SRC-3 PROTAC compound, preparation method thereof and application of SRC-3 PROTAC compound in treatment of breast cancer

Abstract

The invention discloses an SRC-3 PROTAC compound as well as a preparation method and application thereof in treating breast cancer, and relates to the technical field of biological medicines. The SRC-3 PROTAC compound provided by the invention is prepared by carrying out an amide condensation reaction, a carbon-nitrogen coupling reaction or a reductive amination reaction on an SRC-3 inhibitor 20L derivative and an E3 ligase ligand (pomaferamine or VHL) derivative. The SRC-3 PROTAC compound provided by the invention has good SRC-3 degradation biological activity, can be applied to development of anti-cervical cancer drugs, and has a wide application prospect.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a SRC-3 PROTAC compound and a preparation method thereof and application thereof in the treatment of breast cancer. Background Art

[0002] Breast cancer (BCa) is a highly prevalent malignant tumor in women and has become one of the tumors with the highest new incidence and mortality, posing a serious threat to women's life and health. The occurrence and development of breast cancer is closely related to the excessive activation of estrogen signals. Breast cancer can be divided into four different subtypes based on whether estrogen receptors (ER), progesterone receptors (PR) and human epidermal growth factor 2 (HER2) are expressed. Studies have shown that 70% of breast cancer patients are estrogen receptor-positive (ER-positive). + ) patients, ER is in a state of significant overexpression. Therefore, ER has become one of the main targets for the development of targeted therapeutic drugs for breast cancer, and a variety of drugs have been approved for the clinical treatment of breast cancer.

[0003] According to their different mechanisms of action, drugs targeting ER can be divided into two types: selective estrogen receptor modulators (SERMs) and selective estrogen receptor downregulators (SERDs), with representative drugs being tamoxifen (Tam) and fulvestrant (Ful). However, long-term use has led to serious drug resistance problems, especially tamoxifen and other SERMs, which can induce a series of point mutations in the estrogen receptor ligand binding pocket, the most common of which is ER D538G and ER Y537S . ESR1Gene mutations can even directly confer ligand-independent transcriptional activity to the ER, enhance the recruitment of transcriptional coactivators, reduce ER proteolysis, and continuously drive gene transcription and cell proliferation. Long-term use of tamoxifen may lead to endometriosis, endometrial polyps, hyperplasia, and even tumors. In comparison, although Fulvestrant has significant therapeutic effects, its oral bioavailability is extremely low and it can currently only be administered by intramuscular injection. In order to address the problem of resistance to endocrine therapy for breast cancer and reduce the potential toxic side effects of existing therapeutic drugs, it is particularly urgent to develop targeted therapeutic drugs with unique mechanisms of action based on new drug design targets.

[0004] Steroid receptor coactivator 3 (SRC-3) is a multi-effect coactivator that can directly interact with the coactivator binding site (CBS) of ERα to form a transcriptional complex, which plays a very important role in regulating the transcription of downstream target genes. SRC-3 can also enhance the transcriptional activity of ERα and regulate the protein level of ERα. Therefore, it is a key molecule in the occurrence and development of breast cancer and resistance to endocrine therapy, and can be used as a potential target for the development of targeted therapeutic drugs for breast cancer. However, few SRC-3 small molecule inhibitors have been reported, and the reported small molecule inhibitors have poor anti-tumor effects or obvious toxic side effects. Therefore, it is very promising to design new, potent and safe small molecule targeted drugs for this target.

[0005] Proteolysis targeting chimera (PROTAC) technology is a proximity-induced protein degradation strategy developed based on the ubiquitin-proteasome system. PROTAC technology constructs a heterologous bifunctional molecule composed of a protein of interest ligand (POI ligand), a linker (Linker) and an E3 ubiquitin ligase ligand (E3 ligand), inducing the target protein and the E3 ligase to be close to each other in space, so that the former is degraded by ubiquitin and plays a role in treating diseases. PROTACs have great advantages over traditional small molecule inhibitors; for example, they have lower requirements for binding affinity, which enables them to target undruggable proteins and overcome target protein mutations. PROTACs also have the characteristics of low dosage, high selectivity and few side effects.

[0006] In view of this, using PROTAC technology to develop a series of PROTAC molecules that target the degradation of SRC-3 will be a potential targeted treatment strategy for breast cancer. Summary of the invention

[0007] The present invention provides a SRC-3 PROTAC compound and a preparation method thereof and an application thereof in treating breast cancer. The provided SRC-3 PROTAC compound has significant anti-breast cancer activity and can be developed as a new anti-breast cancer drug with broad application prospects. The present invention is specifically implemented by the following technologies.

[0008] In the first aspect of the present invention, a SRC-3 PROTAC compound is provided, the general structural formula of which is shown below: ; Among them, Linker is selected from , , , , , , , , , , or , and the right end of the Linker is connected to the E3 ligand; E3 ligand is selected from , or .

[0009] The present invention found through in vitro anti-breast cancer activity test that the above-mentioned SRC-3 PROTAC compound can significantly degrade SRC-3, and at the same time has strong anti-proliferation activity against a variety of breast cancer cells, and can be used to prepare SRC-3 degraders or drugs for treating breast cancer.

[0010] Furthermore, the SRC-3 PROTAC compound is selected from any one of the compounds in Table 1 below.

[0011] Table 1

[0012]

[0013]

[0014]

[0015]

[0016] In a second aspect of the present invention, a method for preparing a SRC-3 PROTAC compound is provided, wherein the SRC-3 PROTAC compound is any one of the SRC-3 PROTAC compounds described above; a SRC-3 inhibitor 20L derivative and an E3 ligase ligand derivative are subjected to an amide condensation reaction by adding a condensing agent HATU and a base DIPEA, or a carbon-nitrogen coupling reaction by adding a base DIPEA, or a reductive amination reagent NaBH(OAc) is added to react with the SRC-3 inhibitor 20L derivative and the E3 ligase ligand derivative. 3 Reductive amination reaction with acid AcOH to generate the SRC-3 PROTAC compound; The chemical structural formula of the SRC-3 inhibitor 20L derivative is: , Among them, R 2 Selected from , , , or ; The chemical structural formula of the E3 ligase ligand derivative is: , or , where n = 4, 5, 6, 7, 8, 9.

[0017] Furthermore, when the amide condensation reaction is carried out, the chemical structural formula of the SRC-3 inhibitor 20L derivative is: , The chemical structural formula of the E3 ligase ligand derivative is: , The chemical structure of the SRC-3 PROTAC compound generated accordingly is: , where n=4, 5, 6, 7, 8, 9.

[0018] Furthermore, when the carbon-nitrogen coupling reaction is performed, the chemical structural formula of the SRC-3 inhibitor 20L derivative is: , Among them, R 3 Selected from , , or , The chemical structural formula of the E3 ligase ligand derivative is: , The chemical structure of the SRC-3 PROTAC compound generated accordingly is: , Among them, L is selected from , , , , or .

[0019] Furthermore, when the reductive amination reaction is performed, the chemical structural formula of the SRC-3 inhibitor 20L derivative is:

[0020] The chemical structural formula of the E3 ligase ligand derivative is: , The chemical structure of the SRC-3 PROTAC compound generated accordingly is: .

[0021] Furthermore, when the amide condensation reaction is performed, the molar ratio of the SRC-3 inhibitor 20L derivative, the E3 ligase ligand derivative, the condensing agent HATU and the base DIPEA is 1:1.1:1.1:3; When the carbon-nitrogen coupling reaction is performed, the molar ratio of the SRC-3 inhibitor 20L derivative, the E3 ligase ligand derivative and the base DIPEA is 1:1:3; When the reductive amination reaction is performed, the SRC-3 inhibitor 20L derivative, the E3 ligase ligand derivative, the reductive amination reagent NaBH(OAc) 3 The molar ratio of HCl and acid AcOH is 1:1.2:2:1.

[0022] The third aspect of the present invention provides a pharmacologically or physiologically acceptable salt of any one of the above-mentioned SRC-3 PROTAC compounds.

[0023] In a fourth aspect, the present invention provides any one of the above-mentioned SRC-3 PROTAC compounds, or a SRC-3 PROTAC compound prepared by any one of the above-mentioned preparation methods, or a pharmacologically or physiologically acceptable salt of the above-mentioned SRC-3 PROTAC compound, for use in preparing a SRC-3 degrader, or in preparing a drug for treating breast cancer.

[0024] In a fifth aspect, the present invention provides a product, which is a SRC-3 degrader or a drug for treating breast cancer, and the product comprises any one of the above-mentioned SRC-3 PROTAC compounds, or a pharmacologically or physiologically acceptable salt of the above-mentioned SRC-3 PROTAC compound.

