A degrading agent targeting Aurora A, its preparation method and application

Abstract

This invention discloses a degrading agent targeting Aurora A, its preparation method, and its application. The Aurora A-targeting degrading agent consists of three key components: a target protein-binding ligand, an E3 ubiquitin ligase ligand, and a linker connecting the former two. This degrading agent can target Aurora A and exhibits significant degradation activity, thereby effectively inhibiting tumor cell proliferation and formation. Specifically, its inhibitory activity against Jurkat leukemia cells has an IC50 value of [missing information]. 50 It can be as low as 0.89μM.

Description

Technical Field

[0001] This invention belongs to the fields of pharmacology and medicinal chemistry, specifically relating to a degrading agent targeting Aurora A, its preparation method, and its application. Background Technology

[0002] Cancer is one of the major diseases threatening human health. Current mainstream treatments include drug therapy, surgery, and radiation therapy, with drug therapy being the most widely used. Traditional anticancer drugs struggle to distinguish between tumor cells and normal cells, often causing significant side effects. Targeted therapies, on the other hand, target specific sites on tumor cells, precisely acting on the tumor. This not only significantly improves treatment efficacy but also reduces the likelihood of adverse reactions.

[0003] Aurora kinases are a class of serine / threonine kinases, comprising three isoforms: Aurora A, Aurora B, and Aurora C. During cell mitosis, Aurora kinases participate in regulating processes such as spindle checkpoint recognition, centrosome maturation, spindle assembly, chromatin separation, and cytoplasmic segregation. Studies have shown that aberrant expression of Aurora A leads to centrosome expansion, genomic aneuploidy, and chromosomal instability. Because Aurora A is aberrantly expressed in many types of tumors, it is a potential therapeutic target for related cancers.

[0004] The mechanisms by which Aurora A and related tumorigenesis remain unclear in current research. Overexpression of Aurora A kinase easily leads to abnormal cell mitosis and is closely related to tumor formation. Aurora A kinase is overexpressed in many cancer cells (such as lung cancer, breast cancer, and pancreatic cancer), while the proliferation rate of most normal cells in the human body is not fast. Inhibiting the activity of Aurora A kinase can lead to the accumulation of polyploid tumor cells, promote apoptosis, and block cell proliferation. Therefore, anti-tumor drugs targeting Aurora A kinase are receiving increasing attention. Currently, most research focuses on Aurora A kinase inhibitors, while research on Aurora A kinase degrading agents is extremely scarce. Inducing the degradation of Aurora-A protein is the best way to completely eliminate Aurora protein and achieve optimal therapeutic effects. Therefore, it is necessary to research novel degrading agents with good degradation activity that target Aurora A. Summary of the Invention

[0005] To overcome the problems existing in the prior art, one objective of the present invention is to provide a degrading agent targeting Aurora A. A second objective of the present invention is to provide a method for preparing the aforementioned Aurora A-targeting degrading agent. A third objective of the present invention is to provide a composition. A fourth objective of the present invention is to provide the application of the aforementioned composition and the Aurora A-targeting degrading agent. A fifth objective of the present invention is to provide the application of the aforementioned composition and the Aurora A-targeting degrading agent.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] The first aspect of this invention provides a degrading agent targeting Aurora A, wherein the degrading agent targeting Aurora A is a compound with the general structural formula shown in formula (I):

[0008]

[0009] Where Ar is

[0010] Ligand is an E3 ubiquitin ligase ligand.

[0011] Preferably, the linker is any one of the following:

[0012]

[0013] Where n is 1-6.

[0014] Preferably, the E3 ubiquitin ligand is a CRBN ligand or a VHL ligand.

[0015] More preferably, the CRBN ligand is selected from groups with the structure shown in Formula C1 or Formula C2:

[0016]

[0017] Wherein, X1 is selected from C=O or CH2, X2 is a single bond or selected from O; R1 is selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl or cyano.

[0018] More preferably, the structure of the VHL ligand is shown in Formula V:

[0019]

[0020] R2 and R5 are each independently hydrogen or C1-C4 alkyl; R3, R4, R6, and R7 are each independently hydrogen, deuterated, halogenated, nitro, cyano, amino, hydroxyl, or any one of C1-C4 alkyl.

[0021] More preferably, the Aurora A-targeting degrader is one of the following compounds:

[0022]

[0023]

[0024]

[0025]

[0026]

[0027]

[0028] Preferably, the compound further includes: an acceptable salt, enantiomer, hydrate, or prodrug.

[0029] More preferably, the pharmaceutically acceptable salt includes at least one of a metal salt, an ammonium salt, a salt formed with an inorganic acid, a salt formed with an organic base, a salt formed with an organic acid, a salt formed with a basic amino acid, and a salt formed with an acidic amino acid.

[0030] More preferably, the metal salt includes an alkali metal salt and an alkaline earth metal salt. More preferably, the alkali metal salt includes at least one of sodium salt and potassium salt. More preferably, the alkaline earth metal salt includes at least one of calcium salt, magnesium salt, barium salt, and aluminum salt. The salt formed with an organic base includes at least one of the following organic bases: trimethylamine, triethylamine, pyridine, methylpyridine, 2,6-dimethylpyridine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, and N,N'-dibenzylethylenediamine. The salt formed with an inorganic acid includes at least one of the following inorganic acids: hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, and phosphoric acid. The salt formed with an organic acid includes at least one of the following organic acids: formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid. The salts formed with basic amino acids include salts formed with at least one of the following basic amino acids: arginine, lysine, and ornithine. The salts formed with acidic amino acids include salts formed with at least one of the following acidic amino acids: aspartic acid and glutamic acid.

[0031] A second aspect of this invention provides a method for preparing the Aurora A-targeting degrader described in the first aspect, comprising the following steps:

[0032] S1, methyl 5-bromo-2-methyl-3-nitrobenzene reacts with vinyl magnesium bromide to generate intermediate a

[0033] S2. The intermediate a reacts with pinacol diboronate to generate intermediate b.

[0034] S3, intermediate b and compound c The reaction produces intermediate d

[0035] S4. The intermediate d is hydrolyzed to generate intermediate e.

[0036] S5, the intermediate e and compound f intermediate g is formed by amide condensation.

[0037] S6. The intermediate g is deprotected to generate intermediate h.

[0038] S7, intermediate h and The amide condensation produces a degrading agent targeting AuroraA as shown in formula (I).

[0039] Preferably, in step S1, the reaction temperature is -78 to 25°C.

[0040] Preferably, in step S1, the reaction time is 3-12 hours.

[0041] Preferably, the reaction in step S1 is carried out in a solvent; the solvent is ultra-dry tetrahydrofuran.

[0042] Preferably, in step S2, the reaction temperature is 80-100℃.

[0043] Preferably, in step S2, the reaction time is 3-6 hours.

[0044] Preferably, the reaction in step S2 is carried out in a solvent; the solvent is ultradry dimethyl sulfoxide.

[0045] Preferably, the reaction in step S2 is carried out under the action of a palladium catalyst; more preferably, the catalyst is a 1,1-bis(diphenylphosphino)ferrocene palladium dichloromethane complex.

[0046] Preferably, in step S3, the reaction temperature is 110-130℃.

[0047] Preferably, in step S3, the reaction time is 0.5-2 hours.

[0048] Preferably, the reaction in step S3 is carried out in a solvent; the solvent is a mixed solution of ethylene glycol dimethyl ether and water.

[0049] Preferably, the reaction in step S3 is carried out under the action of a palladium catalyst; more preferably, the catalyst is a 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex.

[0050] Preferably, in step S3, the preparation method of compound c includes: reacting 5-bromo-2-chlorophenol with 2-iodopyridine to generate intermediate c.

[0051] More preferably, in the preparation method of compound c, the reaction temperature is 130-150℃ and the reaction time is 12-16h.

[0052] Preferably, in step S4, the reaction temperature is 40-70℃.

[0053] Preferably, in step S4, the reaction time is 5-12 hours.

[0054] Preferably, the reaction in step S4 is carried out in a solvent; the solvent is a mixed solution of tetrahydrofuran-methanol-water.

[0055] Preferably, the reagent used in the hydrolysis reaction of step S4 is lithium hydroxide.

[0056] Preferably, in step S5, the reaction temperature is 45-80℃.

[0057] Preferably, in step S5, the reaction time is 3-12 hours.

[0058] Preferably, the reaction in step S5 is carried out in a solvent; the solvent is tetrahydrofuran.

[0059] Preferably, the reaction in step S5 is carried out under the action of a condensing agent; more preferably, the condensing agent is bicycloamidine and N,N-carbonyldiimidazole.

[0060] Preferably, in step S5, the preparation method of compound f includes: reacting sulfonamide with piperazine-1-carboxylic acid tert-butyl ester to generate compound f.

[0061] Preferably, in step S6, the reaction temperature is 20-35℃.

[0062] Preferably, in step S6, the reaction time is 3-6 hours.

[0063] Preferably, the reaction in step S6 is carried out in a solvent; the solvent is dichloromethane.

[0064] Preferably, the reagent used in the deprotection reaction of step S6 is trifluoroacetic acid.

[0065] Preferably, in step S7, the reaction temperature is 20-35℃.

[0066] Preferably, in step S7, the reaction time is 1-2 hours.

[0067] Preferably, the reaction in step S7 is carried out in a solvent; the solvent is N,N-dimethylformamide.

[0068] Preferably, the condensing agent used in the amide condensation reaction of step S7 is HATU.

[0069] A third aspect of the present invention provides a composition comprising an active molecule and a pharmaceutically acceptable excipient, said active molecule being the Aurora A-targeting degrader described in the first aspect.

[0070] Preferably, the pharmaceutically acceptable excipient refers to a conventional drug carrier in the pharmaceutical field, including at least one of fillers, binders, disintegrants, solubilizers, suspending agents, wetting agents, pigments, fragrances, solvents, surfactants, or flavoring agents.

[0071] Substances that can be used as pharmaceutically acceptable excipients include, but are not limited to, ion exchangers; aluminum; aluminum stearate; lecithin; serum proteins, such as human serum albumin; buffering substances such as phosphates; glycine; sorbic acid; potassium sorbate; mixtures of partial glycerides of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts; colloidal silica; magnesium trisilicate; polyvinylpyrrolidone; polyacrylates; waxes; polyethylene-polyoxypropylene-blocking polymers; lanolin; sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives. Biological agents such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; gum powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic salts; Ringer's solution; ethanol; phosphate buffer solutions; and other non-toxic and suitable lubricants such as sodium lauryl sulfate and magnesium stearate; colorants; release agents; preservatives and antioxidants.

[0072] Preferably, the composition is an injection, an oral preparation, or a mucosal delivery preparation.

