A rar protac compound, pharmaceutical composition, preparation method and application

Abstract

The application provides a RAR PROTAC compound, which has the structure shown in the following general formula: the application is characterized in that a retinoic acid receptor RAR PROTAC molecule with a new structure is designed and synthesized, RAR is targeted and degraded by using the PROTAC technology, the RAR PROTAC molecule has a good inhibitory effect on tumor stem cells, and the RAR PROTAC molecule has good pharmacokinetics, bioavailability and in-vivo drug efficacy, and can be developed into a new type of antitumor drug.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of drug synthesis and application, and particularly relates to a RAR PROTAC compound, a pharmaceutical composition, a preparation method and application. BACKGROUND

[0002] Cancer Stem Cells (CSCs) are a special subpopulation of tumor cells that play a key role in tumor development, invasion, and distant metastasis. CSCs have the ability of unlimited or long-term self-renewal, which means they can continuously divide and produce new tumor cells. Although CSCs may remain undifferentiated in the tumor microenvironment, they have the ability to differentiate into various types of tumor cells. CSCs usually have a high resistance to conventional chemotherapy and radiotherapy, and can also promote immune escape by affecting immune cells in the tumor microenvironment, which allows them to survive after treatment and can lead to tumor recurrence and metastasis.

[0003] The development of drugs targeting CSCs is an important research direction. If CSCs can be effectively targeted and eliminated, it can significantly improve the success rate of cancer treatment and reduce recurrence. WYC-209, as a new synthetic retinoid, shows significant inhibitory effect on CSCs of various types of tumors by targeting Retinoic Acid Receptors (RAR), which provides new possibilities for developing therapeutic strategies targeting CSCs.

[0004] Proteolysis-Targeting Chimeras (PROTACs) technology is a new protein degradation strategy that utilizes the ubiquitin-proteasome system (UPS) in cells to specifically degrade target proteins. The core idea of this technology is to design and develop small molecule compounds that can simultaneously bind two proteins: one is the target protein (POI), and the other is the E3 ubiquitin ligase; through this dual-specific binding, PROTACs promote the ubiquitination of the target protein, which in turn marks the protein for recognition and degradation by the proteasome, making PROTACs show great potential in treating diseases caused by tumors. Given this, the use of PROTAC technology to develop a series of PROTAC molecules targeting the degradation of RAR will be a potential targeted therapy strategy for inhibiting malignant cancer stem cells. SUMMARY

[0005] The purpose of the present application is to provide a RAR PROTAC compound, which at least can solve some defects in the prior art.

[0006] To achieve the above object, the present application adopts the following technical solutions:

[0007] A RAR PROTAC compound has the structure shown in the general formula:

[0008]

[0009] The E3 ligand is selected from an IAP ligand

[0010] One of the above.

[0011] Further, the linker is one of a rigid linker, a flexible linker, or a flexible and rigid combined linker.

[0012] Further, the rigid linker is selected from one of the following structural formulas:

[0013] The flexible linker is selected from one of the following structural formulas:

[0014]

[0015]

[0016] The flexible and rigid combined linker is selected from one of the following structural formulas:

[0017]

[0018] Wherein n = 0-10.

[0019] Further, the RAR PROTAC compound is selected from any one of the following compounds:

[0020]

[0021]

[0022]

[0023]

[0024] In addition, the present application also provides a preparation method of the above RAR PROTAC compound, comprising the following steps:

[0025] S1, synthesizing 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-hydroxy pyrimidine-5-carboxylic acid ethyl ester or 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-bromopyrimidine-5-carboxylic acid ethyl ester;

[0026] S2, using 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-hydroxy pyrimidine-5-carboxylic acid ethyl ester or 2-((4,4-dimethyl-1-oxobenzothiopyran-6-)ethynyl)-4-bromopyrimidine-5-carboxylic acid ethyl ester synthesized in step S1 to prepare a retinoic acid compound;

[0027] S3, reacting an E3 ligand with a linker to obtain an intermediate compound;

[0028] S4, using the retinoic acid compound and the intermediate compound to prepare a RAR PROTAC compound.

[0029] Further, in the step S4, the process of using the retinoic acid compound and the intermediate compound to prepare the RAR PROTAC compound is as follows: the retinoic acid compound and the intermediate compound are dissolved in N,N dimethylformamide, then triacetyl sodium borohydride or HATU is added, and the reaction is stirred at room temperature; after the reaction is completed, saturated ammonium chloride solution is added to quench the reaction, then ethyl acetate and water are added for extraction, saturated brine is used to wash the ethyl acetate phase, and after distillation under reduced pressure, the RAR PROTAC compound is obtained by plate separation.

[0030] The application also provides a use of the above-mentioned RAR PROTAC compound and pharmaceutically acceptable salt thereof in the preparation of an antitumor drug.

[0031] Further, the pharmaceutically acceptable salt of the RAR PROTAC compound is a salt formed by the RAR PROTAC compound and an acid selected from any one of the following: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid or ferulic acid.

[0032] Further, the tumor is melanoma, breast cancer or prostate cancer.

[0033] The application also provides a pharmaceutical composition comprising the above-mentioned RAR PROTAC compound and one or more combinations of pharmaceutically acceptable carriers, excipients, diluents, adjuvants, vehicles of the RAR PROTAC compound.

[0034] Compared with the prior art, the application has the following beneficial effects:

[0035] This invention designs and synthesizes a novel retinoic acid receptor (RAR) PROTAC molecule, and uses PROTAC technology to target and degrade RAR, which has a good inhibitory effect on tumor stem cells. It has good pharmacokinetic, bioavailability and in vivo efficacy properties, and can be developed into a novel anti-tumor drug. Detailed Implementation

[0036] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0037] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This application contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (2)- isomers, racemic mixtures thereof, and other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this application. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this application.

[0038] The compounds and intermediates of this invention may also exist in different tautomer forms, and all such forms are included within the scope of this application. The terms "tautomer" or "tautomer form" refer to structural isomers with different energies that can interconvert via a low energy barrier. The compounds of this application may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compound may be labeled with radioactive isotopes, such as tritium (…). 2 H), Iodine-125 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. All isotopic variations of the compounds in this application, regardless of radioactivity, are included within the scope of this application.

[0039] The term "targeting chimera" refers to a bifunctional molecule containing two small molecule ligands: one with high affinity for the target protein and a second for recruiting an E3 ligase that ubiquitinates the protein and targets it for proteolysis via the proteasome.

[0040] In this embodiment of the invention, the structure of the target compound was determined by high-resolution mass spectrometry (HR-ESI-MS) and proton nuclear magnetic resonance (NMR) spectroscopy. 1 H-NMR,13 Confirmed by C-NMR. NMR was determined using a Bruker Avance III 400 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD) as solvents, and tetramethylsilane (TMS) as the internal standard. MS was determined using an Agilent 6120B (ESI). HPLC was performed using an Agilent 1260DAD high-performance liquid chromatograph (Zorbax SB-C18100×4.6mm, 3.5μM). Thin-layer chromatography (TLC) used Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The silica gel plate size for TLC was 0.15mm-0.20mm, and the size for TLC separation and purification was 0.4mm-0.5mm. Column chromatography generally used Yantai Huanghai 200-400 mesh silica gel as the carrier.