[0025] Furthermore, the product also includes at least one pharmaceutically acceptable carrier or excipient. The product can be prepared according to existing conventional pharmaceutical technology. Compared with the prior art, the present invention is beneficial in that: the present invention provides a SRC-3 PROTAC compound with good anti-breast cancer activity; the inhibitory activity of some compounds reaches the nmol level. The SRC-3 PROTAC compound of the present invention can be developed as a new anti-breast cancer drug and has a very broad application prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Three approaches for preparing the SRC-3 PROTAC compound of the present invention; wherein, approach a) is an amide condensation reaction approach, b) is a carbon-nitrogen coupling reaction approach, and c) is a reductive amination reaction approach.

[0027] Figure 2 and Figure 3 The results of immunoblotting analysis of the in vitro degradation activity of 1 μM SRC-3 PROTAC compounds on SRC-3. DETAILED DESCRIPTION

[0028] The technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] In some embodiments of the present invention, the chemical structure of the provided SRC-3 PROTAC compound is shown in Formula I below.

[0030] Formula Ⅰ.

[0031] In other embodiments of the present invention, a method for preparing the above-mentioned SRC-3 PROTAC compound is provided, which comprises the following steps: S1, through the following chemical reaction formula ( i ) were used as the starting materials for the next reaction.

[0032] ; Among them, R in compounds 2a, 2b, 2c, 2d and compounds 3a, 3b, 3c, 3d 1 Select the following 4 chemical structures respectively: , , , .

[0033] Furthermore, the above step S1 is carried out with (2-(2-chloro-1 H -Benzo[ d ] imidazole-1-yl)ethyl)carbamic acid tert-butyl ester (compound 3a) is taken as an example. The specific preparation method is as follows: commercially available raw materials 2-chlorobenzimidazole 1 (1500 mg, 9.83 mmol) and N -Boc-iodoethylamine 2a (3465 mg, 12.78 mmol) was dissolved in anhydrous DMF (8 mL) and then Cs 2 CO 3 (9609 mg, 29.49 mmol) was added to the reaction solution in batches, and the reaction system was heated to 110°C and stirred overnight. The reaction progress was monitored by TLC. When the raw material 1 completely disappeared, the reaction system was extracted with DCM, and the organic phase after drying with anhydrous sodium sulfate was concentrated to obtain the crude product, which was further purified by column chromatography (mobile phase: V 石油醚（PE） :V 乙酸乙酯（EA） = 3:1) to obtain a white solid product compound 3a (2035 mg, 6.88 mmol).

[0034] The preparation process of compounds 3b-d is similar to that of compound 3a, except that the raw material compound 2a is replaced by compounds 2b, 2c and 2d, respectively.

[0035] S2, through the following formula ( ii ) were used as the starting materials for the next reaction.

[0036] ; R in compounds 4a, 4b, 4c, and 4d 1 and R in compounds 2a, 2b, 2c, 2d and compounds 3a, 3b, 3c, 3d 1 same.

[0037] Furthermore, the above step S2 is carried out with (2-(2-hydrazino-1 H -Benzo[ d] imidazole-1-yl) ethyl) carbamic acid tert-butyl ester (compound 4a) is taken as an example, and the specific preparation method is: react ( i The raw material compound 3a (1000 mg, 3.38 mmol) was dissolved in EtOH (6 mL) and then hydrazine hydrate (N 2 H 4 H 2 O) (3 mL) and dropwise add AcOH (1 mL). Then the reaction system was heated to 130 ° C and stirred for 12 h. The reaction progress was monitored by TLC. When the raw material 3a completely disappeared, the reaction system was concentrated by vacuum rotary evaporator and extracted with ethyl acetate. The organic phase was dried with anhydrous sodium sulfate. After removing the ethyl acetate, a lavender solid product compound 4a (906 mg, 3.11 mmol) was obtained.

[0038] The preparation process of compounds 4b-d is similar to that of compound 4a, except that the raw material compound 3a is replaced by compounds 3b, 3c and 3d, respectively.

[0039] S3, through the following formula ( iii ) is synthesized by the reaction shown in Z )-1-methyl-2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -Benzo[ d ]imidazole derivatives (compounds 6a, 6b, 6c and 6d) and used as raw materials for subsequent reactions.

[0040] ; Among them, R in compounds 6a, 6b, 6c and 6d 2 Select the following 4 chemical structures respectively: , , , .

[0041] Furthermore, the above step S3 is performed with ( Z )-2-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazine)-1 H -Benzo[ d ] imidazole-1-yl)ethane-1-amine (compound 6a) is prepared as an example, and the specific preparation method is: react ( ii) and commercially available 2-acetylpyridine 5 (333 mg, 2.75 mmol) were dissolved in anhydrous methanol (5 mL), and then AcOH (0.5 mL) was slowly added and stirred at room temperature for 6 h. When the raw material compound 4a completely disappeared, saturated brine was slowly added until a large amount of red-brown solid precipitated, filtered, and the filter cake was rinsed with water (3´10 mL) and dried. The filter cake was ground into powder and redissolved in anhydrous DCM (6 mL), and trifluoroacetic acid (TFA) (3 mL) was added dropwise under ice bath conditions and stirred for 2 h. After removing DCM and TFA in the system with a vacuum rotary evaporator, it was extracted with DCM, and the organic phase was respectively treated with saturated NaHCO 3 The solution was washed with saturated brine and dried over anhydrous sodium sulfate, and DCM was removed to obtain a reddish brown solid compound 6a (691 mg, 2.35 mmol).

[0042] The preparation processes of compounds 6b, 6c and 6d are similar to that of compound 6a, except that the raw material compound 4a is replaced by compounds 4b, 4c and 4d, respectively.

[0043] S4, through the following formula ( iv ) is synthesized by the reaction shown in E )- N -(2-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazine)-1 H -Benzo[ d ] imidazol-1-yl) ethyl) -7-azaspiro [3.5] nonane-2-carboxamide (Compound 8) and used as the raw material for the subsequent reaction.

[0044]

[0045] Further, the specific preparation method is: react ( iii ) was dissolved in anhydrous DCM, and DIPEA (264 mg, 2.04 mmol) was added dropwise at room temperature. After the addition was complete, stirring was continued for 6 h. When the raw material compound 6a completely disappeared, the reaction system was extracted with DCM, the organic phase was dried and concentrated, and then redissolved in DCM (4 mL). TFA (2 mL) was added dropwise under ice bath conditions, and stirring was continued for 2 h. After removing DCM and TFA with a vacuum rotary evaporator, the reaction system was extracted with DCM, and the organic phase was respectively treated with saturated NaHCO 3The solution was washed with saturated brine and concentrated to obtain a reddish brown solid product (Compound 8, 259 mg, 0.58 mmol).

[0046] The above formula ( iv ) can also be replaced by compounds 6b, 6c and 6d, and the preparation process is similar to generate the corresponding products.

[0047] S5, through the following formula ( v ) were used as the starting materials for subsequent reactions.

[0048]

[0049] Among them, the difference between compounds 9a-f is that n is 4, 5, 6, 7, 8 and 9, respectively, and compounds 11a-f also correspond to them.

[0050] Further, with 6-(( S )-1-((2 S , 4 R )-4-hydroxy-2-(( S Taking the preparation of tert-butyl 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)hexanoate (Compound 11a) as an example, the specific preparation method is as follows: the commercially available raw material compound 9a (220 mg, 0.88 mmol) is dissolved in MeCN, and KI (6 mg, 0.03 mmol) and K 2 CO 3 (280 mg, 2.02 mmol), stirred at room temperature for 10 min. Then commercially available raw materials (2 S , 4 R )-1-(( S )-2-amino-3,3-dimethylbutyryl)-4-hydroxy- N -(( S )-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Compound 10, 300 mg, 0.67 mmol), and the reaction system was heated to 85°C and stirred overnight. When the raw material compound 10 completely disappeared, the reaction system was cooled to room temperature, water was added to quench the reaction system and extracted with DCM. The organic phase was dried and concentrated to obtain a crude product, which was further purified by column chromatography (mobile phase: V DCM :V MeOH = 30:1) to obtain a white solid product, compound 11a (353 mg, 0.57 mmol).

[0051] The preparation process of compounds 11b-f is similar to that of compound 11a, except that the raw material compound 9a is replaced by compounds 9b-f, respectively.

[0052] S6, through the following formula ( vi ) was used to synthesize 2-(2,6-dioxopiperidine-3-yl)-4-(4-oxopiperidine-1-yl)isoindoline-1,3-dione (Compound 14) and used as the raw material for subsequent reactions.