[0073] More preferably, the dosage forms of oral preparations include granules, tablets, powders, capsules, oral solutions, oral suspensions, dry suspensions, oral emulsions, gels, oral liquids, emulsions, emulsions, colloidal solutions, mixtures, tinctures, drops, suspension drops, syrups, sustained-release tablets, sustained-release coated tablets, controlled-release tablets, sustained-release capsules, and controlled-release capsules.

[0074] More preferably, the dosage form of the injectable preparation includes injections, injection solutions, solutions for injection, injection solutions for intravenous infusion, suspensions for injection, sterile powders for injection, intravenous injections, emulsions for injection, emulsion injections, powder injections, injections, sterile powder injections, lyophilized powder injections, and concentrated solutions for injection.

[0075] The fourth aspect of the present invention provides the use of the Aurora A-targeting degrader described in the first aspect, or the composition described in the third aspect, in the preparation of an Aurora A protein kinase inhibitor.

[0076] The fifth aspect of the present invention provides the use of the Aurora A-targeting degrading agent described in the first aspect, or the composition described in the third aspect, in the preparation of an antitumor drug.

[0077] Preferably, the tumor is selected from breast cancer, leukemia, lung cancer, liver cancer, esophageal cancer, pancreatic cancer, colorectal cancer, stomach cancer, cervical cancer, brain cancer, and nasopharyngeal carcinoma.

[0078] The beneficial effects of this invention are:

[0079] This invention proposes a degrading agent targeting Aurora A, comprising three key components: a target protein-binding ligand, an E3 ubiquitin ligase ligand, and a linker connecting the former two. This degrading agent can target Aurora A and exhibits significant degrading activity, thereby effectively inhibiting tumor cell proliferation and formation. Specifically, its inhibitory activity against Jurkat leukemia cells has an IC50 value of [missing information]. 50 It can be as low as 0.89μM. Detailed Implementation

[0080] The present invention will be further described in detail below through specific embodiments. Unless otherwise specified, the raw materials used in the following embodiments can be obtained from conventional commercial channels or prepared and isolated through simple synthesis; unless otherwise specified, the processes employed are conventional processes in the art.

[0081] Example 1

[0082] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-oxy)phenyl)-N-((4-(6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindololin-5-yl)hex-5-ynyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-1); its structural formula is shown in formula AP-1:

[0083]

[0084] The synthetic route for intermediate (i) is as follows:

[0085]

[0086] The specific synthetic steps for preparing intermediate (i) are as follows:

[0087] 1. Weigh 765.1 mg of 4-bromo-1-butanol and 20 mg of pyridinium 4-methylbenzenesulfonic acid, and add 0.55 mL of 3,4-dihydropyran to a 10 mL dry reaction tube. Replace with N2 three times, and add ultra-dry dichloromethane. React at room temperature for 8-12 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with dichloromethane, combine the organic phases, wash the organic phase with saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (petroleum ether / ethyl acetate) to obtain intermediate (1).

[0088] 2. Weigh 1.05 g of intermediate (1) and add it to a 25 mL reaction flask that has been dried. Replace with N2 three times and dissolve in ultra-dry dimethyl sulfoxide. Then weigh 302 mg of lithium ethynylene ethylenediamine complex and add it to another reaction flask. Replace with N2 three times and dissolve in ultra-dry dimethyl sulfoxide. Slowly add the solution of intermediate (1) dropwise to it and react in an oil bath at 80 °C for 12 h. After the reaction is complete, add water to quench the reaction, adjust to acidity with dilute hydrochloric acid, extract the reaction solution three times with ethyl acetate, combine the organic phases, wash the organic phase with saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (petroleum ether / ethyl acetate) to obtain intermediate (2).

[0089] 3. Weigh 513 mg of intermediate (2) and 10 mg of pyridine 4-methylbenzenesulfonic acid and add them to a 50 mL reaction flask. Add anhydrous ethanol to dissolve and stir. React at room temperature for 12 h. After the reaction is complete, evaporate the solvent under vacuum, add water, extract the reaction solution three times with ethyl acetate, combine the organic phases, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (petroleum ether / ethyl acetate) to obtain intermediate (3).

[0090] 4. Weigh 538 mg of intermediate (3) and 968 mg of intermediate (4) and add them to a 100 mL dried reaction flask. Then add 250 mg of bis(triphenylphosphine)palladium dichloride and 57 mg of cuprous iodide. Replace with N2 three times. Add 1.26 mL of ultra-dry triethylamine to the reaction flask, then add ultra-dry N,N-dimethylformamide to dissolve and stir. React in an oil bath at 80 °C for 8-12 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (5).

[0091] 5. Weigh 370 mg of intermediate (5) and add it to a 100 mL reaction flask that has been dried. Add dichloromethane to dissolve and stir. Add 1.35 g of Des Martin oxidant under ice bath and react for 8-12 h. After the reaction is complete, add saturated sodium sulfite solution to quench the reaction. Adjust the pH to acidity with dilute hydrochloric acid. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase with saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (i).

[0092] The synthetic route for the Aurora A-targeting degrader is as follows:

[0093]

[0094] The specific synthetic steps for the Aurora A-targeting degrader are as follows:

[0095] 1. Weigh 2.74 g of methyl 5-bromo-2-methyl-3-nitrobenzoate and add it to a dry 250 mL round-bottom flask. Replace the flask with N2 three times, add ultra-dry tetrahydrofuran at room temperature, and after dissolving, transfer the flask to a -78 °C cryogenic bath. Slowly add 20 mL of vinyl magnesium bromide and react at -78 °C for 3 h, then raise the temperature to room temperature and react for 8 h. Quench the reaction by adding saturated ammonium chloride solution under an ice-water bath. Extract the reaction solution three times with ethyl acetate, combine the organic phases, wash the organic phase three times with water, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (petroleum ether / ethyl acetate) to obtain intermediate (a).

[0096] 2. Weigh 1.07 g of intermediate (a) and add it to a 100 mL dried round-bottom flask. Add 0.326 g of 1,1-bis(diphenylphosphino)ferrocene palladium dichloride dichloromethane complex, 2.03 g of pinacol diborate and 1.17 g of potassium acetate to the flask. Replace with N2 three times, add ultra-dry dimethyl sulfoxide, and react in an oil bath at 90 °C for 3-6 h. After the reaction is complete, add ethyl acetate to the system, filter the system with diatomaceous earth, wash the organic phase three times with water and saturated sodium chloride solution respectively, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (petroleum ether / ethyl acetate) to obtain intermediate (b).

[0097] 3. Weigh 6.22 g of 5-bromo-2-chlorophenol and 7.17 g of 2-iodopyridine and add them to a 100 mL dried round-bottom flask. Add 13 g of cesium carbonate and N,N-dimethylformamide to the flask and react in an oil bath at 150 °C for 12 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate, combine the organic phases, wash the organic phase with saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (petroleum ether / ethyl acetate) to obtain intermediate (c).

[0098] 4. Weigh 1.26 g of intermediate (b) and 1.25 g of intermediate (c) into a 100 mL round-bottom flask. Add 0.326 g of 1,1-bis(diphenylphosphino)ferrocene palladium dichloromethane complex to the flask. Attach a reflux condenser to the top of the flask, replace with N2 three times, add triethylamine, and then add a mixed solution of ethylene glycol dimethyl ether-water (3:1) and stir. React in an oil bath at 120 °C for 3 h. After the reaction is complete, remove the solvent by vacuum rotary evaporation, extract with dichloromethane, dry the organic phase with anhydrous sodium sulfate, and perform rapid column chromatography (petroleum ether / ethyl acetate) to obtain intermediate (d).

[0099] 5. Weigh 1.57 g of intermediate (d) and add it to a 100 mL round-bottom flask. Add tetrahydrofuran to dissolve and stir. Weigh 0.12 g of lithium hydroxide and dissolve it in water. Slowly add the lithium hydroxide aqueous solution to the round-bottom flask and react in an oil bath at 45 °C for 8 h. After the reaction is complete, remove the organic solvent by vacuum rotary evaporation, add water, adjust to acidity with dilute hydrochloric acid, and filter to obtain intermediate (e).

[0100] 6. Weigh 5g of sulfonamide and 9.7g of piperazine-1-carboxylic acid tert-butyl ester and add them to a 250mL round-bottom flask. Replace with N2 three times, add 1,4-dioxane to the flask, and react in an oil bath at 115℃ for 20h. After the reaction is completed, cool to room temperature and recrystallize. After filtration, the intermediate (f) can be obtained.

[0101] 7. Weigh 0.568 g of intermediate (e) and 0.535 g of N,N-carbonyldiimidazole into a 100 mL round-bottom flask. Attach a reflux condenser to the top of the flask, replace with N2 three times, add ultra-dry tetrahydrofuran to dissolve and stir, and react in an oil bath at 45 °C for 3 h. Then add 0.795 g of tetrahydrofuran solution of intermediate (f) and dicycloamidinium into the flask, and react in an oil bath at 80 °C for 12 h. After the reaction is complete, remove the organic solvent by vacuum rotary evaporation, add water and adjust to acidity with dilute hydrochloric acid, extract three times with ethyl acetate, combine the organic phases, remove the organic solvent by vacuum rotary evaporation, and obtain intermediate (g) by rapid column chromatography (petroleum ether / ethyl acetate).

[0102] 8. Weigh 0.577 g of the intermediate (g) and add it to a 100 mL round-bottom flask. Add dichloromethane to the flask to dissolve and stir. Then add 1 mL of trifluoroacetic acid dropwise to the flask and react at room temperature for 2-3 h. After the reaction is complete, remove the organic solvent by vacuum rotary evaporation and dry to obtain the intermediate (h).

[0103] 9. Weigh 0.102 g of intermediate (h) and 0.709 g of intermediate (i) (n = 1) and add them to a 10 mL reaction tube. Add 0.5 mL of N,N-diisopropylethylamine and 0.114 g of condensing agent (HATU) to the reaction tube. Then add N,N-dimethylformamide to dissolve and stir. React at room temperature for 2 h. After the reaction is complete, add water to quench the reaction and extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution respectively. Dry the organic phase with anhydrous sodium sulfate, remove the organic solvent by vacuum rotary evaporation, and obtain the degradation agent by rapid column chromatography (dichloromethane / methanol), denoted as AP-1.