[0041] This invention provides a RAR PROTAC compound having the structure shown in the following general formula:

[0042]

[0043] Among them, the E3 ligand (E3 ligase ligand) is selected from the IAP ligand.

[0044] One of them.

[0045] The linker can be selected from one of the following: a rigid connecting base, a flexible connecting base, or a combination of flexible and rigid connecting bases. Specifically, the rigid connecting base is selected from one of the following structural formulas:

[0046]

[0047] The flexible connection base is selected from one of the following structural formulas:

[0048] in,

[0049] n = 0-10.

[0050] The flexible and rigid combined connection base is selected from one of the following structural formulas:

[0051]

[0052] Where n = 0-10.

[0053] Example 1: Synthesis of ethyl 2-((4,4-dimethylbenzothiaran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate (i.e., compound 3)

[0054]

[0055] 6-ethynyl-4,4-dimethyldihydrobenzothiaran (compound 1) and ethyl 2-chloro-4-hydroxy-5-pyrimidinecarboxylate (compound 2) were added to a flask, along with Pd(PPh3)2Cl2 (dichlorodi(triphenylphosphine)palladium) and CuI (copper iodide). The mixture was protected with argon and purged three times to remove oxygen. 5 mL of dry THF (anhydrous tetrahydrofuran) and dried Et3N (triethylamine) were added using a syringe. The mixture was refluxed, and the reaction was monitored by TLC until complete. After THF was removed, the mixture was extracted with ethyl acetate. The extract was washed successively with 5.0 eq 2N dilute hydrochloric acid, 2 × 30 mL saturated sodium bicarbonate solution, and 30 mL saturated sodium chloride solution. The organic layer was dried with anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the crude product. After purification by column chromatography, compound 3 was obtained.

[0056] The test results of the products prepared above are as follows: 1 H NMR (400MHz, CDCl3) δ7.81(d,J=8.1Hz,1H),7.47(dd,J=6.2,2.2Hz,1H),7.36(s,0H),7.16(dd,J=8.1,2.1Hz,1H),7.09(d ,J=6.2Hz,1H),7.03(d,J=2.1Hz,1H),4.30(q,J=6.4Hz,2H),3.02–2.96(m,2H),2.07–2.01(m,2H),1.37(t,J=6.4Hz,3H). 13 C NMR (125MHz, Common NMR Solvents) δ 170.08, 164.28, 143.59, 135.91, 131.59, 130.85, 128.73, 127.85, 126.37, 123.09, 120.70, 116.74, 115.41, 89.18, 61.42, 37.38, 37.09, 30.22, 26.90, 14.32; This indicates that compound 3 was synthesized.

[0057] Example 2: Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiaran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate (i.e., compound 4)

[0058]

[0059] Compound 3 prepared in Example 1 was added to a flask, dried dichloromethane (DCM) was added, and m-CPBA (m-chloroperoxybenzoic acid) was added under an ice-water bath. The mixture was stirred and brought to room temperature. After the reaction was monitored by TLC thin-layer chromatography, DCM was removed by evaporation, and ethyl acetate was added for extraction. The mixture was washed successively with 2×30 mL of saturated sodium bicarbonate solution and 30 mL of saturated sodium chloride solution. The organic layer was dried with anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the crude product. After purification by column chromatography, compound 4 was obtained.

[0060] The test results of the products prepared above are as follows: 1 ¹H NMR (400MHz, CDCl₃) δ 8.06 (d, J = 6.4 Hz, 1H), 7.81 (d, J = 8.1 Hz, 1H), 7.63–7.57 (m, 2H), 7.16 (dd, J = 8.1, 2.1 Hz, 1H), 7.03 (s, OH), 4.30 (q, J = 6.4 Hz, 2H), 3.18–3.12 (m, 2H), 2.34–2.28 (m, 2H), 1.37 (d, J = 12.7 Hz, 1H), 1.25 (s, 5H); This indicates that compound 4 was synthesized.

[0061] Example 3: Synthesis of ethyl 2-((4,4-dimethyl-1-benzothiaran-6-)ethynyl)-4-bromopyrimidine-5-carboxylate (i.e., compound 6)

[0062]

[0063] Compound 6-ethynyl-4,4-dimethyldihydrobenzothiaran (i.e., compound 1) and ethyl 2-bromo-4-iodobenzoate (i.e., compound 5) were added to a flask, along with Pd(PPh3)2Cl2 and CuI. The mixture was protected with argon gas and the gas was purged three times to remove oxygen. 5 mL of dry THF and dried Et3N were added using a syringe. The mixture was refluxed, and the reaction was monitored by TLC until complete. After THF was removed by evaporation, the mixture was extracted with ethyl acetate. The extract was washed successively with 5.0 eq 2N dilute hydrochloric acid, 2 × 30 mL saturated sodium bicarbonate solution, and 30 mL saturated sodium chloride solution. The organic layer was dried with anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the crude product. Compound 6 was then purified by column chromatography.

[0064] The test results of the products prepared above are as follows: 1¹H NMR (400MHz, CDCl₃) δ 7.89 (d, J = 7.9Hz, 1H), 7.47 (dd, J = 6.2, 2.2Hz, 1H), 7.36 (d, J = 2.2Hz, 1H), 7.23 (s, OH), 7.12–7.07 (m, 2H), 6.69 (s, 2H), 4.30 (q, J = 6.4Hz, 2H), 3.02–2.96 (m, 2H), 2.07–2.01 (m, 2H), 1.37 (t, J = 6.4Hz, 3H); This indicates that compound 6 was synthesized.

[0065] Example 4: Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiaran-6-)ethynyl)-4-bromopyrimidine-5-carboxylate (i.e., compound 7)

[0066]

[0067] Compound 6, obtained in Example 3 above, was added to a flask, dried dichloromethane was added, m-CPBA was added under an ice-water bath, the mixture was stirred and brought to room temperature, and the reaction was monitored by TLC thin-layer chromatography until complete. DCM was removed by evaporation, and ethyl acetate was added for extraction. The mixture was washed successively with 2×30 mL of saturated sodium bicarbonate solution and 30 mL of saturated sodium chloride solution. The organic layer was dried with anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the crude product. After purification by column chromatography, compound 7 (yield 85%) was obtained.

[0068] The test results of the products prepared above are as follows: 1 H NMR(400MHz, CDCl3)δ8.06(d,J=6.4Hz,1H),7.89(d,J=7.9Hz,1H),7.63-7.57(m,2H),7.23(s,0H),7.10(dd,J=7.9,2.2 Hz,1H),6.69(s,2H),4.30(q,J=6.4Hz,2H),3.18-3.12(m,2H),2.34-2.28(m,2H),1.37(d,J=12.7Hz,1H),1.25(s,5H). 13 C NMR (125MHz, CDCl3) δ 168.47, 150.39, 145.26, 144.99, 132.89, 131.86, 128.63, 128.56, 127.63, 125.81, 119.77, 116.13, 111.80, 89.23, 89.14, 60.25, 49.74, 37.24, 33.34, 30.37, 14.32; This indicates that compound 7 was synthesized.