[0053]

[0054] Further, the commercially available raw materials 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione (compound 12, 500 mg, 1.81 mmol) and 4-piperidone ethylene glycol acetal (compound 13, 285 mg, 1.99 mmol) were dissolved in DMSO (6 mL), DIPEA (705 mg, 5.43 mmol) was added and the reaction system was heated to 100 ° C. When the raw material compound 12 completely disappeared, the reaction system was cooled to room temperature and quenched by adding ice water, followed by extraction with DCM. The organic phase was concentrated and dissolved in a mixed solvent of acetone / H 2 O(6mL, V acetone :V H2O = 1:1), and add p -TsOH (156 mg, 0.91 mmol), and continued stirring at room temperature for 8 h. The reaction system was extracted with DCM, and the organic phase was dried and concentrated to obtain a crude product, which was further purified by column chromatography (mobile phase: V DCM :V MeOH = 50:1) to obtain a bright yellow solid product compound 14 (550 mg, 1.55 mmol).

[0055] S7, through the following formula ( vii ) was synthesized by the reaction shown in FIG. 17 and used as the raw material for the next reaction.

[0056]

[0057] Further, commercially available raw materials tert-butyl bromoacetate (compound 15400 mg, 2.05 mmol), 3,9-diazaspiro[5.5]undecane (compound 16, 949 mg, 6.15 mmol), KI (17 mg, 0.10 mmol) and K 2 CO 3(850 mg, 6.15 mmol) were dissolved in MeCN, and the reaction system was heated to 85 °C and stirred for 10 h. When the raw material compound 15 completely disappeared, the reaction system was cooled to room temperature, and ice water was added dropwise, and then extracted with DCM. The organic phase was dried and concentrated to obtain a crude product, which was further purified by column chromatography (mobile phase: V DCM :V MeOH = 60:1) to obtain a colorless oily liquid product compound 17 (385 mg, 1.44 mmol).

[0058] S8, through the following formula ( viii ) was synthesized by the reaction shown in the figure to obtain tert-butyl 2-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undec-3-yl)acetate (Compound 19) which was used as the raw material for the subsequent reaction.

[0059]

[0060] Furthermore, the commercially available raw material 2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione (compound 18, 300 mg, 1.09 mmol) and the intermediate (compound 17, 321 mg, 1.19 mmol) were dissolved in DMSO, and DIPEA (424 mg, 3.27 mmol) was added dropwise. After the addition was completed, the reaction system was heated to 100 ° C and continued to stir for 6 hours. The reaction progress was monitored by TLC. When the raw material compound 18 completely disappeared, the reaction system was cooled to room temperature and extracted with DCM. The organic phase was dried and concentrated to obtain a crude product, which was further purified by column chromatography (mobile phase: V DCM :V MeOH = 50:1) to obtain a bright yellow solid product compound 19 (513 mg, 0.98 mmol).

[0061] S9, through the following formula ( ix ) to synthesize SRC-3 PROTAC (compounds I-1~I-6).

[0062]

[0063] Among them, n in chemical formulas 11a-f corresponds to 4, 5, 6, 7, 8 and 9 respectively, and n in chemical formulas 11a-f also corresponds thereto.

[0064] Furthermore, the SRC-3 PROTAC compound (2 S , 4 R )-1-(( S)-3,3-dimethyl-2-((6-oxo-6-((2-(( E )-1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -benzimidazol-1-yl)ethyl)amino)hexyl)amino)butyryl)-4-hydroxy- N -(( S )-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide (Compound I-1) is taken as an example. The specific preparation method is as follows: After the intermediate (Compound 11a, 300 mg, 0.49 mmol) is dissolved in anhydrous DCM (2 mL), TFA (2 mL) is added dropwise at room temperature and stirred at room temperature for 3 hours. After the raw material compound 11a completely disappears, DCM and TFA are removed by a vacuum rotary evaporator, the residue is redissolved in DCM, and DIPEA (173 mg, 1.33 mmol), HATU (186 mg, 0.49 mmol) and the intermediate (Compound 6a, 131 mg, 0.44 mmol) are added in sequence, and stirred at room temperature for 6 hours. The reaction system is extracted with DCM, and the organic phase is dried and concentrated to obtain a crude product, which is further purified by column chromatography (mobile phase: V DCM :V MeOH = 20:1) to obtain a yellow-brown solid product compound I-1 (287 mg, 0.34 mmol).

[0065] The preparation process of SRC-3 PROTAC compounds I-2~I-6 is similar to that of compound I-1, with the only difference being that the raw material compound 11a is replaced by compounds 11b-f.

[0066] S10, through the following formula ( x ) were used to synthesize SRC-3 PROTAC compounds II-1, II-2, and compounds III-1~III-4.

[0067]

[0068] Wherein, L in compound II-1 is , L in compound II-2 is , L in compound III-1 is , L in compound III-2 is , L in compound III-3 is , L in compound III-4 is .

[0069] Furthermore, the SRC-3 PROTAC compound ( E)-2-(2,6-dioxopiperidin-3-yl)-4-(3-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazine)-1 H -Benzo[ d ] imidazole-1-yl) azetidine-1-yl) isoindolin-1,3-dione (Compound II-1) is taken as an example. The specific preparation method is as follows: the intermediate (Compound 6b, 150 mg, 0.49 mmol), commercially available raw materials (Compound 12, 135 mg, 0.49 mmol) and DIPEA (191 mg, 1.47 mmol) are dissolved in DMSO (5 mL) in sequence, and then the reaction system is heated to 100 ° C and stirred overnight. The reaction progress is monitored by TLC. When the raw material compound 12 completely disappears, the reaction system is cooled to room temperature, extracted with DCM, and the organic phase is dried over anhydrous sodium sulfate. The crude product is concentrated and further purified by column chromatography (mobile phase: V DCM :V MeOH = 20:1) to obtain a yellow-brown solid product (compound II-1, 221 mg, 0.39 mmol).

[0070] The preparation process of SRC-3 PROTAC compounds II-2, III-1~III-4 is similar to that of compound II-1, the only difference is that the raw material combination "Compound 6b + Compound 12" is replaced by the following combinations: "Compound 6c + Compound 12", "Compound 6b + Compound 18", "Compound 6c + Compound 18", "Compound 6d + Compound 18" and "Compound 8 + Compound 18".

[0071] S11, through the following formula ( xii ) were used to synthesize lenalidomide carboxylic acid derivatives (compounds 17a-f) containing linkers of different lengths, and used as raw materials for the next reaction.

[0072]

[0073] Further, the intermediate (compound 19, 200 mg, 0.38 mmol) was dissolved in anhydrous DCM (2 mL), and TFA (2 mL) was added dropwise at room temperature and stirred at room temperature for 3 h. After the raw material compound 19 completely disappeared, DCM and TFA were removed by vacuum rotary evaporator, the residue was redissolved in DCM, and DIPEA (135 mg, 1.04 mmol), HATU (145 mg, 0.38 mmol) and the intermediate (compound 6a, 102 mg, 0.35 mmol) were added in sequence, and stirred at room temperature for 6 h. The reaction system was extracted with DCM, and the organic phase was dried and concentrated to obtain a crude product, which was further purified by column chromatography (mobile phase: V DCM :VMeOH = 20:1) to obtain a yellow-brown solid product (compound III-5, 209 mg, 0.28 mmol).

[0074] It should be noted that the above-mentioned preparation step S1-11 provided in the specific embodiment of the present invention is not limited to the order of S1-11, nor is it limited to completing all steps S1-11; rather, based on the SRC-3PROTAC compound to be prepared, refer to the above-mentioned step S1-11 to select and design the corresponding appropriate preparation process.

[0075] Example 1: (2 S , 4 R )-1-(( S )-3,3-dimethyl-2-((6-oxo-6-((2-(( E )-1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -benzimidazol-1-yl)ethyl)amino)hexyl)amino)butyryl)-4-hydroxy- N -(( S Preparation of 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide I-1.

[0076] After the intermediate 11a (300 mg, 0.49 mmol) was dissolved in anhydrous DCM (2 mL), TFA (2 mL) was added dropwise at room temperature and stirred at room temperature for 3 h. After the raw material 11a completely disappeared, DCM and TFA were removed by vacuum rotary evaporator, the residue was redissolved in DCM, and DIPEA (173 mg, 1.33 mmol), HATU (186 mg, 0.49 mmol) and intermediate 6a (131 mg, 0.44 mmol) were added in sequence and stirred at room temperature for 6 h. The reaction system was extracted with DCM, and the organic phase was dried and concentrated to obtain a crude product, which was further purified by column chromatography (mobile phase: V DCM :V MeOH = 20:1) to obtain a yellow-brown solid product I-1 (287 mg, 0.34 mmol) with a yield of 77%.

