Aurora A protein degradation agent based on HSP90 as well as preparation method and application of Aurora A protein degradation agent

Abstract

The invention discloses an HSP90-based AuroraA protein degradation agent as well as a preparation method and application thereof, the structural formula of the AuroraA protein degradation agent is # imgabs0 #. According to the scheme, 32 protein degradation agents based on a PROTAC technology are obtained by using an HSP90 recruitment ligand and an AuroraA inhibitor MLN-8237 through an own Linker design technology; the effectiveness of the AuroraA protein degradation agent on inhibition of tumor cell proliferation is proved through a cell common sieve and a CCK-8 experiment; a Western blot experiment proves that the degradation effect of the Aurora A protein degradation agent is achieved; a co-immunoprecipitation experiment proves that the protein degradation agent can be used for inducing the formation of an HSP90 / E3-PROTAC-AuroraA ternary complex; a mouse xenotransplantation model experiment proves that the AuroraA protein degradation agent has an anti-tumor effect in vivo; therefore, the compound has a prospect of being developed into a medicine for treating tumors and a medicine for treating or preventing proliferative diseases, and belongs to the technical field of targeted medicines.

Description

Technical Field

[0001] The present invention relates to an Aurora A protein degrader, specifically, an HSP90-based Aurora A protein degrader. The present invention also relates to an HSP90-based Aurora A protein degrader, a preparation method, and an application thereof, belonging to the technical field of targeted drugs. Background Art

[0002] Aurora kinases are a class of serine / threonine kinases, encompassing three isoforms: Aurora A, B, and C. Aurora kinases are essential for mitosis, participating in processes such as centrosome maturation and separation, spindle assembly and stability, chromosome condensation, and midplate polymerization. Aurora A kinase is highly expressed in breast, colon, pancreatic, bladder, ovarian, and prostate cancers, and is associated with poor prognosis. Designing targeted anti-tumor drugs targeting Aurora A kinase activity holds significant clinical value.

[0003] Aurora A can bind to c-MYC and N-MYC oncoproteins, thereby protecting them from proteasomal degradation in a non-catalytic manner (Otto T et al. Cancer Cell, 2009, 15(1):67; Dauch D et al. Nat Med, 2016, 22(7):744). The Aurora A inhibitor MLN-8237 can slightly reduce N-MYC protein levels in the treatment of N-MYC-expressing neuroblastoma, but this effect is relatively limited. Clinically, MLN-8237 has poor efficacy in patients with N-MYC-amplified neuroblastoma (Gustafson W C et al. Cancer Cell, 2014, 26(3):414).

[0004] In addition, Aurora A can stabilize spindle microtubules and induce a stem-like phenotype in breast cancer cells, promoting cell resistance (Toya M et al. Nat Cell Biol, 2011, 13(6):708; Zheng F et al. Nat Commun, 2016, 7(1):10180), indicating that Aurora A kinase inhibitors may not completely eliminate the kinase-independent oncogenic function. Therefore, it is necessary to develop strategies that can effectively disrupt both the kinase-dependent and -independent effects of Aurora A.

[0005] To overcome this problem, using PROTACs (Proteolysis targeting chimeras) to degrade AuroraA is a feasible method. PROTACs are bifunctional molecules that contain three parts: a ligand for the target protein (POI), a linker, and an E3 ubiquitin ligase that recruits the ligand. PROTACs induce POI to approach the E3 ligase to form a ternary complex of POI-PROTAC-E3 ligase, causing POI to be ubiquitinated and degraded by the proteasome. However, PROTACs may cause POI to be promiscuously degraded in many tissues and organs, resulting in adverse side effects (Bondeson DP et al. Nat Chem Biol, 2015, 11(8):611). Therefore, it is very important to enhance the targeting ability of PROTACs.

[0006] Heat shock protein 90 (HSP90), which is highly expressed in tumors, is a member of the HSP protein family with a molecular weight of 90 kDa. HSP90 is overactivated and abnormally expressed in tumor cells induced by some oncogenes and their products, and is involved in tumor growth, invasion, metastasis, etc. Approximately 30% of E3 ligases in the human body bind to HSP90 (Taipale M et al. Cell, 2012, 150(5):987). This provides a theoretical basis for the strategy of targeting Aurora A kinase degradation mediated by HSP90. At the same time, HSP90 stabilizes Aurora A in tumor cells, maintains high expression of Aurora A, helps tumor cells adapt to adverse environments, and enhances their resistance to treatment. Studies have shown that the HSP90 inhibitor 17-AAG effectively reduces Aurora A levels in HEL and HCT-116 in a dose-dependent manner (Wang F et al. Hematol Oncol, 2023, 41(3):474; Park JH et al. J Mol Med, 2008, 86:117).

[0007] In recent years, researchers have proposed a combined therapy approach that simultaneously inhibits Aurora A and HSP90 to treat breast cancer and acute megakaryocytic leukemia (Fiskus W et al. Breast Cancer Res Tr, 2012, 135:433; Wang Fet al. Hematol Oncol, 2023, 41(3):474). However, combined inhibition of Aurora A and HSP90 can only inhibit protein function, rather than eliminate the protein itself, which can easily lead to drug resistance, especially target mutations or pathway activation. In contrast, PROTACs can mark and completely degrade the target protein, making it less likely to produce residual function or restore activity.

[0008] Chinese patent CN 112062768 A discloses a small molecule with Aurora kinase degradation activity, its preparation method and application. This scheme selects three E3 ubiquitin ligase recruiting ligands, CRBN, cIAP and VHL, and the Aurora A inhibitor MLN-8237, and obtains 12 Aurora-A protein degraders through linker connection. However, the activity of the degraders is still not ideal, and the types of E3 ubiquitin ligase ligands tried are also limited. Summary of the Invention

[0009] In view of the above shortcomings, the first object of the present invention is to provide an Aurora A protein degrader based on HSP90;

[0010] In order to facilitate the preparation of the Aurora A protein degrading agent, the second object of the present invention is to provide a method for preparing the Aurora A protein degrading agent;

[0011] The third object of the present invention is to provide uses of the Aurora A protein degrading agent;

[0012] The fourth object of the present invention is to provide a drug containing the Aurora A protein degrader as an active ingredient.

[0013] To this end, the first technical solution provided by the present invention is as follows:

[0014] Based on existing PROTAC design concepts and the challenges that need to be overcome, this invention utilizes HSP90 recruitment ligands and the known Aurora A inhibitor MLN-8237, and through proprietary linker design technology, 32 PROTAC-based Aurora A protein degraders were obtained. Through CCK-8 and Western blot screening, multiple effective Aurora A protein degraders were ultimately obtained, and this series of compounds was named XH, with the structural formula described in formula (A):

[0015]

[0016] Specifically, the linkers corresponding to compounds XH1-XH32 are as follows:

[0017]

[0018]

[0019] The second technical solution provided by the present invention is the preparation method of the HSP90-based Aurora A protein degrader described in the first technical solution, wherein the target product is prepared by reacting an Aurora A binding ligand, an HSP90 recruiting ligand and a linker compound connecting the two.

[0020] Furthermore, in the preparation method of the above-mentioned HSP90-based Aurora A protein degrader, the Aurora A binding ligand is MLN-8237; the HSP90 recruitment ligand structural formula is HSP90 Ligand-1 or HSP90 Ligand-2, and the structural formulas of HSP90 Ligand-1 and HSP90 Ligand-2 are as follows:

[0021]

[0022] Furthermore, in the preparation method of the above-mentioned HSP90-based Aurora A protein degrader, the linker is one of the compounds having the general formula 2, 4, or 6:

[0023]

[0024] In general formula 2: R 1 C 2-6 Alkyl, -(CH2CH2O) n CH2CH2-, and n is an integer from 1 to 3;

[0025] In general formula 4: R 2 C 1-5 Alkyl, -(CH2CH2O) n CH2CH2-, and n is an integer from 1 to 3;

[0026] In general formula 4: R 3 -CH2-, -(CH2CH2O) m CH2CH2-, and m is an integer from 1 to 2;

[0027] In general formula 6: R 4 C 4-6 Alkyl, -(CH2CH2O) nCH2CH2-, and n is an integer from 1 to 3;

[0028] In general formula 6: R 5 C 2-8 Alkyl, -(CH2CH2O) m CH2CH2-, and m is an integer of 1-2.

[0029] Furthermore, the preparation method of the above-mentioned HSP90-based Aurora A protein degrader comprises the following steps in sequence:

[0030] 1)MLN-8237 and NH2R 1 Boc or NHR 2 C(=O)OC(CH3)3 or NH2R 4 Boc undergoes amide condensation at room temperature to obtain amide condensation product A;

[0031] 2) At room temperature, the amide condensation product A prepared in step 1) is deprotected by trifluoroacetic acid and then reacted with propiolic acid or NH2R 3 C≡C or HOC(=O)R 5 C≡C amide condensation gives amide condensation product B;

[0032] 3) The amide condensation product B prepared in step 2) reacts with an HSP90 recruitment ligand to obtain the target product, an HSP90-based Aurora A protein degrader.

[0033] Furthermore, the preparation method of the above-mentioned HSP90-based Aurora A protein degrader has a synthesis route of the following synthesis route a, synthesis route b, or synthesis route c, which are specifically as follows:

[0034] Synthesis route a:

[0035]

[0036] Synthesis route b:

[0037]

[0038] Synthesis route c:

[0039]

[0040] The R 1 、R 2 、R 3 、R 4 、R 5 As defined in the second technical solution.

[0041] Another technical solution of the present invention is the use of the HSP90-based Aurora A protein degrader described in the first technical solution in the preparation of a drug for treating tumors; or the use of the HSP90-based Aurora A protein degrader in the preparation of a drug for treating or preventing proliferative diseases.

[0042] Preferably, the proliferative disease includes but is not limited to at least one selected from tumors, rheumatic diseases, chronic inflammation, and infectious mononucleosis.

[0043] Preferably, the proliferative disease includes but is not limited to at least one selected from the group consisting of gastric cancer, colorectal cancer, lung cancer (such as lung adenocarcinoma), breast cancer, liver cancer, prostate cancer, thyroid cancer, pancreatic cancer, bladder cancer, kidney cancer, brain tumor, cervical cancer, CNS (central nervous system) cancer, malignant glioma, myeloproliferative disease, atherosclerosis, leukemia, pulmonary fibrosis, lymphoma, rheumatic disease, chronic inflammation, non-lymphoreticular tumors, cryoglobulinemia, papular mucinosis, familial splenic anemia, multiple myeloma, amyloidosis, solitary plasmacytoma, heavy chain disease, light chain disease, malignant lymphoma, chronic lymphocytic leukemia, monocytic leukemia, semi-molecular disease, primary macroglobulinemia, primary macroglobulinemia Purpura aureus, secondary benign monoclonal gammopathy, osteolytic lesions, acute lymphoblastic leukemia, lymphoblastoma, some non-Hodgkin lymphomas, Sézary syndrome, infectious mononucleosis, acute histiocytosis, hairy cell leukemia, Hodgkin lymphoma, colon cancer, rectal cancer, intestinal polyps, diverticulitis, colitis, pancreatitis, hepatitis, small cell lung cancer, neuroblastoma, neuroendocrine cell tumor, islet cell tumor, medullary thyroid carcinoma, melanoma, uterine cancer, chronic hepatitis, cirrhosis, ovarian cancer, retinoblastoma, cholecystitis, head and neck squamous cell carcinoma, digestive tract malignancies, non-small cell lung cancer, cervical cancer, testicular cancer, bladder cancer, myeloma, or bone malignancies (such as osteosarcoma).

[0044] Another technical solution of the present invention is to provide a drug for treating tumors, comprising an active ingredient and a pharmaceutically acceptable carrier;

[0045] The active ingredient is: the HSP90-based Aurora A protein degrader described in the first technical solution, or a pharmaceutically acceptable salt of the HSP90-based Aurora A protein degrader described in the first technical solution; or a hydrate of the HSP90-based Aurora A protein degrader described in the first technical solution; or a solvate of the HSP90-based Aurora A protein degrader described in the first technical solution; or a polymorph of the HSP90-based Aurora A protein degrader described in the first technical solution, or a tautomer of the HSP90-based Aurora A protein degrader described in the first technical solution, or a prodrug of the HSP90-based Aurora A protein degrader described in the first technical solution; the carrier is at least one of a diluent, an excipient, and a filler.

[0046] Another technical solution of the present invention is to provide a drug for treating or preventing proliferative diseases, comprising an active ingredient and a pharmaceutically acceptable carrier;

[0047] The active ingredient is: the HSP90-based Aurora A protein degrader described in the first technical solution, or a pharmaceutically acceptable salt of the HSP90-based Aurora A protein degrader described in the first technical solution; or a hydrate of the HSP90-based Aurora A protein degrader described in the first technical solution; or a solvate of the HSP90-based Aurora A protein degrader described in the first technical solution; or a polymorph of the HSP90-based Aurora A protein degrader described in the first technical solution, or a tautomer of the HSP90-based Aurora A protein degrader described in the first technical solution, or a prodrug of the HSP90-based Aurora A protein degrader described in the first technical solution; and the carrier is at least one of a diluent, an excipient, and a filler.

[0048] In many cases, the compounds of the present invention are capable of forming acid and / or base salts by virtue of the presence of amino and / or carboxyl groups, acid groups or groups similar thereto.