[0069] Example 5: Synthesis of intermediate compound I (i.e., compound 9)

[0070]

[0071] 2.3 g (6.4 mmol) of compound 7 and 2.4 g (12.8 mmol) of 4-(diethoxymethyl)piperidine (compound 8) were added to a flask and reacted with methanol as solvent and TsOH (p-toluenesulfonic acid) as catalyst. After the reaction was completed by TLC thin-layer chromatography, hydrochloric acid was added to give compound 9.

[0072] The test results of the products prepared above are as follows: 1 H NMR (400MHz, CDCl3) δ9.61(dp,J=7.7,0.9Hz,1H),8.06(d,J=6.4Hz,1H),7.83(d,J=8.3Hz,1H),7. 63–7.57(m,2H),7.30(dd,J=8.2,2.2Hz,1H),7.19(s,0H),4.30(q,J=6.4Hz,2H),3.58(ddd,J=12. 1,8.3,5.6Hz,2H),3.45(ddd,J=11.9,8.3,5.6Hz,2H),3.18–3.12(m,2H),2.46(dp,J=7.7,5.4Hz, 1H),2.34–2.28(m,2H),2.12–2.02(m,2H),1.87–1.77(m,2H),1.37(t,J=6.3Hz,3H),1.25(s,5H). 13 C NMR (125MHz, CDCl3) δ 205.56, 167.08, 149.90, 145.26, 144.99, 131.86, 130.41, 128.59, 127.72, 127.63, 125.81, 124.25, 122.39, 117.99, 89.39, 89.24, 61.10, 49.74, 48.97, 45.50, 37.24, 33.34, 30.37, 25.55, 14.32; This indicates the synthesis of compound 9.

[0073] Example 6: Synthesis of compound MJ-1

[0074]

[0075] 1.2 g (1.8 mmol) of compound 9 and 1.2 g (2.7 mmol) of compound 10 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.96 g of pure compound MJ-1, with a yield of 48%.

[0076] The test results of the products prepared above are as follows: 1 H NMR(400MHz, CDCl3)δ8.70(s,1H),7.89(s,1H),7.81–7.75(m,2H),7.60(dq,J=6.2,5.2H z,1H),7.49(d,J=9.0Hz,1H),7.09–7.03(m,2H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41( dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.5 4(t,J=9.1Hz,1H),4.27(q,J=6.4Hz,2H),4.10–4.00(m,2H),3.72–3.64(m,1H),3.59–3.5 0(m,2H),3.42(dddd,J=12.8,4.8,3.6,0.8Hz,1H),3.35(tt,J=8.1,1.8Hz,1H),3.22(dd d,J=8.8,5.7,2.9Hz,4H),3.15(p,J=6.5Hz,1H),2.93–2.78(m,2H),2.73–2.47(m,7H),2. 42(d,J=5.3Hz,3H),2.29–1.99(m,6H),1.95–1.82(m,4H),1.78(ddd,J=12.5,7.9,5.3Hz, 1H),1.66–1.39(m,6H),1.42–1.23(m,12H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13C NMR (125MHz, CDCl3) δ187.22,172.67,171.59,170.54,165.17,163.87,157.51,155.75,153.95,15 1.40,140.94,134.64,132.51,129.31,128.42,123.77,122.95,113.31,109.25,94.16,84.88,66. 20, 63.91, 61.01, 59.05, 55.83, 54.31, 53.13, 49.97, 48.47, 48.16, 47.45, 46.85, 38.58, 36.55, 34.10, 34.03, 33.33, 32.96, 29.89, 29.19, 26.29, 26.10, 25.50, 24.97, 17.21, 14.28; This indicates the synthesis of compound MJ-1.

[0077] Example 7: Synthesis of compound MJ-2

[0078]

[0079] 1.2 g (1.8 mmol) of compound 9 and 1.0 g (2.7 mmol) of compound 11 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.76 g of pure compound MJ-2, with a yield of 48%.

[0080] The test results of the products prepared above are as follows: 1HNMR(400MHz, CDCl3)δ8.70(s,1H),7.31-7.24(m,2H),7.26-7.19(m,1H),7.23-7.17(m ,3H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J= 7.5,2.0Hz,1H),4.47-4.39(m,1H),4.27(q,J=6.4Hz,2H),4.10-4.00(m,2H),3.95(dd, J=6.0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3.64-3.50(m,5H),3.47(dd,J=6.2,3.5Hz,2H ),3.39-3.31(m,1H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.78(m,4H),2.78-2.65(m,2H) ,2.69-2.59(m,1H),2.55(dd,J=6.2,3.5Hz,2H),2.53-2.47(m,2H),2.48(d,J=3.5Hz,1 H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.95-1.82(m,4H),1.82-1.64(m,3H),1.63-1.5 4(m,1H),1.37(t,J=6.4Hz,3H),0.89(d,J=1.5Hz,3H),0.88(s,1H),0.90-0.82(m,8H). 13 C NMR(125MHz,Chloroform-d)δ173.94,171.78,165.17,163.87,157.51,155.75,140 .94,137.18,134.64,129.42,128.79,128.42,127.22,123.77,109.25,94.16,84.88 ,70.54,66.20,63.90,61.01,55.77,53.11,51.55,48.47,48.16,46.85,45.98,40.77,37.51,36.55,34.10,32.96,29.19,26.10,24.29,22.30,14.28; This indicates that compound MJ-2 was synthesized.

[0081] Example 8: Synthesis of compound MJ-3

[0082]

[0083] 1.2 g (1.8 mmol) of compound 9 and 1.0 g (2.7 mmol) of compound 12 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.86 g of pure compound MJ-2, with a yield of 45%.

[0084] The test results of the products prepared above are as follows: 1 H NMR(400MHz, CDCl3)δ8.70(s,1H),7.89(s,1H),7.68-7.56(m,2H),7.52-7.42(m,2H),7.1 3(t,J=2.2Hz,1H),6.73(ddd,J=8.1,2.3,1.3Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6 .41(dtd,J=8.4,2.0,1.0Hz,1H),6.25(t,J=5.7Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26 (dd,J=4.8,3.2Hz,1H),4.54(t,J=9.1Hz,1H),4.27(q,J=6.4Hz,2H),4.08-3.99(m,2H),3 .72-3.64(m,1H),3.60-3.50(m,2H),3.42(dddd,J=12.8,4.8,3.7,0.8Hz,1H),3.35(tt,J =8.1,1.8Hz,1H),3.19(d,J=4.4Hz,1H),3.17(d,J=4.5Hz,1H),3.14(q,J=6.5Hz,1H),2.9 3-2.78(m,2H),2.73-2.65(m,1H),2.42(d,J=5.3Hz,3H),2.29-1.99(m,6H),1.98-1.73(m ,7H),1.66-1.39(m,6H),1.43-1.23(m,12H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13 C NMR (125MHz, Chloroform-d)δ

[0085] 186.74, 172.67, 171.59, 170.54, 165.17, 163.87, 157.51, 155.75, 151.08, 148.68, 140.94, 135.37, 134.64, 131.01, 128.42, 124.83, 123.77, 122.94, 117.88, 115.75, 109.25, 94.16, 84.88 The values ​​are: 66.20, 61.01, 59.05, 55.83, 54.31, 48.47, 48.16, 48.10, 47.45, 46.82, 38.58, 36.55, 34.33, 34.10, 34.03, 33.33, 29.89, 28.93, 26.29, 26.10, 25.50, 24.97, 17.21, 14.28; This indicates that compound MJ-3 was synthesized.

