol 51 (100 mg, 805.55 μmol) was dissolved in DMF (2 mL), cesium carbonate (393.69 mg, 1.21 mmol) was added, and the mixture was stirred at 80 °C for 15 min. Then 50 (805.55 μmol) was added, and the reaction was carried out overnight. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. Then it was separated and purified by column chromatography (DCM / MeOH = 40 / 1). The separated product was dried and weighed to obtain the target product 52 (R = O(CH2)2, 113 mg, 63.5%; R = (CH2)2, 230 mg, 76.8%). 52 was dissolved in anhydrous dichloromethane, and thionyl chloride (0.058 mL, 1.3 equiv) was added dropwise at 0 °C. After the addition, the reaction system was warmed to room temperature and continued to react for 1 h. The reaction was monitored by TLC. After the reaction was complete, an appropriate amount of small ice cubes was added and stirred for 10 min to quench the reaction. The mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, filtered, concentrated in vacuo, and the crude product was purified by silica gel column (PE / EA = 1.5 / 1) to obtain 53 (R = O(CH2)2, 50 mg, 42%; R = (CH2)2, 515 mg, 64%). Compound 53 (50 mg, 137.63 μmol) was dissolved in DMF (2 mL), and sodium hydride (6.06 mg, 1.2 equiv) was added portionwise at 0 °C. After stirring for 0.5 h, 3 (151.39 μmol) was added to the reaction system and reacted overnight. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, concentrated in vacuo, and the crude product was purified by silica gel column (PE / EA = 1.5 / 1) to obtain 54 (R = O(CH2)2, 40 mg, 44%; R = (CH2)2, 45 mg, 45.6%). 54 (20 mg) was dissolved in a mixed solution of tetrahydrofuran and water, lithium hydroxide (3 equiv) was added, and the mixture was stirred at room temperature for 0.5 h. Then the reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, and the filtrate was concentrated under reduced pressure to remove the solvent to obtain the target product D36 in quantitative yield. 1 H NMR (400 MHz, CDCl3) δ 8.84 (d, J = 3.8 Hz, 1H), 8.50 (s, 1H), 7.58 (d, J = 8.1 Hz, 2H), 7.43 (d, J = 8.5 Hz, 1H), 7.34 (d, J = 8.5 Hz, 1H), 7.22 (d, J = 8.5 Hz, 2H), 7.10 (d, J = 8.1 Hz, 2H), 6.83 (d, J = 8.1 Hz, 2H), 5.27 (s, 2H), 4.02 (s, 2H), 3.73 (s, 2H), 3.54 (s, 2H), 3.40 (s, 2H), 1.84 (s, 5H), 1.58 (d, J = 12.0 Hz, 3H), 1.50 (s, 9H); D321 1H NMR (400 MHz, CDCl3) δ 8.81 (d, J = 8.5 Hz, 1H), 8.50 (s, 1H), 7.57 (d, J = 8.5 Hz, 2H), 7.42 (d, J = 8.4 Hz, 1H), 7.33 (d, J = 8.5 Hz, 1H), 7.27–7.16 (m, 3H), 7.08 (d, J = 7.8 Hz, 2H), 6.78 (d, J = 7.9 Hz, 2H), 5.24 (s, 2H), 3.88 (d, J = 5.2 Hz, 2H), 3.23 (d, J = 5.2 Hz, 2H), 1.85 (d, J = 9.9 Hz, 5H), 1.72 (d, J = 6.1 Hz, 2H), 1.60 (d, J = 12.3 Hz, 5H), 1.52 (s, 9H).

[0082] Example 9: Synthesis of Compound D34

[0083]

[0084]

[0085] Reagents and reaction conditions: (a) Pd(PPh3)2Cl2, CuI, DIPEA, THF, rt; (b) TFA, DCM, rt; HATU, DIPEA, DMF; (c) Pd / C, H2, MeOH; (d) LiOH, THF, H2O.