[0049] The compounds and pharmaceutically acceptable salts of the present invention also include solvates or hydrates. Generally speaking, solvates or hydrates are equivalent to unsolvated or unhydrated forms and are encompassed within the scope of the present invention. Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms have equivalent uses and are encompassed within the scope of the present invention.

[0050] The compounds of the present invention also include tautomeric forms, which result from the exchange of a single bond with an adjacent double bond accompanied by the migration of a proton.

[0051] The term "pharmaceutically acceptable carrier" as used herein includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. Unless any conventional media or agent is incompatible with the active ingredient, it shall not be used.

[0052] The composition is preferably formulated into a unit dosage form. This includes pre-filled, pre-measured ampoules or syringes of liquid compositions, or pills, tablets, capsules, etc. in the case of solid compositions. In such compositions, the compound is typically a relatively small component (about 0.1 to about 50 weight %, or preferably about 1 to about 40 weight %), with the remainder being various carriers or excipients and processing aids useful for forming the desired dosage form. An effective amount of the compound provided herein is administered.

[0053] In order to prepare solid compositions such as tablets, the main active ingredient is mixed with a pharmaceutical excipient (or carrier) to form a solid preformulation composition, which contains a uniform mixture of the compound of the present invention. When these preformulation compositions are referred to as uniform, it is meant that the active ingredient is evenly dispersed throughout the composition so that the composition can be easily subdivided into equally effective unit dosage forms such as tablets, pills and capsules.

[0054] Tablet of the present invention or pill can be coated or otherwise be compounded to provide a dosage form with prolongation advantage, or protect tablet or pill from the effect of acidic conditions in the stomach.For example, tablet or pill can comprise inner dosage and outer dosage component, and the latter has the form of the outer skin on the former.Can separate two kinds of compositions with enteric layer, wherein enteric layer is used for stopping the disintegration in the stomach and allows the complete entry of composition in the duodenum or delayed release.Various materials can be used for such enteric layer or coating, and above-mentioned material comprises the mixture of many macromolecular acids and macromolecular acids and such material as shellac, cetyl alcohol and cellulose acetate.

[0055] Injectable compositions are typically based on injectable sterile saline or phosphate buffered saline, or other injectable excipients known in the art. In such compositions, the active compound is typically a minor component, often about 0.05 to 10% by weight, with the remainder being injectable excipients and the like.

[0056] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharmaceutical excipients as described above. Preferably, these compositions are administered by oral or nasal respiratory route to achieve a local or systemic effect. Compositions in preferred pharmaceutically acceptable solvents may be atomized using an inert gas. The atomized solution may be inhaled directly from the atomizing device, or the atomizing device may be connected to a face mask or intermittent positive pressure breathing machine. The solution, suspension, or powder composition may be administered by a device that delivers the dosage form in an appropriate manner, preferably by oral or nasal route.

[0057] The compounds of the invention can also be administered in sustained release form or from a sustained release delivery system. Descriptions of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.

[0058] The present invention also includes prodrugs of the compounds. A prodrug is a pharmacological substance (drug) derived from a parent drug. Once in the body, the prodrug is metabolized and converted into the parent drug. Prodrugs can be prepared by substituting one or more functional groups of the parent drug, where the substituent groups are degraded in the body to release the parent compound. The preparation and use of prodrugs can be found in Higuchi T, Stella V. Pro-drugs as Novel Delivery Systems [M]. American Chemical Society, 1975 and Roche E B. Bioreversible Carriers in Drug Design: theory and application [M]. American Pharmaceutical Association and Pergamon Press, 1987.

[0059] Compared with the prior art, the present invention has the following beneficial effects:

[0060] The compound molecules provided by the present invention can target the active pocket of Aurora A, degrading Aurora A and inhibiting HSP90 function by recruiting the HSP90 protein. Cell screening and CCK-8 assays demonstrate the effectiveness of this class of Aurora A protein degraders in inhibiting tumor cell proliferation. Western blot experiments demonstrate the degradation effect of this class of Aurora A protein degraders. Immunoprecipitation experiments demonstrate that the protein degraders can induce the formation of the HSP90 / E3-PROTAC-Aurora A ternary complex. Mouse xenograft model experiments demonstrate the anti-tumor effect of this class of Aurora A protein degraders in vivo. These compounds have the potential to be developed into Aurora A protein degraders and subsequently into drugs for treating diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] Figure 1 Compound XH-3 provided in Example 3 1 H NMR spectrum;

[0062] Figure 2 Compound XH-3 provided in Example 3 13 C NMR spectrum;

[0063] Figure 3 Compound XH-5 provided in Example 5 1 H NMR spectrum;

[0064] Figure 4 Compound XH-5 provided in Example 5 13 C NMR spectrum;

[0065] Figure 5 Compound XH-11 provided in Example 11 1 H NMR spectrum;

[0066] Figure 6 Compound XH-11 provided in Example 11 13 C NMR spectrum;

[0067] Figure 7 Compound XH-13 provided in Example 13 1 H NMR spectrum;

[0068] Figure 8 Compound XH-13 provided in Example 13 13 C NMR spectrum;

[0069] Figure 9 Compound XH-14 provided in Example 14 1 H NMR spectrum;

[0070] Figure 10 Compound XH-14 provided in Example 14 13 C NMR spectrum;

[0071] Figure 11 Aurora A protein levels in different cells treated with compound XH-14 provided in Example 14;

[0072] (A) Endogenous Aurora A protein levels in MCF7 cells after 6 h; (B) Endogenous Aurora A protein levels in A549 cells after 6 h;

[0073] (C) Correlation between the concentration of Aurora A protein after 6 h of XH-14 treatment of cells;

[0074] Figure 12 The half degradation concentration DC of the XH series compounds provided in Examples 1-32 after treating A549 cells for 6 hours 50 value;

[0075] Figure 13 The maximum degradation capacity of the XH series compounds provided in Examples 1-32 on Aurora A protein in A549 cells is D max value;

[0076] Figure 14 This is a diagram verifying that the compound XH-14 provided in Example 14 induces the formation of an HSP90 / E3-XH-14-AuroraA ternary complex in cells;

[0077] (A) Verification of compound XH-14 inducing the formation of HSP90 / E3-XH-14-Aurora A ternary complex in MCF7 cells;

[0078] (B) Verification of compound XH-14 inducing the formation of HSP90 / E3-XH-14-Aurora A ternary complex in A549 cells;

[0079] Figure 15 This is the experimental flow chart for evaluating the in vivo antitumor activity of XH series compounds against non-resistant / resistant non-small cell lung cancer xenograft tumor nude mouse models;

[0080] Figure 16 The results of nude mouse tumor tissue quality after treatment with solvent or XH series compounds;

[0081] Figure 17 This is the in vivo tumor inhibition rate result diagram of XH series compounds on non-resistant / resistant cell tumors. DETAILED DESCRIPTION

[0082] The technical solutions in the embodiments of the present invention will be described in detail below in conjunction with the embodiments of the present invention and the accompanying drawings. However, the following embodiments and drawings are only used to understand the present invention and cannot limit the present invention. The present invention can be implemented in a variety of different ways as defined and covered by the claims.

[0083] 1. General Methods for Preparing the Compounds of the Invention

[0084] The raw materials used in the synthesis include:

[0085] Aurora A inhibitor (MLN-8237, chemical formula C 27 H 20 ClFN4O4, Chinese name: Alecetin, CAS No. 1028486-01-2);

[0086] HSP90 Ligand-1 Chemical Formula C 15 H 15 ClIN5O, Chinese name: 4-chloro-5-iodo-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine).

[0087] HSP90 Ligand-2 Chemical Formula C 19 H 20 ClN8O, Chinese name: 5-(4-azidobut-1-yn-1-yl)-4-chloro-7-((4-methoxy-3,5-dimethylpyridin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-2-amine).

[0088] The structural formula is as follows:

[0089]

[0090] The synthetic routes of HSP90 Ligand-1 and HSP90 Ligand-2 are as follows:

[0091]

[0092] in:

[0093] Synthesis of Compound A2: 2-Amino-4-chloropyrrolo[2,3-d]pyrimidine (5.00 g, 29.66 mmol, A1) and 40 mL of pyridine were added to a flask, followed by pivaloyl chloride (10.9 mL, 88.89 mmol). The mixture was stirred at room temperature for 7 h. After TLC monitoring, the reaction was complete. The mixture was then dried by spin drying. 40 mL of methanol and 25% aqueous ammonia (6.7 mL) were added, and the mixture was stirred at room temperature for 30 min. Filtered to obtain Compound A2 (6.73 g, 26.71 mmol) as a yellow solid in a 90% yield.

[0094] Synthesis of Compound A3: Compound A2 (6.73 g, 26.71 mmol), N-iodosuccinimide (7.21 g, 32.05 mmol), and 30 mL of dry tetrahydrofuran were added to a flask. The mixture was purged with nitrogen three times and stirred at room temperature for 6 h. TLC monitoring was performed. After the reaction was complete, the mixture was spin-dried to dryness. The solid was dissolved in dichloromethane and washed sequentially with saturated sodium thiosulfate solution (30 mL × 3) and saturated brine (30 mL × 3). The organic phase was collected to obtain a brown solid, Compound A3 (10.01 g, 26.44 mmol), in a yield of 99%.

[0095] Synthesis of Compound A5: Compound A3 (10.01 g, 26.44 mmol), 2-chloromethyl-3,5-dimethyl-4-methoxypyridine hydrochloride (6.46 g, 29.08 mmol, A4), potassium carbonate (10.96 g, 79.32 mmol), and 30 mL of DMF were added to a flask and stirred at 60°C for 6 h. After completion of the reaction, the mixture was diluted with ethyl acetate and washed with saturated brine (30 mL x 5). The organic phase was collected, dried over anhydrous sodium sulfate, and spin-dried. Purification was performed by silica gel column chromatography using a gradient elution ratio of dichloromethane / methanol (200:1 to 100:3). Compound A5 (12.00 g, 22.74 mmol) was obtained as a yellow solid in an 86% yield.

[0096] Synthesis of HSP90 Ligand-1: Compound A5 (12.00 g, 22.74 mmol), zinc chloride (15.50 g, 113.7 mmol), and 30 mL of a 20:1 ethanol / water mixture were added to a flask and heated under reflux at 80°C for 12 h. After completion of the reaction, the mixture was quenched with water and filtered to afford HSP90 Ligand-1 (7.66 g, 17.28 mmol) as a white solid in a 76% yield. 1 HNMR(400MHz,DMSO-d6)δ:8.07(s,1H),7.26(s,1H),6.73(s,2H),5.28(s,2H),3.73(s,3H),2.25(s,3H),2.16(s,3H).MS(ESI+APCI)m / z:444.1[M+H] + .

[0097] Synthesis of Compound A6: 3-Butyn-1-ol (2.00 g, 28.53 mmol) and 30 mL of dichloromethane were added to a flask under nitrogen. Triethylamine (8.66 g, 85.60 mmol) and 4-toluenesulfonyl chloride (6.52 g, 34.20 mmol) dissolved in dichloromethane were added sequentially at 0°C and stirred at room temperature for 12 h. After completion of the reaction, the mixture was spin-dried and purified by silica gel column chromatography using a gradient elution ratio of petroleum ether / ethyl acetate (20:1 to 20:3). Compound A6 (6.27 g, 27.96 mmol) was obtained as a clear oil in a 98% yield.

[0098] Synthesis of Compound A7: White solid HSP90 Ligand-1 (2.00 g, 4.51 mmol), Compound A6 (1.22 g, 5.44 mmol), cuprous iodide (85 mg, 0.45 mmol), bistriphenylphosphine palladium dichloride (247 mg, 0.35 mmol), and dry dichloromethane (30 mL) were added to a flask. The mixture was purged with nitrogen three times, and triethylamine (3.1 mL, 22.58 mmol) was added, followed by stirring at room temperature for 12 h. After completion of the reaction, the mixture was diluted with dichloromethane and washed with saturated brine (30 mL x 3). The organic phase was collected, dried over anhydrous sodium sulfate, and then spin-dried. Purification was performed by silica gel column chromatography using a gradient elution ratio of petroleum ether / ethyl acetate (1:1 to 1:3). Compound A7 (2.11 g, 3.91 mmol) was obtained as a yellow solid in an 87% yield.

[0099] Synthesis of HSP90 Ligand-2: Compound A7 (2.11 g, 3.91 mmol) and 25 mL of DMF were added to a flask. Sodium azide (763 mg, 11.72 mmol) was added with stirring and the mixture was heated at reflux for 4 h at 60°C. After completion of the reaction, the mixture was diluted with ethyl acetate and washed with saturated brine (30 mL x 5). The organic phase was collected, dried over anhydrous sodium sulfate, and spin-dried. Purification was performed by silica gel column chromatography using a gradient elution ratio of petroleum ether / ethyl acetate (1:1 to 1:3). HSP90 Ligand-2 (1.41 g, 3.44 mmol) was obtained as a yellow solid in an 88% yield. 1 HNMR(400MHz,DMSO-d6)δ:8.06(s,1H),7.28(s,1H),6.71(s,2H),5.76(s,2H),3.72(s,3H) ,3.50(t,J=6.8Hz,2H),2.72(t,J=6.8Hz,2H),2.25(s,3H),2.16(s,3H).MS(ESI+APCI)m / z 411.1[M+H] + .