[0077] 1 H NMR (400 MHz, Chloroform- d ) δ 8.68 (d, J = 5.5 Hz, 1H), 8.57 (d, J =4.8 Hz, 1H), 7.93 (d, J= 7.7 Hz, 1H), 7.83 – 7.74 (m, 2H), 7.63 (t, J = 4.0 Hz,1H), 7.44 – 7.37 (m, 5H), 7.28 (d, J = 3.3 Hz, 1H), 7.11 – 6.99 (m, 4H), 5.12 –5.06 (m, 1H), 4.96 (t, J = 8.0 Hz, 1H), 4.77 (s, 1H), 4.34 (d, J = 11.0 Hz, 1H),4.13 (dd, J = 14.3, 6.3 Hz, 1H), 3.90 (d, J = 11.1 Hz, 1H), 3.75 – 3.53 (m, 4H),3.03 (s, 1H), 2.53 (d, J = 8.1 Hz, 7H), 2.30 (dt, J = 9.9, 5.2 Hz, 1H), 2.23 –2.11 (m, 2H), 2.06 – 1.99 (dd, J = 12.0, 5.0 Hz, 2H), 1.54 – 1.37 (m, 7H), 1.32– 1.23 (m, 2H), 0.97 (s, 9H). 13 C NMR (151 MHz, CDCl 3 ) δ 176.58, 174.03,169.71, 150.29, 149.00, 148.49, 143.35, 138.39, 132.12, 131.67, 130.83,129.59, 129.54, 127.99, 127.12, 126.47, 122.85, 121.94, 121.59, 121.57,107.43, 69.99, 67.55, 58.15, 56.95, 49.01, 48.92, 48.62, 40.70, 40.45, 36.96,35.27, 35.11, 34.81, 30.03, 26.85, 26.80, 26.49, 25.60, 22.39, 16.10, 13.28.HRMS (ESI): m / z calcd for C 45 H59 N 10 O 4 S [M + H] + : 835.4441; found, 835.4442.

[0078] Example 2: (2 S , 4 R )-1-((S)-3,3-dimethyl-2-((7-oxo-7-((2-((2-(( E )-1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -benzimidazol-1-yl)ethyl)amino)heptyl)amino)butyryl)-4-hydroxy- N -(( S Preparation of 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide I-2.

[0079] The preparation method is basically the same as that in Example 1, except that the raw material compound 11a is replaced by the raw material compound 11b to obtain a yellow-brown solid product compound I-2 with a yield of 81%.

[0080] 1 H NMR (400 MHz, Chloroform- d ) δ 8.67 (s, 1H), 8.56 (d, J = 4.8 Hz,1H), 7.91 (t, J = 7.2 Hz, 2H), 7.69 (td, J = 7.8, 1.8 Hz, 1H), 7.64 (d, J = 4.5 Hz,1H), 7.42 – 7.36 (m, 4H), 7.26 – 7.14 (m, 2H), 7.04 (dq, J = 11.5, 6.3, 5.8 Hz,4H), 5.07 (p, J = 7.0 Hz, 1H), 4.89 (dd, J = 8.4, 6.5 Hz, 1H), 4.66 (q, J = 4.0 Hz,1H), 4.22 (qd, J = 15.0, 12.2, 8.0 Hz, 2H), 3.74 – 3.55 (m, 5H), 3.07 (d, J= 5.8Hz, 1H), 2.72 – 2.62 (m, 1H), 2.52 (d, J = 5.7 Hz, 6H), 2.42 – 2.36 (m, 1H), 2.28 – 2.21 (m, 1H), 2.09 – 2.01 (t, J = m, 3H), 1.43 (dd, J = 24.6, 7.0 Hz, 7H),1.11 (t, J = 6.5 Hz, 4H), 0.98 (s, 9H). 13 C NMR (151 MHz, CDCl 3 ) δ 176.12,174.17, 169.84, 156.36, 150.30, 148.59, 148.46, 143.36, 142.61, 136.89,132.27, 131.70, 130.79, 129.56, 127.85, 126.46, 122.85, 121.83, 121.59,121.13, 120.97, 107.40, 69.91, 67.00, 58.29, 56.51, 48.94, 48.72, 48.60,40.73, 40.45, 36.32, 35.26, 34.96, 29.50, 28.15, 26.80, 26.48, 26.29, 25.15,22.35, 22.24, 16.09, 13.21. HRMS (ESI): m / z Calculate for C 46 H 61 N 10 O 4 S [M + H] + :849.4598; found, 849.4590.

[0081] Example 3: (2 S , 4 R )-1-(( S )-3,3-dimethyl-2-((8-oxo-8-((2-(( E )-1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -benzimidazol-1-yl)ethyl)amino)octyl)amino)butyryl)-4-hydroxy- N -(( SPreparation of 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide I-3.

[0082] The preparation method is basically the same as that in Example 1, except that the raw material compound 11a is replaced by the raw material compound 11c to obtain a yellow-brown solid product compound I-3 with a yield of 78%.

[0083] 1 H NMR (400 MHz, Chloroform- d ) δ 8.66 (s, 1H), 8.58 (d, J = 4.8 Hz,1H), 7.88 (dd, J = 16.0, 7.9 Hz, 2H), 7.74 – 7.66 (m, 1H), 7.52 (s, 1H), 7.38(s, 4H), 7.28 – 7.13 (m, 2H), 7.10 – 6.97 (m, 4H), 5.07 (p, J = 7.0 Hz, 1H),4.89 (dt, J = 14.9, 6.8 Hz, 1H), 4.68 (dt, J = 9.0, 4.5 Hz, 1H), 4.31 – 4.14 (m,2H), 3.76 (d, J = 8.4 Hz, 1H), 3.65 (d, J = 6.6 Hz, 4H), 3.12 (d, J = 5.7 Hz, 1H),2.64 (tt, J = 12.1, 4.9 Hz, 1H), 2.52 (d, J = 3.8 Hz, 6H), 2.42 (dt, J = 9.6, 5.4Hz, 1H), 2.28 – 2.21 (m, 1H), 2.06 (t, J = 7.8 Hz, 3H), 1.46 (d, J = 6.9 Hz, 2H),1.39 (t, J = 7.5 Hz, 3H), 1.29 – 1.22 (m, 2H), 1.10 – 0.98 (m, 15H). 13 C NMR (151MHz, CDCl 3) δ 176.23, 172.82, 169.78, 156.22, 150.31, 148.52, 148.47, 143.34,142.59, 137.10, 132.36, 131.71, 131.66, 130.81, 129.58, 127.84, 126.45,122.85, 121.84, 121.56, 121.27, 120.98, 107.46, 69.83, 67.30, 58.33, 56.50,48.96, 48.85, 48.73, 40.71, 40.27, 36.76, 35.20, 35.05, 29.70, 28.57, 28.47,26.85, 26.44, 25.32, 22.36, 22.25, 16.09, 13.18. HRMS (ESI): m / z Calculate for C 47 H 63 N 10 O 4 S [M + H] + : 863.4754; found, 863.4759.

[0084] Example 4: (2 S , 4 R )-1-(( S )-3,3-dimethyl-2-((9-oxo-9-((2-((2-(( E )-1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -benzimidazol-1-yl)ethyl)amino)nonyl)amino)butyryl)-4-hydroxy- N -(( S Preparation of 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide I-4.

[0085] The preparation method is basically the same as that in Example 1, except that the raw material compound 11a is replaced by the raw material compound 11d to obtain a yellow-brown solid product compound I-4 with a yield of 73%.

[0086] 1 H NMR (400 MHz, Chloroform- d ) δ 8.65 (s, 1H), 8.55 (d, J = 4.8 Hz,1H), 7.93 (d, J= 7.8 Hz, 1H), 7.87 (d, J = 8.2 Hz, 1H), 7.66 (t, J = 7.8 Hz, 1H),7.51 (s, 1H), 7.36 (s, 4H), 7.25 – 7.11 (m, 2H), 7.01 (q, J = 6.0 Hz, 4H), 5.05(p, J = 7.0 Hz, 1H), 4.87 (ddd, J = 15.9, 8.1, 5.8 Hz, 1H), 4.65 (q, J = 4.3 Hz,1H), 4.19 (t, J = 6.6 Hz, 2H), 3.73 – 3.61 (m, 4H), 3.21 (p, J = 6.7 Hz, 1H),3.11 (s, 1H), 2.66 – 2.60 (m, 1H), 2.51 (d, J = 8.8 Hz, 6H), 2.47 – 2.41 (m,1H), 2.31 (td, J = 10.8, 10.3, 5.4 Hz, 1H), 2.08 – 1.97 (m, 3H), 1.45 (d, J = 6.9Hz, 3H), 1.14 (d, J = 6.5 Hz, 4H), 1.07 (s, 8H), 1.00 (s, 9H). 13 C NMR (151 MHz,CDCl 3) δ 176.39, 174.19, 169.81, 157.20, 156.37, 151.96, 150.31, 148.60,148.45, 143.37, 142.61, 136.86, 132.30, 131.69, 130.77, 130.41, 129.56,127.83, 126.44, 122.78, 121.76, 121.52, 121.11, 120.96, 107.35, 69.74, 67.19,58.28, 56.18, 48.93, 48.79, HRMS (ESI): m / z Calculate for C 48 H 65 N 10 O 4 S [M + H] + : 877.4911; found, 877.4907.