[0086] Under an argon atmosphere, 7 (200 mg, 345.24 μmol), tert-butyl (2-(2-(prop-2-yn-1-yloxy)ethoxy)ethyl)carbamate 55 (100.8 mg, 414.28 μmol), bis(triphenylphosphine)palladium(II) dichloride (24.24 mg, 34.52 μmol, 0.1 equiv), copper(I) iodide (6.58 mg, 34.52 μmol, 0.1 equiv), diisopropylethylamine (0.18 mL, 1.03 mmol, 3 equiv), and 4 mL of tetrahydrofuran were added successively, and the mixture was stirred at room temperature for 12 h. The reaction was monitored by TLC. After the reaction was complete, it was concentrated under vacuum, and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain 56 (92 mg, 38%). 56 (100 mg, 143.95 μmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (0.4 mL) was slowly added dropwise, and the mixture was stirred at room temperature for 0.5 h. After the reaction was complete as monitored by TLC on a plate, it was concentrated under vacuum and directly used for the next step. Under an argon atmosphere, adamantaneacetic acid 46 (31.37 mg, 161.46 μmol), HATU (102.33 mg, 269.09 μmol), DIPEA (0.117 mL, 672.74 μmol), and DMF (2 mL) were added successively, and the mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After the reaction was complete, it was extracted with ethyl acetate three times, concentrated under vacuum, and the crude product was purified by silica gel column chromatography (DCM / CH3OH = 40 / 1) to obtain 57 (62 mg, 60%). 2-30 (40 mg, 57.08 μmol) was added to a solution of palladium on carbon (4 mg) in methanol, and the reaction was carried out under a hydrogen atmosphere. After the reaction was complete as detected by TLC, it was purified by silica gel column chromatography. The obtained product was dissolved in a mixed solution of tetrahydrofuran and water (tetrahydrofuran / water = 1 mL / 1 mL), and the mixture was stirred at room temperature for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, it was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation under reduced pressure to obtain the target product D34 in quantitative yield. 1 H NMR (500 MHz, CDCl3) δ 7.56 (d, J = 8.6 Hz, 2H), 7.42–7.24 (m, 2H), 7.20 (d, J = 7.1 Hz, 2H), 7.14–6.95 (m, 7H), 5.18 (d, J = 12.0 Hz, 2H), 3.54–3.46 (m, 7H), 3.40–3.35 (m, 4.6 Hz, 5H), 2.60–2.55 (m, 2H), 1.88 (d, J = 13.5 Hz, 6H), 1.65–1.57 (m, 11H).

[0087] Example 10: Synthesis of Compounds D35 and D37

[0088]

[0089]

[0090] Reagents and reaction conditions: (a) Pd(PPh3)4, CuI, DIPEA, THF, 100 °C; (b) TFA, DCM, rt; (c) HATU, DIPEA, DMF; (d) Pd / C, H2, MeOH; (e) LiOH, THF, H2O.

[0091] For the specific synthetic route and operation process of compounds D35 and D37, refer to that of compound D34, and replace the intermediate tert-butyl (2-(2-(prop-2-ynyloxy)ethoxy)ethyl)carbamate and adamantaneacetic acid with ethynyl-triglycol-tert-butyl ester and adamantanemethylamine.

[0092] Example 11: Synthesis of Compounds D30 and D31

[0093]

[0094]

[0095] Reagents and reaction conditions: (a) EDCI, DCM, rt; (b) Pd(PPh3)4, CuI, DIPEA, THF, 100 °C; (c) Pd / C, H2, MeOH; (d) LiOH, THF / H2O.