[0100] Protein degrader synthesis scheme:

[0101] Synthesis route a: (Compounds XH-1 to XH-6)

[0102]

[0103] Synthesis route b: (Compounds XH-7 to XH-12, XH-22 to XH-23, XH-26 to XH-27, XH-30 to XH-31)

[0104]

[0105] Synthesis route c: (Compounds XH-13 to XH-21, XH-24 to XH-25, XH-28 to XH-29, XH-32)

[0106]

[0107] Compounds XH-1 to XH-32 were prepared by synthetic routes a, b, and c, wherein R 1 、R 2 、R 3 、R 4 、R 5 MLN-8237 and NH2R 1 NHBoc or NHR 2 C(=O)OC(CH3)3 or NH2R 4 NHBoc amide condensation at room temperature gave compound 1, 3 or 5. Then, trifluoroacetic acid was used for deprotection and propiolic acid or NH2R 3 C≡C or HOC(=O)R 5 C≡C amide condensation gave compounds 2, 4, or 6. Compound 2 reacted with HSP90 Ligand-2 via click reaction at 40°C to generate XH-1 to XH-6, and compound 4 or 6 reacted with HSP90 Ligand-1 via Sonogashira coupling at room temperature to give compounds XH-7 to XH-32.

[0108] Example 1 Synthesis of Aurora A protein degrader XH-1

[0109]

[0110] Synthesis of B1: MLN-8237 (100 mg, 0.19 mmol), tert-butyl (2-aminoethyl)carbamate (37 mg, 0.23 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 87 mg, 0.23 mmol), and N,N-diisopropylethylamine (DIPEA, 123 mg, 0.95 mmol) were dissolved in 6 mL of N,N-dimethylformamide (DMF) and stirred at room temperature for 12 h. After completion of the reaction, the mixture was diluted with ethyl acetate and washed with saturated brine (30 mL x 5). The organic phase was collected, dried over anhydrous sodium sulfate, and spin-dried. Purification was performed by silica gel column chromatography using a gradient elution ratio of dichloromethane / methanol (100:1 to 50:2). B1 (122 mg, 0.18 mmol) was obtained as a yellow solid in a 95% yield. 1 H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.21 (d, J = 8.4Hz, 1H), 8.16 (d, J = 8.6Hz ,1H),8.09(s,1H),7.95(s,1H),7.88(s,1H),7.54(dd,J=8.5,2.3Hz,1H),7. 33-7.30(m,2H),7.11(d,J=8.7Hz,1H),6.69(brs,2H),5.18(s,1H),4.85(s, 1H),3.98(s,3H),3.58(q,J=6.1Hz,2H),3.38(q,J=6.1Hz,2H),1.43(s,9H). 13 CNMR(101MHz, CDCl3)δ:166.0,161.9,161.6,159.3,159.2,158.5,158.2,156.7,156.4,144.2,138.0,136.0,135.2,132.9,130.9,13 0.3,130.1,128.6,123.9,118.3,114.7,110.9,108.5,107.1,101.2,79.3,56.1,55.8,50.4,40.8,40.0,28.4(3C).MS(ESI+APCI)m / z 661.2[M+H] + .

[0111] Synthesis of C1: Dissolve the above-prepared B1 (122 mg, 0.18 mmol) in 4 mL of dichloromethane, then add 2 mL of trifluoroacetic acid. Stir at room temperature for 2 h until the reaction is complete. The reaction product is dried by rotary evaporation, dissolved in dichloromethane, and the pH is adjusted to alkaline with saturated sodium bicarbonate solution. Stir at room temperature for 30 minutes, extract with dichloromethane (30 mL × 3), and wash with saturated brine (30 mL × 3). The organic phase is collected and dried by rotary evaporation to obtain a yellow solid, which is directly used in the next step without purification.

[0112] The entire yellow solid prepared in the previous step, propiolic acid (15 mg, 0.22 mmol), HATU (84 mg, 0.22 mmol), and DIPEA (116 mg, 0.90 mmol) were dissolved in DMF and stirred at room temperature for 12 h. The reaction was complete. The mixture was diluted with ethyl acetate and washed with saturated brine (30 mL x 5). The organic phase was collected, dried over anhydrous sodium sulfate, and spin-dried. Purification was performed by silica gel column chromatography using a gradient elution ratio of dichloromethane / methanol (25:1 to 40:3). C1 (64 mg, 0.10 mmol) was obtained as a yellow solid in a 58% yield. 1 H NMR (400MHz, CDCl3) δ: 8.52 (s, 1H), 8.21 (d, J = 8.4Hz, 1H), 8.17-8.14 (m, 2H), 7.98(d,J=2.2Hz,1H),7.87(s,1H),7.55(dd,J=8.5,2.1Hz,1H),7.44(t,J=5.1 Hz,1H),7.32-7.27(m,2H),7.11(dd,J=8.7,2.1Hz,1H),6.67(brs,2H),4.86( s,1H),3.99(s,3H),3.63(q,J=5.6Hz,2H),3.54(q,J=5.5Hz,2H),2.79(s,1H). 13 C NMR(101MHz,DMSO-d6)δ:165.2,161.4,160.7,159.9,158.8,158.4,158.3,157.5,152.5,145.1,138.1,135.6,135.1,132.0,131.6 ,131.3,130.7,127.9,123.8,118.5,118.3,114.9,110.9,108.4,101.8,79.6,78.7,76.3,56.7,56.1,50.0,29.5.MS(ESI+APCI)m / z 613.2[M+H] + .

[0113] Synthesis of XH-1: Dissolve C1 (64 mg, 0.10 mmol) and HSP90 Ligand-2 (49 mg, 0.12 mmol) prepared above in 5 mL of a mixed solvent of ethanol / dichloromethane (v:v = 1:1) and stir at 40°C for 15 minutes. Dissolve ascorbic acid (70 mg, 0.40 mmol) in 0.5 mL of water, add anhydrous copper sulfate (24 mg, 0.15 mmol), and the solution, which turns brown, is then added dropwise to the reaction system. The reaction is allowed to proceed at 40°C for 3 hours. TLC monitoring indicates that the reaction is complete. The reaction solution is dried by vortexing, dissolved in 2M ammonia (30 mL), extracted with dichloromethane (30 mL × 3), and washed with saturated brine (30 mL × 3). The organic phase is collected, dried by vortexing, and purified by silica gel column chromatography using a gradient elution of dichloromethane / methanol (20:1 to 10:1). A yellow solid XH-1 (46 mg, 0.05 mmol) was obtained with a yield of 45%. 1 HNMR(400MHz, CDCl3)δ:8.43(s,1H),8.25(s,1H),8.13-7.99(m,5H),7.69(t ,J=5.9Hz,1H),7.48-7.45(m,2H),7.24-7.22(m,2H),7.18-7.17(m,1H),7.01 (s,1H),6.63(brs,2H),5.22(s,2H),5.18(s,2H),4.53(t,J=6.5Hz,2H),3.7 1(s,3H),3.66-3.63(m,7H),2.97(t,J=6.4Hz,2H),2.13(s,3H),2.12(s,3H). 13 C NMR(101MHz, CDCl3)δ:165.8,164.7,161.9,161.6,160.6,159.4,159.2,156.0,158.4,158.2,15 6.8,153.1,152.7,149.1,144.1,143.1,137.9,135.9,135.2,132.8,130.9,130.8,130.5,130.3, 130.2,128.6,126.4,126.0,125.5,123.7,118.3,114.6,110.8,109.3,108.2,107.1,101.1,96. 2,86.4,76.1,60.0,56.2,55.7,50.3,49.4,47.0,39.8,39.0,21.7,13.4,11.0.MS(ESI+APCI)m / z 1023.3[M+H] + .

[0114] Example 2 Synthesis of Aurora A protein degrader XH-2

[0115] The synthesis conditions of XH-2, including the steps, reaction parameters, and solvents, were similar to those of XH-1 in Example 1, except that tert-butyl (4-aminobutyl)carbamate was used instead of tert-butyl (2-aminoethyl)carbamate. The total yield was 27%. 1 H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.27 (s, 1H), 8.21 (d, J = 8.4Hz, 1H), 8.18-8.14 (m, 2H), 7.92-7 .87(m,2H),7.75(d,J=2.0Hz,1H),7.56(dd,J=8.5,2.2Hz,1H),7.34-7.30(m,3H),7.19(dd,J=8.6, 1.9Hz,1H),7.07(s,1H),6.73(brs,2H),5.26(s,2H),5.22(s,2H),4.60(t,J=6.4Hz,2H),3.90(s, 3H),3.72(s,3H),3.52-3.49(m,4H),3.04(t,J=6.4Hz,2H),2.21(s,3H),2.19(s,3H),1.72(m,4H). 13 C NMR(101MHz, CDCl3)δ:165.2,164.3,161.9,161.6,160.1,159.4,159.2,158.9,158.3,158.2,156.7 ,153.1,152.9,149.5,143.9,143.4,138.0,135.9,135.2,132.9,130.9,130.8,130.4,130.3,130.1 ,128.6,126.2,125.8,125.2,123.8,118.3,115.1,110.9,109.4,108.5,107.1,101.2,96.2,86.3,7 6.1,60.0,56.2,55.8,50.4,49.4,47.3,39.2,38.9,27.3,27.1,21.7,13.3,10.9.MS(ESI+APCI)m / z 1051.3[M+H] + .

[0116] Example 3 Synthesis of Aurora A protein degrader XH-3

[0117] The synthesis steps, reaction parameters, solvents, and other conditions of XH-3 were similar to those of XH-1 in Example 1, except that (6-aminohexyl)carbamic acid tert-butyl ester was used instead of (2-aminoethyl)carbamic acid tert-butyl ester. The total yield was 34%. Figure 1 and Figure 2 , 1 H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.32 (s, 1H), 8.22-8.15 (m, 4H), 7.89 (d, J=1.9Hz, 1H), 7.84 (t, J= 5.6Hz,1H),7.55(dd,J=8.5,2.1Hz,1H),7.32-7.31(m,2H),7.29-7.25(m,1H),7.16(dd,J=8.6,1.8Hz ,1H),7.05(s,1H),6.71(brs,2H),5.34(s,2H),5.25(s,2H),4.60(t,J=6.8Hz,2H),3.95(s,3H),3.72 (s,3H),3.48-3.41(m,4H),3.03(t,J=6.6Hz,2H),2.21(s,3H),2.18(s,3H),1.62(m,4H),1.43(m,4H). 13 C NMR(101MHz, CDCl3)δ:165.2,164.3,161.8,161.6,160.1,159.4,159.1,159.0,158.3,158.1,156.7,1 53.1,153.1,152.9,149.5,144.0,143.4,137.9,135.9,135.2,132.8,130.9,130.8,130.3(2C),130.1 ,128.5,126.2,125.8,125.2,123.7,118.3,115.1,111.0,109.3,108.2,107.1,101.3,96.2,86.3,76. 1,60.0,56.2,55.9,50.4,49.3,47.2,39.5,39.0,29.6,26.7,26.6,21.7,13.3,10.9.MS(ESI+APCI)m / z 1079.4[M+H] + .

[0118] Example 4 Synthesis of Aurora A Protein Degrader XH-4

[0119] The synthesis conditions for XH-4, including the steps, reaction parameters, and solvents, were similar to those for XH-1 in Example 1, except that tert-butyl (2-aminoethyl)carbamate was replaced with [2-(2-aminoethoxy)ethyl]carbamate. The total yield was 33%. 1 H NMR (400MHz, CDCl3) δ: 8.52 (s, 1H), 8.32 (s, 2H), 8.21 (d, J = 7.6Hz, 3H), 8.14 (d, J = 8. 7Hz,1H),7.79(s,1H),7.62(s,1H),7.56(d,J=8.4Hz,1H),7.32(s,1H),7.28-7.21(m, 2H),7.04(s,1H),6.69(brs,2H),5.44(s,2H),5.24(s,2H),4.59(t,J=6.6Hz,2H),3. 92(s,3H),3.70(s,3H),3.68(m,8H),3.01(t,J=6.4Hz,2H),2.21(s,3H),2.19(s,3H). 13 C NMR (101MHz, CDCl3) δ: 165.3, 164.3, 161.8, 161.6, 160.2, 159.4, 159.1, 159.0, 158.5, 158.2, 156. 8,153.0,152.9,149.5,144.1,143.2,137.9,135.9,135.2,132.8,130.9,130.8,130.4(2C),130.1, 128.5,126.3,125.9,125.1,123.7,118.3,114.8,110.8,109.2,108.4,107.1,101.3,96.1,86.3,7 6.1,70.2,69.6,59.9,56.2,56.0,50.4,49.3,47.3,39.5,39.1,21.7,13.3,10.9.MS(ESI+APCI)m / z 1067.3[M+H] + .