[0087] Example 5: (2 S , 4 R )-1-(( S )-3,3-dimethyl-2-((10-oxo-10-((2-((2-(( E )-1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -benzimidazol-1-yl)ethyl)amino)decyl)amino)butyryl)-4-hydroxy- N -(( S Preparation of 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide I-5.

[0088] The preparation method is basically the same as that in Example 1, except that the raw material compound 11a is replaced by the raw material compound 11e to obtain a yellow-brown solid product compound I-5 with a yield of 80%.

[0089] 1 H NMR (400 MHz, Chloroform- d ) δ 8.66 (s, 1H), 8.56 (d, J = 4.8 Hz,1H), 7.90 (t,J = 8.3 Hz, 2H), 7.68 (td, J = 7.8, 1.8 Hz, 1H), 7.48 (d, J = 6.0 Hz,1H), 7.45 – 7.33 (m, 4H), 7.27 – 7.11 (m, 2H), 7.10 – 6.97 (m, 4H), 5.05 (p, J = 7.0 Hz, 1H), 4.87 (dt, J = 7.8, 3.7 Hz, 1H), 4.64 (dt, J = 8.5, 4.4 Hz, 1H),4.25 – 4.16 (m, 2H), 3.65 (dt, J = 10.3, 4.4 Hz, 4H), 3.12 (d, J = 5.5 Hz, 1H),2.64 – 2.57 (m, 1H), 2.51 (d, J = 8.2 Hz, 7H), 2.33 (td, J = 10.2, 9.1, 5.1 Hz,1H), 2.08 – 2.04 (m, 2H), 1.99 (dq, J = 8.7, 4.2 Hz, 1H), 1.46 (s, 4H), 1.37(d, J = 6.5 Hz, 4H), 1.18 – 1.06 (m, 10H), 1.00 (s, 9H). 13 C NMR (151 MHz, CDCl 3)δ 175.88, 173.39, 168.32, 156.64, 150.30, 148.52, 148.46, 143.35, 142.60,136.91, 132.43, 131.70, 131.67, 130.79, 129.57, 127.82, 126.43, 122.83,121.77, 121.57, 121.04, 120.97, 107.45, 69.81, 67.21, 58.33, 55.98, 54.19,48.91, 48.89, 48.69, 42.46, 40.81, 40.13, 36.62, 35.24, 35.05, 29.88, 28.88,28.81, 28.69, 27.00, 26.84, 25.39, 22.38, 22.27, 18.11, 16.09, 13.10. HRMS(ESI): m / z Calculate for C 49 H 67 N 10 O 4 S [M + H] + : 891.5067; found, 891.5071.

[0090] Example 6: (2 S , 4 R )-1-(( S )-3,3-dimethyl-2-((11-oxo-11-((2-(( E )-1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -benzimidazol-1-yl)ethyl)amino)undecyl)amino)butyryl)-4-hydroxy- N -(( S Preparation of 1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide I-6.

[0091] The preparation method is basically the same as that in Example 1, except that the raw material compound 11a is replaced by the raw material compound 11f to obtain a yellow-brown solid product compound I-6 with a yield of 75%.

[0092] 1 H NMR (400 MHz, Chloroform- d ) δ 8.66 (s, 1H), 8.56 (d, J= 4.8 Hz,1H), 7.93 (d, J = 7.8 Hz, 1H), 7.88 (d, J = 7.9 Hz, 1H), 7.67 (t, J = 7.8 Hz, 1H),7.54 – 7.30 (m, 5H), 7.27 – 7.12 (m, 2H), 7.02 (d, J = 5.4 Hz, 4H), 5.04 (p, J =7.0 Hz, 1H), 4.86 (ddd, J = 15.6, 8.2, 5.3 Hz, 1H), 4.66 (q, J = 6.6, 5.7 Hz,1H), 4.20 (t, J = 5.3 Hz, 2H), 3.66 (q, J = 5.2 Hz, 4H), 3.11 (d, J = 6.0 Hz, 2H),2.71 – 2.61 (m, 1H), 2.52 (d, J = 10.2 Hz, 6H), 2.49 – 2.43 (m, 1H), 2.34 (q, J =6.4, 4.7 Hz, 1H), 2.07 (t, J = 7.5 Hz, 2H), 1.96 (dq, J = 12.2, 3.4 Hz, 1H), 1.45(d, J = 7.0 Hz, 3H), 1.26 – 1.05 (m, 16H), 1.01 (s, 9H). 13 C NMR (151 MHz, CDCl 3)δ 176.40, 176.39, 174.09, 169.77, 157.09, 156.44, 150.32, 148.60, 148.45,143.34, 142.58, 136.74, 132.39, 131.70, 131.67, 130.78, 130.39, 129.56,127.81, 126.43, 122.76, 121.68, 121.52, 121.00, 107.38, 69.77, 67.31, 58.29,55.81, 48.91, 48.69, 40.76, 40.18, 36.77, 35.21, 35.01, 30.27, 29.25, 29.17,28.98, 28.95, 27.07, 26.82, 25.49, 22.39, 22.28, 16.07, 13.16. HRMS (ESI): m / z Calculate for C 50 H 69 N 10 O 4 S [M + H] + : 905.5224; found, 905.5223.

[0093] Example 7: E )-2-(2,6-dioxopiperidin-3-yl)-4-(3-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazine)-1 H -Benzo[ d ] imidazol-1-yl) azetidin-1-yl) isoindoline-1,3-dione II-1.

[0094] The intermediate (compound 6b, 150 mg, 0.49 mmol), commercially available raw material (compound 12, 135 mg, 0.49 mmol) and DIPEA (191 mg, 1.47 mmol) were dissolved in DMSO (5 mL) in sequence, and then the reaction system was heated to 100 °C and stirred overnight. The reaction progress was monitored by TLC. When the raw material compound 12 completely disappeared, the reaction system was cooled to room temperature, extracted with DCM, and the organic phase was dried over anhydrous sodium sulfate. The crude product was concentrated and further purified by column chromatography (mobile phase: V DCM :V MeOH = 20:1) to obtain a yellow-brown solid product (compound II-1, 221 mg, 0.39 mmol) with a yield of 80%.

[0095] 1 H NMR (600 MHz, DMSO- d 6 ) δ 11.20 (s, 1H), 11.08 (s, 1H), 8.50 (dt, J =4.8, 1.3 Hz, 1H), 8.44 (d, J = 8.1 Hz, 1H), 7.75 (td, J = 7.7, 1.8 Hz, 1H), 7.63(dd, J = 8.5, 7.0 Hz, 1H), 7.38 (dd, J = 6.2, 3.0 Hz, 1H), 7.26 (ddd, J = 7.3, 4.9,1.2 Hz, 1H), 7.20 (d, J = 7.0 Hz, 1H), 7.15 (dd, J = 5.7, 3.2 Hz, 1H), 7.07 –7.01 (m, 2H), 6.95 (d, J = 8.5 Hz, 1H), 5.60 (q, J = 8.2, 7.1 Hz, 1H), 5.09 (dd, J = 12.9, 5.4 Hz, 1H), 4.98 (q, J = 7.5, 6.8 Hz, 2H), 4.70 (t, J = 8.8 Hz, 2H),2.88 (ddd, J = 13.8, 10.4, 6.8 Hz, 1H), 2.62 – 2.55 (m, 2H), 2.07 – 2.01 (m,1H), 1.99 (s, 3H). 13 C NMR (151 MHz, DMSO- d 6) δ 173.28, 170.48, 167.77, 167.05,157.26, 155.15, 153.72, 148.68, 136.03, 135.57, 133.78, 131.78, 131.28,122.94, HRMS (ESI): m / z Calculate for C 30 H 27 N 8 O 4 [M + H] + : 563.2155; found, 563.2160.

[0096] Example 8: E )-2-(2,6-dioxopiperidin-3-yl)-4-(3-((2-(2-(1-(pyridin-2-yl)ethylidene)hydrazine)-1 H -Benzo[ d ] imidazol-1-yl) methyl) azetidin-1-yl) isoindoline-1,3-dione II-2.

[0097] The preparation method is basically the same as that of Example 7, except that the raw material compound 6b is replaced by the raw material compound 6c to obtain a yellow-brown solid product compound II-2 with a yield of 85%.