[0086] Example 9: Synthesis of compound MJ-4

[0087]

[0088] 1.2 g (1.8 mmol) of compound 9 and 0.6 g (2.7 mmol) of compound 13 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.56 g of pure compound MJ-4, with a yield of 33%.

[0089] The test results of the products prepared above are as follows: 1H NMR (400MHz, Chloroform-d) δ8.70(s,1H),7.33-7.17(m,6H),6.73(t,J=5.9Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1. 0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.35-4.26(m,2H),4.26(d,J=6.4Hz,1H),4.04(ddd,J=12.5,8.9,6.3Hz,2H),3.97(dd,J=6.1,4.5Hz,1H), 3.87(d,J=4.5Hz,1H),3.58(ddd,J=12.5,8.8,6.2Hz,3H),3.39-3.31(m,1H),3.11(dd,J=5.9,4.9Hz,2H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.7 8(m,4H),2.78-2.65(m,2H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.94-1. 71(m,7H),1.65-1.51(m,2H),1.37(t,J=6.4Hz,3H),0.91-0.82(m,12H). 13 C NMR(125MHz,Chloroform-d)δ173.89,173.67,165.17,163.87,157.51,155.75, 140.94,137.18,134.64,129.42,128.79,128.42,127.22,123.77,109.25,94.16 The values ​​are: 84.88, 70.54, 66.20, 61.01, 55.77, 52.62, 48.47, 48.16, 46.82, 44.88, 40.73, 37.51, 36.55, 34.77, 34.10, 28.74, 26.10, 24.99, 22.40, 14.28; This indicates that compound MJ-4 was synthesized.

[0090] Example 10: Synthesis of compound MJ-5

[0091]

[0092] 1.2 g (1.8 mmol) of compound 9 and 0.8 g (2.7 mmol) of compound 14 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.75 g of pure compound MJ-5, with a yield of 36%.

[0093] The test results of the products prepared above are as follows: 1 H NMR(400MHz,Chloroform-d)δ8.70(s,1H),7.89(s,1H),7.81-7.75(m,2H),7.60(dq,J=6.2,5.2Hz,1H),7.4 9(d,J=9.0Hz,1H),7.09-7.03(m,2H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H), 6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.54(t,J=9.1Hz,1H),4.27(q,J=6.4Hz,2H),4.10 -4.00(m,2H),3.72-3.64(m,1H),3.59-3.50(m,4H),3.50(dd,J=5.4,2.7Hz,2H),3.42(dddd,J=12.8,4.8,3. 6,0.8Hz,1H),3.35(tt,J=8.1,1.8Hz,1H),3.15(p,J=6.5Hz,1H),2.88(ddd,J=12.3,7.9,5.4Hz,1H),2.83( ddd,J=12.5,7.9,5.4Hz,1H),2.79(s,1H),2.76(s,1H),2.72-2.63(m,4H),2.57-2.51(m,1H),2.42(d,J=5. 3Hz,3H),2.29-2.20(m,1H),2.20-2.03(m,4H),1.99-1.81(m,7H),1.81-1.73(m,1H),1.74(d,J=2.6Hz,1H) ,1.73(d,J=2.7Hz,1H),1.66-1.39(m,6H),1.43-1.23(m,12H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13C NMR(125MHz,Chloroform-d)δ187.22,172.67,171.59,170.54,165.17,163.87,157.51,155.75,152.96, 151.40,140.94,134.64,132.50,129.31,128.42,123.77,122.95,113.30,109.25,94.16,84.88,66.20, The values ​​are 63.91, 61.01, 59.97, 59.05, 55.83, 54.31, 48.47, 48.16, 47.45, 47.41, 46.85, 38.58, 36.55, 34.16, 34.10, 34.03, 33.33, 31.93, 31.62, 29.89, 29.19, 26.29, 26.10, 25.50, 24.97, 17.21, 14.28; this indicates the synthesis of compound MJ-5.

[0094] Example 11: Synthesis of compound MJ-6

[0095]

[0096] 1.2 g (1.8 mmol) of compound 9 and 0.8 g (2.7 mmol) of compound 15 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.84 g of pure compound MJ-6, with a yield of 38%.

[0097] The test results of the products prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.70(s,1H),7.31-7.24(m,2H),7.26-7.17(m,4H ),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt ,J=7.5,2.0Hz,1H),4.47-4.39(m,1H),4.27(q,J=6.4Hz,2H),4.09-4.00(m,2H ),3.95(dd,J=6.0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3.59(dt,J=12.7,6.3Hz,1 H),3.58-3.52(m,1H),3.55-3.50(m,1H),3.47(ddd,J=12.5,5.7,3.1Hz,2H),3 .41-3.31(m,3H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.63(m,11H),2.57-2.51(m ,1H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.95-1.64(m,12H),1.63-1.54(m,1H ),1.37(t,J=6.4Hz,3H),0.89(d,J=1.5Hz,3H),0.88(s,1H),0.90-0.82(m,8H). 13 C NMR(125MHz,Chloroform-d)δ173.94,171.90,165.17,163.87,157.51,155.75,140.94 ,137.18,134.64,129.42,128.79,128.42,127.22,123.77,109.25,94.16,84.88,70.54 The values ​​are: 66.20, 63.93, 61.01, 59.97, 55.77, 51.55, 48.47, 48.16, 46.85, 42.73, 40.77, 37.51, 36.55, 34.38, 34.10, 31.93, 31.06, 29.19, 26.10, 24.29, 22.30, 14.28; This indicates that the compound MJ-6 was synthesized.

[0098] Example 12: Synthesis of compound MJ-7

[0099]

[0100] 1.2 g (1.8 mmol) of compound 9 and 0.9 g (2.7 mmol) of compound 16 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.63 g of pure compound MJ-7, with a yield of 33%.