[0120] Example 5 Synthesis of Aurora A protein degrader XH-5

[0121] The synthesis steps, reaction parameters, solvents, and other conditions of XH-5 were similar to those of XH-1 in Example 1, except that tert-butyl [2-[2-(2-aminoethoxy)ethoxy]ethyl]carbamate was used instead of tert-butyl (2-aminoethyl)carbamate. The total yield was 34%. Figure 3 and Figure 4 , 1H NMR (400MHz, CDCl3) δ: 10.18 (s, 1H), 8.68 (s, 1H), 8.66 (s, 1H), 8.42 (t, J = 5.8Hz, 1H), 8.29 (d, J = 8.5Hz, 1H), 8.19 (t, J=5.1Hz,1H),8.05(s,1H),7.99(s,1H),7.91(d,J=8.6Hz,1H),7.78(dd,J=8.5,2.2Hz,1H),7.45(dd,J=8.6,1.8Hz,1 H),7.42-7.35(m,J=8.4,6.8Hz,1H),7.25-7.24(m,2H),6.85(brs,2H),6.71(s,2H),5.25(s,2H),4.64(t,J=6.6Hz,2 H),3.95(s,3H),3.69(s,3H),3.59-3.55(m,8H),3.51-3.46(m,4H),3.11(t,J=6.7Hz,2H),2.23(s,3H),2.13(s,3H). 13 C NMR(101MHz,DMSO-d6)δ:164.8,163.8,161.3,160.7,160.3,160.0,159.9,158.8,158.4,158.3,158.2, 157.5,153.8,153.8,152.2,149.3,145.2,143.2,138.1,135.6,135.1,132.1,131.8,131.6,131.5,131 .3,130.6,127.9,126.9,125.5,123.9,118.5,114.8,111.0,108.4,107.9,101.9,95.1,87.7,75.9,70. 1,70.1,69.6,69.3,60.2,56.7,56.2,50.0,49.1,46.9,39.4,38.7,21.2,13.3,10.7.MS(ESI+APCI)m / z 1111.4[M+H] + .

[0122] Example 6 Synthesis of Aurora A Protein Degrader XH-6

[0123] The synthesis conditions for XH-6, including the steps, reaction parameters, and solvents, were similar to those for XH-1 in Example 1, except that tert-butyl [2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethyl]carbamate was used instead of tert-butyl (2-aminoethyl)carbamate. The total yield was 33%. 1H NMR(400MHz, CDCl3)δ:8.51(s,1H),8.43(s,1H),8.36(s,1H),8.24-8.14(m,4H),7.83(d,J =2.0Hz,1H),7.64(t,J=5.6Hz,1H),7.54(dd,J=8.5,2.2Hz,1H),7.31-7.24(m,3H),7.03(s, 1H),6.69(brs,2H),5.44(s,2H),5.24(s,2H),4.60(t,J=6.7Hz,2H),3.96(s,3H),3.71(s,3 H),3.69(m,4H),3.67(m,6H),3.64(m,6H),3.00(t,J=6.7Hz,2H),2.20(s,3H),2.17(s,3H). 13 C NMR (101MHz, CDCl3) δ: 165.3, 164.2, 161.7, 161.6, 160.2, 159.5, 159.1, 159.0, 158.4, 158.1, 156.7, 153. 2,153.0,152.9,149.4,144.2,143.2,137.9,135.8,135.2,132.7,130.9,130.4,130.3,130.1,128.5,126 .1,126.0,125.1,123.6,118.3,114.8,110.8,109.1,108.2,107.1,101.4,96.1,86.3,76.1,70.7,70.5,7 0.5,70.4,70.1,69.8,59.9,56.2,55.9,50.3,49.3,47.2,39.4,38.9,21.7,13.3,10.9.MS(ESI+APCI)m / z 1155.4[M+H] + .

[0124] Example 7 Synthesis of Aurora A Protein Degrader XH-7

[0125]

[0126] Synthesis of B2: MLN-8237 (100 mg, 0.19 mmol), glycine tert-butyl ester (30 mg, 0.23 mmol), HATU (87 mg, 0.23 mmol), and DIPEA (123 mg, 0.95 mmol) were dissolved in 6 mL of DMF and stirred at room temperature for 12 h. After the reaction was complete, the mixture was diluted with ethyl acetate and washed with saturated brine (30 mL x 5). The organic phase was collected, dried over anhydrous sodium sulfate, and spin-dried. Purification was performed by silica gel column chromatography using a gradient elution ratio of dichloromethane / methanol (100:1 to 50:2). B2 (101 mg, 0.16 mmol) was obtained as a yellow solid in an 84% yield. 1 H NMR (400MHz, CDCl3) δ: 8.52 (s, 1H), 8.39 (t, J = 4.8Hz, 1H), 8.22 (d, J = 8.4Hz, 1H), 8.16 (d, J = 8.6Hz, 1H), 7.96 (d, J = 2.0Hz, 1H), 7.82 (s, 1H), 7.54 (d d,J=8.8,2.4Hz,1H),7.33-7.28(m,2H),7.09(dd,J=8.4,2.0Hz,1H),6.7 0(brs,2H),4.86(s,1H),4.18(d,J=4.8Hz,2H),4.00(s,3H),1.51(s,9H). 13 C NMR(101MHz, CDCl3)δ:169.6,165.1,161.9,161.6,159.4,159.2,158.8,158.2,156.7,144.3,138.0,136.0,135.2,133.0,130.9, 130.3,130.1,128.6,123.9,118.3,114.4,110.8,108.5,107.1,101.2,82.0,56.1,55.9,50.4,42.7,28.1(3C).MS(ESI+APCI)m / z 632.2[M+H] + .

[0127] Synthesis of C2: Dissolve the above-prepared B2 (101 mg, 0.16 mmol) in 4 mL of dichloromethane, add 2 mL of trifluoroacetic acid, and stir at room temperature for 2 h. After the reaction is complete, dissolve it in dichloromethane and spin dry. Repeat this three times to obtain a yellow oil, which is directly used in the next step without purification.

[0128] The yellow oil from the previous step, propargylamine (11 mg, 0.19 mmol), HATU (72 mg, 0.19 mmol), and DIPEA (123 mg, 0.95 mmol) were dissolved in DMF and stirred at room temperature for 12 h. Once the reaction was complete, the mixture was diluted with ethyl acetate and washed with saturated brine (30 mL x 5). The organic phase was collected, dried over anhydrous sodium sulfate, and spin-dried. Purification was performed by silica gel column chromatography using a gradient elution ratio of dichloromethane / methanol (50:1 to 20:1). C2 (69 mg, 0.09 mmol) was obtained as a yellow solid in a 49% yield. 1 H NMR (400MHz, DMSO-d6) δ: 10.22 (s, 1H), 8.72 (s, 1H), 8.48 (t, J = 5.4Hz, 1H), 8.4 3(t,J=5.6Hz,1H),8.31(d,J=8.6Hz,1H),8.01(s,1H),7.90(d,J=8.7Hz,1H),7 .82(dd,J=8.4,2.2Hz,1H),7.46-7.39(m,2H),7.23(s,1H),6.90(brs,2H),4.8 7(s,1H),3.98-3.96(m,5H),3.93(dd,J=5.6,2.6Hz,2H),3.13(t,J=2.5Hz,1H). 13 C NMR(101MHz,DMSO-d6)δ:169.3,164.8,161.2,160.7,159.8,158.8,158.6,158.2,157.6,145.4,138.1,135.6,135.1,132.2,131.7 ,131.3,130.7,127.9,123.9,118.5,118.3,114.3,110.9,108.5,101.9,81.5,73.5,56.7,56.2,49.9,43.1,28.4.MS(ESI+APCI)m / z 611.2[MH] - .

[0129] Synthesis of XH-7: C2 (69 mg, 0.09 mmol), HSP90 Ligand-1 (44 mg, 0.10 mmol), CuI (2 mg, 0.01 mmol), and Pd(PPh3)2Cl2 (5 mg, 0.007 mmol) prepared above were dissolved in DCM. Triethylamine (45 mg, 0.45 mmol) was added and stirred at room temperature under nitrogen for 12 h. After the reaction was complete, the mixture was diluted with dichloromethane, washed with saturated brine (30 mL x 3), dried over anhydrous sodium sulfate, and spin-dried. Purification was performed by silica gel column chromatography using a gradient elution ratio of dichloromethane / methanol (25:1 to 40:3). XH-7 (48 mg, 0.05 mmol) was obtained as a yellow solid in a 58% yield. 1 H NMR(400MHz, DMSO-d6)δ:10.23(s,1H),8.73(s,1H),8.54-8.51(m,2H),8.32(d,J=8.5 Hz,1H),8.07(s,1H),8.03(s,1H),7.93(d,J=8.6Hz,1H),7.82(dd,J=8.5,2.2Hz,1H), 7.48-7.36(m,3H),7.25(s,1H),6.89(brs,2H),6.76(s,2H),5.31(s,2H),4.22(d,J=5 .4Hz,2H),4.03(d,J=5.2Hz,2H),3.99(s,3H),3.73(s,3H),2.26(s,3H),2.16(s,3H). 13 C NMR(101MHz,DMSO-d6)δ:169.2,164.8,163.8,161.2,160.7,160.1,159.8,158.8,158.6,1 58.2,157.6,153.9,153.8,152.2,149.3,145.4,138.1,135.6,135.1,132.3,132.2,131.7, 131.5,131.3,130.7,127.9,125.5,123.9,123.9,118.5,114.3,110.9,108.5,107.9,101. 8,94.8,88.6,75.5,60.3,56.7,56.2,50.0,46.9,43.2,29.5,13.3,10.7.MS(ESI+APCI)m / z 928.3[M+H] + .

[0130] Example 8 Synthesis of Aurora A Protein Degrader XH-8

[0131] The synthesis steps, reaction parameters, solvents, and other conditions of XH-8 were similar to those of XH-7 in Example 7, except that tert-butyl 4-aminobutyrate was used instead of tert-butyl glycine. The total yield was 20%. 1 HNMR (400MHz, CDCl3) δ: 8.49 (s, 1H), 8.24 (s, 1H), 8.19-8.18 (m, 2H), 8.11 (d, J = 8.6Hz, 1H), 8.00 (t ,J=5.9Hz,1H),7.93(s,1H),7.53(d,J=8.8Hz,1H),7.31(s,2H),7.27-7.25(m,1H),7.03(d,J=8.7Hz ,1H),6.98(s,1H),6.68(brs,2H),5.26(s,2H),5.24(s,2H),4.24(d,J=5.0Hz,2H),3.94(s,3H),3.7 1(s,3H),3.52(q,J=6.3Hz,2H),2.33(t,J=7.0Hz,2H),2.20(s,3H),2.14(s,3H),1.99-1.92(m,2H). 13 C NMR (101MHz, CDCl3) δ: 172.6, 165.9, 164.3, 161.8, 161.6, 159.4, 159.1, 158.9, 158.4, 158. 2,156.7,153.2,153.1,152.9,149.4,144.2,137.9,135.7,135.2,132.7,130.9,130.3(2C), 130.1,128.5,126.3,125.2,123.6,118.1,114.5,110.8,109.3,108.4,107.1,101.1,96.4,8 7.5,75.4,59.9,56.1,55.8,50.3,47.1,39.0,33.7,30.1,26.1,13.3,10.8.HRMS(ESI+):m / z calcd.for C 49 H 44 Cl2FN 11 O5[M+H] + :956.2922; found:956.2933.

[0132] Example 9 Synthesis of Aurora A Protein Degrader XH-9

[0133] The synthesis steps, reaction parameters, solvents, and other conditions of XH-9 were similar to those of XH-7 in Example 7, except that tert-butyl 6-aminohexanoate was used instead of tert-butyl glycine. The total yield was 27%.1 HNMR (400MHz, CDCl3) δ: 8.50 (s, 1H), 8.20 (s, 1H), 8.18 (s, 1H), 8.13 (d, J = 8.4Hz, 1H), 8.10-8.05 (m, 1H), 7. 91-7.87(m,2H),7.54(dd,J=8.4,2.3Hz,1H),7.32-7.29(m,2H),7.27-7.26(m,1H),7.04(d,J=8.8Hz,1H),7. 00(m,1H),6.66-6.52(m,3H),5.25(s,2H),5.18(s,2H),4.24(d,J=5.1Hz,2H),3.95(s,3H),3.72(s,3H),3.4 5(q,J=6.6Hz,2H),2.26-2.21(m,5H),2.16(s,3H),1.74-1.67(m,2H),1.64-1.58(m,2H),1.45-1.37(m,2H). 13 C NMR(101MHz, CDCl3)δ:172.8,165.4,164.4,161.8,161.6,159.4,159.1,158.9,158.3,158.1,15 6.7,153.1,152.9,149.5,144.0,137.9,135.9,135.2,132.7,130.9,130.3(2C),130.1,128.6,1 26.3,125.3,123.7,118.3,114.8,110.9,109.3,108.5,107.1,101.2,96.4,87.4,75.6,60.0,56 .2,55.9,50.3,47.2,39.4,36.2,30.1,29.4,26.6,25.2,13.3,10.7.HRMS(ESI+):m / zcalcd.for C 51 H 48 Cl2FN 11 O5[M+H] + :984.3235; found:984.3244.