[0096] Under an argon atmosphere, add 46 adamantaneacetic acid (400 mg, 2.06 mmol), 63 propargylamine (136.09 mg, 2.47 mmol), EDCI (473.65 mg, 2.47 mmol), and dichloromethane (10 mL) to a 50 mL round-bottom flask in sequence, stir overnight at room temperature, extract with dichloromethane three times, concentrate under vacuum, and purify the crude product by silica gel column chromatography to obtain product 64 (290 mg, 61%). Under an argon atmosphere, add 7 (100 mg, 172.62 μmol), tetrakis(triphenylphosphine)palladium (3.99 mg, 34.52 μmol, 0.02 equiv), copper(I) iodide (1.32 mg, 6.9 μmol, 0.1 equiv), diisopropylethylamine (0.08 mL, 0.517 mmol, 3 equiv), 4 mL of tetrahydrofuran to a 50 mL round-bottom flask in sequence, and react at 100 °C for 12 h. Monitor the reaction by TLC. After the reaction is complete, concentrate under vacuum, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain 65 (80 mg, 68%). Dissolve 65 (20 mg) in an aqueous solution of tetrahydrofuran, add lithium hydroxide (3 equiv), and stir at room temperature for 0.5 h. Extract with ethyl acetate three times, dry with anhydrous magnesium sulfate, evaporate the solvent under reduced pressure from the filtrate, and obtain the target product D30 in quantitative yield.1 1H NMR (500 MHz, CDCl3) δ 8.93 (s, 1H), 8.51 (d, J = 1.4 Hz, 1H), 7.58 (d, J = 8.7 Hz, 2H), 7.43 (dd, J = 8.5, 1.6 Hz, 1H), 7.35–7.18 (m, 6H), 7.07 (d, J = 8.2 Hz, 2H), 5.34 (s, 2H), 4.18 (d, J = 5.2 Hz, 2H), 1.89 (s, 5H), 1.62 (d, J = 12.1 Hz, 3H), 1.54 (d, J = 11.6 Hz, 9H). 65 (60 mg) was dissolved in methanol (2 mL), palladium carbon (6 mg) was added, and the reaction was carried out in a hydrogen atmosphere. The reaction was monitored by TLC. After the reaction was complete, it was concentrated under vacuum. The crude product was purified by silica gel column (PE / EA = 3 / 1) to obtain the product 66 quantitatively. 66 was dissolved in a mixed solution of tetrahydrofuran and water (2 mL, tetrahydrofuran / water = 1 mL / 1 mL), and stirred at room temperature for 0.5 h. Extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation under reduced pressure to obtain the target product D31 in quantitative yield. 1 1H NMR (400 MHz, CDCl3) δ 8.64 (d, J = 0.9 Hz, 1H), 8.45 (s, 1H), 7.68–7.62 (m, 2H), 7.50–7.45 (m, 1H), 7.36 (d, J = 8.6 Hz, 1H), 7.32–7.25 (m, 2H), 7.13 (q, J = 8.0 Hz, 4H), 5.34 (s, 2H), 3.24 (dd, J = 13.4, 6.7 Hz, 2H), 2.62 (t, J = 7.6 Hz, 2H), 1.94 (s, 3H), 1.88 (s, 2H), 1.78 (d, J = 7.4 Hz, 2H), 1.68 (d, J = 12.1 Hz, 3H), 1.60 (t, J = 5.0 Hz, 9H).

[0097] Example 12: Synthesis of Compounds D38 and D39

[0098]

[0099] Reagents and reaction conditions: (a) HATU, DIPEA, DMF; (b) HATU, DIPEA, DMF; (c) LiOH, THF, H2O.

[0100] Under an argon atmosphere, 67 (500 mg, 1.66 mmol, 1 equiv), adamantanemethylamine 60 (300 mg, 1.99 mmol, 1.2 equiv), HATU (1.26 g, 3.32 mmol, 2 equiv), DIPEA (0.866 mL, 5 mmol), and DMF (6 mL) were added to a 50 mL beaker at once, and the mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, concentrated in vacuo, and the crude product was purified by silica gel column chromatography (DCM / CH3OH = 40 / 1) to obtain the target product (280 mg, 19.4%). 80 mg of the target product was dissolved in a mixed solution of dichloromethane and trifluoroacetic acid and stirred for 0.5 h. After the reaction was complete, the solvent was evaporated, and 69 (89.56 mg, 1180.9 μmol, 1 equiv), HATU (137.59 mg, 361.86 μmol, 2 equiv), DIPEA (0.94 mL, 542.79 μmol, 3 equiv), and DMF (2 mL) were added in turn under an argon atmosphere, and the mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, concentrated in vacuo, and the crude product was purified by silica gel column chromatography (DCM / MeOH = 40 / 1) to obtain D38 (99 mg, 67%). 1 1H NMR (400 MHz, CDCl3) δ 8.56 (d, J = 1.4 Hz, 1H), 8.39 (s, 1H), 7.69 (d, J = 8.2 Hz, 2H), 7.57 (d, J = 8.7 Hz, 2H), 7.38 (dd, J = 8.5, 1.6 Hz, 1H), 7.24–7.20 (m, 2H), 7.14 (d, J = 8.1 Hz, 2H), 6.41 (d, J = 4.7 Hz, 1H), 5.35 (s, 2H), 5.11 (s, 1H), 3.86 (s, 3H), 3.33 (d, J = 6.4 Hz, 2H), 2.71 (s, 3H), 1.95 (d, J = 7.4 Hz, 4H), 1.89 (d, J = 2.4 Hz, 6H), 1.57 (s, 5H), 1.48 (s, 5H), 1.17 (s, 9H). 55 mg of D38 was dissolved in a mixed solution of tetrahydrofuran and water, lithium hydroxide (3 equiv) was added, and the mixture was stirred for 0.5 h. The reaction was monitored by TLC. After the reaction was complete, the mixture was extracted with ethyl acetate three times, dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation under reduced pressure to obtain the target product D39 in quantitative yield. 11H NMR (400 MHz, CDCl3) δ 8.56 (d, J = 1.4 Hz, 1H), 8.39 (s, 1H), 7.69 (d, J = 8.2 Hz, 2H), 7.57 (d, J = 8.7 Hz, 2H), 7.38 (dd, J = 8.5, 1.6 Hz, 1H), 7.24–7.20 (m, 2H), 7.14 (d, J = 8.1 Hz, 2H), 6.41 (d, J = 4.7 Hz, 1H), 5.35 (s, 2H), 5.11 (s, 1H), 3.33 (d, J = 6.4 Hz, 2H), 2.71 (s, 3H), 1.95 (d, J = 7.4 Hz, 4H), 1.89 (d, J = 2.4 Hz, 6H), 1.57 (s, 5H), 1.48 (s, 5H), 1.17 (s, 9H).