[0101] The test results of the products prepared above are as follows: 1 H NMR(400MHz,Chloroform-d)δ8.70(s,1H),7.89(s,1H),7.81-7.75(m,2H),7.60(dq,J=6.2,5.2Hz,1H ),7.49(d,J=9.0Hz,1H),7.09-7.03(m,2H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1 .0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.54(t,J=9.1Hz,1H),4.27(q,J=6 .4Hz,2H),4.10-4.00(m,2H),3.72-3.64(m,1H),3.59-3.38(m,7H),3.35(tt,J=8.1,1.8Hz,1H),3.15( p,J=6.5Hz,1H),2.93-2.78(m,2H),2.73-2.65(m,2H),2.65(d,J=3.7Hz,1H),2.61(dd,J=11.4,4.6Hz ,1H),2.56-2.50(m,1H),2.47(dd,J=6.5,3.7Hz,2H),2.42(d,J=5.3Hz,3H),2.29-2.20(m,1H),2.20-2 .03(m,4H),1.99-1.72(m,9H),1.69(dd,J=6.1,3.4Hz,2H),1.63(d,J=3.7Hz,1H),1.64-1.56(m,3H),1 .59-1.55(m,2H),1.55-1.45(m,3H),1.48-1.23(m,13H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13C NMR(125MHz,Chloroform-d)δ187.22,172.67,171.59,170.54,165.17,163.87,157.51,155.75,152.83,1 51.40,140.94,134.64,132.50,129.31,128.42,123.77,122.95,113.30,109.25,94.16,84.88,66.20,63. 93,61.01,59.05,55.83,54.31,50.71,48.47,48.16,47.45,46.85,45.64,38.58,36.55,36.23,35.16,34.10,34.03,33.38,33.33,32.97,29.89,29.19,26.29,26.10,25.50,24.97,17.21,14.28; This indicates the synthesis of compound MJ-7.

[0102] Example 13: Synthesis of compound MJ-8

[0103]

[0104] 1.2 g (1.8 mmol) of compound 9 and 0.9 g (2.7 mmol) of compound 17 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (3.6 mmol) of sodium triacetylborohydride was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.58 g of pure compound MJ-8, with a yield of 30%.

[0105] The test results of the products prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.70(s,1H),7.41(d,J=9.3Hz,1H),7.31-7.17(m,5H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.35-4.26(m,2H),4.26(d,J=6.4Hz,1H),4.10-4.00(m,2H),3.95(dd,J=6.0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3.64-3.43(m,5H),3.42-3.35(m,2H),3.38-3.31(m,1H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.78(m,4H),2.78-2.70(m,1H),2.74-2.66(m,1H),2.67(d,J=3.7Hz,1H),2.65(d,J=3.7Hz,1H),2.61(dd,J=11.4,4.6Hz,1H),2.56-2.50(m,1H),2.47(dd,J=6.5,3.8Hz,2H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.95-1.70(m,8H),1.69(dd,J=13.7,6.6Hz,1H),1.62(ddd,J=6.6,3.7,0.9Hz,4H),1.56(dd,J=6.6,3.7Hz,2H),1.41-1.30(m,5H),0.94(t,J=7.3Hz,3H),0.89(d,J=1.5Hz,3H),0.84(d,J=1.6Hz,3H). 13 C NMR(125MHz,Chloroform-d)δ

[0106] 173.90, 172.38, 165.17, 163.87, 157.51, 155.75, 140.94, 137.18, 134.64, 129.42, 128.79, 128.42, 127.22, 123.77, 109.25, 94.16, 84.88, 71.30, 66.20, 63.93, 61 .01,55.77,52.71,50.71,48.47,48.16,46.85,42.81,37.51,36.55,36.23,35.46,34.10,33.85,33.40,32.97,29.19,26.10,18.85,14.28,13.44; This indicates the synthesis of compound MJ-8.

[0107] Example 14: Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiaran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate (i.e., compound 19)

[0108]

[0109] 1.2 g (1.2 mmol) of compound 4 and 0.7 g (2.4 mmol) of compound 20 were dissolved in 30 mL of N,N-dimethylformamide. K₂CO₃ was added and the mixture was stirred at 90 °C. After the reaction was monitored by TLC until complete, a saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate was added and water was used for extraction. The ethyl acetate phase was washed with saturated brine. After vacuum distillation, dichloromethane was added to dissolve the compound. 1 M HCl was added to remove the protecting group. The mixture was stirred at room temperature for 8–10 hours. After the reaction was monitored by TLC until complete, dichloromethane was removed by rotary distillation. Ethyl acetate was added and water was used for extraction. The ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate separation to give 1.1 g of pure compound 19, with a yield of 68%.

[0110] The test results of the products prepared above are as follows: 1 HNMR(400MHz,Chloroform-d)δ8.06(d,J=6.4Hz,1H),7.86(d,J=8.0Hz,1H),7.63–7.57(m,2H),7.23(dd,J=8.0,2.1Hz,1H),7.10(s,0H),4. 30(q,J=6.4Hz,2H),4.16(t,J=4.0Hz,2H),3.21–3.12(m,4H),2.34–2.28(m,2H),1.73(d,J=13.7Hz,1H),1.37(t,J=6.3Hz,3H),1.25(s,5H). 13C NMR (125MHz, Chloroform-d) δ 166.09, 159.34, 145.26, 144.99, 131.86, 131.39, 128.62, 127.63, 127.35, 125.82, 125.57, 120.84, 114.53, 89.25, 89.20, 70.94, 61.11, 49.74, 41.81, 37.24, 33.34, 30.37, 14.32; This indicates the synthesis of compound 19.

[0111] Example 15: Synthesis of ethyl 2-((4,4-dimethyl-1-oxobenzothiaran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate (i.e., compound 21)

[0112]

[0113] 1.2 g (1.2 mmol) of compound 4 and 0.8 g (2.4 mmol) of compound 20 were dissolved in 30 mL of N,N-dimethylformamide. K₂CO₃ was added and the mixture was stirred at 90 °C. After the reaction was monitored by TLC until complete, a saturated ammonium chloride solution was added to quench the reaction. Ethyl acetate was added and water was used for extraction. The ethyl acetate phase was washed with saturated brine. After vacuum distillation, dichloromethane was added to dissolve the compound. 1 M HCl was added to remove the protecting group. The mixture was stirred at room temperature for 8–10 hours. After the reaction was monitored by TLC until complete, dichloromethane was removed by rotary distillation. Ethyl acetate was added and water was used for extraction. The ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate separation to obtain 1.1 g of pure compound 21, with a yield of 75%.

[0114] The test results of the products prepared above are as follows: 1 HNMR(500MHz,Chloroform-d)δ8.06(d,J=6.4Hz,1H),7.86(d,J=8.0Hz,1H),7.63– 7.57(m,2H),7.23(dd,J=8.0,2.1Hz,1H),7.11(s,0H),4.30(q,J=6.4Hz,2H),4.05( t,J=5.7Hz,2H),3.18–3.12(m,2H),2.85(tt,J=6.3,5.6Hz,2H),2.34–2.28(m,2H) ,2.16(d,J=12.8Hz,1H),1.93(p,J=5.6Hz,2H),1.37(t,J=6.3Hz,3H),1.25(s,5H). 13C NMR (125MHz, Chloroform-d) δ 166.26, 159.65, 145.26, 144.99, 131.86, 131.45, 128.62, 127.63, 127.38, 125.82, 125.57, 121.02, 114.36, 89.25, 89.20, 67.70, 61.11, 49.74, 37.24, 36.42, 33.34, 32.70, 30.37, 14.32; This indicates the synthesis of compound 21.

[0115] Example 16: Synthesis of compound MJ-9

[0116]

[0117] 1.1 g (1.2 mmol) of compound 19 and 0.9 g (1.3 mmol) of compound 22 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (2.4 mmol) of HATU (N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea) was added. The mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC to ensure complete reaction, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.8 g of pure compound MJ-9, with a yield of 43%.