[0104] White solid, yield: 15%. 1 H NMR(500MHz,Acetone-d6)δ10.87(s,1H),10.68(br s,1H),9.85(s,1H),8.12(dd,J=4.9,1.3Hz,1H),7.91–7.85(m,1H),7.68–7.64(m,4H),7.60(t,J=2.8Hz,1H),7 .55(s,1H),7.51–7.47(m,2H),7.15–7.11(m,2H),6.75(dd,J=2.9,2.0Hz,1H),5.16(dd,J=13.4,5.1Hz,1H),4.3 8(dd,J=17.0,14.0Hz,2H),3.72–3.68(m,4H),3.58–3.41(m,4H),2.96–2.90(m,1H),2.76–2.72(m,1H),2.71(s ,3H),2.59(t,J=7.2Hz,2H),2.52(t,J=7.0Hz,2H),2.50–2.40(m,1H),2.16–2.10(m,1H),1.89(p,J=7.1Hz,2H). 13C NMR (126MHz, Acetone-d6) δ171.93,170.42,170.39,168.10,167.86,163.02,150.01,147.2 6,142.43,140.80,140.04,136.56,131.19,130.43,129.01,128.56,127.52,127.25,126.25 ,126.11,126.02,125.53,124.14,123.20,121.65,119.18,118.97,111.07,101.42,92.19,80.65,51.95,46.90,46.84,46.40,44.95,40.96,31.28,31.20,24.05,22.90,18.45,13.63.

[0105] Example 2

[0106] This embodiment provides a degrading agent targeting Aurora A, which is prepared using a method similar to that of Example 1, except that the starting material for intermediate (i) is changed to 6-bromo-1-hexanol. Its chemical name and structural formula are as follows:

[0107] 4-(4-chloro-3-(pyridin-2-oxy)phenyl)-N-((4-(8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)oct-7-ynyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-2)

[0108]

[0109] White solid, yield: 15%. 1H NMR(500MHz,Acetone-d6)δ10.85–10.77(s,1H),10.57(br s,1H),9.78(s,1H),8.14(dd,J=4.9,1.3Hz,1H),7.93–7.88(m,1H),7.71–7.68(m,1H),7.68–7.64(m,3H),7.62- 7.59(m,1H),7.58(s,1H),7.53–7.50(m,1H),7.49(s,1H),7.16–7.12(m,2H),6.78–6.74(m,1H),5.19(dd,J=13. 4,5.1Hz,1H),4.46(dd,J=16.8,14.0Hz,2H),3.74–3.64(m,4H),3.58–3.38(m,4H),3.05–2.95(m,1H),2.77(m,1 H),2.72(s,3H),2.60–2.50(m,1H),2.49–2.42(m,4H),2.23–2.16(m,1H),1.71–1.62(m,4H),1.59–1.49(m,2H). 13 C NMR (126MHz, Acetone-d6) δ171.91,170.91,170.38,167.98,167.85,163.02,150.01,147.27 ,142.47,140.82,140.03,136.57,131.16,130.42,129.04,128.53,127.56,127.43,126.21,1 26.10,125.53,124.14,123.19,121.66,119.13,118.97,111.06,101.40,92.65,80.27,51.97,46.96,46.87,46.41,45.00,40.87,32.39,31.30,28.39,28.24,24.47,22.94,18.80,13.57.

[0110] Example 3

[0111] This embodiment provides a degrading agent targeting Aurora A, which is prepared using a method similar to that of Example 1, except that the starting material for intermediate (i) is changed to 8-bromo-1-octanol. Its chemical name and structural formula are as follows:

[0112] 4-(4-chloro-3-(pyridin-2-oxy)phenyl)-N-((4-(10-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)dec-9-ynyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-3)

[0113]

[0114] White solid, yield 15%. 1 H NMR(500MHz,Acetone-d6)δ10.76(s,1H),10.61(br s,1H),9.80(s,1H),8.13(dd,J=4.9,1.9Hz,1H),7.92–7.87(m,1H),7.70(d,J=7.8Hz,1H),7.68–7.62(m,3H),7.60–7.56(m ,2H),7.53(s,1H),7.50(d,J=7.8Hz,1H),7.16–7.11(m,2H),6.75–6.73(m,1H),5.19(dd,J=13.4,5.1Hz,1H),4.46(dd,J=16 .9,14.0Hz,2H),3.71–3.61(m,4H),3.52–3.36(m,4H),3.03–2.93(m,1H),2.80–2.74(m,1H),2.73(s,3H),2.59–2.49(m,1H ),2.45(t,J=7.0Hz,2H),2.39(t,J=7.5Hz,2H),2.23–2.14(m,1H),1.65–1.57(m,4H),1.53–1.45(m,2H),1.42–1.34(m,4H). 13 C NMR (126MHz, Acetone-d6) δ171.93,171.05,170.39,168.20,167.89,163.02,150.01,147.27,1 42.48,140.85,140.02,136.59,131.15,131.12,130.42,128.98,128.49,127.50,127.45,126.2 3,126.11,125.50,124.13,123.20,121.65,119.23,118.97,111.06,101.40,92.77,80.24,51.97,46.97,46.89,46.42,45.00,40.87,32.50,31.30,28.51,28.35,24.92,22.94,18.84,13.63.

[0115] Example 4

[0116] This embodiment provides a degrading agent targeting Aurora A, which is prepared using a method similar to that of Example 1, except that the starting material for intermediate (i) is changed to 10-bromo-1-decyl alcohol. Its chemical name and structural formula are as follows:

[0117] 4-(4-chloro-3-(pyridin-2-oxy)phenyl)-N-((4-(12-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)dodecyl-11-ynyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-4)

[0118]

[0119] White solid, yield 15%. 1 H NMR (500MHz, Acetone-d6) δ10.86(s,1H),10.65(s,1H),9.83(s,1H),8.13(dd,J=4.8,1.4Hz,1H),7.92–7.86(m,1H),7.72–7.68(m,1H),7.6 7–7.64(m,3H),7.61(s,1H),7.60(d,J=2.9Hz,1H),7.57(s,1H),7.52–7.48(m,2H),7.16–7.10(m,2H),6.76(dd,J=2.8,2.0Hz,1H),5.18(dd, J=13.4,5.1Hz,1H),4.45(dd,J=16.9,14.0Hz,2H),3.72–3.63(m,4H),3.57–3.40(m,4H),3.00–2.90(m,6H),2.79–2.73(m,1H),2.72(s,3H), 2.57–2.46(m,1H),2.44(t,J=7.0Hz,2H),2.37(t,J=7.5Hz,2H),2.21– 2.14(m,1H),1.64–1.54(m,4H),1.51–1.44(m,2H),1.38–1.29(m,8H). 13C NMR (126MHz, Acetone-d6) δ205.54,205.39,205.23,171.88,170.98,170.36,168.11,167.84,163.02,1 50.01,147.27,142.50,140.85,140.01,136.59,131.17,131.14,130.41,129.00,128.48,127.51,127. 46,126.21,126.09,125.51,124.14,123.20,121.64,119.22,118.96,111.06,101.41,92.75,80.22,51.97,46.97,46.87,46.42,44.98,40.84,32.54,31.30,29.21,28.58,28.38,25.02,22.94,18.85,13.60.

[0120] Example 5

[0121] This embodiment provides a degrading agent targeting Aurora A, which is prepared using a method similar to that of Example 1, except that the starting material for intermediate (i) is changed to 12-bromo-1-dodecanool. Its chemical name and structural formula are as follows:

[0122] 4-(4-chloro-3-(pyridin-2-oxy)phenyl)-N-((4-(14-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)tetradecane-13-ynyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-5)

[0123]

[0124] White solid, yield: 15%. 1H NMR (500MHz, Acetone-d6) δ10.83(s,1H),10.62(s,1H),9.81(s,1H),8.13(d,J=4.0Hz,1H),7.93–7.87(m,1H),7.71(d,J=7.8Hz,1H),7.6 8–7.63(m,3H),7.63–7.60(m,1H),7.59(s,1H),7.51(d,J=8.0Hz,1H),7.47(s,1H),7.17–7.11(m,2H),6.79–6.75(m,1H),5.19(dd,J=13.4 ,5.1Hz,1H),4.48(dd,J=16.8,14.0Hz,2H),3.73–3.62(m,4H),3.57–3.41(m,4H),2.98–2.92(m,1H),2.82–2.74(m,1H),2.72(s,3H),2.6 0–2.50(m,1H),2.45(t,J=7.0Hz,2H),2.38(t,J=7.5Hz,2H),2.24–2.16(m,1H),1.65–1.53(m,4H),1.51–1.44(m,2H),1.39–1.27(s,12H). 13 C NMR (126MHz, Acetone-d6) δ171.94,171.16,170.39,167.94,163.01,150.03,147.26,142.53,1 40.80,140.04,136.56,131.15,130.42,129.06,128.59,127.59,127.53,126.22,126.08,125. 89,125.54,124.14,123.21,121.65,119.11,119.00,111.07,101.43,92.81,80.20,52.03,46.98,46.91,46.46,45.01,40.88,32.56,31.29,29.34,28.59,28.38,25.03,22.90,18.85,13.54.

[0125] Example 6

[0126] This embodiment provides a degrading agent targeting Aurora A, which is prepared using a method similar to that of Example 1, except that the starting material for intermediate (i) is changed to 14-bromo-1-tetradecaneol. Its chemical name and structural formula are as follows:

[0127] 4-(4-chloro-3-(pyridin-2-oxy)phenyl)-N-((4-(16-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)hexadecane-15-ynyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-6)

[0128]

[0129] White solid, yield: 15%. 1 H NMR(500MHz,Acetone-d6)δ10.86(s,1H),10.62(br s,1H),9.82(s,1H),8.16–8.11(m,1H),7.92–7.87(m,1H),7.70(d,J=7.8Hz,1H),7.68–7.64(m,3H),7.61(t,J=2.9Hz,1H),7.5 8(s,1H),7.51(d,J=7.9Hz,1H),7.48(s,1H),7.17–7.12(m,2H),6.78–6.75(m,1H),5.19(dd,J=13.4,5.1Hz,1H),4.47(dd,J=1 6.9,14.0Hz,2H),3.71–3.65(m,4H),3.49(d,J=50.9Hz,4H),3.00–2.96(m,1H),2.80–2.74(m,1H),2.73(s,3H),2.57–2.49(m, 1H),2.46(t,J=7.0Hz,2H),2.37(t,J=7.5Hz,2H),2.21–2.15(m,1H),1.66–1.56(m,4H),1.53–1.45(m,2H),1.38–1.24(m,16H). 13C NMR (126MHz, Acetone-d6) δ171.91,171.01,170.36,168.20,167.85,163.03,150.01,147.27,142. 51,140.87,140.01,136.59,131.17,131.15,130.41,129.00,128.45,127.50,127.48,126.20,126. 09,125.49,124.13,123.21,121.60,119.21,118.96,111.06,101.38,92.74,80.20,51.97,46.97,46.87,46.42,45.00,40.84,32.54,31.30,29.38,28.87,28.61,28.40,25.02,22.95,18.84,13.57.