[0134] Example 10 Synthesis of Aurora A protein degrader XH-10

[0135] The synthesis steps, reaction parameters, solvents, and other conditions of XH-10 were similar to those of XH-7 in Example 7, except that tert-butyl 3-(2-aminoethoxy)propionate was used instead of tert-butyl glycine. The total yield was 29%. 1H NMR (400MHz, CDCl3) δ: 8.49 (s, 1H), 8.19-8.16 (m, 4H), 8.10 (d, J = 8.4Hz, 1H), 7.81 (d, J = 2.2Hz, 1 H),7.53(dd,J=8.5,2.1Hz,1H),7.31(s,1H),7,27-7.26(m,1H),7.07(dd,J=8.7,1.9Hz,1H),6.95 -6.92(m,2H),6.71(brs,2H),5.21(m,4H),4.24(d,J=5.2Hz,2H),3.93(d,J=2.1Hz,3H),3.78(t,J =5.9Hz,2H),3.71(d,J=2.1Hz,3H),3.66(m,4H),2.53(t,J=5.8Hz,2H),2.20(s,3H),2.15(s,3H). 13 C NMR(101MHz, CDCl3)δ:170.9,165.5,164.3,161.8,161.6,159.4,159.2,158.9,158.4,158.2, 156.7,153.1,153.0,152.9,149.5,144.2,137.9,135.9,135.2,132.7,130.9,130.3(2C),130 .1,128.6,126.2,125.1,123.6,118.3,114.6,110.8,109.3,108.3,107.3,101.2,96.2,87.2, 75.6,70.9,66.9,60.0,56.2,55.9,50.4,47.1,39.4,36.9,30.1,13.3,10.8.HRMS(ESI+):m / z calcd.for C 50 H 46 Cl2FN 11 O6[M+H] + :986.3027; found:986.3039.

[0136] Example 11 Synthesis of Aurora A protein degrader XH-11

[0137] The synthesis steps, reaction parameters, solvents, and other conditions of XH-11 were similar to those of XH-7 in Example 7, except that tert-butyl 3-[2-(2-aminoethoxy)ethoxy]propionate was used instead of tert-butyl glycine. The total yield was 33%. Figure 5 and Figure 6 , 1H NMR (400MHz, CDCl3) δ: 8.50 (s, 1H), 8.21-8.17 (m, 4H), 8.11 (d, J = 8.6Hz, 1H), 7.81 (d, J = 2.0Hz, 1H), 7.5 5(dd,J=8.5,2.2Hz,1H),7.32(d,J=2.3Hz,1H),7.27-7.25(m,1H),7.11(dd,J=8.4,2.0Hz,1H),7.06(t, J=5.3Hz,1H),6.99(s,1H),6.68(brs,2H),5.24(s,2H),5.22(s,2H),4.24(d,J=5.2Hz,2H),3.92(s,3H) ,3.76(t,J=5.8Hz,2H),3.71(s,3H),3.67-3.65(m,8H),2.52(t,J=5.8Hz,2H),2.21(s,3H),2.15(s,3H). 13 C NMR(101MHz, CDCl3)δ:171.1,165.4,164.3,161.8,161.6,159.4,159.1,158.9,158.4,158.2,156 .7,153.1,153.0,152.9,149.5,144.1,137.9,135.9,135.2,132.7,130.9,130.4,130.3,130.1,1 28.6,126.2,125.1,123.7,118.3,114.7,110.8,109.3,108.4,107.1,101.2,96.3,87.4,75.5,70 .4,70.1,70.0,67.1,60.0,56.2,55.8,50.4,47.1,39.4,36.7,30.1,13.3,10.8.MS(ESI+APCI)m / z 1030.3[M+H] + .

[0138] Example 12 Synthesis of Aurora A Protein Degrader XH-12

[0139] The synthesis of XH-12 was similar to that of XH-7 in Example 7, including the steps, reaction parameters, and solvents. The difference was that tert-butyl 3-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]propionate was used instead of tert-butyl glycine. The total yield was 30%. 1HNMR(400MHz, CDCl3)δ:8.50(s,1H),8.29(s,1H),8.20(d,J=8.1Hz,2H),8.15(s,1H),8.12(d,J=8.5Hz,1H),7 .81(d,J=2.0Hz,1H),7.54(dd,J=8.5,2.2Hz,1H),7.32(s,1H),7.29-7.25(m,1H),7.21(t,J=5.3Hz,1H),7.16- 7.14(m,1H),7.00(s,1H),6.69(brs,2H),5.41(s,2H),5.23(s,2H),4.25(d,J=5.2Hz,2H),3.91(s,3H),3.74( t,J=5.8Hz,2H),3.70(s,3H),3.66-3.65(m,9H),3.63(m,5H),2.51(t,J=5.8Hz,2H),2.19(s,3H),2.15(s,3H). 13 C NMR (101MHz, CDCl3) δ: 171.2, 165.3, 164.2, 161.8, 161.6, 159.4, 159.1, 159.0, 158.4, 158.1, 156. 7,153.1,153.1,153.0,149.5,144.2,137.9,135.9,135.2,132.7,130.9,130.4,130.3,130.1,128 .5,126.2,125.1,123.6,118.3,114.8,110.9,109.2,108.4,107.1,101.3,96.3,87.4,75.5,70.6, 70.3(3C),70.2,67.3,59.9,56.2,55.8,50.3,47.2,39.5,36.7,30.0,13.3,10.8.MS(ESI+APCI)m / z 1074.4[M+H] + .

[0140] Example 13 Synthesis of Aurora A Protein Degrader XH-13

[0141]

[0142] The steps, reaction parameters, solvents and other conditions for the synthesis of B3 and C3 are similar to those of B1 and C1 in Example 1, except that (4-aminobutyl)carbamic acid tert-butyl ester is used instead of (2-aminoethyl)carbamic acid tert-butyl ester, and 7-octynoic acid is used instead of propiolic acid.

[0143] The steps, reaction parameters, solvents and other conditions for the synthesis of XH-13 are similar to those of XH-7 in Example 7, except that C3 is used instead of C2 prepared in Example 7. The total yield is 40%. Figure 7 and Figure 8 , 1 H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.22 (s, 1H), 8.20 (s, 1H), 8.15 (d, J = 8.6Hz, 1H), 7.94-7.89 (m, 2H), 7.83(s,1H),7.56(dd,J=8.5,2.1Hz,1H),7.33-7.28(m,2H),7.10(d,J=8.0Hz,1H),6.99(s,1H),6.70(b rs,2H),6.11(t,J=8.4Hz,1H),5.28(s,2H),5.08(s,2H),3.98(s,3H),3.73(s,3H),3.47(q,J=6.4Hz,2H ),3.30(q,J=6.3Hz,2H),2.39(t,J=6.8Hz,2H),2.24-2.17(m,8H),1.69-1.56(m,8H),1.52-1.45(m,2H). 13 C NMR(101MHz, CDCl3)δ:173.2,165.5,164.3,161.9,161.6,159.4,159.1,158.8,158.4,158.2,156.7 ,153.3,153.1,152.8,149.5,144.1,138.0,135.9,135.2,132.8,130.9,130.3,130.1,129.3,128.6, 126.3,125.3,123.7,118.1,114.8,111.0,109.6,108.4,107.1,101.4,97.6,92.4,72.6,60.0,56.2 ,55.9,50.3,47.3,39.1,36.6,29.7,28.6,28.3,27.4,26.7,25.4,19.5,13.3,10.9.HRMS(ESI+):m / z calcd.for C 54 H 54 Cl2FN 11 O5[M+H] + :1026.3704; found:1026.3717.

[0144] Example 14 Synthesis of Aurora A Protein Degrader XH-14

[0145] The synthesis steps, reaction parameters, solvents, and other conditions of XH-14 were similar to those of Example 13, except that tert-butyl (4-aminobutyl)carbamate was replaced with [2-(2-aminoethoxy)ethyl]carbamate. The total yield was 47%. Figure 9 and Figure 10 , 1 H NMR (400MHz, CDCl3) δ: 8.50 (s, 1H), 8.31 (s, 1H), 8.21-8.14 (m, 4H), 7.88 (d, J = 2.0Hz, 1H), 7.54 (dd, J = 8.5, 2.2Hz, 1 H),7.32-7.31(m,1H),7.29-7.25(m,1H),7.20(dd,J=8.7,2.0Hz,1H),7.00(s,1H),6.70(brs,2H),6.44(t,J=5.2Hz, 1H),5.40(s,2H),5.26(s,2H),3.94(s,3H),3.71(s,3H),3.68-3.65(m,4H),3.59(t,J=5.2Hz,2H),3.46(q,J=5.1Hz, 2H),2.37(t,J=6.8Hz,2H),2.21-2.17(m,5H),2.16(s,3H),1.69-1.62(m,2H),1.61-1.53(m,2H),1.51-1.45(m,2H). 13 C NMR(101MHz, CDCl3)δ:173.3,165.6,164.3,161.8,161.6,159.4,159.1,159.0,158.4,158.2,156.7 ,153.3,153.1,152.9,149.5,144.3,138.0,135.9,135.1,132.8,130.9,130.3,130.1,129.3,128.5, 126.2,125.2,123.8,118.1,114.6,111.1,109.5,108.4,107.1,101.4,97.6,92.4,72.7,70.1,69.4 ,60.0,56.2,56.0,50.3,47.3,39.4,39.2,36.4,28.5,28.3,25.3,19.5,13.3,10.9.HRMS(ESI+):m / z calcd.for C 54 H 54 Cl2FN 11 O6[M+H] + :1042.3653;found:1042.3665.

[0146] Example 15 Synthesis of Aurora A Protein Degrader XH-15

[0147] The synthesis of XH-15 was similar to that of Example 13, including the steps, reaction parameters, and solvents, except that tert-butyl (6-aminohexyl)carbamate was used instead of tert-butyl (4-aminobutyl)carbamate. The total yield was 35%. 1 H NMR(400MHz, CDCl3)δ:8.51(s,1H),8.22-8.19(m,2H),8.15(d,J=8.6Hz,1H),7.95-7.92(m,2H),7.84(t,J=5.7Hz,1 H),7.55(dd,J=8.5,2.2Hz,1H),7.35-7.32(m,1H),7.28-7.26(m,1H),7.12(dd,J=8.8,2.0Hz,1H),6.99(s,1H),6.7 3(brs,2H),5.92(t,J=5.8Hz,1H),5.27(s,2H),5.16(s,2H),3.97(s,3H),3.72(s,3H),3.45(q,J=7.2Hz,2H),3.22( q,J=6.6Hz,2H),2.39(t,J=6.8Hz,2H),2.23-2.16(m,8H),1.71-1.56(m,6H),1.52-1.44(m,4H),1.40-1.34(m,4H). 13 C NMR(101MHz, CDCl3)δ:173.1,165.3,164.4,161.9,161.6,159.4,159.2,158.8,158.3,158.2,156.7,15 3.3,153.2,152.8,149.4,144.0,138.0,135.9,135.2,132.8,130.9,130.3,130.1,129.2,128.6,126.3 ,125.4,123.8,118.3,115.0,111.0,109.6,108.3,107.1,101.4,97.6,92.4,72.6,60.0,56.1,55.9,50 .3,47.3,39.2,39.1,36.7,29.6,29.4,28.5,28.3,26.3,26.2,25.4,19.5,13.4,10.9.HRMS(ESI+):m / z calcd.for C 56 H 58 Cl2FN 11 O5[M+H]+ :1054.4017; found:1054.4029.

[0148] Example 16 Synthesis of Aurora A Protein Degrader XH-16

[0149] The synthesis conditions for XH-16, including the steps, reaction parameters, and solvents, were similar to those for Example 13, except that tert-butyl [2-[2-(2-aminoethoxy)ethoxy]ethyl]carbamate was used instead of tert-butyl (4-aminobutyl)carbamate, and 5-hexynoic acid was used instead of 7-octynoic acid. The total yield was 58%. 1 H NMR (400MHz, CDCl3) δ: 8.50 (s, 1H), 8.22-8.10 (m, 5H), 7.85 (s, 1H), 7.55 (dd, J = 8.5, 2.1Hz, 1H), 7.32 -7.31(m,1H),7.27-7.25(m,1H),7.15(d,J=8.6Hz,1H),6.99(s,1H),6.67(brs,2H),6.32(t,J=5.6Hz, 1H),5.29-5.25(m,4H),3.94(s,3H),3.72(s,3H),3.67-3.64(m,8H),3.56(t,J=5.2Hz,2H),3.46(q,J= 5.2Hz,2H),2.44(t,J=6.7Hz,2H),2.37(t,J=7.4Hz,2H),2.22(s,3H),2.17(s,3H),1.94-1.87(m,2H). 13 C NMR(101MHz, CDCl3)δ:172.6,165.3,164.3,161.9,161.6,159.4,159.1,158.8,158.4,158.2,156.7,1 53.2,153.0,152.8,149.6,144.2,138.0,135.9,135.2,132.8,130.9,130.3,130.1,129.5,128.6,126. 3,125.3,123.8,118.3,114.8,111.0,109.5,108.2,107.1,101.3,97.4,91.4,73.3,70.4,70.2,70.1,6 9.9,59.9,56.2,55.9,50.4,47.4,39.5,39.2,35.2,24.5,19.1,13.3,10.9.HRMS(ESI+):m / zcalcd.for C 54 H 54 Cl2FN 11O7[M+H] + :1058.3602;found:1058.3618.