[0098] 1 H NMR (600 MHz, DMSO- d 6 ) δ 11.14 (s, 1H), 11.06 (s, 1H), 8.54 (d, J =4.5 Hz, 1H), 8.48 (d, J = 8.1 Hz, 1H), 7.77 (td, J = 7.7, 1.8 Hz, 1H), 7.58 –7.54 (m, 1H), 7.33 – 7.29 (m, 1H), 7.28 (dd, J = 7.4, 5.0 Hz, 1H), 7.15 – 7.09(m, 2H), 7.03 (td, J= 6.8, 5.6, 3.9 Hz, 2H), 6.77 (d, J = 8.5 Hz, 1H), 5.04 (dd, J = 12.7, 5.5 Hz, 1H), 4.31 (d, J = 7.3 Hz, 4H), 4.28 – 4.20 (m, 2H), 3.32 –3.26 (m, 1H), 2.85 (ddd, J = 16.7, 13.7, 5.4 Hz, 1H), 2.57 – 2.53 (m, 1H), 2.47(dd, J = 13.3, 4.6 Hz, 1H), 2.38 (s, 3H), 2.00 – 1.95 (m, 1H). 13 C NMR (151 MHz, DMSO- d 6 ) δ 173.24, 170.48, 167.70, 166.92, 157.52, 155.87, 153.19, 148.73,148.52, 135.97, 135.41, 133.76, 132.82, 131.26, 122.82, 121.50, 121.28,120.46, 120.30, 112.11, 110.49, 109.31, 108.07, 57.34, 49.07, 44.82, 31.41,29.07, 22.57, 12.67. HRMS (ESI): m / z Calculate for C 31 H 29 N 8 O 4 [M + H] + : 577.2312; found, 577.2317.

[0099] Example 9: E )-2-(2,6-dioxopiperidin-3-yl)-5-(3-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazine)-1 H -Benzo[ d Preparation of 1-imidazol-1-yl)azetidin-1-yl)isoindoline-1,3-dione III-1 The preparation method is basically the same as that of Example 7, except that the raw material compound 12 is replaced by the raw material compound 18 to obtain a yellow-brown solid product compound III-1 with a yield of 79%.

[0100] 1 H NMR (400 MHz, DMSO- d 6 ) δ 11.20 (s, 1H), 11.09 (s, 1H), 8.50 (dt, J =4.6, 1.5 Hz, 1H), 8.47 – 8.42 (m, 1H), 7.76 (td, J = 7.7, 1.8 Hz, 1H), 7.64(dd, J = 8.5, 7.0 Hz, 1H), 7.41 – 7.36 (m, 1H), 7.27 (ddd, J = 7.4, 4.9, 1.2 Hz,1H), 7.21 (d, J = 7.0 Hz, 1H), 7.18 – 7.14 (m, 1H), 7.05 (dd, J = 5.8, 3.2 Hz,2H), 6.96 (d, J = 8.5 Hz, 1H), 5.61 (tt, J = 8.4, 5.9 Hz, 1H), 5.09 (dd, J = 12.6,5.4 Hz, 1H), 5.05 – 4.94 (m, 2H), 4.70 (t, J = 8.7 Hz, 2H), 2.89 (td, J = 12.5,11.3, 6.5 Hz, 1H), 2.60 (d, J = 3.4 Hz, 1H), 2.56 (d, J = 2.4 Hz, 1H), 2.07 –2.01 (m, 1H), 2.00 (s, 3H). 13 C NMR (151 MHz, DMSO- d 6) δ 173.28, 170.48,167.77, 167.05, 157.30, 155.15, 153.76, 148.70, 148.69, 135.99, 135.57,133.78, 131.78, 131.28, HRMS (ESI): m / z Calculate for C 30 H 27 N 8 O 4 [M + H] + : 563.2155; found, 563.2155.

[0101] Example 10: E )-2-(2,6-dioxopiperidin-3-yl)-5-(3-((2-(2-(1-(pyridin-2-yl)ethylidene)hydrazine)-1 H -Benzo[ d ] imidazol-1-yl) methyl) azetidin-1-yl) isoindoline-1,3-dione III-2.

[0102] The preparation method is basically the same as that of Example 7, except that the raw material compound 6b and compound 12 are replaced by the raw material compound 6c and compound 18, respectively, to obtain a yellow-brown solid product compound III-2 with a yield of 86%.

[0103] 1 H NMR (400 MHz, Chloroform- d ) δ 9.54 (s, 1H), 8.54 (d, J = 4.8 Hz,1H), 7.97 (s, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.16 – 6.93 (m, 4H), 6.69 (s, 1H), 6.40 (d, J = 8.4 Hz, 1H), 4.92 (dd, J = 11.9, 5.5 Hz, 1H), 4.29 (s, 1H), 4.01(dt, J= 17.8, 8.7 Hz, 4H), 3.40 (p, J = 6.7 Hz, 1H), 2.89 – 2.63 (m, 3H), 2.54(s, 3H), 2.07 (dt, J = 10.7, 5.0 Hz, 1H), 0.87 (q, J = 7.7, 6.0 Hz, 1H). 13 C NMR (151 MHz, DMSO- d 6 ) δ 173.28, 170.58, 167.95, 167.65, 157.49, 155.87, 155.73,153.17, 148.73, 135.98, 134.29, 132.81, 131.25, 125.29, 122.84, 121.53,121.30, 120.46, 117.33, 114.65, 109.32, 108.13, 104.89, 55.38, 49.18, 44.76,31.45, 29.24, 26.81, 22.68, 12.71. HRMS (ESI): m / z Calculate for C 31 H 29 N 8 O 4 [M + H] + :577.2312; found, 577.2316. Example 11: E )-2-(2,6-dioxopiperidin-3-yl)-5-(4-((2-(2-(1-(pyridin-2-yl)ethylidene)hydrazine)-1 H -Benzo[ d ] Preparation of (1-imidazol-1-yl)methyl)piperidin-1-yl)isoindole-1,3-dione III-3.

[0104] The preparation method is basically the same as that in Example 1, except that the raw material compound 6b and compound 12 are replaced by the raw material compound 6d and compound 18, respectively, to obtain a yellow-brown solid product compound III-3 with a yield of 76%.

[0105] 1 H NMR (400 MHz, Chloroform- d ) δ 8.82 (s, 1H), 8.59 (d, J= 4.8 Hz,1H), 8.01 (d, J = 8.1 Hz, 1H), 7.67 (d, J = 8.3 Hz, 2H), 7.27 (s, 1H), 7.22 –7.17 (m, 1H), 7.02 (h, J = 8.0, 7.3 Hz, 4H), 4.96 (dd, J = 12.1, 5.4 Hz, 1H),3.94 (d, J = 9.4 Hz, 2H), 3.06 – 2.67 (m, 6H), 2.58 (s, 3H), 2.34 (ddt, J = 12.1,8.3, 4.2 Hz, 1H), 2.13 (dt, J = 10.3, 6.7 Hz, 1H), 1.86 (d, J = 13.0 Hz, 2H),1.51 (q, J = 11.8, 11.0 Hz, 2H), 1.28 (s, 1H). 13 C NMR (151 MHz, DMSO- d 6 ) δ173.28, 170.59, 168.10, 167.43, 157.58, 155.70, 155.32, 152.90, 148.71,135.93, 134.55, 133.20, 131.20, 125.48, 122.75, 121.26, 121.14, 120.39,118.09, 117.87, 109.20, 108.32, 108.23, 49.20, 47.44, 46.67, 35.20, 31.46,29.20, 22.66, 12.66. HRMS (ESI): m / z Calculate for C 33 H 33 N 8 O 4 [M + H] + : 605.2625; found, 605.2621.

[0106] Example 12: E )-7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-N -(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -Benzo[ d ] imidazol-1-yl) ethyl) -7-azaspiro [3.5] nonane-2-carboxamide III-4.

[0107] The preparation method is basically the same as that of Example 7, except that the raw material compound 6b and compound 12 are replaced by raw material compound 8 and compound 18, respectively, to obtain a yellow-brown solid product compound III-4 with a yield of 69%.