[0130] Example 7

[0131] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-((4-(2-(2-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisodihydroindole-5-yl)prop-2-yn-1-yl)oxy)ethoxy)ethyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-7); its structural formula is shown in formula AP-7:

[0132]

[0133] The synthetic route for AP-7 and intermediate (i) is as follows:

[0134]

[0135] The specific synthesis steps are as follows:

[0136] 1. Weigh 646.2 mg of methyl methacrylate, 353.8 mg of bis(triphenylphosphine)palladium dichloride, and 38 mg of copper iodide into a 100 mL dried reaction flask. Purge with nitrogen three times. Add 20 mL of ultra-dry DMF to the reaction flask to dissolve and stir. Then add 432.5 mg of 2-[2-(2-propynoxy)ethoxy]ethanol and 0.84 mL of triethylamine to the reaction and react in an oil bath at 80 °C for 8 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (1).

[0137] 2. Weigh 205 mg of intermediate (1), 330 mg of carbon tetrabromide, and 262 mg of triphenylphosphine and add them to a 50 mL reaction flask. Add 20 mL of tetrahydrofuran to dissolve and stir at room temperature for 1 h. After the reaction is complete, directly evaporate the system to dryness, add dichloromethane to dissolve and mix with silica gel, and perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (i).

[0138] 3. Weigh 45 mg of intermediate (i), 78.9 mg of intermediate (h), 31.7 mg of sodium carbonate, and 8 mg of sodium iodide into a 25 mL reaction flask. Add DMF to dissolve and stir. React in an oil bath at 80 °C for 4 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (I).

[0139] 4. Using a method similar to that in Example 1, intermediate (h) and intermediate (i) are reacted to obtain the degradation agent, denoted as AP-7.

[0140] White solid, yield 15% 1 H NMR (500MHz, Acetone-d6) δ10.85(s,1H),9.82(s,1H),8.12(ddd,J=5.0,2.0,0.8Hz,1H),7.88(ddd,J= 8.4,7.2,2.0Hz,1H),7.71(d,J=7.8Hz,1H),7.65(s,3H),7.60–7.54(m,2H),7.46(s,1H),7.15–7.09(m ,2H),5.15(dd,J=13.4,5.2Hz,1H),4.49–4.39(m,4H),3.74–3.71(m,2H),3.65–3.59(m,4H),3.46(t,J =4.9Hz,4H),2.78–2.73(m,1H),2.71(s,3H),2.65–2.57(m,6H),2.53–2.41(m,1H),2.19–2.11(m,1H). 13C NMR (126MHz, Acetone-d6) δ172.74,171.22,168.55,163.92,150.90,148.15,143.41,141.76 ,140.89,137.45,132.84,132.18,131.29,129.89,129.29,128.31,127.34,126.98,126.80,1 26.37,125.02,124.20,122.35,119.98,119.83,111.94,102.24,88.82,85.95,70.94,69.92,69.85,59.25,58.23,53.79,52.86,47.78,47.58,32.61,32.16,23.77,23.31,14.45,14.34.

[0141] Example 8

[0142] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-((4-(2-(2-(2-((3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisodihydroindole-5-yl)prop-2-yn-1-yl)oxy)ethoxy)ethoxy)ethyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-8); its structural formula is shown in formula AP-8:

[0143]

[0144] The synthetic route for AP-8 and intermediate (i) is as follows:

[0145]

[0146] The specific synthesis steps are as follows:

[0147] 1. Weigh 646.2 mg of methyl methacrylate, 353.8 mg of bis(triphenylphosphine)palladium dichloride, and 38 mg of copper iodide into a 100 mL dried reaction flask. Purge with nitrogen three times. Add 20 mL of ultra-dry DMF to the reaction flask to dissolve and stir. Then add 451.7 mg of 2-(2-(2-(prop-2-yn-1-yloxy)ethoxy)ethoxy)ethanol and 0.84 mL of triethylamine to the reaction and react in an oil bath at 80 °C for 8 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (1).

[0148] 2. Weigh 430 mg of intermediate (1) and 1 g of Dys-Martin oxidant (DMP) and add them to a 50 mL reaction flask. Add 20 mL of ultra-dry dichloromethane solvent to dissolve and stir at room temperature for 5 h. After the reaction is complete, add saturated sodium bisulfite solution to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate and remove the solvent by vacuum rotary evaporation to obtain a pale yellow solid (i).

[0149] 3. Weigh 100 mg of intermediate (i), 121 mg of intermediate (h), 75 mg of sodium triacetoxyborohydride, and 20 mg of anhydrous sodium sulfate and add them to a 10 mL reaction flask. Add 3 mL of ultra-dry dichloromethane and 3 mL of ultra-dry methanol mixed solvent to dissolve and stir at room temperature for 3 h. After the reaction is completed, add ice water to the system under ice bath conditions to quench the reaction. Extract the reaction solution three times with ethyl acetate, combine the organic phases, wash the organic phase three times with water and saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (I).

[0150] 4. Using a method similar to that in Example 1, intermediate (h) and intermediate (i) are reacted to obtain the degradation agent, denoted as AP-8.

[0151] White solid, yield 15% 1 H NMR (500MHz, Acetone-d6) δ10.84(s,1H),9.82(s,1H),8.13(dd,J=4.8,1.4Hz,1H),7.95–7.84(m,1H),7.73(d,J=7.8Hz,1H) ,7.68–7.62(m,4H),7.62–7.56(m,2H),7.49(s,1H),7.14(dd,J=10.2,4.2Hz,2H),6.84–6.70(m,1H),5.18(dd,J=13.4,5.1Hz ,1H),4.71–4.25(m,4H),3.75–3.72(m,2H),3.68–3.65(m,2H),3.64–3.60(m,4H),3.60–3.57(m,2H),3.50–3.43(m,4H),3.0 2–2.98(m,2H),2.77(dd,J=4.4,2.4Hz,1H),2.74(s,3H),2.66–2.57(m,6H),2.51(dd,J=13.2,4.5Hz,1H),2.24–2.14(m,1H). 13CNMR (126MHz, Acetone-d6) δ171.94,170.35,167.76,163.03,150.01,147.28,142.52,140. 92,140.03,136.59,131.91,131.32,130.42,128.96,128.37,127.38,126.47,126.11,125. 97,125.45,124.12,123.33,121.47,119.17,118.96,111.07,101.32,87.97,85.08,70.33,70.23,70.16,69.14,68.93,58.39,57.38,52.89,52.00,46.93,46.67,31.28,22.90,13.61.

[0152] Example 9

[0153] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-((4-(10-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisodihydroindole-4-yl)dec-9-ynyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-9); its structural formula is as follows:

[0154]

[0155] The linker is connected at position 4 using a synthesis method similar to that in Example 3. The synthesis route is as follows:

[0156]

[0157] The specific preparation method is as follows:

[0158] 1. Weigh 538 mg of intermediate (1) and 968 mg of intermediate (2) (Br at position 4), and add them to a 100 mL dried reaction flask. Then add 250 mg of bis(triphenylphosphine)palladium dichloride and 57 mg of cuprous iodide. Replace with N2 three times, add 1.26 mL of ultra-dry triethylamine to the reaction flask, then add ultra-dry N,N-dimethylformamide to dissolve and stir. React in an oil bath at 80 °C for 8-12 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate, combine the organic phases, wash the organic phase three times with water and saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (3).

[0159] 2. Weigh 370 mg of intermediate (3) and add it to a 100 mL reaction flask that has been dried. Add dichloromethane to dissolve and stir. Add 1.35 g of Des Martin oxidant under ice bath and react for 8-12 h. After the reaction is complete, add saturated sodium sulfite solution to quench the reaction. Adjust the pH to acidity with dilute hydrochloric acid. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase with saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (i).

[0160] 3. Weigh 0.102 g of intermediate (h) and 0.709 g of intermediate (i) (n = 3) and add them to a 10 mL reaction tube. Add 0.5 mL of N,N-diisopropylethylamine and 0.114 g of condensing agent (HATU) to the reaction tube. Then add N,N-dimethylformamide to dissolve and stir. React at room temperature for 2 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution respectively. Dry the organic phase with anhydrous sodium sulfate. Remove the organic solvent by vacuum rotary evaporation. Obtain the degradation agent by rapid column chromatography (dichloromethane / methanol), denoted as AP-9.

[0161] White solid, yield 15% 1H NMR (500MHz, Acetone-d6) δ10.81(s,1H),9.87(s,1H),8.12(dd,J=5.0,1.9Hz,1H),7.88(ddd,J=8.9,7.2,2.0Hz,1H),7.69(d, J=7.5Hz,1H),7.65–7.63(m,2H),7.61–7.56(m,2H),7.52–7.45(m,2H),7.15–7.09(m,2H),6.74(s,1H),5.21(dd,J=13.4,5.1Hz ,1H),4.46(dd,2H),3.71–3.63(m,4H),3.54–3.48(m,2H),3.45–3.39(m,1H),3.02–2.91(m,1H),2.86–2.73(m,1H),2.70(s,3H ),2.46(t,J=6.9Hz,2H),2.38(t,J=7.4Hz,2H),2.25–2.17(m,1H),1.64–1.54(m,4H),1.51–1.44(m,2H),1.33–1.25(m,4H).13C NMR (126MHz, Acetone-d6) δ172.84,171.98,171.31,168.93,163.88,150.88,148.13,144.91 ,141.71,140.87,137.44,134.83,133.18,131.27,129.88,129.34,129.19,128.38,126.96,1 26.37,125.00,123.40,122.48,120.44,120.05,119.82,111.92,102.27,96.95,77.21,52.83,49.76,47.82,47.79,47.26,45.87,41.74,33.37,32.17,29.19,25.79,23.86,19.73,14.46.

[0162] Example 10

[0163] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl-N-[(4-{10-[2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindoline-5-yl]dec-9-ethynyl}piperazin-1-yl)sulfonyl]-7-methyl-1H-indole-6-carboxamide (AP-10); its structural formula is as follows:

[0164]

[0165] The linker is connected at position 6 using a synthesis method similar to that in Example 9. The synthesis route is as follows:

[0166]

[0167] The specific preparation method is as follows:

[0168] 1. Weigh 403 mg of intermediate (1) and 726 mg of intermediate (2) (Br at position 6), and add them to a 100 mL dried reaction flask. Then add 187.5 mg of bis(triphenylphosphine)palladium dichloride and 42.5 mg of cuprous iodide. Replace with N2 three times, add 0.94 mL of ultra-dry triethylamine to the reaction flask, then add ultra-dry N,N-dimethylformamide to dissolve and stir. React in an oil bath at 80 °C for 8-12 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate, combine the organic phases, wash the organic phase three times with water and saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (3).

[0169] 2. Weigh 410 mg of intermediate (3) and add it to a 100 mL reaction flask that has been dried. Add dichloromethane to dissolve and stir. Add 1.70 g of Des Martin oxidant under ice bath and react for 2-4 h. After the reaction is complete, add saturated sodium sulfite solution to quench the reaction. Adjust the pH to acidity with dilute hydrochloric acid. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase with saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (i).