[0118] The test results of the products prepared above are as follows: 1HNMR(400MHz,Chloroform-d)δ8.99(s,1H),7.45(t,J=4.8Hz,1H),7.33-7.17(m,6H),7.03(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7, 1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.66(d,J=1.3Hz,2H),4.36-4.25(m,3H),3.95(dd,J=6. 0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3.59(h,J=6.3Hz,1H),3.40-3.25(m,5H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.78(m,4H),2.78 -2.65(m,2H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.87-1.73(m,2H),1.66-1.51(m,2H),1.38(t,J=6.3Hz,3H),0.91-0.82(m,11H). 13 C NMR(125MHz,Chloroform-d)δ173.89,173.82,169.69,165.10,163.40,157.21, 156.74,140.80,137.18,134.65,129.42,128.79,128.42,127.22,123.80,109.7 3,94.22,84.89,70.54,67.37,66.20,61.01,55.77,52.57,48.47,48.16,40.73,39.52,39.07,37.51,36.55,34.10,26.10,24.99,22.40,14.28; This indicates the synthesis of compound MJ-9.

[0119] Example 17: Synthesis of compound MJ-10

[0120]

[0121] 1.1 g (1.2 mmol) of compound 19 and 1.4 g (1.3 mmol) of compound 22 were dissolved in 30 mL of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (2.4 mmol) of HATU was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.86 g of pure compound MJ-10, with a yield of 42%.

[0122] The test results of the products prepared above are as follows: 1HNMR(400MHz,Chloroform-d)δ8.99(s,1H),8.20(t,J=2.2Hz,1H),7.94(ddd,J=8.2,2.2,1.1Hz,1H),7.91(ddd,J=7.9,2.2,1.2Hz,1H),7.90-7.84(m,2H),7.67(t,J=8.0Hz,1H),7.60(dq,J=6.2,5.2Hz,1H),7.52-7.44(m,2H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.66(d,J=1.3Hz,2H),4.54(t,J=9.1Hz,1H),4.32(q,J=6.3Hz,2H),3.72-3.64(m,1H),3.53(dtd,J=5.3,4.3,2.2Hz,2H),3.46-3.38(m,3H),3.35(tt,J=8.1,1.8Hz,1H),3.15(p,J=6.5Hz,1H),2.93-2.78(m,2H),2.73-2.65(m,1H),2.42(d,J=5.3Hz,3H),2.29-2.20(m,1H),2.20-2.03(m,4H),1.95(ddd,J=12.5,7.8,5.4Hz,1H),1.77(ddd,J=12.5,7.9,5.3Hz,1H),1.59(dddd,J=11.2,8.6,5.7,4.0Hz,2H),1.55-1.47(m,2H),1.51-1.46(m,1H),1.49-1.39(m,1H),1.39(s,1H),1.38(s,2H),1.38-1.31(m,3H),1.34-1.23(m,6H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13CNMR(125MHz,Chloroform-d)δ187.05,172.67,171.59,170.54,169.69,167.85,165.10,163.40,157.21 ,156.74,151.55,140.80,135.49,134.65,134.23,133.57,131.03,130.73,129.85,128.42,123.80,122 .94,109.73,94.22,84.89,67.37,66.20,61.01,59.05,55.83,54.31,48.47,48.16,47.45,39.46,39.25,38.58,36.55,34.10,34.03,33.33,29.89,26.29,26.10,25.50,24.97,17.21,14.28; This indicates the synthesis of compound MJ-10.

[0123] Example 18: Synthesis of compound MJ-11

[0124]

[0125] 1.1 g (1.2 mmol) of compound 19 and 0.9 g (1.3 mmol) of compound 23 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (3.6 mmol) of HATU was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.66 g of pure compound MJ-11, with a yield of 32.0%.

[0126] The test results of the products prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.99(s,1H),7.33-7.17(m,6H),6.92(t,J=4.8Hz,1H),6.82(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.4 1(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.66(d,J=1. 3Hz,2H),4.33(d,J=6.3Hz,1H),4.33-4.25(m,2H),3.95(dd,J=6.0,4.4Hz ,1H),3.87(d,J=4.6Hz,1H),3.59(h,J=6.3Hz,1H),3.39-3.31(m,1H),3. 24-3.06(m,4H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.78(m,4H),2.78-2. 65(m,2H),2.03(ddd,J=12.5,7.9,5.4Hz,1H),1.87-1.73(m,2H),1.65-1 .51(m,2H),1.50-1.41(m,4H),1.38(t,J=6.3Hz,3H),0.91-0.82(m,12H). 13 C NMR(125MHz,Chloroform-d)δ173.89,172.55,169.83,165.10,163.40,157.21,156. 74,140.80,137.18,134.65,129.42,128.79,128.42,127.22,123.80,109.73,94.22 ,84.89,70.54,67.37,66.20,61.01,55.77,52.57,48.47,48.16,40.73,40.12,39.66,37.51,36.55,34.10,26.71,26.61,26.10,24.99,22.40,14.28; This indicates that compound MJ-11 was synthesized.

[0127] Example 19: Synthesis of compound MJ-12

[0128]

[0129] 1.1 g (1.2 mmol) of compound 19 and 1.0 g (1.2 mmol) of compound 22 were dissolved in 30 mL of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (2.4 mmol) of HATU was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.85 g of pure compound MJ-12, with a yield of 45%.

[0130] The test results of the products prepared above are as follows: 1 HNMR(400MHz,Chloroform-d)δ8.99(s,1H),8.20(t,J=2.2Hz,1H),7.92(dddd,J=15.4,7.9,2.2,1.2Hz,2H ),7.89(s,1H),7.73(t,J=5.0Hz,1H),7.67(t,J=8.0Hz,1H),7.60(dq,J=6.2,5.2Hz,1H),7.49(d,J=9.0Hz ,1H),6.92(t,J=4.8Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt ,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.66(d,J=1.3Hz,2H),4.54(t,J=9.1Hz,1H),4.32(q,J=6. 3Hz,2H),3.72-3.64(m,1H),3.46-3.37(m,1H),3.40–3.29(m,3H),3.19(qd,J=5.1,2.4Hz,2H),3.18-3.10 (m,1H),2.93-2.78(m,2H),2.73-2.65(m,1H),2.42(d,J=5.3Hz,3H),2.29-2.20(m,1H),2.20-2.03(m,4H), 1.95(ddd,J=12.5,7.8,5.4Hz,1H),1.77(ddd,J=12.5,7.9,5.3Hz,1H),1.66-1.58(m,1H),1.61-1.57(m,2 H),1.58-1.54(m,1H),1.57-1.40(m,6H),1.43-1.23(m,12H),0.89(d,J=1.4Hz,3H),0.84(d,J=1.6Hz,3H). 13C NMR(125MHz,Chloroform-d)δ187.05,172.67,171.59,170.54,169.83,167.97,165.10,163.40,157.21,15 6.74,151.55,140.80,135.48,134.65,134.22,133.57,131.03,130.75,129.85,128.42,123.80,122.94,10 The values ​​are 9.73, 94.22, 84.89, 67.37, 66.20, 61.01, 59.05, 55.83, 54.31, 48.47, 48.16, 47.45, 39.75, 39.66, 38.58, 36.55, 34.10, 34.03, 33.33, 29.89, 26.86, 26.72, 26.29, 26.10, 25.50, 24.97, 17.21, 14.28; this indicates the synthesis of compound MJ-12.