[0150] Example 17 Synthesis of Aurora A Protein Degrader XH-17

[0151] The synthesis steps, reaction parameters, solvents, and other conditions of XH-17 were similar to those of Example 13, except that 10-undecynoic acid was used instead of 7-octynoic acid. The total yield was 67%. 1 H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.22-8.20 (m, 2H), 8.15 (d, J = 8.6Hz, 1H), 7.94-7.89 (m, 3H), 7.55 (dd, J = 8 .5,2.2Hz,1H),7.33-7.32(m,1H),7.28-7.26(m,1H),7.13(d,J=8.5Hz,1H),6.99(s,1H),6.75(brs,2H),6.13( t,J=5.8Hz,1H),5.27(s,2H),5.18(s,2H),3.98(s,3H),3.72(s,3H),3.48(q,J=6.4Hz,2H),3.30(q,J=6.4Hz, 2H), 2.37(t,J=6.9Hz,2H),2.23(s,3H),2.18-2.15(m,5H),1.68-1.52(m,8H),1.46-1.41(m,2H),1.30(m,6H). 13 C NMR(101MHz, CDCl3)δ:173.4,165.5,164.4,161.9,161.6,159.4,159.2,158.8,158.4,158.2,156.7,153 .3,153.2,152.8,149.4,144.1,138.0,136.0,135.2,132.8,131.0,130.3,130.1,129.2,128.5,126.3,12 5.4,123.8,118.3,114.8,111.0,109.6,108.5,107.0,101.3,97.7,92.7,72.4,60.0,56.1,55.9,50.4,4 7.4,39.2,39.1,36.8,29.3,29.2,29.0,28.8,28.6,27.5,26.7,25.8,19.6,13.4,10.9.MS(ESI+APCI)m / z 1068.4[M+H] + .

[0152] Example 18 Synthesis of Aurora A Protein Degrader XH-18

[0153] The synthesis of XH-18 was similar to that of Example 13, including the steps, reaction parameters, and solvents. The differences were that tert-butyl [2-(2-aminoethoxy)ethyl]carbamate was used instead of tert-butyl (4-aminobutyl)carbamate, and 10-undecynoic acid was used instead of 7-octynoic acid. The total yield was 61%. 1 H NMR (400MHz, CDCl3) δ: 8.50 (s, 1H), 8.27 (s, 1H), 8.21-8.14 (m, 4H), 7.94 (d, J = 2.0Hz, 1H), 7.54 (dd, J = 8.5, 2.1Hz, 1H) ,7.32(m,1H),7.29-7.25(m,1H),7.15(dd,J=8.6,2.0Hz,1H),6.99(s,1H),6.69(brs,2H),6.46(t,J=5.4Hz,1H),5.37 (s,2H),5.26(s,2H),3.95(s,3H),3.71(s,3H),3.68-3.64(m,4H),3.59(t,J=5.2Hz,2H),3.47(q,J=5.2Hz,2H),2.35( t,J=6.9Hz,2H),2.21(s,3H),2.18-2.15(m,5H),1.64-1.57(m,2H),1.55-1.50(m,2H),1.44-1.39(m,2H),1.27(m,6H). 13 CNMR(101MHz, CDCl3)δ:173.5,165.5,164.3,161.8,161.6,159.4,159.1,158.9,158.4,158.2,156.7,15 3.3,153.1,152.8,149.4,144.3,137.9,135.9,135.2,132.8,130.8,130.3,130.1,129.2,128.6,126.2, 125.3,123.8,118.3,114.6,111.0,109.5,108.4,107.1,101.3,97.6,92.6,72.5,70.0,69.2,59.9,56.1 ,55.9,50.3,47.3,39.3,39.2,36.6,29.2(2C),28.9,28.7,28.6,25.8,19.6,13.3,10.9.HRMS(ESI+):m / z calcd.forC 57 H 60 Cl2FN11 O6[M+H] + :1084.4123; found:1084.4152.

[0154] Example 19 Synthesis of Aurora A Protein Degrader XH-19

[0155] The synthesis of XH-19 was similar to that of Example 13, including the steps, reaction parameters, and solvents. The difference was that tert-butyl (4-aminobutyl)carbamate was replaced with tert-butyl [2-[2-(2-aminoethoxy)ethoxy]ethyl]carbamate. The total yield was 46%. 1 H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.24-8.14 (m, 5H), 7.90 (d, J = 2.0Hz, 1H), 7.55 (dd, J = 8.5, 2.2Hz, 1H) ,7.32-7.32(m,1H),7.29-7.25(m,1H),7.16(dd,J=8.7,1.9Hz,1H),6.99(s,1H),6.69(brs,2H),6.34(t,J =5.6Hz,1H),5.33(s,2H),5.26(s,2H),3.94(s,3H),3.71(s,3H),3.69-3.62(m,8H),3.55(t,J=5.1Hz,2H) ,3.44(q,J=5.6Hz,2H),2.36(t,J=6.8Hz,2H),2.21-2.15(m,8H),1.68-1.1.53(m,4H),1.48-1.41(m,2H). 13 C NMR (101MHz, CDCl3) δ: 173.2, 165.4, 164.3, 161.8, 161.6, 159.4, 159.1, 158.9, 158.4, 158.2, 156.7, 153. 3,153.1,152.9,149.4,144.3,137.9,135.9,135.2,132.8,130.9,130.3,130.1,129.3,128.5,126.2,125 .2,123.7,118.3,114.7,111.0,109.5,108.4,107.1,101.4,97.6,92.3,72.6,70.4,70.2,70.1,69.9,59. 9,56.1,55.9,50.3,47.3,39.5,39.2,36.5,28.5,28.3,25.2,19.5,13.3,10.9.HRMS(ESI+):m / zcalcd.for C 56 H58 Cl2FN 11 O7[M+H] + :1086.3915;found:1086.3914.

[0156] Example 20 Synthesis of Aurora A Protein Degrader XH-20

[0157] The synthesis steps, reaction parameters, solvents, and other conditions for XH-20 were similar to those for XH-13 in Example 13, except that tert-butyl [2-[2-(2-aminoethoxy)ethoxy]ethyl]carbamate was used instead of tert-butyl (4-aminobutyl)carbamate, and 6-heptynoic acid was used instead of 7-octynoic acid. The total yield was 61%. 1 H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.22-8.15 (m, 4H), 7.94 (s, 1H), 7.88 (d, J = 1.9Hz, 1H), 7.56 (dd, J =8.4,2.2Hz,1H),7.32-7.28(m,2H),7.15(d,J=7.2Hz,1H),6.99(s,1H),6.77(brs,2H),6.28(t,J=5.6 Hz,1H),5.27(s,2H),5.13(s,2H),3.96(s,3H),3.73(s,3H),3.69-3.64(m,8H),3.55(t,J=5.0Hz,2H), 3.43(q,J=4.4Hz,2H),2.39(t,J=6.9Hz,2H),2.23-2.17(m,8H),1.82-1.75(m,2H),1.62-1.57(m,2H). 13C NMR(101MHz, CDCl3)δ:172.9,165.4,164.8,161.9,161.6,159.4,159.2,158.8,158.5,158.2,156.7,1 53.2,153.0,152.8,149.0,144.1,138.0,136.0,135.2,132.9,130.9,130.3,130.1,129.3,128.6,126 .5,125.6,123.8,118.1,114.8,111.0,109.6,108.5,107.1,101.4,97.7,92.1,72.8,70.5,70.2,70.1 ,70.0,60.1,56.2,55.9,50.3,47.0,39.5,39.2,36.1,28.2,25.0,19.4,13.4,11.0.MS(ESI+APCI)m / z 1072.4[M+H] + .

[0158] Example 21 Synthesis of Aurora A Protein Degrader XH-21

[0159] The synthesis of XH-21, including the steps, reaction parameters, and solvents, was similar to that of Example 13, except that tert-butyl [2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethyl]carbamate was used instead of tert-butyl (4-aminobutyl)carbamate, and 5-hexynoic acid was used instead of 7-octynoic acid. The total yield was 32%. 1 H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.22-8.13 (m, 4H), 8.07 (s, 1H), 7.85 (d, J = 1.9Hz, 1H), 7.55 (dd, J = 8.5 ,2.2Hz,1H),7.32-7.28(m,2H),7.16(d,J=8.7Hz,1H),6.99(s,1H),6.72(brs,2H),6.35(t,J=5.5Hz,1H), 5.26(m,4H),3.94(s,3H),3.72(s,3H),3.67-3.63(m,8H),3.63-3.59(m,4H),3.54(t,J=5.2Hz,2H),3.44( q,J=5.2Hz,2H),2.45(t,J=6.8Hz,2H),2.38(t,J=7.4Hz,2H),2.22(s,3H),2.16(s,3H),1.95-1.88(m,2H). 13C NMR(101MHz, CDCl3)δ:172.6,165.3,164.3,161.9,161.6,159.4,159.1,158.9,158.4,158.1,156.7,15 3.3,153.1,152.9,149.5,144.1,138.0,135.9,135.2,132.8,130.8,130.3,130.1,129.5,128.6,126.2 ,125.2,123.8,118.1,114.8,110.9,109.4,108.2,107.1,101.3,97.3,91.4,73.3,70.6,70.5,70.3,70 .2,70.1,69.9,59.9,56.2,55.8,50.3,47.3,39.5,39.2,35.3,24.6,19.2,13.3,10.9.HRMS(ESI+):m / z calcd.for C 56 H 58 Cl2FN 11 O8[M+H] + :1102.3864; found:1102.3892.

[0160] Example 22 Synthesis of Aurora A Protein Degrader XH-22

[0161] The synthesis steps, reaction parameters, solvents, and other conditions for XH-22 were similar to those for XH-7 in Example 7, except that tert-butyl 4-aminobutyrate was used instead of tert-butyl glycine, and 2-(but-3-yn-1-yloxy)ethanamine was used instead of propargylamine. The total yield was 50%. 1H NMR (400MHz, CDCl3) δ: 8.52 (s, 1H), 8.50 (s, 1H), 8.03 (s, 1H), 7.99 (t, J = 5.0Hz, 1H), 7.93-7.87 (m, 2H), 7. 67(t,J=0.9Hz,1H),7.63(d,J=2.4Hz,1H),7.52(dd,J=9.0,2.6Hz,1H),7.38(dd,J=8.2,2.2Hz,1H),7.32-7 .28(m,1H),7.11-1.08(m,1H),6.85-6.79(m,3H),5.61(s,2H),4.89(s,2H),3.89(s,3H),3.71(s,3H),3.66 -3.57(m,4H),3.42-3.34(m,4H),2.69(t,J=6.5Hz,2H),2.24-2.17(m,5H),2.16(s,3H),1.89-1.84(m,2H). 13 C NMR(101MHz, CDCl3)δ:174.7,171.0,167.5,164.0,161.8,160.9,160.2,159.6,159.1,154.9,154 .7,153.0,151.4,150.9,148.3,145.1,134.5,134.1,133.8,133.5,130.8,130.3,130.0,129.8,1 29.1,124.8,123.4,120.9,118.2,117.3,111.1,110.3,108.4,104.2,101.4,97.0,96.4,79.9,69 .7,69.3,60.0,56.1,55.8,53.9,47.8,40.7,39.5,33.6,24.3,19.8,13.6,10.9.MS(ESI+APCI)m / z 1014.3[M+H] + .

[0162] Example 23 Synthesis of Aurora A Protein Degrader XH-23

[0163] The synthesis of XH-23 was similar to that of XH-7 in Example 7, including the steps, reaction parameters, and solvents. The difference was that 2-[2-(but-3-yn-1-yloxy)ethoxy]ethanamine was used instead of propargylamine. The total yield was 44%. 1H NMR(400MHz, CDCl3)δ:8.51(s,1H),8.48(s,1H),8.03(dd,J=12.5,6.3Hz,2H),7.96-7.90(m,2H),7.69(t,J=0.9Hz, 1H),7.63(d,J=2.4Hz,1H),7.53(dd,J=9.0,2.6Hz,1H),7.41(dd,J=8.2,2.2Hz,1H),7.33-7.27(m,1H),7.12-7.05(m ,1H),6.99(t,J=4.9Hz,1H),6.82(brs,2H),5.55(s,2H),4.96(s,2H),3.93(d,J=6.2Hz,2H),3.92(s,3H),3.71(s,3 H),3.65-3.61(m,6H),3.54(t,J=4.3Hz,2H),3.31(q,J=4.4Hz,2H),2.72(t,J=6.4Hz,2H),2.21(s,3H),2.16(s,3H). 13 C NMR(101MHz, CDCl3)δ:170.9,170.7,168.0,163.6,161.9,160.9,160.1,159.5,159.0,154.9,154 .7,153.0,151.4,151.0,148.2,145.2,134.4,134.1,133.9,133.4,130.8,130.3,129.9,129.7,1 29.2,124.8,123.2,120.8,118.1,116.9,111.2,110.0,108.0,104.1,101.5,97.3,96.1,80.2,70 .0,69.7,69.5,69.3,59.9,56.2,55.5,53.8,48.1,44.6,39.7,21.1,12.4,10.3.MS(ESI+APCI)m / z 1030.3[M+H] + .