[0170] 3. Weigh 0.051 g of intermediate (h) and 0.095 g of intermediate (i) (n = 3) and add them to a 10 mL reaction tube. Add 0.5 mL of N,N-diisopropylethylamine and 0.114 g of condensing agent (HATU) to the reaction tube. Then add N,N-dimethylformamide to dissolve and stir. React at room temperature for 2 h. After the reaction is complete, add water to quench the reaction and extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution respectively. Dry the organic phase with anhydrous sodium sulfate and remove the organic solvent by vacuum rotary evaporation. Obtain the degradation agent by rapid column chromatography (dichloromethane / methanol), denoted as AP-10.

[0171] White solid, yield 15% 1H NMR (500MHz, Acetone-d6) δ10.90(s,1H),9.84(s,1H),8.11(dd,J=4.9,2.0Hz,1H),7.87(t,J=2.0Hz,1H),7.68(s,1H),7.65–7 .62(m,3H),7.57(dd,J=6.0,2.4Hz,2H),7.53(s,1H),7.45(s,1H),7.12(d,J=7.6Hz,2H),6.75–6.71(m,1H),5.18(dd,J=13.4, 5.1Hz,1H),4.45(q,J=17.1Hz,2H),3.66(t,J=5.1Hz,4H),3.47(dt,J=47.5,5.1Hz,4H),3.01–2.90(m,1H),2.83–2.71(m,1H), 2.70(s,3H),2.56–2.44(m,1H),2.46–2.36(m,4H),2.21–2.12(m,1H),1.63–1.54(m,4H),1.52–1.43(m,2H),1.39–1.33(m,4H). 13 C NMR (126MHz, Acetone-d6) δ172.81,171.96,171.22,168.80,168.70,163.87,150.86,148.12, 142.30,141.67,140.89,137.43,135.46,133.21,131.28,129.87,128.41,126.96,126.75,12 6.63,126.36,124.98,124.79,124.42,122.55,119.97,119.83,111.92,102.23,91.95,80.63,52.85,48.00,47.81,47.29,45.88,41.73,33.35,32.14,29.25,25.81,23.77,19.62,14.44.

[0172] Example 11

[0173] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 5-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-((4-(10-(2-(2,6-dioxopiperidin-3-yl)-3-oxoisoindoline-4-yl)dec-9-ynyl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-11); its structural formula is as follows:

[0174]

[0175] The linker is connected at position 7 using a synthesis method similar to that in Example 9. The synthesis route is as follows:

[0176]

[0177] The specific preparation method is as follows:

[0178] 1. Weigh 462 mg of intermediate (1) and 646 mg of intermediate (2) (Br at position 7), and add them to a 100 mL dried reaction flask. Then add 353.8 mg of bis(triphenylphosphine)palladium dichloride and 38 mg of cuprous iodide. Replace with N2 three times. Add 0.84 mL of ultra-dry triethylamine to the reaction flask, then add ultra-dry N,N-dimethylformamide to dissolve and stir. React in an oil bath at 80 °C for 8-12 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (3).

[0179] 2. Weigh 615 mg of intermediate (3) and add it to a 100 mL reaction flask that has been dried. Add dichloromethane to dissolve and stir. Add 2.52 g of Des Martin oxidant under ice bath and react for 2-4 h. After the reaction is complete, add saturated sodium sulfite solution to quench the reaction. Adjust the pH to acidity with dilute hydrochloric acid. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase with saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (i).

[0180] 3. Weigh 0.191 g of intermediate (h) and 0.093 g of intermediate (i) (n = 3) and add them to a 10 mL reaction tube. Add 0.5 mL of N,N-diisopropylethylamine and 0.194 g of condensing agent (HATU) to the reaction tube. Then add N,N-dimethylformamide to dissolve and stir. React at room temperature for 2 h. After the reaction is complete, add water to quench the reaction and extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution respectively. Dry the organic phase with anhydrous sodium sulfate and remove the organic solvent by vacuum rotary evaporation. Obtain the degradation agent by rapid column chromatography (dichloromethane / methanol), denoted as AP-11.

[0181] White solid, yield 15% 1H NMR (500MHz, Acetone-d6) δ10.89(s,1H),10.63(s,1H),9.82(s,1H),8.28–8.11(m,1H),7.98–7.85(m,1H),7.71 –7.63(m,3H),7.60(t,J=2.8Hz,1H),7.56–7.47(m,3H),7.45(d,J=7.2Hz,1H),7.19–7.09(m,2H),5.21(dd,J=13. 3,5.0Hz,1H),4.51–4.33(m,2H),3.73–3.64(m,4H),3.60–3.39(m,4H),3.01(m,1H),2.79–2.74(m,1H),2.72(s, 3H),2.54(m,1H),2.47–2.35(m,4H),2.25–2.18(m,1H),1.66–1.57(m,4H),1.55–1.47(m,2H),1.40–1.33(m,4H). 13 C NMR (126MHz, Acetone-d6) δ171.99,170.51,167.86,163.02,150.00,147.28,143.22,14 0.81,140.03,136.60,132.24,131.42,131.14,130.40,129.03,128.56,127.59,126.11 ,125.84,125.50,124.13,122.46,121.75,120.90,119.14,118.97,111.06,101.39,96.54,77.19,51.91,46.97,46.46,45.03,40.85,32.52,31.35,24.95,22.95,19.22,13.59.

[0182] Example 12

[0183] This embodiment provides a degrading agent targeting Aurora A, which is prepared using a method similar to that of Example 3, except that the piperazine ring in the intermediate (h) of Example 3 is replaced with cyclobutanediamine. Its chemical name and structural formula are as follows:

[0184] 4-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-(N-((1r,3r)-3-(10-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)dec-9-ynylamide)cyclobutyl)aminosulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-12)

[0185]

[0186] Preparation of intermediate (h):

[0187]

[0188] The specific synthesis steps are as follows:

[0189] 1. Intermediate (e) was obtained using a method similar to that in Example 1, and the synthesis method was the same.

[0190] 2. Weigh 960 mg of sulfonamide and 2 g of tert-butyl (trans-3-aminocyclobutyl)carbamate, and add them to a 100 mL dried reaction flask. Purge with nitrogen three times. Add 100 mL of ultra-dry 1,4-dioxane to the reaction flask to dissolve and stir. Heat to 115 °C and stir overnight. After the reaction is complete, cool the reaction solution to room temperature, filter, and obtain a white solid. Wash the white solid with 1,4-dioxane, and vacuum dry the solid to obtain intermediate (f).

[0191] 3. The compound is then cleaved with trifluoroacetic acid to remove the Boc, yielding the intermediate (h).

[0192] 4. Using a method similar to that of Example 3, the intermediate (h) is reacted with the intermediate (i) of Example 3 to obtain the degradation agent, denoted as AP-12.

[0193] White solid, yield: 15% 1 H NMR (500MHz, CD3OD) δ8.17–8.10(m,1H),7.91–7.82(m,1H),7.70(d,J=7.9Hz,1H),7.66–7.62(m,2H),7.61–7.58(m,1H),7. 52(s,1H),7.49–7.44(m,2H),7.29(s,1H),7.17–7.11(m,1H),7.07(d,J=8.3Hz,1H),6.68(d,J=3.1Hz,1H),5.13(dd,1H),4. 50–4.38(m,2H),4.26–4.16(m,2H),2.93–2.86(m,1H),2.83–2.75(m,1H),2.70(s,3H),2.54–2.45(m,3H),2.43(t,J=6.9Hz ,2H),2.37–2.28(m,2H),2.17(t,J=7.4Hz,2H),2.15–2.11(m,1H),1.66–1.55(m,4H),1.51–1.43(m,2H),1.41–1.35(m,4H). 13CNMR(126MHz,CD3OD)δ174.86,173.40,170.79,169.56,163.04,149.73,146.83,142.34,14 1.15,140.34,136.42,131.11,130.43,130.11,129.06,128.14,128.10,127.47,126.12,125 .81,125.32,123.43,122.91,121.30,118.89,118.64,110.97,100.84,92.85,79.85,52.26,45.90,41.31,36.59,35.50,30.93,28.59,28.33,28.27,28.07,25.47,22.62,18.56,12.95.

[0194] Example 13

[0195] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-((4-(10-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindololin-5-yl)dec-9-yn-1-yl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-13); its structural formula is shown in formula AP-13:

[0196]

[0197] Synthetic route of intermediate (i):

[0198]

[0199] The specific synthetic route is similar to that in Example 7 to obtain AP-13.

[0200] White solid, yield 15% 1H NMR (500MHz, Acetone-d6) δ10.88(s,1H),9.83(s,1H),8.15–8.10(m,1H),7.96–7.85(m,1H),7.71(d,J=7.9Hz,1H),7.69–7 .64(m,3H),7.62–7.57(m,2H),7.52(d,J=7.9Hz,1H),7.48(s,1H),7.22–7.10(m,2H),6.76(s,1H),5.19(dd,J=13.4,5.1Hz, 1H),4.48(q,J=16.9Hz,2H),3.51–3.45(m,4H),2.79(d,J=4.0Hz,1H),2.74(s,3H),2.56(d,J=4.6Hz,1H),2.55–2.50(m,4H ),2.47(t,J=7.0Hz,2H),2.37(t,J=7.3Hz,2H),2.25–2.16(m,1H),1.56–1.48(m,4H),1.41–1.35(m,4H),1.34–1.28(m,4H). 13 CNMR(126MHz,Acetone-d6)δ171.89,170.35,167.82,163.04,150.03,147.27,142.51,14 0.88,140.01,136.59,131.19,131.15,130.41,129.04,128.40,127.46,126.32,126.19,1 26.09,125.50,124.15,123.21,121.47,119.08,118.95,111.05,101.37,92.71,80.18,57.92,52.54,51.97,46.86,46.68,31.30,29.71,28.39,27.10,26.53,22.94,18.83,13.54.

[0201] Example 14

[0202] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-((4-(4-(4-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)prop-2-yn-1-yl)piperazin-1-yl)-4-oxobutyryl)piperazin-1-yl)sulfonyl)-7-methyl-1H-indole-6-carboxamide (AP-14); its structural formula is shown in formula AP-14:

[0203]

[0204] The synthetic route for AP-14 and intermediate (i) is as follows:

[0205]

[0206] The specific steps for preparing AP-14 are as follows:

[0207] 1. Intermediate (e) was obtained using a method similar to that in Example 1, and the synthesis method was the same.

[0208] 2. Weigh 526 mg of (e) and 100 mg of succinic anhydride, add them to a dried 50 mL reaction flask, purge with nitrogen three times, add 100 mL of a mixed solution of ultra-dry dichloromethane and tetrahydrofuran to the reaction flask to dissolve and stir, then add 0.14 mL of triethylamine to the reaction system and stir overnight at room temperature. After the reaction is complete, concentrate the reaction solution under vacuum and use it directly in the next step without separation and purification.