[0108] 1 H NMR (600 MHz, DMSO- d 6 ) δ 11.09 (d, J = 7.7 Hz, 2H), 8.52 (d, J = 4.5Hz, 1H), 8.47 (s, 1H), 7.87 (t, J = 5.8 Hz, 1H), 7.75 (td, J = 7.7, 1.8 Hz, 1H),7.63 (d, J = 8.5 Hz, 1H), 7.26 (p, J = 3.5 Hz, 2H), 7.18 (dd, J = 8.7, 2.2 Hz, 1H),7.12 (td, J = 6.9, 2.0 Hz, 2H), 7.00 (ddd, J = 6.4, 4.0, 1.6 Hz, 2H), 5.07 (dd, J =12.8, 5.5 Hz, 1H), 4.08 (t, J = 5.9 Hz, 2H), 3.47 (q, J = 5.9 Hz, 2H), 3.36 (d, J =4.6 Hz, 2H), 3.25 (t, J = 5.6 Hz, 2H), 2.91 – 2.87 (m, 1H), 2.60 (dd, J = 4.5,2.6 Hz, 1H), 2.59 – 2.52 (m, 2H), 2.41 (s, 3H), 2.02 (ddd, J = 9.5, 6.7, 3.8Hz, 1H), 1.81 (d,J = 8.7 Hz, 4H), 1.56 (t, J = 5.6 Hz, 2H), 1.48 – 1.44 (m, 2H). 13 C NMR (151 MHz, DMSO- d 6 ) δ 175.13, 173.29, 170.59, 168.09, 167.43, 161.47,157.60, 155.38, 152.73, 148.71, 135.92, 134.46, 133.11, 132.42, 131.28,125.42, 122.73, 121.78, 121.22, 120.38, 118.09, 109.19, 108.26, 49.20, 44.87,44.57, 41.78, 41.10, 37.52, 37.44, 35.69, 34.46, 33.98, 32.71, 31.46, 22.66,12.68. HRMS (ESI): m / z Calculate for C 38 H 40 N 9 O 5 [M + H] + : 702.3152; found, 702.3157.

[0109] Example 13: E )-2-(2,6-dioxopiperidin-3-yl)-4-(3-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazine)-1 H -Benzo[ d ] imidazol-1-yl) azetidin-1-yl) piperidin-1-yl) preparation of isoindoline-1,3-dione II-3.

[0110] The intermediate (compound 6b, 150 mg, 0.49 mmol) and (compound 14, 183 mg, 0.51 mmol) were dissolved in anhydrous DCM (6 mL), purged with argon (Ar) and stirred at room temperature for 1 h. NaBH(OAc) was then added in portions. 3 (208 mg, 0.98 mmol), stirring was continued overnight, and the reaction progress was monitored by TLC. After the raw material completely disappeared, ice water was added to quench the reaction, and DCM was used for extraction. The organic phase was dried and concentrated to obtain a crude product, which was further purified by column chromatography (mobile phase: V DCM :VMeOH = 20:1), a yellow-brown solid product (compound II-3, 247 mg, 0.38 mmol) was obtained with a yield of 78%.

[0111] 11 H NMR (400 MHz, DMSO- d 6 ) δ 11.14 (s, 1H), 11.12 (s, 1H), 8.55 (dt, J =4.7, 1.5 Hz, 1H), 8.52 – 8.44 (m, 1H), 7.82 – 7.74 (m, 1H), 7.70 (dd, J = 8.5,7.1 Hz, 1H), 7.52 (dd, J = 6.0, 3.1 Hz, 1H), 7.36 (dd, J = 9.4, 7.7 Hz, 2H), 7.29(ddd, J = 7.4, 4.8, 1.2 Hz, 1H), 7.18 – 7.11 (m, 1H), 7.07 – 7.01 (m, 2H), 5.11(dt, J = 13.4, 6.2 Hz, 2H), 3.81 (q, J = 6.4 Hz, 4H), 3.62 (d, J = 11.6 Hz, 2H),3.03 (t, J = 10.7 Hz, 2H), 2.94 – 2.86 (m, 1H), 2.64 – 2.54 (m, 3H), 2.44 (s,3H), 2.07 – 2.01 (m, 1H), 1.93 – 1.82 (m, 2H), 1.53 – 1.46 (m, 2H). 13 C NMR (151 MHz, DMSO- d 6) δ 173.29, 170.51, 167.57, 166.79, 157.47, 155.63, 153.35,150.46, 148.74, 136.22, 135.99, 134.14, 131.79, 131.36, 124.41, 122.88,121.61, 121.15, 120.53, 116.92, 114.99, 109.25, 62.70, 56.26, 49.26, 43.33,34.66, 31.43, 26.81, 25.25, 22.54, 14.44, 13.05, 11.72. HRMS (ESI): m / z calcdfor C 35 H 36 N 9 O 4 [M + H] + : 646.2890; found, 646.2894.

[0112] Example 14: E )-2-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undec-3-yl)- N -(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1 H -Benzo[ d ] Preparation of (1-imidazol-1-yl)ethyl)acetamide III-5.

[0113] The preparation method of this example is basically the same as that of Example 1, except that the raw material 11a is replaced by the raw material 19 to obtain a yellow-brown solid product III-5 with a yield of 75%.

[0114] 1 H NMR (400 MHz, Chloroform- d ) δ 9.10 (s, 1H), 8.50 – 8.41 (m, 1H), 7.85 (d, J = 8.1 Hz, 2H), 7.47 (dd, J = 13.7, 7.9 Hz, 2H), 7.17 (s, 1H), 7.12 –7.04 (m, 4H), 6.98 (d, J = 2.3 Hz, 1H), 6.76 (dd, J= 8.6, 2.3 Hz, 1H), 4.99 (dd, J = 12.2, 5.3 Hz, 1H), 4.32 (s, 2H), 3.81 (t, J = 5.4 Hz, 2H), 3.67 (p, J = 6.7Hz, 1H), 3.22 – 3.11 (m, 1H), 3.08 (t, J = 5.3 Hz, 4H), 3.00 (s, 2H), 2.92 –2.75 (m, 3H), 2.52 (s, 3H), 2.42 (t, J = 5.1 Hz, 4H), 2.14 (dt, J = 10.0, 3.6 Hz,1H), 1.51 – 1.35 (m, 5H), 1.29 – 1.25 (m, 1H). 13 C NMR (151 MHz, DMSO- d 6 ) δ173.30, 170.61, 168.00, 167.44, 157.27, 155.81, 155.54, 152.88, 148.60,135.80, 134.19, 132.67, 131.35, 125.17, 122.68, 121.42, 121.20, 120.28,118.87, 118.10, 109.34, 108.26, 108.05, 54.06, 52.73, 49.24, 46.78, 42.31,37.92, 31.47, 22.67, 18.55, 17.19, 12.96, 12.79. HRMS (ESI): m / z Calculate for C 40 H 45 N 10 O 5 [M + H] + : 745.3574; found, 745.3568.

[0115] The chemical structures of the target compounds I-1-I-6, II-1-II-3 and III-1-III-5 of the present invention synthesized above are shown in Table 1. Experimental Example 1: In vitro SRC-3 degradation activity of SRC-3 PROTAC compounds.

[0116] Western blot analysis steps: (1) MCF-7 cells were treated with different concentrations of SRC-3 PROTAC compounds for 10 h, and whole cell lysates were obtained using RIPA buffer and SDS-PAGE protein loading buffer. (2) The protein concentration of the samples was analyzed using the BCA protein analysis kit (KR0008), and the volume of the samples for electrophoresis was adjusted according to the standard protein curve. (3) The proteins were separated using 7.5% or 10% SDS-PAGE gel and transferred to a 0.45 μM thick PVDE membrane (Millipore, 000027346). (4) The membrane was blocked with 5% bovine serum albumin (BSA, KR9048-466-8) or 5% skim milk at room temperature for 2 h, the primary antibody was incubated at 4°C for more than 12 h, and the secondary antibody was incubated at room temperature on a shaker for more than 1 h. (5) The protein was detected using ultrasensitive enhanced chemiluminescence (ECL, meilunbio, MAO186-2) reagent. SRC-3 antibody (4877S) was purchased from Cell Signaling Technology (CST), and GAPDH (60004-1-Ig) antibody was purchased from Proteintech.

[0117] The results of compounds Ⅰ-1~Ⅰ-6, compounds Ⅱ-1~Ⅱ-3, and compounds Ⅲ-1~Ⅲ-5 are as follows Figure 2 and 3 As shown, it was shown that most SRC-3 PROTAC compounds can induce SRC-3 degradation, such as I-2-I-6, II-3, III-1 and III-2. Experimental Example 2: In vitro anti-breast cancer activity of SRC-3 PROTAC compounds.

[0118] CCK-8 is used to test the anti-proliferative activity of compounds. Principle: CCK-8 (Cell Counting Kit-8) is a WST-8-based detection reagent widely used in cell proliferation and cytotoxicity. WST-8 (chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid benzene)-2 H -tetrazolyl monosodium salt), is a compound similar to MTT (chemical name: 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide), which is reduced to a highly water-soluble orange-yellow formazan product (Formazan) by dehydrogenase in mitochondria under the action of electron carrier 1-methoxy-5-methylphenazinium dimethyl sulfate. The more cells proliferate and the faster the speed, the darker the color; the greater the cell toxicity, the lighter the color. For the same cells, the depth of the color is proportional to the number of living cells, so this characteristic can be used to directly analyze cell proliferation and toxicity.