[0164] Example 24 Synthesis of Aurora A Protein Degrader XH-24

[0165] The synthesis steps, reaction parameters, solvents, and other conditions for XH-24 were similar to those for XH-13 in Example 13, except that tert-butyl [2-[2-(2-aminoethoxy)ethoxy]ethyl]carbamate was used instead of tert-butyl (4-aminobutyl)carbamate, and 4-pentynoic acid was used instead of 7-octynoic acid. The total yield was 48%. 1H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.22-8.15 (m, 4H), 7.94 (s, 1H), 7.88 (d, J = 1.9Hz, 1H), 7.56 (dd, J=8.4,2.2Hz,1H),7.32-7.28(m,2H),7.15(d,J=7.2Hz,1H),6.99(s,1H),6.77(brs,2H),6.28(t,J= 5.6Hz,1H),5.27(s,2H),5.13(s,2H),3.96(s,3H),3.73(s,3H),3.69-3.64(m,8H),3.55(t,J=5.0Hz ,2H),3.43(q,J=4.4Hz,2H),2.71(t,J=8.7Hz,2H),2.39(t,J=6.9Hz,2H),2.23(s,3H),2.16(s,3H). 13 C NMR (101MHz, CDCl3) δ: 173.0, 170.9, 167.3, 163.6, 160.9, 160.2, 159.9, 159.6, 159.1, 155.0, 154. 7,152.9,151.5,150.8,148.2,145.1,134.5,134.1,133.7,133.4,131.0,130.5,130.1,129.9,129. 0,124.7,123.5,121.2,118.2,117.1,111.2,110.2,108.1,103.9,100.9,99.3,96.5,78.6,69.7,6 9.6,69.4,69.0,60.1,56.3,56.0,54.0,47.2,40.7,40.3,36.4,15.1,13.1,10.8.MS(ESI+APCI)m / z 1044.3[M+H] + .

[0166] Example 25 Synthesis of Aurora A Protein Degrader XH-25

[0167] The synthesis of XH-25 was similar to that of Example 13, including the steps, reaction parameters, and solvents, except that 3-[2-(but-3-yn-1-yloxy)ethoxy]propionic acid was used instead of 7-octynoic acid. The total yield was 52%. 1HNMR(400MHz, CDCl3)δ:8.51(s,1H),8.23(s,1H),8.03-8.00(m,2H),7.94-7.88(m,2H),7.65(t,J=0.9Hz,1H),7.57(d,J= 2.4Hz,1H),7.45(dd,J=9.0,2.6Hz,1H),7.38(dd,J=8.2,2.2Hz,1H),7.34-7.28(m,1H),7.10(m,1H),6.83(brs,2H),5.88 (t,J=4.9Hz,1H),5.60(s,2H),4.93(s,2H),3.91(s,3H),3.72(s,3H),3.71-3.61(m,8H),3.37(q,J=5.1Hz,2H),3.15(q,J =5.0Hz,2H),2.69(t,J=6.4Hz,2H),2.47(t,J=6.0Hz,2H),2.22(s,3H),2.16(s,3H),1.62-1.57(m,2H),1.51-1.46(m,2H). 13 C NMR(101MHz, CDCl3)δ:173.1,171.1,167.5,163.8,161.0,160.2,159.8,159.6,158.9,155.1,154.5,1 52.9,151.6,151.1,147.9,145.2,134.5,134.0,133.7,133.3,131.0,130.3,130.1,129.6,129.2,124 .7,123.5,121.2,118.2,117.2,111.0,110.1,108.2,104.3,101.7,97.0,96.2,80.1,70.4,69.6,69.4 ,66.4,59.7,56.1,55.5,54.0,47.5,42.0,40.9,37.4,27.3,26.4,21.2,13.1,10.6.MS(ESI+APCI)m / z 1072.4[M+H] + .

[0168] Example 26 Synthesis of Aurora A Protein Degrader XH-26

[0169] The synthesis steps, reaction parameters, solvents, and other conditions for XH-26 were similar to those for XH-7 in Example 7, except that tert-butyl 6-aminohexanoate was used instead of tert-butyl glycine, and 2-(but-3-yn-1-yloxy)ethanamine was used instead of propargylamine. The total yield was 38%. 1H NMR(400MHz, CDCl3)δ:8.51(s,1H),8.48(s,1H),8.03-7.96(m,2H),7.94-7.90(m,2H),7.66-7.62(m,2H),7.5 1(dd,J=9.2,2.6Hz,1H),7.48(dd,J=8.2,2.2Hz,1H),7.34-7.27(m,1H),7.13(m,1H),6.82(brs,2H),6.67(s, 1H),5.54(s,2H),4.90(s,2H),3.95(s,3H),3.78(s,3H),3.62-3.55(m,4H),3.40(dt,J=5.1,4.2Hz,2H),3.33 (q,J=5.3Hz,2H),2.65(t,J=6.5Hz,2H),2.24(s,3H),2.19-2.14(m,5H),1.60-1.53(m,2H),1.50-1.37(m,4H). 13 C NMR(101MHz, CDCl3)δ:173.3,170.9,167.6,163.6,161.9,160.8,160.0,159.4,159.1,155.0,154.7 ,152.9,151.6,150.5,147.9,145.2,134.5,134.1,133.7,133.4,130.8,130.2,130.0,129.8,129.1, 124.7,123.4,121.0,118.0,117.1,111.1,110.3,108.4,104.4,101.6,97.1,96.3,79.9,69.5,69.1, 59.9,56.1,55.8,53.5,47.5,40.4,39.9,35.2,28.9,25.9,24.7,20.4,13.0,10.8.MS(ESI+APCI)m / z 1042.4[M+H] + .

[0170] Example 27 Synthesis of Aurora A Protein Degrader XH-27

[0171] The synthesis of XH-27 was similar to that of Example 7, including the steps, reaction parameters, and solvents. The differences were that tert-butyl 3-(2-aminoethoxy)propionate was used instead of tert-butyl glycine, and 2-(but-3-yn-1-yloxy)ethanamine was used instead of propargylamine. The overall yield was 43%. 1H NMR (400MHz, CDCl3) δ: 8.52 (s, 1H), 8.52 (s, 1H), 8.02-7.87 (m, 3H), 7.72 (t, J = 0.9Hz, 1H), 7.62 (d, J = 2.4Hz ,1H),7.51(dd,J=8.4,2.6Hz,1H),7.44(dd,J=8.0,2.2Hz,1H),7.31-7.26(m,2H),7.13(m,1H),6.82(brs,2H ),6.60(t,J=5.0Hz,1H),5.63(s,2H),4.88(s,2H),3.93(s,3H),3.70(s,3H),3.72-3.68(m,4H),3.61-3.57 (m,6H),3.42(dt,J=5.2,4.2Hz,2H),2.68(t,J=6.4Hz,2H),2.45(t,J=6.0Hz,2H),2.23(s,3H),2.16(s,3H). 13 C NMR (101MHz, CDCl3) δ: 172.9, 170.9, 167.4, 163.6, 161.8, 160.6, 159.9, 159.3, 158.8, 154.8, 154. 7,153.0,151.6,150.8,148.2,145.3,134.4,134.1,133.8,133.5,130.9,130.2,129.9,129.7,129. 0,124.8,123.5,121.0,118.3,117.2,111.1,110.2,108.4,104.2,101.4,97.1,96.3,80.1,69.8,6 9.5,69.2,66.3,59.8,56.0,55.9,53.5,47.6,40.6,40.3,37.4,19.8,13.0,10.7.MS(ESI+APCI)m / z 1044.3[M+H] + .

[0172] Example 28 Synthesis of Aurora A Protein Degrader XH-28

[0173] The synthesis of XH-28 was similar to that of Example 13, including the steps, reaction parameters, and solvents. The differences were that (6-aminohexyl)carbamic acid tert-butyl ester was used instead of (4-aminobutyl)carbamate, and 3-(but-3-yn-1-yloxy)propionic acid was used instead of 7-octynoic acid. The total yield was 41%. 1H NMR (400MHz, CDCl3) δ: 8.51 (s, 1H), 8.45 (s, 1H), 8.12-8.04 (m, 2H), 7.87-7.85 (m, 2H), 7.65 (t, J = 0.9Hz, 1H), 7.52 (d, J = 2.4Hz, 1 H),7.44(dd,J=8.4,2.6Hz,1H),7.38(dd,J=8.0,2.2Hz,1H),7.30-7.26(m,1H),7.15-7.12(m,1H),6.77(brs,2H),5.74(t,J=4.9 Hz,1H),5.47(s,2H),4.82(s,2H),3.98(s,3H),3.72(s,3H),3.70(t,J=6.0Hz,2H),3.61(t,J=6.5Hz,2H),3.32(q,J=5.2Hz,2H), 3.15-3.09(m,2H),2.69(t,J=6.5Hz,2H),2.47(t,J=6.0Hz,2H),2.23(s,3H),2.16(s,3H),1.59-1.51(m,4H),1.40-1.33(m,4H). 13 C NMR(101MHz, CDCl3)δ:173.3,170.9,167.7,163.9,161.0,160.2,159.8,159.6,159.1,155.0,154.6,1 53.0,151.3,150.7,148.3,145.4,134.6,134.1,133.7,133.4,130.8,130.1,130.0,129.7,129.1,124 .7,123.3,121.0,118.4,117.3,111.0,110.3,108.1,103.8,101.5,97.1,96.4,80.2,69.0,66.8,60.4 ,56.7,56.1,53.9,47.1,40.9,39.8,38.5,37.0,29.3,26.8,26.2,20.9,13.2,10.9.MS(ESI+APCI)m / z 1056.4[M+H] + .

[0174] Example 29 Synthesis of Aurora A Protein Degrader XH-29

[0175] The synthesis of XH-29 was similar to that of Example 13, including the following steps, reaction parameters, and solvents. The differences were that tert-butyl [2-(2-aminoethoxy)ethyl]carbamate was used instead of tert-butyl (4-aminobutyl)carbamate, and 3-(but-3-yn-1-yloxy)propionic acid was used instead of 7-octynoic acid. The total yield was 49%. 1 H NMR(400MHz, CDCl3)δ:8.52(s,1H),8.46(s,1H),8.01(s,1H),7.98-7.91(m,2H),7.66(t,J=0.9Hz,1 H),7.55(d,J=2.4Hz,1H),7.49-7.40(m,3H),7.35-7.29(m,1H),7.09(m,1H),6.84-6.75(m,3H),5.6 0(s,2H),4.90(s,2H),3.97(s,3H),3.70(s,3H),3.72-3.65(m,4H),3.63-3.59(m,4H),3.52(t,J=4. 2Hz,2H),3.41-3.37(m,2H),2.53(t,J=6.0Hz,2H),2.47(t,J=6.2Hz,2H),2.23(s,3H),2.16(s,3H). 13 C NMR (101MHz, CDCl3) δ: 173.0, 170.9, 167.3, 163.7, 161.0, 160.2, 159.7, 159.4, 159.0, 154.9, 154. 5,153.0,151.7,151.0,148.3,145.1,134.8,134.1,133.8,133.4,130.9,130.4,130.0,129.8,129. 2,124.6,123.4,121.0,118.6,117.3,111.3,110.3,108.5,104.4,101.5,97.1,96.4,80.2,69.6,6 9.5,69.1,66.5,59.9,56.4,55.7,54.5,47.5,40.7,40.2,37.6,20.7,13.0,10.9.MS(ESI+APCI)m / z 1044.3[M+H] + .

[0176] Example 30 Synthesis of Aurora A Protein Degrader XH-30

[0177] The synthesis steps, reaction parameters, solvents, and other conditions for XH-30 were similar to those for XH-7 in Example 7, except that tert-butyl 4-aminobutyrate was used instead of tert-butyl glycine, and 2-[2-(but-3-yn-1-yloxy)ethoxy]ethanamine was used instead of propargylamine. The overall yield was 35%. 1 H NMR (400MHz, CDCl3) δ: 8.49 (s, 1H), 8.46 (s, 1H), 8.03 (s, 1H), 7.97 (t, J = 5.0Hz, 1H), 7.95-7.88 (m, 2H), 7.63 (t, J = 0.9Hz, 1H),7.55(d,J=2.4Hz,1H),7.49(dd,J=8.6,2.6Hz,1H),7.40(dd,J=8.2,2.2Hz,1H),7.33(dd,J=8.1,5.1Hz,1H),7.10-7.0 7(m,1H),6.83-6.75(m,3H),5.55(s,2H),5.00(s,2H),3.98(s,3H),3.74(s,3H),3.66-3.60(m,6H),3.58(t,J=4.3Hz,2H) ,3.48(dt,J=5.1,4.3Hz,2H),3.38-3.34(m,2H),2.70(t,J=6.4Hz,2H),2.20-2.17(m,5H),2.15(s,3H),1.83-1.79(m,2H). 13 CNMR(101MHz, CDCl3)δ:174.6,170.7,167.5,163.6,161.8,160.8,160.1,159.6,159.1,154.8,154.7 ,153.0,151.7,150.7,148.2,145.3,134.4,134.1,133.7,133.4,130.8,130.3,130.0,129.5,129.0, 124.5,123.4,121.0,118.2,117.3,111.0,110.3,108.4,104.1,101.5,97.1,96.4,80.2,69.9,69.7, 69.6,69.2,59.9,56.2,56.0,53.8,47.6,40.5,39.6,34.2,24.0,20.8,12.8,10.7.MS(ESI+APCI)m / z 1058.4[M+H] + .