[0209] 3. Weigh 1.9 g of bromopropyne, 2 g of tert-butylpiperazine carboxylate, and 3.4 g of sodium carbonate, and add them to a 50 mL dried reaction flask. Purge the flask with nitrogen three times. Add 100 mL of ultra-dry acetonitrile to the reaction flask to dissolve and stir. React at 50 °C for 5 h. After the reaction is complete, cool the reaction solution to room temperature and concentrate the reaction solution under vacuum. Directly pass the reaction solution through trifluoroacetic acid to remove Boc. Concentrate the reaction solution under vacuum and perform rapid column chromatography (dichloromethane / methanol) to obtain the corresponding intermediate. Then, perform a Sonogashira coupling reaction with a hydroxylamine. The specific synthetic route is the same as the previous hydroxylamine coupling reaction to obtain intermediate (i).

[0210] 4. Using a method similar to that in Example 1, intermediate (h) and intermediate (i) are reacted to obtain the degradation agent, denoted as AP-14.

[0211] White solid, yield 16%. 1H NMR (500MHz, Acetone-d6) δ10.84(s,1H),10.59(s,1H),9.79(s,1H),8.26–8.09(m,1H),8.01–7.88(m ,1H),7.76–7.72(m,1H),7.70–7.63(m,4H),7.64–7.56(m,2H),7.49(s,1H),7.18–7.09(m,2H),6.87– 6.70(m,1H),5.20(dd,J=13.3,5.1Hz,1H),3.85–3.50(m,14H),3.45(s,2H),3.08–2.93(m,2H),2.80– 2.75(m,2H),2.73(s,3H),2.67–2.61(m,5H),2.60–2.52(m,3H),2.47–2.37(m,1H),2.24–2.17(m,2H). 13 C NMR (126MHz, Acetone-d6) δ171.80,170.32,169.84,167.65,150.04,147.28,14 2.55,140.82,140.00,136.57,131.34,130.41,128.48,127.56,126.47,126.37, 126.09,125.98,125.53,124.16,123.27,119.14,118.95,111.05,101.42,51.99,51.75,46.98,46.89,44.93,41.21,41.11,31.30,27.68,27.50,22.94,13.60.

[0212] Example 15

[0213] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl-N-[(4-{5-[4-(3-{2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl}prop-2-yn-1-yl)piperazin-1-yl]-5-oxopentanoyl}piperazin-1-yl)sulfonyl]-7-methyl-1H-indole-6-carboxamide (AP-15); its structural formula is shown in formula AP-15:

[0214]

[0215] The specific synthetic route is similar to that of Example 14, yielding Example 15, except that the intermediate succinic anhydride is replaced with glutaric anhydride. White solid, yield 16%. 11H NMR (500 MHz, Acetone-d6) 1 1H NMR (500 MHz, Acetone) δ 10.86 (s, 1H), 10.86 (s, 1H), 9.82 (s, 1H), 9.82 (s, 1H), 8.14 (dd, J = 4.8, 1.1 Hz, 1H), 8.14 (dd, J = 4.8, 1.1 Hz, 1H), 7.95–7.85 (m, 1H), 7.73 (d, J = 7.8 Hz, 1H), 7.69–7.63 (m, 5H), 7.62–7.58 (m, 1H), 7.56 (d, J = 7.8 Hz, 1H), 7.49 (s, 1H), 7.19–7.09 (m, 3H), 6.79–6.72 (m, 1H), 5.19 (dd, J = 13.4, 5.1 Hz, 1H), 4.60–4.42 (m, 3H), 3.72–3.67 (m, 4H), 3.64–3.52 (m, 10H), 3.44 (s, 3H), 3.04–3.02 (m, 1H), 2.75–2.73 (m, 1H), 2.72 (s, 3H), 2.65–2.57 (m, 4H), 2.60–2.51 (m, 4H), 2.44 (t, J = 7.2 Hz, 3H), 2.40 (t, J = 7.2 Hz, 3H), 2.25–2.20 (m, 1H), 1.90–1.81 (m, 2H). 13 13C NMR (151 MHz, Acetone-d6) δ 171.81, 170.67, 170.32, 167.89, 167.67, 163.03, 150.03, 147.27, 142.55, 140.82, 140.01, 136.57, 131.75, 131.31, 130.41, 129.05, 128.49, 127.55, 126.45, 126.37, 126.10, 126.00, 125.53, 124.16, 123.27, 121.63, 119.15, 118.95, 111.05, 101.42, 86.87, 84.72, 51.99, 51.89, 51.41, 46.97, 46.89, 46.42, 45.04, 44.91, 41.06, 40.85, 31.95, 31.88, 31.30, 22.94, 20.71, 13.54。

[0216] Example 16

[0217] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-[(4-{4-[(5-{2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl}pent-4-yn-1-yl)amino]-4-oxobutyryl}piperazin-1-yl)sulfonyl]-7-methyl-1H-indole-6-carboxamide (AP-16); its structural formula is shown in formula AP-16:

[0218]

[0219] The synthetic route for AP-16 and intermediate (i) is as follows:

[0220]

[0221] The specific synthesis steps of the preparation method are as follows:

[0222] 1. Weigh 646.2 mg of methyl methacrylate, 140.2 mg of bis(triphenylphosphine)palladium dichloride, and 38 mg of copper iodide and add them to a 100 mL dried reaction flask. Purge with nitrogen three times. Add 20 mL of ultra-dry DMF to the reaction flask to dissolve and stir. Then add 550 mg of N-(4-pentynyl)carbamate tert-butyl ester and 1.6 mL of triethylamine to the reaction and react in an oil bath at 80 °C for 8 h. After the reaction is complete, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (1).

[0223] 2. Weigh 100 mg of intermediate (1) and add it to a 100 mL reaction flask that has been dried. Add 5 mL of ultra-dry dichloromethane and then add 5 mL of trifluoroacetic acid to the reaction solution. After the reaction is completed, directly concentrate the reaction solution under vacuum. Then add dichloromethane to dissolve and mix with silica gel. Perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (i).

[0224] 3. Weigh 100 mg of intermediate (i) and 193 mg of intermediate (h) from Example 14, and add them to a 10 mL reaction tube. Add 0.5 mL of N,N-diisopropylethylamine and 176 mg of condensing agent (HATU) to the reaction tube, then add 1 mL of N,N-dimethylformamide to dissolve and stir. React at room temperature for 1 h. After the reaction is complete, add water to quench the reaction and extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution, respectively. Dry the organic phase with anhydrous sodium sulfate, remove the organic solvent by vacuum rotary evaporation, and obtain AP-16 by rapid column chromatography (dichloromethane / methanol).

[0225] White solid, yield 16%. 1 H NMR (500MHz, Acetone-d6) δ10.96(s,1H),9.87(s,1H),8.13(d,J=3.9Hz,1H),7.98–7.86(m,1H),7.76–7.62 (m,4H),7.59(dd,J=10.1,7.5Hz,2H),7.53–7.47(m,2H),7.33(s,1H),7.18–7.11(m,2H),6.80–6.73(m,1H) ,5.18(dd,J=13.3,5.0Hz,1H),4.53–4.36(m,2H),3.73–3.64(m,4H),3.60–3.45(m,4H),3.36–3.31(m,4H), 2.95–2.93(m,1H),2.80–2.62(m,7H),2.73(s,3H),2.57–2.44(m,5H),2.23–2.15(m,1H),1.83–1.71(m,2H). 13 C NMR (151MHz, Acetone-d6) δ171.89,170.38,167.82,163.03,150.02,147.28,142.46,140 .88,140.01,136.60,131.21,131.18,130.41,128.97,128.50,127.52,127.31,126.24,12 6.10,125.48,124.14,123.16,121.73,119.21,118.96,111.06,101.34,92.15,80.35,51.97,46.86,46.75,46.42,44.86,41.07,38.11,31.30,30.80,28.04,22.94,16.45,13.61.

[0226] Example 17

[0227] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-[(4-{3-[6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)hex-5-ynylamido]propionyl}piperazin-1-yl)sulfonyl]-7-methyl-1H-indole-6-carboxamide (AP-17); its structural formula is shown in formula AP-17:

[0228]

[0229] The synthesis route is as follows:

[0230]

[0231] The specific synthesis steps are as follows:

[0232] 1. Weigh 526.1 mg of intermediate (h), 283.8 mg of Boc-β-alanine, and 260.4 mg of HATU into a 50 mL reaction flask. Add DMF to dissolve and stir. Add 1 mL of DIPEA to the system and react at room temperature for 2 h. After the reaction is complete, quench the reaction with water. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain the compound. The compound is then subjected to trifluoroacetic acid to remove Boc to obtain intermediate (1).

[0233] 2. Weigh 105.9 mg of intermediate (1), 63 mg of intermediate (i), and 114 mg of HATU into a 50 mL reaction flask. Add DMF to dissolve and stir. Add 1 mL of DIPEA to the system and react at room temperature for 2 h. After the reaction is complete, quench the reaction with water. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain compound AP-17.

[0234] White solid, yield 15% 11H NMR (500 MHz, Acetone-d6) δ 10.90 (s, 1H), 9.84 (s, 1H), 8.11 (ddd, J = 4.8, 2.0, 0.9 Hz, 1H), 7.88 (ddd, J = 8.3, 7.2, 2.0 Hz, 1H), 7.67 (d, J = 7.8 Hz, 1H), 7.66–7.62 (m, 3H), 7.58 (t, J = 2.9 Hz, 1H), 7.55 (s, 1H), 7.49 (dd, J = 7.9, 1.3 Hz, 1H), 7.46 (s, 1H), 7.19 (t, J = 5.7 Hz, 1H), 7.14–7.10 (m, 2H), 6.74 (dd, J = 3.2, 1.8 Hz, 1H), 5.17 (dd, J = 13.4, 5.1 Hz, 1H), 4.43 (q, J = 16.9 Hz, 2H), 3.76–3.62 (m, 4H), 3.60–3.51 (m, 2H), 3.47–3.41 (m, 4H), 2.78–2.72 (m, 1H), 2.70 (s, 3H), 2.58 (t, J = 6.4 Hz, 2H), 2.56–2.47 (m, 1H), 2.45 (t, J = 7.0 Hz, 2H), 2.32 (t, J = 7.3 Hz, 2H), 2.21–2.14 (m, 1H). 13 13C NMR (126 MHz, Acetone-d6) δ 172.80, 171.27, 168.75, 163.90, 150.89, 148.14, 143.32, 141.70, 140.91, 137.45, 132.09, 132.07, 131.30, 129.91, 129.42, 128.44, 128.18, 127.15, 126.99, 126.79, 126.40, 125.02, 124.04, 122.57, 120.00, 119.85, 111.94, 102.26, 92.95, 81.44, 52.86, 47.76, 47.65, 47.23, 45.75, 41.75, 36.01, 35.41, 33.60, 32.17, 25.38, 23.80, 19.30, 14.45。

[0235] Example 18 ​This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl)-N-{[4-(3-{4-[3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl)prop-2-yn-1-yl]piperazin-1-yl}propionyl)piperazin-1-yl]sulfonyl}-7-methyl-1H-indole-6-carboxamide (AP-18); its structural formula is shown in formula AP-18:

[0237]

[0238] The synthesis route is as follows:

[0239]

[0240] The specific steps of the preparation method are as follows:

[0241] 1. Weigh 2.0 g of bromopropyne, 1.9 g of tert-butylpiperazine carboxylic acid ester, and 3.4 g of sodium carbonate into a 100 mL dried reaction flask. Purge with nitrogen three times. Add 100 mL of ultra-dry acetonitrile to the reaction flask to dissolve and stir. React the reaction solution at 50 °C for 5 h. After the reaction is complete, add water to the system to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate and remove the solvent by vacuum rotary evaporation to obtain intermediate (a).