[0119] Procedure: (1) Prepare cell suspension: breast cancer cells (MCF-7, LCC2, MCF-7 D538G or MCF-7 Y537S ) When the growth density reaches 80%-90%, digest the cells with 0.25% Trypsin-EDTA, add fresh culture medium, mix the cell suspension, count the cells using a cell counting plate, and dilute the cells to 5×10 4 (2)Plate: 100 μL of single cell suspension was inoculated into a microplate (tissue culture grade, 96-well, flat bottom). (3)Pre-culture: 37°C, 5% CO 2 Incubate the cells in the incubator for about 24 hours. (4) Add drugs: Aspirate the original culture medium, add 200 μL of culture medium containing different concentrations (100 μM-0.01 μM) of the drug to be tested to each well of the culture plate in turn (set up 3 replicates for each concentration, and set up a blank (DMSO) group, a tamoxifen control group, and a fulvestrant control group), and place in the incubator for incubation for 72 hours. (5) Collect the plates: Prepare CCK-8 working solution at a ratio of 10 μL of culture medium plus 10 μL of CCK-8 reagent, add 100 μL of CCK-8 working solution to each well, and place the culture plate in the incubator for incubation for 1-2 hours. (6) Measure the plate: Use an ELISA reader to measure the absorbance (OD) at 450 nm.

[0120] Table 2. Anti-breast cancer cell proliferation activity of SRC-3 PROTAC compounds (I-1-I-6, II-1-II-3 and III-1-III-5) a .

[0121]

[0122] a All inhibitory concentration values ​​(IC 50 ± SD) are the results of three independent repeated experiments. b 4-OHT: 4-hydroxytamoxifen; c Ful:Fulvestrant.

[0123] The experimental results in Table 2 above show that most of the synthesized SRC-3 PROTAC compounds were active in the wild-type (MCF-7) and drug-resistant mutant cells (LCC2, MCF-7 D538G and MCF-7 Y537S ) showed good anti-breast cancer activity, generally better than the positive control drug 4-hydroxytamoxifen (IC 50 = 200-1290nM, among which some compounds showed inhibitory activity at the nanomolar level, such as compound I-4 (MCF-7, IC 50= 0.09±0.018 µM), I-5 (MCF-7, IC 50 = 0.08±0.15 µM) I-6 (MCF-7, IC 50 = 0.08±0.03 µM) and II-3 (MCF-7, IC 50 = 0.01±0.01 µM;MCF-7 Y537S , IC 50 = 0.003 ± 0.002 µM).

[0124] The above specific embodiments describe the implementation of the present invention in detail, but the present invention is not limited to the specific details in the above embodiments. Within the scope of the claims and technical concept of the present invention, the technical solution of the present invention can be modified and changed in many simple ways, and these simple modifications all belong to the protection scope of the present invention.

Claims

1. A SRC-3 PROTAC compound, characterized in that The general structural formula is shown below: ； Among them, Linker is selected from , , , , , , , , , , or , and the right end of the Linker is connected to the E3 ligand; E3 ligand is selected from , or .

2. The SRC-3 PROTAC compound according to claim 1, characterized in that Any one of the following compounds: (2S,4R)-1-((S)-3,3-dimethyl-2-((6-oxo-6-((2-((E)-1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzimidazol-1-yl)ethyl)amino)hexyl)amino)butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, (2S,4R)-1-((S)-3,3-dimethyl-2-((7-oxo-7-((2-((2-((E)-1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzimidazol-1-yl)ethyl)amino)heptyl)amino)butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, (2S,4R)-1-((S)-3,3-dimethyl-2-((8-oxo-8-((2-((E)-1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzimidazol-1-yl)ethyl)amino)octyl)amino)butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, (2S,4R)-1-((S)-3,3-dimethyl-2-((9-oxo-9-((2-((2-((E)-1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzimidazol-1-yl)ethyl)amino)nonyl)amino)butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, (2S,4R)-1-((S)-3,3-dimethyl-2-((10-oxo-10-((2-((2-((E)-1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzimidazol-1-yl)ethyl)amino)decyl)amino)butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, (2S,4R)-1-((S)-3,3-dimethyl-2-((11-oxo-11-((2-((E)-1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzimidazol-1-yl)ethyl)amino)undecyl)amino)butanoyl)-4-hydroxy-N-((S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide, (E)-2-(2,6-dioxopiperidin-3-yl)-4-(3-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzo[d]imidazol-1-yl)azetidin-1-yl)isoindoline-1,3-dione, (E)-2-(2,6-dioxopiperidin-3-yl)-4-(3-((2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzo[d]imidazol-1-yl)methyl)azetidin-1-yl)isoindoline-1,3-dione, (E)-2-(2,6-dioxopiperidin-3-yl)-4-(3-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzo[d]imidazol-1-yl)azetidin-1-yl)piperidin-1-yl)isoindoline-1,3-dione, (E)-2-(2,6-dioxopiperidin-3-yl)-5-(3-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzo[d]imidazol-1-yl)azetidin-1-yl)isoindoline-1,3-dione, (E)-2-(2,6-dioxopiperidin-3-yl)-5-(3-((2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzo[d]imidazol-1-yl)methyl)azetidin-1-yl)isoindoline-1,3-dione, (E)-2-(2,6-dioxopiperidin-3-yl)-5-(4-((2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzo[d]imidazol-1-yl)methyl)piperidin-1-yl)isoindole-1,3-dione, (E)-7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-N-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzo[d]imidazol-1-yl)ethyl)-7-azaspiro[3.5]nonane-2-carboxamide, (E)-2-(9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undecan-3-yl)-N-(2-(2-(1-(pyridin-2-yl)ethylidene)hydrazino)-1H-benzo[d]imidazol-1-yl)ethyl)acetamide.

3. A method for preparing a SRC-3 PROTAC compound, characterized in that: The SRC-3 PROTAC compound is the SRC-3 PROTAC compound according to claim 1 or 2; the SRC-3 inhibitor 20L derivative and the E3 ligase ligand derivative are subjected to an amide condensation reaction by adding a condensing agent HATU and a base DIPEA, or a carbon-nitrogen coupling reaction by adding a base DIPEA, or a reductive amination reaction by adding a reductive amination agent NaBH(OAc)3 and an acid AcOH to generate the SRC-3PROTAC compound; The chemical structure formula of the SRC-3 inhibitor 20L derivative is: ， Among them, R 2 Selected from , , , or ; The chemical structural formula of the E3 ligase ligand derivative is: , or , where n=4, 5, 6, 7, 8, 9.

4. The method for preparing the SRC-3 PROTAC compound according to claim 3, characterized in that When the amide condensation reaction is carried out, the chemical structural formula of the SRC-3 inhibitor 20L derivative is: ， The chemical structural formula of the E3 ligase ligand derivative is: ， The chemical structure of the SRC-3 PROTAC compound generated accordingly is: , where n=4, 5, 6, 7, 8, 9; When the carbon-nitrogen coupling reaction is carried out, the chemical structure of the SRC-3 inhibitor 20L derivative is: ， Among them, R 3 Selected from , , or , The chemical structural formula of the E3 ligase ligand derivative is: ， The chemical structure of the SRC-3 PROTAC compound generated accordingly is: ， Among them, L is selected from , , , , or ; When the reductive amination reaction is performed, the chemical structural formula of the SRC-3 inhibitor 20L derivative is: The chemical structural formula of the E3 ligase ligand derivative is: ， The chemical structure of the SRC-3 PROTAC compound generated accordingly is: 。 5. The method for preparing the SRC-3 PROTAC compound according to claim 3 or 4, characterized in that When the amide condensation reaction is performed, the molar ratio of the SRC-3 inhibitor 20L derivative, the E3 ligase ligand derivative, the condensing agent HATU and the base DIPEA is 1:1.1:1.1:3; When the carbon-nitrogen coupling reaction is performed, the molar ratio of the SRC-3 inhibitor 20L derivative, the E3 ligase ligand derivative and the base DIPEA is 1:1:3; When the reductive amination reaction is performed, the molar ratio of the SRC-3 inhibitor 20L derivative, the E3 ligase ligand derivative, the reductive amination agent NaBH(OAc)3 and the acid AcOH is 1:1.2:2:

1.

6. A pharmacologically or physiologically acceptable salt of the SRC-3 PROTAC compound of claim 1 or 2.

7. Use of a SRC-3 PROTAC compound according to claim 1 or 2, or a SRC-3 PROTAC compound prepared by the preparation method according to any one of claims 3 to 5, or a pharmacologically or physiologically acceptable salt of the SRC-3 PROTAC compound according to claim 6 in the preparation of a SRC-3 degrader, or a drug for treating breast cancer.

8. A product, characterized in that The product is a SRC-3 degrader or a drug for treating breast cancer, and the product comprises the SRC-3 PROTAC compound according to claim 1 or 2, or a pharmacologically or physiologically acceptable salt of the SRC-3 PROTAC compound according to claim 6.

9. The product according to claim 8, characterized in that Also included is at least one pharmaceutically acceptable carrier or excipient.

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