[0101] Example 13: Detection of the degradation effect of the compound on SRPINE1 protein by Western Blot

[0102] (1) Extraction of total protein: Transfer the cells treated with DMSO or the drug to a centrifuge tube, centrifuge to collect the cells, wash them twice with PBS, and transfer them to a 1.5 mL EP tube. Add 200 μL of lysis buffer to each EP tube according to the cell number, shake on ice for 20 minutes, centrifuge at 12,000 g for 10 minutes in a low-temperature centrifuge to collect the supernatant, and measure the protein concentration. Then add the protein supernatant to 5x loading buffer and boil it at 95 °C for 10 minutes in a constant-temperature metal bath.

[0103] (2) Preparation of SDS-PAGE gel: Place the cleaned glass plates to dry, and prepare 10% separating gel and 5% stacking gel successively according to the formula. Add 20 μL of protein sample to each well equally. Electrophoresis: Set the voltage for stacking gel electrophoresis at 100 V and the voltage for separating gel electrophoresis at 150 V.

[0104] (3) Transfer: Prepare a PVDF membrane soaked in methanol, filter paper soaked in transfer buffer, black sponge, separating gel, transfer clip, and filter paper, and place them in a tray filled with transfer buffer. Place the black side of the transfer clip down, and successively place the sponge, filter paper, separating gel, PVDF membrane, filter paper, and sponge on the transfer clip. After removing the air bubbles, clamp it and place it in the transfer tank for fixation. Use the wet transfer method and transfer at a constant current of 0.2 A for 2 hours.

[0105] (4) Immunoblotting blocking: Place the PVDF membrane after transfer into a plastic box containing 5% non-fat milk powder and block at room temperature for 1 hour. Incubate with primary antibody: Dilute the primary antibody with the primary antibody diluent from Beyotime Biotechnology according to the instructions. Place the transfer membrane into the diluted primary antibody and incubate overnight on a shaker at 4°C. Incubate with secondary antibody: Wash the membrane with TBST for 10 minutes × 3 times the next day. Then transfer the transfer membrane into the diluted secondary antibody and incubate at room temperature for 1 hour. Then wash it again with TBST for 10 minutes × 3 times. ECL development: Evenly apply the prepared ECL luminescent solution onto the strip to be detected. After placing it in the dark for 1 minute, use a chemiluminescence imaging system for development and photography.