[0242] 2. Weigh 660 mg of intermediate (a), 1.05 g of methylbenzylamine, 210.6 mg of palladium dichloride (triphenylphosphine), and 57 mg of copper iodide into a 100 mL dried reaction flask. Purge with nitrogen three times. Add 20 mL of ultra-dry DMF to the reaction flask to dissolve and stir. Then add 1.6 mL of triethylamine to the reaction mixture and react in an oil bath at 80 °C for 8 h. After the reaction is complete, quench the reaction with water. Extract the reaction solution three times with ethyl acetate. Combine the organic phases, wash the organic phase three times with water and saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (b).

[0243] 3. Weigh 100 mg of intermediate (1) and add it to a 20 mL reaction flask that has been dried. Add 5 mL of ultra-dry dichloromethane and then add 5 mL of trifluoroacetic acid to the reaction solution. After the reaction is completed, directly concentrate the reaction solution under vacuum. Then add dichloromethane to dissolve and mix with silica gel. Perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (c).

[0244] 4. Weigh 183 mg of intermediate (c), 117 mg of tert-butyl bromopropionate, 8.3 mg of potassium iodide, and 104 mg of potassium carbonate. Add 20 mL of ultra-dry DMF and react overnight at 80 °C. After the reaction is complete, add water to quench the reaction mixture. Extract the reaction mixture three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution, respectively. Dry the organic phase with anhydrous sodium sulfate, remove the organic solvent by vacuum rotary evaporation, and obtain intermediate (d) by rapid column chromatography (dichloromethane / methanol).

[0245] 5. Weigh 100 mg of intermediate (d) and add it to a 20 mL reaction flask that has been dried. Add 5 mL of ultra-dry dichloromethane and then add 5 mL of trifluoroacetic acid to the reaction solution. After the reaction is complete, directly concentrate the reaction solution under vacuum. Then add dichloromethane to dissolve and mix with silica gel. Perform rapid column chromatography (dichloromethane / methanol) to obtain intermediate (1).

[0246] 6. Weigh 66 mg of intermediate (1) and 79 mg of intermediate (e) and add them to a 10 mL reaction tube. Add 0.5 mL of N,N-diisopropylethylamine and 86 mg of condensing agent (HATU) to the reaction tube. Then add 1 mL of N,N-dimethylformamide to dissolve and stir. React at room temperature for 1 h. After the reaction is complete, add water to the reaction solution to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution respectively. Dry the organic phase with anhydrous sodium sulfate. Remove the organic solvent by vacuum rotary evaporation. Obtain Example 18 by rapid column chromatography (dichloromethane / methanol).

[0247] White solid, yield 15% 1 H NMR (500MHz, DMSO-d6) δ11.58(s,1H),11.01(s,1H),8.15(d,J=3.3Hz,1H),7.95–7.89(m,1H),7. 74–7.66(m,3H),7.66–7.61(m,2H),7.60–7.53(m,2H),7.34(s,1H),7.22–7.13(m,2H),6.61(s,1H ),5.11(dd,J=13.2,5.0Hz,1H),4.44(d,J=17.6Hz,1H),4.33(d,J=17.6Hz,1H),3.60–3.54(m,6H) ,3.32–3.20(m,6H),2.95–2.86(m,1H),2.80–2.59(m,14H),2.45–2.34(m,1H),2.06–1.97(m,1H). 13C NMR(126MHz,DMSO-d6)δ173.34,171.39,169.85,167.86,162.90,149.86,147.71,14 2.86,141.30,140.93,136.70,131.71,130.96,129.13,128.01,126.97,126.51,126. 07,125.12,124.15,123.68,121.72,120.09,119.69,114.47,111.48,101.03,88.09,85.08,53.71,52.67,52.16,50.94,47.53,46.94,46.61,45.08,31.66,22.88,14.86.

[0248] Example 19

[0249] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl-N-[(4-{10-[2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1-oxoisoindololin-5-yl]dec-9-ethynyl}piperazin-1-yl)sulfonyl]-7-methyl-1H-indole-6-carboxamide (AP-19); its structural formula is shown in formula AP-19:

[0250]

[0251] The synthesis method is the same as in Example 3, except that the CRBN ligand is changed to obtain AP-19.

[0252] White solid, yield 15%, ¹H NMR (500 MHz, Acetone-d6) δ 10.81 (s, ¹H), 10.58 (s, ¹H), 9.80 (s, ¹H), 8.14 (d, J = 3.5 Hz, ¹H), 7.97–7.86 (m, ¹H), 7.76–7.58 (m, 5H), 7.52–7.42 (m, 2H), 7.21–7.08 (m, 2H), 6.84–6.65 (m, ¹H), 5.19 (dd, J = 13.4, 5.1 Hz, ¹H), 4.46 (q ,J=16.7Hz,2H),3.76–3.63(m,4H),3.58–3.40(m,4H),3.01–2.95(m,1H),2.79–2.70(m,1H),2.69(s,3H),2.59– 2.46(m,3H),2.41(t,J=7.4Hz,2H),2.24–2.15(m,2H),1.69–1.56(m,5H),1.57–1.47(m,3H),1.44–1.37(m,4H). 19 F NMR(471MHz,Acetone-d6)δ-112.96(s). 13 CNMR(126MHz,Acetone-d6)δ205.28,205.12,171.77,170.85,170.20,166.95,163.54,163.03,161.58,15 0.02,147.27,140.85,140.00,137.67,136.58,133.10,133.03,130.40,129.01,128.44,128.38,127.51,1 26.09,125.52,124.15,121.63,121.59,119.20,118.95,111.05,109.70,109.51,101.41,101.35,98.38,73.72,52.15,46.98,46.64,46.44,44.98,40.82,32.47,31.27,28.43,28.17,24.90,22.89,18.98,13.55.

[0253] Example 20

[0254] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl-N-[(4-{3-[4-({3-[2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl]prop-2-yn-1-yl}oxy)piperidin-1-yl]propionyl}piperazin-1-yl)sulfonyl]-7-methyl-1H-indole-6-carboxamide (AP-20); its structural formula is shown in formula AP-20:

[0255]

[0256] Synthetic route of intermediate (i):

[0257]

[0258] The specific steps for preparing AP-20 are as follows:

[0259] 1. Weigh 2g of 1-tert-butyloxycarbonyl-4-hydroxypiperidine and add it to a 100mL reaction flask that has been dried. Add tetrahydrofuran to dissolve and stir. Add NaH to the system. Bubbles are generated in the system. After 0.5h, add bromopropyne to the reaction system. After 4h of reaction, add water to quench the reaction. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (petroleum ether / ethyl acetate) to obtain intermediate (1).

[0260] 2. Weigh 1.16 g of intermediate (1), 1.45 g of methyl methacrylate, 353.8 mg of palladium dichloride bis(triphenylphosphine) and 95 mg of cuprous iodide into a dried reaction flask. Purge with nitrogen three times, add ultra-dry DMF to dissolve and stir. Add 0.84 mL of triethylamine dropwise to the system and react in an oil bath at 80 °C for 8 h. After the reaction is complete, quench the reaction with water, extract the reaction solution three times with ethyl acetate, combine the organic phases, wash the organic phase three times with water and saturated sodium chloride solution, dry the organic phase with anhydrous sodium sulfate, remove the solvent by vacuum rotary evaporation, and then perform rapid column chromatography (dichloromethane / methanol) to obtain the compound. The compound is then cleaved with trifluoroacetic acid to remove tert-butyl ester to obtain intermediate (2).

[0261] 3. Weigh 1.25 g of intermediate (2), 691.1 mg of potassium carbonate, and 41.5 mg of potassium iodide into a 50 mL reaction flask. Add DMF to dissolve and stir. Add 0.5 mL of tert-butyl 3-bromopropionate dropwise to the system and react in an oil bath at 80 °C for 10 h. After the reaction is complete, quench the reaction with water. Extract the reaction solution three times with ethyl acetate. Combine the organic phases and wash the organic phase three times with water and saturated sodium chloride solution. Dry the organic phase with anhydrous sodium sulfate. Remove the solvent by vacuum rotary evaporation and then perform rapid column chromatography (dichloromethane / methanol) to obtain the compound. The compound is then cleaved by trifluoroacetic acid to remove tert-butyl ester to obtain intermediate (I).

[0262] 4. AP-20 was obtained by amide condensation using a method similar to that in Example 1.

[0263] White solid, yield 15% 1 H NMR (500MHz, DMSO-d6) δ11.55(s,1H),11.01(s,1H),8.14(dd,J=5.1,1.9Hz,1H),7.90(ddd,J=8.9,7.2,2.0Hz,1H),7 .72(d,J=7.9Hz,1H),7.71–7.68(m,2H),7.66–7.60(m,2H),7.59–7.54(m,2H),7.34(s,1H),7.19–7.13(m,2H),6.62– 6.59(m,1H),5.11(dd,J=13.3,5.1Hz,1H),4.51–4.28(m,4H),3.70(s,1H),3.60–3.50(m,4H),3.37–3.21(m,4H),3.1 1–2.94(m,4H),2.94–2.87(m,1H),2.72(s,4H),2.63(s,3H),2.61–2.56(m,1H),2.45–2.29(m,1H),2.11–1.86(m,2H). 13C NMR(126MHz,DMSO-d6)δ172.91,170.96,168.77,167.37,162.48,149.44,147.27,142.45,140 .89,140.50,136.28,131.68,131.29,130.53,128.66,127.59,126.58,126.38,126.08,125.12 ,124.69,123.72,123.32,121.23,119.67,119.26,111.05,100.61,88.66,84.91,55.45,54.94,52.64,51.74,49.68,47.13,46.57,46.15,44.57,40.85,31.23,28.85,28.62,22.44,14.41.