[0178] Example 31 Synthesis of Aurora A Protein Degrader XH-31

[0179] The synthesis of XH-31 was similar to that of Example 7 for XH-7, including the following steps, reaction parameters, and solvents. The differences were that tert-butyl 3-[2-(2-aminoethoxy)ethoxy]propionate was used instead of tert-butyl glycine, and 2-(but-3-yn-1-yloxy)ethanamine was used instead of propargylamine. The overall yield was 46%. 1 H NMR(400MHz, CDCl3)δ:8.50(s,1H),8.22(s,1H),8.02(s,1H),7.96-7.87(m,2H),7.66(t,J=0.9Hz ,1H),7.60(d,J=2.4Hz,1H),7.49-7.40(m,3H),7.30-7.26(m,1H),7.10-7.07(m,2H),6.78(brs,2 H),5.37(s,2H),5.25(s,2H),3.97(s,3H),3.73(s,3H),3.70-3.65(m,4H),3.63-3.55(m,10H),3. 42(dt,J=5.1,4.2Hz,2H),2.64(t,J=6.5Hz,2H),2.45(t,J=6.0Hz,2H),2.21(s,3H),2.17(s,3H). 13 C NMR(101MHz, CDCl3)δ:172.7,171.4,167.3,163.6,160.8,160.2,159.8,159.5,159.1,155.1,154.6,1 53.0,151.6,150.9,148.1,145.3,134.6,134.1,133.8,133.5,130.7,130.4,130.0,129.8,129.1,124 .8,123.5,121.1,118.5,117.3,111.1,110.2,108.4,104.2,101.5,96.8,96.2,80.1,69.8,69.7,69.6 ,69.5,69.2,66.4,59.8,56.5,55.9,54.0,47.9,40.8,40.1,37.7,20.3,13.2,11.0.MS(ESI+APCI)m / z 1088.4[M+H] + .

[0180] Example 32 Synthesis of Aurora A Protein Degrader XH-32

[0181] The synthesis of XH-32, including the steps, reaction parameters, and solvents, was similar to that of Example 13, except that tert-butyl [2-(2-aminoethoxy)ethyl]carbamate was used instead of tert-butyl (4-aminobutyl)carbamate, and 3-[2-(but-3-yn-1-yloxy)ethoxy]propionic acid was used instead of 7-octynoic acid. The total yield was 43%. 1 H NMR(400MHz, CDCl3)δ:8.50(s,1H),8.23(s,1H),8.00(s,1H),7.96-7.86(m,2H),7.67(t,J=0.9Hz,1 H),7.56(d,J=2.4Hz,1H),7.53-7.40(m,3H),7.32-7.25(m,1H),7.10(m,1H),6.80(brs,2H),6.70(t ,J=5.1Hz,1H),5.38(s,2H),5.24(s,2H),3.97(s,3H),3.73(s,3H),3.72-3.60(m,12H),3.60(t,J=4 .1Hz,2H),3.40-3.37(m,2H),2.69(t,J=6.0Hz,2H),2.46(t,J=6.2Hz,2H),2.24(s,3H),2.15(s,3H). 13 C NMR(101MHz, CDCl3)δ:172.6,171.2,167.4,163.6,160.8,160.3,159.9,159.5,158.9,154.9,154.4,1 52.9,151.5,150.9,148.3,145.4,134.7,134.1,133.9,133.3,131.1,130.5,130.0,129.8,129.1,124 .8,123.2,121.1,118.4,117.0,111.4,110.3,108.3,104.4,101.2,97.3,96.2,80.2,70.0,69.6,69.5 ,69.4,69.3,66.4,59.9,56.1,55.9,53.8,47.8,40.6,40.1,37.3,20.5,13.1,10.9.MS(ESI+APCI)m / z 1088.4[M+H] + .

[0182] In order to prove the effect of the technical solution provided by this application, the following is the activity verification experiment and results of the compound provided by this application.

[0183] 1. Antiproliferative activity against tumor cells

[0184] The anti-tumor effects of the compounds of the present invention were evaluated by screening activity at a concentration of 10 μM in five malignant tumor cell lines (including the cervical cancer cell line Hela, the breast cancer cell line MCF7, the chronic myeloid leukemia K562, the T lymphocytic leukemia Jurkat, and the lung cancer cell line A549) (Table 1). The compounds showed inhibitory advantages in the A549 (lung cancer) and MCF7 (breast cancer) cell lines, with flexible linkers containing 14-15 atoms showing better inhibitory effects.

[0185] Table 1 Antitumor cell activity of the compounds of the present invention a (% of Control, 10 μM)

[0186]

[0187]

[0188] (a) Mean, n=3; (b) MLN-8237 and BIIB021 were mixed at a ratio of 0.5:0.5 to obtain a mixture of 1 for combined use.

[0189] 2. Inhibitory effect on tumor cells and drug-resistant tumor cells

[0190] The half-maximal inhibitory concentration (IC50) of XH series compounds on non-resistant tumor cells MCF7 and A549, as well as resistant tumor cells A549 / PTX was evaluated. 50 ), the results are shown in Table 2. The anti-tumor activity of the XH series compounds is generally better than that of the positive drug MLN-8237. Among them, XH-14 has the best inhibitory effect on A549, IC 50 The value is 0.294μM, which is nearly 3 times higher than that of MLN-8237.

[0191] Phase II clinical drug MLN-8237 showed certain activity in some clinical trials, but had a low objective response rate (ORR), especially in children and adolescents, where the ORR was less than 5% for monotherapy; in the treatment of resistant breast cancer, despite partial remission, disease progression (PD) was still the main reason for discontinuation. Existing PROTACs targeting Aurora A have a good inhibitory effect on non-resistant tumors, but their efficacy in resistant cells has not yet been clarified. The inventors evaluated the IC of a new PROTAC designed based on the HSP90 / E3 complex on resistant cells using a paclitaxel-resistant NSCLC cell model. 50The XH series of protein degraders demonstrated superior antitumor activity compared to the active drug MLN-8237 in the A549 / PTX-resistant tumor model, suggesting that protein degradation strategies have more potent tumor-killing effects than simple inhibition. However, degraders did not show significant advantages over combination therapy, possibly due to limited bioavailability caused by their inherent physicochemical properties and impaired drug uptake and enhanced efflux caused by overexpression of ABC transporters in drug-resistant tumor cells.

[0192] Table 2 IC values of preferred compounds against non-resistant and resistant tumor cells 50 Value a

[0193]

[0194]

[0195] (a) mean ± SEM, n = 3; (b) MLN-8237 and BIIB021 were mixed at a ratio of 0.5:0.5 to obtain a mixture of 1 for combined drug administration.

[0196] 3. Determination of Aurora A kinase degradation effect of XH-14

[0197] The degradation effect of XH-14 series compounds on Aurora A kinase in MCF7 and A549 cells was determined by Western blot. Figure 11 ,in, Figure 11 (A) Endogenous Aurora A protein levels in MCF7 cells after 6 hours; (B) Endogenous Aurora A protein levels in A549 cells after 6 hours; (C) Concentration-dependent expression of Aurora A protein in cells after 6 hours of XH-14 treatment; Half-maximal degradation concentration (DC) of A549 cells after 6 hours of XH series compound treatment 50 Value Reference Figure 12 It can be seen that in A549 cells, XH-14 has the best degradation effect on Aurora A at 0.5 μM. The maximum degradation ability of XH series compounds on Aurora A protein in A549 cells is D max Value Reference Figure 13 , it can be seen that DC 50 120nM, D max It is 91.5%.

[0198] 4. Verification of XH-14-induced ternary complex formation

[0199] The core mechanism of the ubiquitination process lies in the formation of a stable ternary complex, in which the synergistic binding of the target protein, PROTAC, and E3 ligase is the key to achieving selective degradation. Taking XH-14 as an example, the endogenous Aurora A protein complex was successfully captured through immunoprecipitation experiments with alpaca-derived nanomagnetic beads, and HSP90 protein was detected in the complex precipitate. The specific coprecipitation of HSP90 confirmed that XH-14 can induce the formation of the HSP90 / E3-XH-14-Aurora A ternary complex ( Figure 14 ), where (A) Compound XH-14 induces the formation of an HSP90 / E3-XH-14-Aurora A ternary complex in MCF7 cells; (B) Compound XH-14 induces the formation of an HSP90 / E3-XH-14-Aurora A ternary complex in A549 cells. It can be seen that this complex recruits E3 ubiquitin ligases to promote ubiquitination of Aurora A, ultimately leading to its degradation by the proteasome. This reveals the dynamic process of PROTAC-mediated target protein degradation at the molecular level.

[0200] 5. Effects of XH series compounds on A549 and drug-resistant A549 xenografted nude mice

[0201] The in vivo antitumor activity of the XH series compounds was evaluated using non-resistant A549 and resistant A549 / PTR xenograft tumor models. For a flowchart of the in vivo antitumor activity evaluation of the XH series compounds against non-resistant / resistant non-small cell lung cancer xenograft tumor nude mice models, please refer to Figure 15 ; Refer to the results of nude mouse tumor tissue quality after treatment with solvent or XH series compounds Figure 16 , refer to the results of in vivo tumor inhibition rate of non-resistant / resistant cell tumors Figure 17 At a dose of 30 mg / kg, all experimental treatment groups demonstrated significant tumor growth inhibition. XH-14 demonstrated significant tumor inhibition in both models, with TGIs of 59.6% and 56.6%, respectively.

Claims

1. An Aurora A protein degrader based on HSP90, characterized in that: A compound having the structural formula (A): The linker is selected from one of the following substituents:

2. The method for preparing the HSP90-based Aurora A protein degrader according to claim 1, characterized in that: The target product is prepared by reacting the AuroraA binding ligand, the HSP90 recruiting ligand and the linker compound connecting the two.

3. The method for preparing an HSP90-based Aurora A protein degrader according to claim 2, wherein: The AuroraA binding ligand is MLN-8237; The HSP90 recruitment ligand structural formula is HSP90 Ligand-1 or HSP90 Ligand-2. The structural formulas of HSP90Ligand-1 and HSP90Ligand-2 are as follows:

4. The method for preparing an HSP90-based Aurora A protein degrader according to claim 2, wherein: The linker is one of the compounds having the general formula 2, 4, or 6: In general formula 2: R 1 C 2-6 Alkyl, -(CH2CH2O) n CH2CH2-, and n is an integer from 1 to 3; In general formula 4: R 2 C 1-5 Alkyl, -(CH2CH2O) n CH2CH2-, and n is an integer from 1 to 3; In general formula 4: R 3 -CH2-, -(CH2CH2O) m CH2CH2-, and m is an integer from 1 to 2; In general formula 6: R 4 C 4-6 Alkyl, -(CH2CH2O) n CH2CH2-, and n is an integer from 1 to 3; In general formula 6: R 5 C 2-8 Alkyl, -(CH2CH2O) m CH2CH2-, and m is an integer of 1-2.

5. The method for preparing the HSP90-based Aurora A protein degrader according to any one of claims 2 to 4, characterized in that: The method includes the following steps in sequence: 1)MLN-8237 and NH2R 1 Boc or NHR 2 C(=O)OC(CH3)3 or NH2R 4 Boc undergoes amide condensation at room temperature to obtain amide condensation product A; 2) At room temperature, the amide condensation product A prepared in step 1) is deprotected by trifluoroacetic acid and then reacted with propiolic acid or NH2R 3 C≡C or HOC(=O)R 5 C≡C amide condensation gives amide condensation product B; 3) The amide condensation product B prepared in step 2) reacts with the HSP90 recruitment ligand to obtain the target product, an HSP90-based Aurora A protein degrader.

6. The method for preparing an HSP90-based Aurora A protein degrader according to claim 5, characterized in that: The synthetic route is the following synthetic route a or synthetic route b or synthetic route c, as follows: Synthesis route a: Synthesis route b: Synthesis route c: The R 1 、R 2 、R 3 、R 4 、R 5 As defined in claim 4.

7. Use of the HSP90-based Aurora A protein degrader according to claim 1 in the preparation of drugs for treating tumors.

8. Use of the HSP90-based Aurora A protein degrader according to claim 1 in the preparation of a drug for treating or preventing proliferative diseases.

9. A drug for treating tumors, characterized in that: comprising an active ingredient and a pharmaceutically acceptable carrier; The active ingredient is: the HSP90-based Aurora A protein degrader according to claim 1, or a pharmaceutically acceptable salt of the HSP90-based Aurora A protein degrader according to claim 1; or a hydrate of the HSP90-based Aurora A protein degrader according to claim 1; or a solvate of the HSP90-based Aurora A protein degrader according to claim 1; or a polymorph of the HSP90-based Aurora A protein degrader according to claim 1, or a tautomer of the HSP90-based Aurora A protein degrader according to claim 1, or a prodrug of the HSP90-based Aurora A protein degrader according to claim 1; the carrier is at least one of a diluent, an excipient, and a filler.

10. A drug for treating or preventing a proliferative disease, characterized in that: comprising an active ingredient and a pharmaceutically acceptable carrier; The active ingredient is: the HSP90-based Aurora A protein degrader according to claim 1, or a pharmaceutically acceptable salt of the HSP90-based Aurora A protein degrader according to claim 1; or a hydrate of the HSP90-based Aurora A protein degrader according to claim 1; or a solvate of the HSP90-based Aurora A protein degrader according to claim 1; or a polymorph of the HSP90-based Aurora A protein degrader according to claim 1, or a tautomer of the HSP90-based Aurora A protein degrader according to claim 1, or a prodrug of the HSP90-based Aurora A protein degrader according to claim 1; the carrier is at least one of a diluent, an excipient, and a filler.

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