[0131] Example 20: Synthesis of compound MJ-13

[0132]

[0133] 1.1 g (1.2 mmol) of compound 19 and 0.9 g (1.2 mmol) of compound 25 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (2.4 mmol) of HATU was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.72 g of pure compound MJ-13, with a yield of 39.0%.

[0134] The test results of the products prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.99(s,1H),7.33-7.17(m,6H),6.92(t,J=4.8Hz,1H),6.82(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H), 6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.66(d,J=1.3Hz,2H),4.33(d,J=6.3Hz,1H),4.33-4.25(m,2H),3.97(dd,J=6.1 ,4.5Hz,1H),3.87(d,J=4.5Hz,1H),3.59(h,J=6.3Hz,1H),3.39-3.31(m ,1H),3.24-3.02(m,5H),2.93-2.78(m,4H),2.78-2.65(m,2H),2.03(dd d,J=12.5,7.9,5.4Hz,1H),1.87-1.73(m,2H),1.66-1.51(m,2H),1.54- 1.48(m,1H),1.51-1.42(m,3H),1.41-1.29(m,7H),0.91-0.82(m,11H). 13 CNMR(125MHz,Chloroform-d)δ173.89,172.55,169.83,165.10,163.40,157.21,156.74 ,140.80,137.18,134.65,129.42,128.79,128.42,127.22,123.80,109.73,94.22,84.8 9,70.54,67.37,66.20,61.01,55.77,52.57,48.47,48.16,40.73,40.29,39.84,37.51,36.55,34.10,31.38,28.94,26.54,26.52,26.10,24.99,22.40,14.28; This indicates the synthesis of compound MJ-13.

[0135] Example 21: Synthesis of compound MJ-14

[0136]

[0137] 1.1 g (1.2 mmol) of compound 19 and 0.9 g (1.3 mmol) of compound 26 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (2.4 mmol) of HATU was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.72 g of pure compound MJ-14, with a yield of 43%.

[0138] The test results of the products prepared above are as follows: 1 HNMR(400MHz,Chloroform-d)δ8.99(s,1H),8.20(t,J=2.2Hz,1H),7.92(dddd,J=15.4,7.9,2.2,1.2Hz ,2H),7.89(s,1H),7.74(t,J=5.0Hz,1H),7.67(t,J=8.0Hz,1H),7.60(dq,J=6.2,5.2Hz,1H),7.49(d,J= 9.0Hz,1H),6.92(t,J=4.8Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2.0,1.0Hz,1H) ,5.94(dt,J=7.6,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.66(d,J=1.3Hz,2H),4.54(s,0H),4.32(q,J =6.3Hz,2H),3.72-3.64(m,1H),3.42(dddd,J=12.1,4.1,3.3,1.0Hz,1H),3.42-3.31(m,1H),3.30(qd, J=5.4,1.2Hz,2H),3.19(tdd,J=5.7,4.8,2.5Hz,2H),3.19-3.10(m,1H),2.93-2.78(m,2H),2.73-2.65( m,1H),2.42(d,J=5.3Hz,3H),2.29-2.03(m,5H),1.95(ddd,J=12.5,7.8,5.4Hz,1H),1.77(ddd,J=12.5 ,7.9,5.3Hz,1H),1.66-1.40(m,10H),1.43-1.23(m,16H),0.89(d,J=1.5Hz,3H),0.84(d,J=1.7Hz,3H). 13CNMR(125MHz,Chloroform-d)δ187.05,172.67,171.59,170.54,169.83,167.96,165.10,163.40,157.21,156.7 4,151.55,140.87,135.48,134.65,134.21,133.57,131.03,130.75,129.85,128.42,123.80,122.94,109.73,9 4.22, 84.89, 67.37, 66.20, 61.01, 59.05, 55.83, 54.31, 48.47, 48.16, 47.45, 39.84, 39.74, 38.58, 36.55, 34.10, 34.03, 33.33, 31.38, 29.89, 28.95, 26.55, 26.52, 26.29, 26.10, 25.50, 24.97, 17.21, 14.28; This indicates the synthesis of compound MJ-14.

[0139] Example 22: Synthesis of compound MJ-15

[0140]

[0141] 1.1 g (1.2 mmol) of compound 19 and 1.2 g (1.3 mmol) of compound 27 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (2.4 mmol) of HATU was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.54 g of pure compound MJ-15, with a yield of 36%.

[0142] The test results of the products prepared above are as follows: 1H NMR(400MHz,Chloroform-d)δ8.98(s,1H),7.33-7.14(m,7H),6.91(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H),6.41(dtd,J=8.4,2 .0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),4.36-4.23(m,5H),4.05(s,1H),3.95(dd,J=6.0,4.4Hz,1H),3.87(d,J=4.6Hz,1H),3.78-3.67( m,2H),3.67-3.51(m,9H),3.44-3.30(m,3H),3.24(td,J=5.9,4.8Hz,2H),3.05(dd,J=7.3,6.6Hz,1H),2.93-2.85(m,1H),2.88-2.78(m,3H) ,2.78-2.65(m,2H),2.11-2.04(m,2H),2.07-1.99(m,1H),1.87-1.73 (m,2H),1.66-1.51(m,2H),1.38(t,J=6.3Hz,3H),0.91-0.82(m,11H). 13 C NMR(125MHz,Chloroform-d)δ173.89,173.87,171.75,165.09,164.08,156.92,156.71,140. 80,137.18,134.65,129.42,128.79,128.42,127.22,123.80,110.16,94.22,84.89,70.59,70 .54,70.30,69.67,69.61,69.57,69.48,67.03,66.20,61.01,55.77,52.57,48.47,48.16,40.73,40.36,37.51,37.43,36.55,34.10,29.01,26.10,24.99,22.40,14.28; This indicates the synthesis of compound MJ-15.

[0143] Example 21: Synthesis of compound MJ-16

[0144]

[0145] 1.2 g (1.2 mmol) of compound 19 and 1.3 g (1.3 mmol) of compound 22 were dissolved in 30 ml of N,N-dimethylformamide. After stirring at room temperature for 1 hour, 0.6 g (2.4 mmol) of HATU was added, and the mixture was stirred at room temperature for 8-10 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. After extraction with ethyl acetate and water, the ethyl acetate phase was washed with saturated brine. After vacuum distillation, the mixture was separated by preparative plate to obtain 0.52 g of pure compound MJ-16, with a yield of 34%.