[0264] Example 21

[0265] This embodiment provides a degrading agent targeting Aurora A, with the chemical name: 4-(4-chloro-3-(pyridin-2-yloxy)phenyl-N-[(4-{3-[4-(3-{2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-yl}prop-2-yn-1-yl)piperazin-1-yl]-3-oxopropyl}piperazin-1-yl)sulfonyl]-7-methyl-1H-indole-6-carboxamide (AP-21); its structural formula is shown in formula AP-21:

[0266]

[0267] The prepared linker was first linked to the inhibitor using a method similar to that in Example 18 to obtain AP-21.

[0268] White solid, yield: 15%, ¹H NMR (500MHz, DMSO-d6) δ 10.87 (s, ¹H), 9.83 (s, ¹H), 8.20–8.12 (m, ¹H), 7.90 (m, J = 1.9 Hz, ¹H), 7.73 (d, J = 7.9 Hz, ¹H), 7.67 (m, 2H), 7.64 (s, ¹H), 7.60 (t, J = 2.9 Hz, ¹H), 7.57–7.54 (m, ¹H), 7.48 (s, ¹H), 7.17–7.11 (m, 2H), 6.79–6.72 ( m,1H),5.19(dd,J=13.4,5.1Hz,1H),4.53–4.42(m,2H),3.64–3.56(m,6H),3.50–3.43(m,4H),3.01(d,J=5.4Hz,1H ),2.99–2.98(m,1H),2.78–2.73(m,4H),2.70(t,J=7.3Hz,2H),2.64(s,2H),2.61–2.49(m,9H),2.22–2.16(m,1H). 13 C NMR (126MHz, DMSO-d6) δ173.34,171.39,169.85,167.86,162.90,149.86,147.71,142. 86,141.30,140.93,136.70,131.71,130.96,129.13,128.01,126.97,126.51,126.07, 125.12,124.15,123.68,121.72,120.09,119.69,114.47,111.48,101.03,88.09,85.08,53.71,52.67,52.16,50.94,47.53,47.08,46.94,46.61,45.08,31.66,22.88,14.86.

[0269] Experimental characterization

[0270] 1. Western blotting method for detecting the degradation activity of the compound on Aurora-A protein.

[0271] Method: 1.5×10 6 Jurkat cells were seeded into 6-well plates and incubated overnight. Different concentrations of the compound (0, 30, 300, 3000 nM) were then added and incubated for another 24 hours. Cells were then collected by centrifugation, washed twice with cold phosphate-buffered saline (PBS), and centrifuged again to obtain the cell pellet. The pellet was then purified using RIPA (Thermo Scientific) containing 2.5% protease and phosphatase inhibitor (Beyotime, P1046). TMProteins were extracted using lysis buffer (#89900). Protein concentrations were determined using a BCA protein assay kit (Beyotime, P0009) and a Synergy H1 multi-functional microplate reader (BioTek).

[0272] Protein samples were separated by SDS-polyacrylamide gel electrophoresis and then electrotransferred to PDVF membranes. The membranes were blocked for 1 hour at room temperature in TBST (0.1% Tween in Tris buffer) with 5% skim milk. After blocking, the membranes were incubated overnight at 4°C with the appropriate primary antibody. After incubation, the membranes were washed with TBST (10 min × 3), and then incubated with a suitable secondary antibody for 1–4 hours. The membranes were washed again with TBST 3–5 times, and finally developed using the Super Signal Sidurah kit (Thermo Scientific) and imaged using an Amersham Imager 600 system (GE Healthcare). Grayscale analysis and calculation of the degradation rate of Aurka-A were performed using software.

[0273] Results: As shown in Table 1, the compounds of the present invention exhibited excellent protein degradation ability in the Aurora A protein degradation experiment. Among them, the AP-3 compound achieved a degradation rate of 80% at a concentration of 300 nM, which is much higher than that of the commercially available Aurora A degrader JB170.

[0274] Table 1

[0275]

[0276]

[0277] 2. CCK-8 assay to detect the inhibitory activity of the compound on the proliferation of Jurkat leukemia cells.

[0278] Methods: 15,000 Jurkat cells were seeded in 96-well plates and cultured overnight. Different concentrations of the compound (0 μM–10 μM) were added to each well, while maintaining a final DMSO concentration of 0.1% in each well. After co-incubation for 72 hours, 10 μL of CCK-8 reagent (CCK-8, DOJINDO, #CK04) was added to each well and incubated for 1–4 hours. The absorbance at 450 nm and 650 nm was then measured using a microplate reader (Synergy HI, BioTek Instruments, Inc., Vermont, US). Cell viability was calculated as VR = (A-A0) / (A...). s -A0)×100%, where A is the absorbance of the experimental group, A sA1 represents the absorbance of the control group (using DMSO as a control), and A0 represents the absorbance of the blank group (cell-free group). Finally, the IC50 was calculated using GraphPad Prism 8.0 software. 50 value.

[0279] Results: As shown in Table 2 below, AP-2 / 3 / 4 / 6 / 9 showed good cell proliferation inhibition activity, among which AP3 had the best activity.

[0280] Table 2. Inhibitory activity of compounds against Jurkat leukemia cell proliferation.

[0281]

[0282] Experimental results show that the compound of the present invention has an inhibitory effect on human T-cell lymphoblastic leukemia cells JURKAT.

[0283] 3. CCK-8 assay to detect the inhibitory activity of compound AP-3 on the proliferation of different tumor cells.

[0284] Methods: The cell lines used in this experiment, namely AGS (human gastric adenocarcinoma cells), KATOⅢ (human gastric cancer cells), Mia PACA-2 (human pancreatic cancer cells), MDA-MB-453 (human breast cancer cells), MDA-MB-231 (human breast cancer cells), SKBR3 (human breast cancer cells), SKOV3 (human ovarian cancer cells), A549 (human non-small cell lung cancer cells), NCI-H446 (human small cell lung cancer cells), HeLa (human cervical cancer cells), SK-N-SH (human neuroblastoma cells), IMR-32 (human neuroblastoma cells), CCRF-CEM (human acute lymphoblastic leukemia cells), MV4-11 (human acute myeloid leukemia cells), K562 (human chronic myeloid leukemia cells), MOLT4 (human acute lymphoblastic leukemia cells), and JURKAT (human acute T-cell leukemia cells), were obtained from ATCC and the Shanghai Cell Bank, respectively. 3000-20000 cells / well were seeded into 96-well plates and incubated overnight. Then, different concentrations of the compound (0-10 μM) were added for continuous treatment for 72 hours. CCK8 reagent was then added, and incubation continued for 1-3 hours. The absorbance at 450 nm and 650 nm was then measured using a microplate reader. The half-maximal inhibitory concentration (IC50) was calculated using GrapPad Prism 8.0 software. 50 ).

[0285] Results: As shown in Table 3 below, AP-3 showed good inhibitory activity against the proliferation of HeLa, NCI-H446, Molt4, CCRF-CEM, and Jurkat cells, with the best activity against Jurkat cells.

[0286] Table 3. Inhibitory activity of compound AP-3 against different tumor cell lines.

[0287] <![CDATA[IC 50 (μM)]]> AI AP-3 Alisertib AGS >10 >10 - Kato III >10 >10 - Mia Paca2 >10 >10 - MDA-MB-453 >10 >10 - MDA-MB-231 >10 >10 - SKBR3 >10 >10 - SKOV3 >10 >10 3.565±0.896 A549 >10 >10 - Hela 9.936±0.80 6.025±2.28 - SK-N-SH >10 >10 9.528 NCI-H446 >10 9.253±0.29 0.129±0.02 K562 >10 >10 - MV4-11 >10 >10 0.194±0.11 Molt4 >10 6.391±1.607 2.467±0.258 CCRF-CEM >10 3.329±0.887 0.147±0.037 Jurkat 9.562±0.428 0.918±0.075 0.040±0.011 IMR-32 3.156±1.953 1.953±1.368 0.022±0.002

[0288] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A degrading agent targeting Aurora A, characterized in that, The Aurora A-targeting degrader is a compound with the general structural formula shown in formula (Ⅰ): Where Ar is The linker is a connecting unit, and the linker can be either an aliphatic chain or an aromatic chain. Ligand is an E3 ubiquitin ligand.

2. The Aurora A-targeting degrader according to claim 1, characterized in that, The linker can be any of the following: Where n is 1-6.

3. The Aurora A-targeting degrader according to claim 1, characterized in that, The E3 ubiquitin ligand is either a CRBN ligand or a VHL ligand. Preferably, the CRBN ligand is selected from groups with the structure shown in formula C1 or C2: Wherein, X1 is selected from C=O or CH2, X2 is a single bond or selected from O or NH; R1 is selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 haloalkyl or cyano; Preferably, the structure of the VHL ligand is shown in Formula V: R2 and R5 are each independently hydrogen or C1-C4 alkyl; R3, R4, R6, and R7 are each independently hydrogen, deuterated, halogenated, nitro, cyano, amino, hydroxyl, or any one of C1-C4 alkyl.

4. The Aurora A-targeting degrading agent according to any one of claims 1-3, wherein the Aurora A-targeting degrading agent is one of the following compounds or an acceptable salt, enantiomer, hydrate, or prodrug:

5. The method for preparing the Aurora A-targeting degrader according to claims 1-4, characterized in that, Includes the following steps: S1, methyl 5-bromo-2-methyl-3-nitrobenzene reacts with vinyl magnesium bromide to generate intermediate a S2, intermediate a reacts with pinacol diboronate to generate intermediate b. S3, intermediate b and compound c The reaction produces intermediate d S4. The intermediate d is hydrolyzed to generate intermediate e. S5, the intermediate e and compound f intermediate g is formed by amide condensation. S6. The intermediate g is deprotected to generate intermediate h. S7, intermediate h and The amide condensation produces a degrading agent targeting Aurora A as shown in formula (I).

6. A composition, characterized in that, It includes an active molecule and a pharmaceutically acceptable excipient, wherein the active molecule is a degrading agent targeting Aurora A as described in any one of claims 1-4.

7. The composition according to claim 6, characterized in that, The composition is an injection, an oral preparation, or a mucosal delivery preparation.

8. The use of the Aurora A-targeting degrader according to any one of claims 1-4, or the composition according to any one of claims 6-7, in the preparation of an Aurora A protein kinase inhibitor.

9. The use of the Aurora A-targeting degrader according to any one of claims 1-4, or the composition according to any one of claims 6-7, in the preparation of an antitumor drug.

10. The application according to claim 9, characterized in that, The tumors are selected from breast cancer, leukemia, lung cancer, liver cancer, esophageal cancer, pancreatic cancer, colorectal cancer, stomach cancer, cervical cancer, brain cancer, and nasopharyngeal carcinoma.

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