[0106] Example 14: Detection of the killing effect of compounds on tumor cells by MTS

[0107] Taking pancreatic cancer cell line AsPC-1 and intestinal cancer cell line Rko as examples, digest and centrifuge the cells in the logarithmic growth phase, discard the supernatant, wash twice with PBS and then resuspend and count. Uniformly seed 3000 cells / 100 μL per well on a 96-well plate. Set up duplicate wells and blank controls. Dilute the drug with fresh medium according to the required concentration and add 100 μL per well to the 96-well plate seeded with cells. Incubate overnight in an incubator at 37°C with CO2. The next day, add DMSO or the drug for treatment and then incubate in an incubator at 37°C with CO2 for 2 days. On the fourth day, add 20 μL of MTS solution to the 96-well plate and react for 4 hours. Use an enzyme-linked immunosorbent assay (ELISA) reader to measure the absorbance (OD value) of different wells at a wavelength of 490 nm.

[0108] Example 15: Antitumor growth and metastasis effect of compound D10 tested by animal experiment of footpad-popliteal lymph node metastasis model

[0109] (1) Collect tumor cells in the logarithmic growth phase, wash twice with PBS solution, resuspend the cells with serum-free RPMI-1640 medium, calculate the number of cells to be inoculated, AsPC-1 model cells: 1×10 5 / 50 μL;

[0110] (2) Inject 50 μL of cell suspension into the left footpad of 6-week-old nude mice in the prone position;

[0111] (3) One week later, randomly divide the mice into 3 groups, namely the solvent control group and 2 drug treatment groups;

[0112] (4) Administration: Drug treatment groups Tiplaxtinin (30 mg / kg), D10 (60 mg / kg), solvent control group DMSO (10%, DMSO:castor oil:PBS buffer = 1:1:8), intraperitoneally inject the drug or solvent 200 μL per mouse every 3 days. After the administration is completed, measure the body weight of the mice and draw a body weight measurement curve graph;

[0113] (5) After 30 days of continuous administration, the experiment was terminated on the 31st day, and nude mice were sacrificed by dislocation. The in-situ plantar tumor tissues and popliteal lymph node tissues were taken. The weight of the in-situ tumor tissues was measured with the contralateral plantar tissues as a control. After the popliteal lymph node tissues were paraffin-embedded, HE staining was performed to identify the tumor invasion situation.

[0114] To screen compounds that target the degradation of SERPINE1 protein. In this invention, Rko and AsPC-1 cells were treated with 10 μM of the compound for 48 h, and 10 μM of Tiplaxtinin or the same volume of DMSO solution was used as a control. The expression level of SERPINE1 was measured by Western blot experiment. Through two rounds of compound screening, the results are shown in Table 1, Table 2 and Figure 1 as follows.

[0115] Table 1. Screening results of different SERPINE1 protein degraders in Rko and AsPC-1 cells

[0116]

[0117] Table 2. Screening results of different SERPINE1 protein degraders in Rko and AsPC-1 cells

[0118]

[0119] From the data in Table 1 and Table 2, it can be seen that:

[0120] 1) When the length of the main chain is 3 to 6 atoms, the degradation activity increases with the extension of the chain length. When the chain length is 6 atoms, the activity reaches the maximum;

[0121] 2) When R is hydrogen or a methyl group, the two have little effect on the activity. The reason may be that the methyl ester will be hydrolyzed by hydrolase to formic acid in the cell;

[0122] 3) When the length of the main chain is 7 to 13 atoms, the degradation activity shows a downward trend with the extension of the chain.

[0123] The above is a further detailed description of the present invention, and it cannot be regarded as a limitation to the specific implementation of the present invention. For those of ordinary skill in the technical field to which the present invention pertains, any simple deduction or substitution without departing from the concept of the present invention is within the protection scope of the present invention.

[0124] As Figure 1 shown, compound D10 can significantly reduce the level of SERPINE1 protein in both Rko and AsPC-1 cells.

[0125] Figure 2Demonstrate the degradation effect of compound D10 of the present invention on SERPINE1 protein. After treating Rko cells with compound Tiplaxtinin and D10 at concentration gradients for 48 hours, the expression level of SERPINE1 protein was determined by Western blot assay. After treating AsPC-1 cells with compound D10 at concentration gradients for 48 hours, the expression level of intracellular SERPINE1 protein was determined by Western blot assay, and the content of secreted SERPINE1 protein in the culture medium was detected by ELISA assay. The above results indicate that some compounds including D10 can significantly reduce the expression of SERPINE1 protein.