[0146] The test results of the products prepared above are as follows: 1 HNMR(400MHz,Chloroform-d)δ8.98(s,1H),8.20(t,J=2.2Hz,1H),7.97-7.83(m,4H),7.67(t,J=8.0Hz,1H) ,7.60(dq,J=6.2,5.2Hz,1H),7.49(d,J=9.0Hz,1H),6.91(t,J=4.9Hz,1H),6.48(ddd,J=8.4,7.7,1.8Hz,1H ),6.41(dtd,J=8.4,2.0,1.0Hz,1H),6.11(dt,J=7.5,2.0Hz,1H),5.26(dd,J=4.8,3.2Hz,1H),4.54(s,0H), 4.36-4.23(m,4H),4.05(s,1H),3.72(td,J=4.9,2.8Hz,2H),3.72-3.65(m,1H),3.68-3.60(m,6H),3.63-3. 53(m,4H),3.42(dddd,J=12.8,4.8,3.6,0.8Hz,1H),3.35(tt,J=8.1,1.8Hz,1H),3.24(td,J=5.9,4.8Hz,2H ),3.15(p,J=6.5Hz,1H),2.93-2.78(m,2H),2.73-2.65(m,1H),2.42(d,J=5.3Hz,3H),2.29-2.20(m,1H),2. 20-2.03(m,6H),1.95(ddd,J=12.5,7.8,5.4Hz,1H),1.77(ddd,J=12.5,7.9,5.3Hz,1H),1.59(dddd,J=11.2 ,8.6,5.7,4.0Hz,2H),1.55-1.45(m,3H),1.48-1.23(m,12H),0.89(d,J=1.5Hz,3H),0.84(d,J=1.7Hz,3H). 13C NMR(125MHz,Chloroform-d)δ187.05,172.67,171.75,171.59,170.54,167.73,165.09,164.08,156.92,156.71,151 .55,140.80,135.49,134.65,134.27,133.57,131.03,130.73,129.85,128.42,123.80,122.94,110.16,94.22,84.8 9,70.59,70.30,69.67,69.57,69.54,69.48,67.03,66.20,61.01,59.05,55.83,54.31,48.47,48.16,47.45,40.60,38.58,37.43,36.55,34.10,34.03,33.33,29.89,29.01,26.29,26.10,25.50,24.97,17.21,14.28; This indicates the synthesis of compound MJ-16.

[0147] Example 22:

[0148] This embodiment tests the bioactivity of the target compounds MJ-1 to MJ-16 obtained in the above embodiments. Specifically, the in vitro growth inhibition activity study of breast cancer regenerative cells (TRCs) was conducted by culturing human breast cancer MCF-7 cells for 5 days in DMEM medium containing 10% fetal bovine serum and 3D fibrin glue (90 Pa). The desired melanoma tumor regenerative cells were then cultured and screened to obtain them (Nat. Mater. 2012, 11, 734). Subsequently, collagenase and neutral protease II were added to the medium to digest the fibrin glue, thereby freeing the breast cancer tumor regenerative cells. The obtained breast cancer tumor regenerative cells were then transferred to freshly prepared medium for resuspension and kept in a single-cell state. Single-cell breast cancer MCF-7 tumor regeneration cells were seeded in 3D fibrin glue (90 Pa) and incubated for 5 days. 0.1% DMSO was used as a negative control, and all-trans retinoic acid (All-trans RA), tazarotene, and WYC-209 were used as positive controls. Drugs were added at a concentration of 10 μM and incubated for 5 days. The tumor clone size was observed and measured using fluorescence microscopy, and the inhibition rate of each drug on breast cancer MCF-7 tumor regeneration cells was calculated. The test results are shown in Table 1.

[0149] Table 1: Results of the MCF-7 breast cancer cell regeneration inhibition activity assay

[0150]

[0151] As shown in Table 1, the target compounds MJ-1 to MJ-16 all have a certain degradation effect on retinoic acid receptors and exhibit a certain inhibitory effect on breast cancer MCF-7 tumor regeneration cells. Furthermore, some of the RAR PROTAC compounds of this invention show better inhibitory effects on breast cancer MCF-7 tumor regeneration cells than commonly used positive control drugs such as all-trans retinoic acid (All-trans RA), tazarotene, and WYC-209.

[0152] In addition, it should be noted that, besides the target compounds MJ-1 to MJ-16 mentioned above showing a certain inhibitory effect on tumor stem cells, other RAR PROTAC compounds synthesized using linker groups have also shown an inhibitory effect on tumor stem cells in experiments; for example, linker groups with the following structures can also be used:

[0153]

[0154] The above examples are merely illustrative of the present invention and do not constitute a limitation on the scope of protection of the present invention. All designs that are the same as or similar to the present invention are within the scope of protection of the present invention.

Claims

1. A RAR PROTAC compound, characterized in that, It has the structure shown in the following general formula: Among them, E3 ligand is selected from IAP ligand. One of them; The Linker is one of a rigid connector, a flexible connector, or a combination of flexible and rigid connectors; the rigid connector is selected from one of the following structural formulas: The flexible connection base is selected from one of the following structural formulas: The flexible and rigid combined connection base is selected from one of the following structural formulas: Where n = 0-10.

2. The RAR PROTAC compound as claimed in claim 1, characterized in that, Selected from any one of the following compounds:

3. A method for preparing the RAR PROTAC compound as described in claim 1 or 2, characterized in that, Includes the following steps: S1. Synthesize ethyl 2-((4,4-dimethyl-1-oxobenzothiaran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate, or synthesize ethyl 2-((4,4-dimethyl-1-oxobenzothiaran-6-)ethynyl)-4-bromopyrimidine-5-carboxylate; S2. Using the ethyl 2-((4,4-dimethyl-1-oxobenzothiaran-6-)ethynyl)-4-hydroxypyrimidine-5-carboxylate or ethyl 2-((4,4-dimethyl-1-oxobenzothiaran-6-)ethynyl)-4-bromopyrimidine-5-carboxylate synthesized in step S1, prepare retinoid compounds; Among them, retinoids are one of the following compounds: S3. Obtain the intermediate compound; the intermediate compound is one of the following compounds: S4. Prepare RAR PROTAC compounds using retinoic acid compounds and intermediate compounds.

4. The method for preparing the RAR PROTAC compound as described in claim 3, characterized in that, In step S4, the process of preparing the RAR PROTAC compound from the retinoic acid compound and intermediate compound is as follows: Retinoids and intermediates were dissolved in N,N-dimethylformamide, and then sodium triacetylborohydride or HATU was added. The mixture was stirred at room temperature. After the reaction was complete, a saturated ammonium chloride solution was added to quench the reaction. Then, ethyl acetate was added and water was used for extraction. The ethyl acetate phase was washed with saturated brine and separated by vacuum distillation to obtain the RAR PROTAC compound.

5. The use of the RAR PROTAC compound of claim 1 or 2 and its pharmaceutically acceptable salt in the preparation of an antitumor drug.

6. The application as described in claim 5, characterized in that, The pharmaceutically acceptable salt of the RAR PROTAC compound is a salt formed by the RAR PROTAC compound and an acid selected from any of the following: Hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, or ferulic acid.

7. The application as described in claim 5, characterized in that, The tumor is melanoma, breast cancer, or prostate cancer.

8. A pharmaceutical composition, characterized in that, Includes the RAR PROTAC compound of claim 1 or 2, and one or more combinations of pharmaceutically acceptable carriers, excipients, diluents, adjuvants, and mediators of the RAR PROTAC compound.

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