[0126] Furthermore, after treating Rko and AsPC-1 cells with compound D10 for 6, 12, 24, 36 and 48 hours respectively, the expression level of intracellular SERPINE1 protein was determined by Western blot assay, and the content of secreted SERPINE1 protein in the cell culture medium was detected by ELISA assay. The results are as Figure 2 shown that compound D10 can effectively inhibit the expression of SERPINE1 protein after treating cells for 48 hours.

[0127] Figure 3 Demonstrate the effects of compound D10 of the present invention and the existing SERPINE1 inhibitor Tiplaxtinin on the proliferation of Rko and AsPC-1 cells. Treat Rko and AsPC-1 cells with compounds at different concentration gradients respectively, and determine the cell viability by MTS after 48 hours. The results show that the killing effect of compound D10 of the present invention on tumor cells is significantly better than that of Tiplaxtinin.

[0128] Figure 4 The results demonstrate that the therapeutic effect of compound D10 of the present invention on tumors is better than that of Tiplaxtinin. The nude mouse plantar-popliteal metastasis animal experimental model was used to evaluate the inhibitory effects of compound D10 and Tiplaxtinin on tumor growth and metastasis. As shown in the schematic diagram of the animal experiment, 8-week-old BALB / c mice were injected with AsPC-1 cells into the plantar surface. One week after transplantation, they were randomly divided into groups to start intraperitoneal injection of compound D10 and Tiplaxtinin, and the compounds were converted to the same concentration according to the molecular weight and administered once every 3 days. The results show that compared with the existing Tiplaxtinin, compound D10 significantly reduces the lymph node metastasis of tumors and effectively inhibits the growth of tumors. In addition, the body weight monitoring data of the mice show that compound D10 has no toxic side effects and good safety.

[0129] The above is a further detailed description of the present invention, and it should not be regarded as a limitation to the specific implementation of the present invention. For those of ordinary skill in the technical field to which the present invention pertains, any simple deduction or substitution without departing from the concept of the present invention falls within the protection scope of the present invention.

[0130] The above is a further detailed description of the present invention, and it should not be regarded as a limitation to the specific implementation of the present invention. For those of ordinary skill in the technical field to which the present invention pertains, any simple deduction or substitution without departing from the concept of the present invention falls within the protection scope of the present invention.

Claims

1. A SERPINE1 protein degrader, characterized in that, Comprising: 1) A structural fragment that inhibits SERPINE1 protein, the structural formula of which is as well as 2) An E3 ligase VHL ligand structural fragment or a hydrophobic tag structural fragment, The structural fragment 1) and the structural fragment 2) are connected by a linker whose main chain contains 1 to 14 atoms, and the general structural formula of the degrader is shown in Formula I or II: In Formula I and Formula II, R is selected from H, an alkyl group of C1-C4.

2. The SERPINE1 protein degrader according to claim 1, wherein The main chain of the linker is an alkyl chain, a polyethylene glycol chain, an ether-containing alkyl chain, an alkyne-containing alkyl chain or an alkyne- and ether-containing alkyl chain.

3. The SERPINE1 protein degrader according to claim 1 or 2, characterized in that, The main chain of the linker contains 4 to 8 atoms.

4. The SERPINE1 protein degrader according to claim 1, wherein The structural formula I is selected from one of the formulas shown in X1-X7: The structural formula II is selected from one of the formulas shown in X8-X18:

5. The SERPINE1 protein degrader according to claim 1, wherein Its structural formula is one of the formulas D1-D39: In Formulas D1 to D39, 6. A pharmaceutical composition comprising an active ingredient and an excipient, characterized in that: Its active ingredient comprises the SERPINE1 protein degrader according to any one of claims 1-5, or a pharmaceutically acceptable salt, crystal, or solvate thereof.

7. The pharmaceutical composition according to claim 6, characterized in that, Its dosage form is an injection, an oral preparation, or a transdermal absorbent.

8. Use of the SERPINE1 protein degrader according to any one of claims 1-5 in the preparation of an anti-tumor drug.

9. The application according to claim 8, characterized in that The tumor is a solid tumor or a hematological tumor.

10. The application according to claim 9, wherein The solid tumor is selected from colorectal cancer, pancreatic cancer, bladder cancer, or cervical cancer; the hematological tumor is selected from leukemia or lymphoma.

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